ASU Scholarship Showcase

Human protein diversity arises as a result of alternative splicing, single nucleotide polymorphisms (SNPs) and posttranslational modifications. Because of these processes, each protein can exists as multiple variants in vivo. Tailored strategies are needed to study these protein variants and understand their role in health and disease. In this work we utilized quantitative mass spectrometric immunoassays to determine the protein variants concentration of beta-2-microglobulin, cystatin C, retinol binding protein, and transthyretin, in a population of 500 healthy individuals. Additionally, we determined the longitudinal concentration changes for the protein variants from four individuals over a 6 month period. Along with the native forms of the four proteins, 13 posttranslationally modified variants and 7 SNP-derived variants were detected and their concentration determined. Correlations of the variants concentration with geographical origin, gender, and age of the individuals were also examined. This work represents an important step toward building a catalog of protein variants concentrations and examining their longitudinal changes.

Proteins can exist as multiple proteoforms in vivo, as a result of alternative splicing and single-nucleotide polymorphisms (SNPs), as well as posttranslational processing. To address their clinical significance in a context of diagnostic information, proteoforms require a more in-depth analysis. Mass spectrometric immunoassays (MSIA) have been devised for studying structural diversity in human proteins. MSIA enables protein profiling in a simple and high-throughput manner, by combining the selectivity of targeted immunoassays, with the specificity of mass spectrometric detection. MSIA has been used for qualitative and quantitative analysis of single and multiple proteoforms, distinguishing between normal fluctuations and changes related to clinical conditions. This mini review offers an overview of the development and application of mass spectrometric immunoassays for clinical and population proteomics studies. Provided are examples of some recent developments, and also discussed are the trends and challenges in mass spectrometry-based immunoassays for the next-phase of clinical applications.

Introduction: Apolipoprotein C-III (apoC-III) regulates triglyceride (TG) metabolism. In plasma, apoC-III exists in non-sialylated (apoC-III_0a without glycosylation and apoC-III[subscript 0b] with glycosylation), monosialylated (apoC-III₁) or disialylated (apoC-III₂) proteoforms. Our aim was to clarify the relationship between apoC-III sialylation proteoforms with fasting plasma TG concentrations.

Methods: In 204 non-diabetic adolescent participants, the relative abundance of apoC-III plasma proteoforms was measured using mass spectrometric immunoassay.

Results: Compared with the healthy weight subgroup (n = 16), the ratios of apoC-III_0a, apoC-III_0b, and apoC-III₁ to apoC-III₂ were significantly greater in overweight (n = 33) and obese participants (n = 155). These ratios were positively correlated with BMI z-scores and negatively correlated with measures of insulin sensitivity (S[subscript i]). The relationship of apoC-III₁ / apoC-III₂ with S_i persisted after adjusting for BMI (p = 0.02). Fasting TG was correlated with the ratio of apoC-III_0a / apoC-III₂ (r = 0.47, p<0.001), apoC-III_0b / apoC-III₂ (r = 0.41, p<0.001), apoC-III₁ / apoC-III₂ (r = 0.43, p<0.001). By examining apoC-III concentrations, the association of apoC-III proteoforms with TG was driven by apoC-III_0a (r = 0.57, p<0.001), apoC-III_0b (r = 0.56. p<0.001) and apoC-III₁ (r = 0.67, p<0.001), but not apoC-III₂ (r = 0.006, p = 0.9) concentrations, indicating that apoC-III relationship with plasma TG differed in apoC-III₂ compared with the other proteoforms.

Conclusion: We conclude that apoC-III_0a, apoC-III_0b, and apoC-III₁, but not apoC-III₂ appear to be under metabolic control and associate with fasting plasma TG. Measurement of apoC-III proteoforms can offer insights into the biology of TG metabolism in obesity.

