ASU Scholarship Showcase

High proportions of autistic children suffer from gastrointestinal (GI) disorders, implying a link between autism and abnormalities in gut microbial functions. Increasing evidence from recent high-throughput sequencing analyses indicates that disturbances in composition and diversity of gut microbiome are associated with various disease conditions. However, microbiome-level studies on autism are limited and mostly focused on pathogenic bacteria. Therefore, here we aimed to define systemic changes in gut microbiome associated with autism and autism-related GI problems. We recruited 20 neurotypical and 20 autistic children accompanied by a survey of both autistic severity and GI symptoms. By pyrosequencing the V2/V3 regions in bacterial 16S rDNA from fecal DNA samples, we compared gut microbiomes of GI symptom-free neurotypical children with those of autistic children mostly presenting GI symptoms. Unexpectedly, the presence of autistic symptoms, rather than the severity of GI symptoms, was associated with less diverse gut microbiomes. Further, rigorous statistical tests with multiple testing corrections showed significantly lower abundances of the genera Prevotella, Coprococcus, and unclassified Veillonellaceae in autistic samples. These are intriguingly versatile carbohydrate-degrading and/or fermenting bacteria, suggesting a potential influence of unusual diet patterns observed in autistic children. However, multivariate analyses showed that autism-related changes in both overall diversity and individual genus abundances were correlated with the presence of autistic symptoms but not with their diet patterns. Taken together, autism and accompanying GI symptoms were characterized by distinct and less diverse gut microbial compositions with lower levels of Prevotella, Coprococcus, and unclassified Veillonellaceae.

Throughout the long history of virus-host co-evolution, viruses have developed delicate strategies to facilitate their invasion and replication of their genome, while silencing the host immune responses through various mechanisms. The systematic characterization of viral protein-host interactions would yield invaluable information in the understanding of viral invasion/evasion, diagnosis and therapeutic treatment of a viral infection, and mechanisms of host biology. With more than 2,000 viral genomes sequenced, only a small percent of them are well investigated. The access of these viral open reading frames (ORFs) in a flexible cloning format would greatly facilitate both in vitro and in vivo virus-host interaction studies. However, the overall progress of viral ORF cloning has been slow. To facilitate viral studies, we are releasing the initiation of our panviral proteome collection of 2,035 ORF clones from 830 viral genes in the Gateway® recombinational cloning system. Here, we demonstrate several uses of our viral collection including highly efficient production of viral proteins using human cell-free expression system in vitro, global identification of host targets for rubella virus using Nucleic Acid Programmable Protein Arrays (NAPPA) containing 10,000 unique human proteins, and detection of host serological responses using micro-fluidic multiplexed immunoassays. The studies presented here begin to elucidate host-viral protein interactions with our systemic utilization of viral ORFs, high-throughput cloning, and proteomic technologies. These valuable plasmid resources will be available to the research community to enable continued viral functional studies.

Rationale: Cell-free protein microarrays display naturally-folded proteins based on just-in-time in situ synthesis, and have made important contributions to basic and translational research. However, the risk of spot-to-spot cross-talk from protein diffusion during expression has limited the feature density of these arrays.

Methods: In this work, we developed the Multiplexed Nucleic Acid Programmable Protein Array (M-NAPPA), which significantly increases the number of displayed proteins by multiplexing as many as five different gene plasmids within a printed spot.

Results: Even when proteins of different sizes were displayed within the same feature, they were readily detected using protein-specific antibodies. Protein-protein interactions and serological antibody assays using human viral proteome microarrays demonstrated that comparable hits were detected by M-NAPPA and non-multiplexed NAPPA arrays. An ultra-high density proteome microarray displaying > 16k proteins on a single microscope slide was produced by combining M-NAPPA with a photolithography-based silicon nano-well platform. Finally, four new tuberculosis-related antigens in guinea pigs vaccinated with Bacillus Calmette-Guerin (BCG) were identified with M-NAPPA and validated with ELISA.

Conclusion: All data demonstrate that multiplexing features on a protein microarray offer a cost-effective fabrication approach and have the potential to facilitate high throughput translational research.

