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 Quadrangles Av-11 and Av-12 on Vesta are located at the northern rim of the giant Rheasilvia south polar impact basin. The primary geologic units in Av-11 and Av-12 include material from the Rheasilvia impact basin formation, smooth material and different types of impact crater structures (such as bimodal craters, dark and bright crater ray material and dark ejecta material). Av-11 and Av-12 exhibit almost the full range of mass wasting features observed on Vesta, such as slump blocks, spur-and-gully morphologies and landslides within craters. Processes of collapse, slope instability and seismically triggered events force material to slump down crater walls or scarps and produce landslides or rotational slump blocks. The spur-and-gully morphology that is known to form on Mars is also observed on Vesta; however, on Vesta this morphology formed under dry conditions.

A variety of geologic landforms and features are observed within quadrangle Av-13 Tuccia in the southern hemisphere of Vesta. The quadrangle covers parts of the highland Vestalia Terra as well as the floors of the large Rheasilvia and Veneneia impact basins, which results in a substantial elevation difference of more than 40 km between the northern and the southern portions of the quadrangle. Measurements of crater size–frequency distributions within and surrounding the Rheasilvia basin indicate that gravity-driven mass wasting in the interior of the basin has been important, and that the basin has a more ancient formation age than would be expected from the crater density on the basin floor alone. Subsequent to its formation, Rheasilvia was superimposed by several mid-sized impact craters. The most prominent craters are Tuccia, Eusebia, Vibidia, Galeria, and Antonia, whose geology and formation ages are investigated in detail in this work. These impact structures provide a variety of morphologies indicating different sorts of subsequent impact-related or gravity-driven mass wasting processes. Understanding the geologic history of the relatively young craters in the Rheasilvia basin is important in order to understand the even more degraded craters in other regions of Vesta.

In this paper we present a time-stratigraphic scheme and geologic time scale for the protoplanet Vesta, based on global geologic mapping and other analyses of NASA Dawn spacecraft data, complemented by insights gained from laboratory studies of howardite–eucrite–diogenite (HED) meteorites and geophysical modeling. On the basis of prominent impact structures and their associated deposits, we propose a time scale for Vesta that consists of four geologic time periods: Pre-Veneneian, Veneneian, Rheasilvian, and Marcian. The Pre-Veneneian Period covers the time from the formation of Vesta up to the Veneneia impact event, from 4.6 Ga to >2.1 Ga (using the asteroid flux-derived chronology system) or from 4.6 Ga to 3.7 Ga (under the lunar-derived chronology system). The Veneneian Period covers the time span between the Veneneia and Rheasilvia impact events, from >2.1 to 1 Ga (asteroid flux-derived chronology) or from 3.7 to 3.5 Ga (lunar-derived chronology), respectively. The Rheasilvian Period covers the time span between the Rheasilvia and Marcia impact events, and the Marcian Period covers the time between the Marcia impact event until the present. The age of the Marcia impact is still uncertain, but our current best estimates from crater counts of the ejecta blanket suggest an age between ∼120 and 390 Ma, depending upon choice of chronology system used. Regardless, the Marcia impact represents the youngest major geologic event on Vesta. Our proposed four-period geologic time scale for Vesta is, to a first order, comparable to those developed for other airless terrestrial bodies.

We report on a preliminary global geologic map of Vesta, based on data from the Dawn spacecraft’s High-Altitude Mapping Orbit (HAMO) and informed by Low-Altitude Mapping Orbit (LAMO) data. This map is part of an iterative mapping effort; the geologic map has been refined with each improvement in resolution. Vesta has a heavily-cratered surface, with large craters evident in numerous locations. The south pole is dominated by an impact structure identified before Dawn’s arrival. Two large impact structures have been resolved: the younger, larger Rheasilvia structure, and the older, more degraded Veneneia structure. The surface is also characterized by a system of deep, globe-girdling equatorial troughs and ridges, as well as an older system of troughs and ridges to the north. Troughs and ridges are also evident cutting across, and spiraling arcuately from, the Rheasilvia central mound.