The impetus for discovery and evaluation of protein biomarkers has been accelerated by recent development of advanced technologies for rapid and broad proteome analyses. Mass spectrometry (MS)-based protein assays hold great potential for in vitro biomarker studies. Described here is the development of a multiplex mass spectrometric immunoassay (MSIA) for quantification of apolipoprotein C-I (apoC-I), apolipoprotein C-II (apoC-II), apolipoprotein C-III (apoC-III) and their proteoforms. The multiplex MSIA assay was fast (∼40 min) and high-throughput (96 samples at a time). The assay was applied to a small cohort of human plasma samples, revealing the existence of multiple proteoforms for each apolipoprotein C. The quantitative aspect of the assay enabled determination of the concentration for each proteoform individually. Low-abundance proteoforms, such as fucosylated apoC-III, were detected in less than 20% of the samples. The distribution of apoC-III proteoforms varied among samples with similar total apoC-III concentrations. The multiplex analysis of the three apolipoproteins C and their proteoforms using quantitative MSIA represents a significant step forward toward better understanding of their physiological roles in health and disease.

Serum Amyloid A (SAA) is an acute phase protein complex consisting of several abundant isoforms. The N- terminus of SAA is critical to its function in amyloid formation. SAA is frequently truncated, either missing an arginine or an arginine-serine dipeptide, resulting in isoforms that may influence the capacity to form amyloid. However, the relative abundance of truncated SAA in diabetes and chronic kidney disease is not known.

Methods: Using mass spectrometric immunoassay, the abundance of SAA truncations relative to the native variants was examined in plasma of 91 participants with type 2 diabetes and chronic kidney disease and 69 participants without diabetes.

Results: The ratio of SAA 1.1 (missing N-terminal arginine) to native SAA 1.1 was lower in diabetics compared to non-diabetics (p = 0.004), and in males compared to females (p<0.001). This ratio was negatively correlated with glycated hemoglobin (r = −0.32, p<0.001) and triglyceride concentrations (r = −0.37, p<0.001), and positively correlated with HDL cholesterol concentrations (r = 0.32, p<0.001).

Conclusion: The relative abundance of the N-terminal arginine truncation of SAA1.1 is significantly decreased in diabetes and negatively correlates with measures of glycemic and lipid control.

The oxidative modification of apolipoprotein A-I’s methionine148 (M148) is associated with defective HDL function in vitro. Multiple reaction monitoring (MRM) is a mass spectrometric technique that can be used to quantitate post-translational modifications. In this study, we developed an MRM assay to monitor the abundance ratio of the peptide containing oxidized M148 to the native peptide in ApoA-I. Measurement of the oxidized-to-unoxidized-M148 ratio was reproducible (CV < 5%). The extent of methionine M148 oxidation in the HDL of healthy controls, and type 2 diabetic participants with and without prior cardiovascular events (CVD) were then examined. The results suggest a significant increase in the relative ratio of the peptide containing oxidized M148 to the unmodified peptide in the HDL of participants with diabetes and CVD (p < 0.001), compared to participants without CVD. Monitoring the abundance ratio of the peptides containing oxidized and unoxidized M148 by MRM provides a means of examining the relationship between M148 oxidation and vascular complications in CVD.

Previous studies in building energy assessment clearly state that to meet sustainable energy goals, existing buildings, as well as new buildings, will need to improve their energy efficiency. Thus, meeting energy goals relies on retrofitting existing buildings. Most building energy models are bottom-up engineering models, meaning these models calculate energy demand of individual buildings through their physical properties and energy use for specific end uses (e.g., lighting, appliances, and water heating). Researchers then scale up these model results to represent the building stock of the region studied.

Studies reveal that there is a lack of information about the building stock and associated modeling tools and this lack of knowledge affects the assessment of building energy efficiency strategies. Literature suggests that the level of complexity of energy models needs to be limited. Accuracy of these energy models can be elevated by reducing the input parameters, alleviating the need for users to make many assumptions about building construction and occupancy, among other factors. To mitigate the need for assumptions and the resulting model inaccuracies, the authors argue buildings should be described in a regional stock model with a restricted number of input parameters. One commonly-accepted method of identifying critical input parameters is sensitivity analysis, which requires a large number of runs that are both time consuming and may require high processing capacity.