Sera from patients with ovarian cancer contain autoantibodies (AAb) to tumor-derived proteins that are potential biomarkers for early detection. To detect AAb, we probed high-density programmable protein microarrays (NAPPA) expressing 5177 candidate tumor antigens with sera from patients with serous ovarian cancer (n = 34 cases/30 controls) and measured bound IgG. Of these, 741 antigens were selected and probed with an independent set of ovarian cancer sera (n = 60 cases/60 controls). Twelve potential autoantigens were identified with sensitivities ranging from 13 to 22% at >93% specificity. These were retested using a Luminex bead array using 60 cases and 60 controls, with sensitivities ranging from 0 to 31.7% at 95% specificity. Three AAb (p53, PTPRA, and PTGFR) had area under the curve (AUC) levels >60% (p < 0.01), with the partial AUC (SPAUC) over 5 times greater than for a nondiscriminating test (p < 0.01). Using a panel of the top three AAb (p53, PTPRA, and PTGFR), if at least two AAb were positive, then the sensitivity was 23.3% at 98.3% specificity. AAb to at least one of these top three antigens were also detected in 7/20 sera (35%) of patients with low CA 125 levels and 0/15 controls. AAb to p53, PTPRA, and PTGFR are potential biomarkers for the early detection of ovarian cancer.

To address the need to study frozen clinical specimens using next-generation RNA, DNA, chromatin immunoprecipitation (ChIP) sequencing and protein analyses, we developed a biobank work flow to prospectively collect biospecimens from patients with renal cell carcinoma (RCC). We describe our standard operating procedures and work flow to annotate pathologic results and clinical outcomes. We report quality control outcomes and nucleic acid yields of our RCC submissions (N=16) to The Cancer Genome Atlas (TCGA) project, as well as newer discovery platforms, by describing mass spectrometry analysis of albumin oxidation in plasma and 6 ChIP sequencing libraries generated from nephrectomy specimens after histone H3 lysine 36 trimethylation (H3K36me3) immunoprecipitation. From June 1, 2010, through January 1, 2013, we enrolled 328 patients with RCC. Our mean (SD) TCGA RNA integrity numbers (RINs) were 8.1 (0.8) for papillary RCC, with a 12.5% overall rate of sample disqualification for RIN <7. Banked plasma had significantly less albumin oxidation (by mass spectrometry analysis) than plasma kept at 25°C (P<.001). For ChIP sequencing, the FastQC score for average read quality was at least 30 for 91% to 95% of paired-end reads. In parallel, we analyzed frozen tissue by RNA sequencing; after genome alignment, only 0.2% to 0.4% of total reads failed the default quality check steps of Bowtie2, which was comparable to the disqualification ratio (0.1%) of the 786-O RCC cell line that was prepared under optimal RNA isolation conditions. The overall correlation coefficients for gene expression between Mayo Clinic vs TCGA tissues ranged from 0.75 to 0.82. These data support the generation of high-quality nucleic acids for genomic analyses from banked RCC. Importantly, the protocol does not interfere with routine clinical care. Collections over defined time points during disease treatment further enhance collaborative efforts to integrate genomic information with outcomes.

The urban heat island (UHI) phenomenon is a significant worldwide problem caused by rapid population growth and associated urbanization. The UHI effect exacerbates heat waves during the summer, increases energy and water consumption, and causes the high risk of heat-related morbidity and mortality. UHI mitigation efforts have increasingly relied on wisely designing the urban residential environment such as using high albedo rooftops, green rooftops, and planting trees and shrubs to provide canopy coverage and shading. Thus, strategically designed residential rooftops and their surrounding landscaping have the potential to translate into significant energy, long-term cost savings, and health benefits. Rooftop albedo, material, color, area, slope, height, aspect and nearby landscaping are factors that potentially contribute. To extract, derive, and analyze these rooftop parameters and outdoor landscaping information, high resolution optical satellite imagery, LIDAR (light detection and ranging) point clouds and thermal imagery are necessary. Using data from the City of Tempe AZ (a 2010 population of 160,000 people), we extracted residential rooftop footprints and rooftop configuration parameters from airborne LIDAR point clouds and QuickBird satellite imagery (2.4 m spatial resolution imagery). Those parameters were analyzed against surface temperature data from the MODIS/ASTER airborne simulator (MASTER). MASTER images provided fine resolution (7 m) surface temperature data for residential areas during daytime and night time. Utilizing these data, ordinary least squares (OLS) regression was used to evaluate the relationships between residential building rooftops and their surface temperature in urban environment. The results showed that daytime rooftop temperature was closely related to rooftop spectral attributes, aspect, slope, and surrounding trees. Night time temperature was only influenced by rooftop spectral attributes and slope.

