ASU Scholarship Showcase

Autoantibodies refer to antibodies that target self-antigens, which can play pivotal roles in maintaining homeostasis, distinguishing normal from tumor tissue and trigger autoimmune diseases. In the last three decades, tremendous efforts have been devoted to elucidate the generation, evolution and functions of autoantibodies, as well as their target autoantigens. However, reports of these countless previously identified autoantigens are randomly dispersed in the literature. Here, we constructed an AAgAtlas database 1.0 using text-mining and manual curation. We extracted 45 830 autoantigen-related abstracts and 94 313 sentences from PubMed using the keywords of either ‘autoantigen’ or ‘autoantibody’ or their lexical variants, which were further refined to 25 520 abstracts, 43 253 sentences and 3984 candidates by our bio-entity recognizer based on the Protein Ontology. Finally, we identified 1126 genes as human autoantigens and 1071 related human diseases, with which we constructed a human autoantigen database (AAgAtlas database 1.0). The database provides a user-friendly interface to conveniently browse, retrieve and download human autoantigens as well as their associated diseases. The database is freely accessible at http://biokb.ncpsb.org/aagatlas/. We believe this database will be a valuable resource to track and understand human autoantigens as well as to investigate their functions in basic and translational research.

There are many proteomic applications that require large collections of purified protein, but parallel production of large numbers of different proteins remains a very challenging task. To help meet the needs of the scientific community, we have developed a human protein production pipeline. Using high-throughput (HT) methods, we transferred the genes of 31 full-length proteins into three expression vectors, and expressed the collection as N-terminal HaloTag fusion proteins in Escherichia coli and two commercial cell-free (CF) systems, wheat germ extract (WGE) and HeLa cell extract (HCE). Expression was assessed by labeling the fusion proteins specifically and covalently with a fluorescent HaloTag ligand and detecting its fluorescence on a LabChip[superscript ®] GX microfluidic capillary gel electrophoresis instrument. This automated, HT assay provided both qualitative and quantitative assessment of recombinant protein. E. coli was only capable of expressing 20% of the test collection in the supernatant fraction with ≥20 μg yields, whereas CF systems had ≥83% success rates. We purified expressed proteins using an automated HaloTag purification method. We purified 20, 33, and 42% of the test collection from E. coli, WGE, and HCE, respectively, with yields ≥1 μg and ≥90% purity. Based on these observations, we have developed a triage strategy for producing full-length human proteins in these three expression systems.

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.

We report a device to fill an array of small chemical reaction chambers (microreactors) with reagent and then seal them using pressurized viscous liquid acting through a flexible membrane. The device enables multiple, independent chemical reactions involving free floating intermediate molecules without interference from neighboring reactions or external environments. The device is validated by protein expressed in situ directly from DNA in a microarray of ~10,000 spots with no diffusion during three hours incubation. Using the device to probe for an autoantibody cancer biomarker in blood serum sample gave five times higher signal to background ratio compared to standard protein microarray expressed on a flat microscope slide. Physical design principles to effectively fill the array of microreactors with reagent and experimental results of alternate methods for sealing the microreactors are presented.

The purpose of the United Nations-guided process to establish Sustainable Development Goals is to galvanize governments and civil society to rise to the interlinked environmental, societal, and economic challenges we face in the Anthropocene. We argue that the process of setting Sustainable Development Goals should take three key aspects into consideration. First, it should embrace an integrated social-ecological system perspective and acknowledge the key dynamics that such systems entail, including the role of ecosystems in sustaining human wellbeing, multiple cross-scale interactions, and uncertain thresholds. Second, the process needs to address trade-offs between the ambition of goals and the feasibility in reaching them, recognizing biophysical, social, and political constraints. Third, the goal-setting exercise and the management of goal implementation need to be guided by existing knowledge about the principles, dynamics, and constraints of social change processes at all scales, from the individual to the global. Combining these three aspects will increase the chances of establishing and achieving effective Sustainable Development Goals.

Introduction: Juvenile idiopathic arthritis (JIA) is a heterogeneous disease characterized by chronic joint inflammation of unknown cause in children. JIA is an autoimmune disease and small numbers of autoantibodies have been reported in JIA patients. The identification of antibody markers could improve the existing clinical management of patients.

Methods: A pilot study was performed on the application of a high-throughput platform, the nucleic acid programmable protein array (NAPPA), to assess the levels of antibodies present in the systemic circulation and synovial joint of a small cohort of juvenile arthritis patients. Plasma and synovial fluid from 10 JIA patients was screened for antibodies against 768 proteins on NAPPAs.

Results: Quantitative reproducibility of NAPPAs was demonstrated with > 0.95 intra-array and inter-array correlations. A strong correlation was also observed for the levels of antibodies between plasma and synovial fluid across the study cohort (r = 0.96). Differences in the levels of 18 antibodies were revealed between sample types across all patients. Patients were segregated into two clinical subtypes with distinct antibody signatures by unsupervised hierarchical cluster analysis.

Conclusion: The NAPPAs provide a high-throughput quantitatively reproducible platform to screen for disease-specific autoantibodies at the proteome level on a microscope slide. The strong correlation between the circulating antibody levels and those of the inflamed joint represents a novel finding and provides confidence to use plasma for discovery of autoantibodies in JIA, thus circumventing the challenges associated with joint aspiration. We expect that autoantibody profiling of JIA patients on NAPPAs could yield antibody markers that can act as criteria to stratify patients, predict outcomes and understand disease etiology at the molecular level.