This paper utilizes the Energy, Carbon and Cost Assessment for Buildings Stocks (ECCABS) model, which calculates the net energy demand of buildings and presents aggregated and individual- building-level, demand for specific end uses, e.g., heating, cooling, lighting, hot water and appliances. The model has already been validated using the Swedish, Spanish, and UK building stock data. This paper discusses potential improvements to this model by assessing the feasibility of using stepwise regression to identify the most important input parameters using the data from UK residential sector. The paper presents results of stepwise regression and compares these to sensitivity analysis; finally, the paper documents the advantages and challenges associated with each method.

Construction waste management has become extremely important due to stricter disposal and landfill regulations, and a lesser number of available landfills. There are extensive works done on waste treatment and management of the construction industry. Concepts like deconstruction, recyclability, and Design for Disassembly (DfD) are examples of better construction waste management methods. Although some authors and organizations have published rich guides addressing the DfD's principles, there are only a few buildings already developed in this area. This study aims to find the challenges in the current practice of deconstruction activities and the gaps between its theory and implementation. Furthermore, it aims to provide insights about how DfD can create opportunities to turn these concepts into strategies that can be largely adopted by the construction industry stakeholders in the near future.

The United State generates the most waste among OECD countries, and there are adverse effects of the waste generation. One of the most serious adverse effects is greenhouse gas, especially CH4, which causes global warming. However, the amount of waste generation is not decreasing, and the United State recycling rate, which could reduce waste generation, is only 26%, which is lower than other OECD countries. Thus, waste generation and greenhouse gas emission should decrease, and in order for that to happen, identifying the causes should be made a priority. The research objective is to verify whether the Environmental Kuznets Curve relationship is supported for waste generation and GDP across the U.S. Moreover, it also confirmed that total waste generation and recycling waste influences carbon dioxide emissions from the waste sector. The annual-based U.S. data from 1990 to 2012 were used. The data were collected from various data sources, and the Granger causality test was applied for identifying the causal relationships. The results showed that there is no causality between GDP and waste generation, but total waste and recycling generation significantly cause positive and negative greenhouse gas emissions from the waste sector, respectively. This implies that the waste generation will not decrease even if GDP increases. And, if waste generation decreases or recycling rate increases, the greenhouse gas emission will decrease. Based on these results, it is expected that the waste generation and carbon dioxide emission from the waste sector can decrease more efficiently.

As the construction continue to be a leading industry in the number of injuries and fatalities annually, several organizations and agencies are working avidly to ensure the number of injuries and fatalities is minimized. The Occupational Safety and Health Administration (OSHA) is one such effort to assure safe and healthful working conditions for working men and women by setting and enforcing standards and by providing training, outreach, education and assistance. Given the large databases of OSHA historical events and reports, a manual analysis of the fatality and catastrophe investigations content is a time consuming and expensive process. This paper aims to evaluate the strength of unsupervised machine learning and Natural Language Processing (NLP) in supporting safety inspections and reorganizing accidents database on a state level. After collecting construction accident reports from the OSHA Arizona office, the methodology consists of preprocessing the accident reports and weighting terms in order to apply a data-driven unsupervised K-Means-based clustering approach. The proposed method classifies the collected reports in four clusters, each reporting a type of accident. The results show the construction accidents in the state of Arizona to be caused by falls (42.9%), struck by objects (34.3%), electrocutions (12.5%), and trenches collapse (10.3%). The findings of this research empower state and local agencies with a customized presentation of the accidents fitting their regulations and weather conditions. What is applicable to one climate might not be suitable for another; therefore, such rearrangement of the accidents database on a state based level is a necessary prerequisite to enhance the local safety applications and standards.