This study examines the spatial and temporal patterns of the surface urban heat island (SUHI) intensity in the Phoenix metropolitan area and the relationship with land use land cover (LULC) change between 2000 and 2014. The objective is to identify specific regions in Phoenix that have been increasingly heated and cooled to further understand how LULC change influences the SUHI intensity. The data employed include MODerate-resolution Imaging Spectroradiometer (MODIS) land surface temperature (LST) 8-day composite June imagery, and classified LULC maps generated using 2000 and 2014 Landsat imagery. Results show that the regions that experienced the most significant LST changes during the study period are primarily on the outskirts of the Phoenix metropolitan area for both daytime and nighttime. The conversion to urban, residential, and impervious surfaces from all other LULC types has been identified as the primary cause of the UHI effect in Phoenix. Vegetation cover has been shown to significantly lower LST for both daytime and nighttime due to its strong cooling effect by producing more latent heat flux and less sensible heat flux. We suggest that urban planners, decision-makers, and city managers formulate new policies and regulations that encourage residential, commercial, and industrial developers to include more vegetation when planning new construction.

This paper reviews how remotely sensed data have been used to understand the impact of urbanization on global environmental change. We describe how these studies can support the policy and science communities’ increasing need for detailed and up-to-date information on the multiple dimensions of cities, including their social, biological, physical, and infrastructural characteristics. Because the interactions between urban and surrounding areas are complex, a synoptic and spatial view offered from remote sensing is integral to measuring, modeling, and understanding these relationships. Here we focus on three themes in urban remote sensing science: mapping, indices, and modeling. For mapping we describe the data sources, methods, and limitations of mapping urban boundaries, land use and land cover, population, temperature, and air quality. Second, we described how spectral information is manipulated to create comparative biophysical, social, and spatial indices of the urban environment. Finally, we focus how the mapped information and indices are used as inputs or parameters in models that measure changes in climate, hydrology, land use, and economics.

Deforestation in Myanmar has recently attracted much attention worldwide. This study examined spatio-temporal patterns of deforestation and forest carbon flux in Myanmar from 2001 to 2010 and environmental impacts at the regional scale using land products of the Moderate Resolution Imaging Spectroradiometer (MODIS). The results suggest that the total deforestation area in Myanmar was 21,178.8 km², with an annual deforestation rate of 0.81%, and that the total forest carbon release was 20.06 million tons, with an annual rate of 0.37%. Mangrove forests had the highest deforestation and carbon release rates, and deciduous forests had both the largest deforestation area and largest amount of carbon release. During the study period, the south and southwestern regions of Myanmar, especially Ayeyarwady and Rakhine, were deforestation hotspots (i.e., the highest deforestation and carbon release rates occurred in these regions). Deforestation caused significant carbon release, reduced evapotranspiration (ET), and increased land surface temperatures (LSTs) in deforested areas in Myanmar during the study period. Constructive policy recommendations are put forward based on these research results.

Land-use mapping is critical for global change research. In Central Arizona, U.S.A., the spatial distribution of land use is important for sustainable land management decisions. The objective of this study was to create a land-use map that serves as a model for the city of Maricopa, an expanding urban region in the Sun Corridor of Arizona. We use object-based image analysis to map six land-use types from ASTER imagery, and then compare this with two per-pixel classifications. Our results show that a single segmentation, combined with intermediary classifications and merging, morphing, and growing image-objects, can lead to an accurate land-use map that is capable of utilizing both spatial and spectral information. We also employ a moving-window diversity assessment to help with analysis and improve post-classification modifications.