However, no volcanic features have been unequivocally identified. Vesta can be divided very broadly into three terrains: heavily-cratered terrain; ridge-and-trough terrain (equatorial and northern); and terrain associated with the Rheasilvia crater. Localized features include bright and dark material and ejecta (some defined specifically by color); lobate deposits; and mass-wasting materials. No obvious volcanic features are evident. Stratigraphy of Vesta’s geologic units suggests a history in which formation of a primary crust was followed by the formation of impact craters, including Veneneia and the associated Saturnalia Fossae unit. Formation of Rheasilvia followed, along with associated structural deformation that shaped the Divalia Fossae ridge-and-trough unit at the equator. Subsequent impacts and mass wasting events subdued impact craters, rims and portions of ridge-and-trough sets, and formed slumps and landslides, especially within crater floors and along crater rims and scarps. Subsequent to the formation of Rheasilvia, discontinuous low-albedo deposits formed or were emplaced; these lie stratigraphically above the equatorial ridges that likely were formed by Rheasilvia. The last features to be formed were craters with bright rays and other surface mantling deposits.

Executed progressively throughout data acquisition, the iterative mapping process provided the team with geologic proto-units in a timely manner. However, interpretation of the resulting map was hampered by the necessity to provide the team with a standard nomenclature and symbology early in the process. With regard to mapping and interpreting units, the mapping process was hindered by the lack of calibrated mineralogic information. Topography and shadow played an important role in discriminating features and terrains, especially in the early stages of data acquisition.

Vesta is a unique, intermediate class of rocky body in the Solar System, between terrestrial planets and small asteroids, because of its size (average radius of ∼263 km) and differentiation, with a crust, mantle and core. Vesta’s low surface gravity (0.25 m/s²) has led to the continual absence of a protective atmosphere and consequently impact cratering and impact-related processes are prevalent. Previous work has shown that the formation of the Rheasilvia impact basin induced the equatorial Divalia Fossae, whereas the formation of the Veneneia impact basin induced the northern Saturnalia Fossae. Expanding upon this earlier work, we conducted photogeologic mapping of the Saturnalia Fossae, adjacent structures and geomorphic units in two of Vesta’s northern quadrangles: Caparronia and Domitia. Our work indicates that impact processes created and/or modified all mapped structures and geomorphic units. The mapped units, ordered from oldest to youngest age based mainly on cross-cutting relationships, are: (1) Vestalia Terra unit, (2) cratered highlands unit, (3) Saturnalia Fossae trough unit, (4) Saturnalia Fossae cratered unit, (5) undifferentiated ejecta unit, (6) dark lobate unit, (7) dark crater ray unit and (8) lobate crater unit. The Saturnalia Fossae consist of five separate structures: Saturnalia Fossa A is the largest (maximum width of ∼43 km) and is interpreted as a graben, whereas Saturnalia Fossa B-E are smaller (maximum width of ∼15 km) and are interpreted as half grabens formed by synthetic faults. Smaller, second-order structures (maximum width of <1 km) are distinguished from the Saturnalia Fossae, a first-order structure, by the use of the general descriptive term ‘adjacent structures’, which encompasses minor ridges, grooves and crater chains. For classification purposes, the general descriptive term ‘minor ridges’ characterizes ridges that are not part of the Saturnalia Fossae and are an order of magnitude smaller (maximum width of <1 km vs. maximum width of ∼43 km). Shear deformation resulting from the large-scale (diameter of <100 km) Rheasilvia impact is proposed to form minor ridges (∼2 km to ∼25 km in length), which are interpreted as the surface expression of thrust faults, as well as grooves (∼3 km to ∼25 km in length) and pit crater chains (∼1 km to ∼25 km in length), which are interpreted as the surface expression of extension fractures and/or dilational normal faults. Secondary crater material, ejected from small-scale and medium-scale impacts (diameters of <100 km), are interpreted to form ejecta ray systems of grooves and crater chains by bouncing and scouring across the surface. Furthermore, seismic shaking, also resulting from small-scale and medium-scale impacts, is interpreted to form minor ridges because seismic shaking induces flow of regolith, which subsequently accumulates as minor ridges that are roughly parallel to the regional slope. In this work we expand upon the link between impact processes and structural features on Vesta by presenting findings of a photogeologic, structural mapping study which highlights how impact cratering and impact-related processes are expressed on this unique, intermediate Solar System body.