The Montreal Protocol is generally credited as a successful example of international cooperation in response to a global environmental problem. As a result, the production and consumption of ozone-depleting substances has declined rapidly, and it is expected that atmospheric ozone concentrations will return to their normal ranges toward the end of this century. This paper applies the social-ecological system framework and common-pool resource theory to explore the congruence between successful resolution of small-scale appropriation problems and ozone regulation, a large-scale pollution problem. The results of our analysis correspond closely to past studies of the Protocol that highlight the importance of attributes such as a limited number of major industrial producers, advances in scientific knowledge, and the availability of technological substitutes. However, in contrast to previous theoretical accounts that focus on one or a few variables, our analysis suggests that its success may have been the result of interactions between a wider range of SES attributes, many of which are associated with successful small-scale environmental governance. Although carefully noting the limitations of drawing conclusions from the analysis of a single case, our analysis reveals the potential for fruitful interplay between common-pool resource theory and large-scale pollution problems.

As part of an international collaboration to compare large-scale commons, we used the Social-Ecological Systems Meta-Analysis Database (SESMAD) to systematically map out attributes of and changes in the Great Barrier Reef Marine Park (GBRMP) in Australia. We focus on eight design principles from common-pool resource (CPR) theory and other key social-ecological systems governance variables, and explore to what extent they help explain the social and ecological outcomes of park management through time. Our analysis showed that commercial fisheries management and the re-zoning of the GBRMP in 2004 led to improvements in ecological condition of the reef, particularly fisheries. These boundary and rights changes were supported by effective monitoring, sanctioning and conflict resolution. Moderate biophysical connectivity was also important for improved outcomes. However, our analysis also highlighted that continued challenges to improved ecological health in terms of coral cover and biodiversity can be explained by fuzzy boundaries between land and sea, and the significance of external drivers to even large-scale social-ecological systems (SES). While ecological and institutional fit in the marine SES was high, this was not the case when considering the coastal SES. Nested governance arrangements become even more important at this larger scale. To our knowledge, our paper provides the first analysis linking the re-zoning of the GBRMP to CPR and SES theory. We discuss important challenges to coding large-scale systems for meta-analysis.

The mission of the DNASU Plasmid Repository is to accelerate research by providing high-quality, annotated plasmid samples and online plasmid resources to the research community through the curated DNASU database, website and repository (http://dnasu.asu.edu or http://dnasu.org). The collection includes plasmids from grant-funded, high-throughput cloning projects performed in our laboratory, plasmids from external researchers, and large collections from consortia such as the ORFeome Collaboration and the NIGMS-funded Protein Structure Initiative: Biology (PSI:Biology). Through DNASU, researchers can search for and access detailed information about each plasmid such as the full length gene insert sequence, vector information, associated publications, and links to external resources that provide additional protein annotations and experimental protocols. Plasmids can be requested directly through the DNASU website. DNASU and the PSI:Biology-Materials Repositories were previously described in the 2010 NAR Database Issue (Cormier, C.Y., Mohr, S.E., Zuo, D., Hu, Y., Rolfs, A., Kramer, J., Taycher, E., Kelley, F., Fiacco, M., Turnbull, G. et al. (2010) Protein Structure Initiative Material Repository: an open shared public resource of structural genomics plasmids for the biological community. Nucleic Acids Res., 38, D743–D749.). In this update we will describe the plasmid collection and highlight the new features in the website redesign, including new browse/search options, plasmid annotations and a dynamic vector mapping feature that was developed in collaboration with LabGenius. Overall, these plasmid resources continue to enable research with the goal of elucidating the role of proteins in both normal biological processes and disease.

A significant challenge of our time is conserving biological diversity while maintaining economic development and cultural values. The United Nations Educational, Scientific and Cultural Organization has established biosphere reserves within its Man and the Biosphere program as a model means for accomplishing this very challenge. The East Carpathians Biosphere Reserve (ECBR), spreading across Poland, Slovakia, and Ukraine, represents a large social-ecological system (SES) that has been protected under the biosphere reserve designation since 1998. We have explored its successes and failures in improving human livelihoods while safeguarding its ecosystems. The SES framework, which includes governance system, actors, resources, and external influences, was used as a frame of analysis. The outcomes of this protected area have been mixed; its creation led to national and international collaboration, yet some actor groups remain excluded. Implementation of protocols arising from the Carpathian Convention has been slow, while deforestation, hunting, erosion, temperature extremes, and changes in species behavior remain significant threats but have also been factors in ecological adaptation. The loss of cultural links and traditional knowledge has also been significant. Nevertheless, this remains a highly biodiverse area. Political barriers and institutional blockages will have to be removed to ensure this reserve fulfills its role as a model region for international collaboration and capacity building. These insights drawn from the ECBR demonstrate that biosphere reserves are indeed learning sites for sustainable development and that this case is exemplary in illustrating the challenges, but more importantly, the opportunities that arise when ensuring parallel care and respect for people and ecosystems through the model of transboundary protected areas around the world.