Devin Bowes

SARS-CoV-2 Monitoring in Wastewater as an Early Warning Signal for COVID-19 Presence in Communities

Presented By Devin Bowes, Arizona State University

The COVID-19 global pandemic caused by the virus Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has highlighted several challenges within our healthcare system, such as accessible testing and rapid reporting. Wastewater-based Epidemiology (WBE) has been proven to serve as an effective, near real-time tool to understand human health and behavior at population-scale by analyzing raw sewage for human biomarkers excreted via urine and/or feces. The study within showcases the first ever longitudinal investigation of monitoring SARS-CoV-2 in municipal wastewater at the neighborhood level to track trends throughout the COVID-19 global pandemic. Untreated wastewater samples were collected from within the sewer collection system throughout Tempe, Arizona and analyzed for SARS-CoV-2 using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). Preliminary results indicate peak loadings of the virus observed in wastewater precede reported peaks of positive new clinical cases, COVID-related hospitalizations, and COVID-related deaths. This study holds great promise for the future of population-level diagnostics; acting as an early warning system for emerging infectious diseases and allowing more time for hospital preparation, resource deployment, and targeted educational interventions throughout communities.

Prasiddhi Gyawali

An R-Shiny app for trait data processing

Presented by Prasiddhi Gyawali, Sonoran Science Academy – Tucson

The Functional Trait Resource for Environmental Studies (FuTRES) project works with vertebrate trait data from individuals intending to make data more accessible and interoperable. Since data are collected by different people using different methods, it can be hard for everyone to understand exactly what the data mean. To solve this problem, FuTRES works to write programs that can be used to easily standardize datasets. Standardization is done by getting the data from the CyVerse Discovery Environment and transferring them into RStudio where functions are applied. Code is written for a specific dataset then generalized so that it functions with other datasets. Code can always be extended or further generalized to fit another need. An R-Shiny app contains the code, providing a user-friendly interface.

Eli Lefkowitz

Automated physiological analysis of Engineered Heart Tissues from Human Induced Pluripotent Stem Cells

Presented by Eli Lefkowitz, Catalina Foothills High School

Cardiovascular disease is responsible for one in every four deaths. To understand these deaths, cardiomyocytes can be generated from a patient’s blood by first converting blood into a stem cell state called induced pluripotent stem cells (iPSCs). Cardiomyocyte derived iPSCs (hiPSC-CM) can be assembled into 3D constructs that resemble the heart called engineered heart tissues (EHTs). While it is possible to perform cardiovascular disease modeling in a dish, the tools to analyze the function of hiPSC-CMs remains laborious. To meet this need, our lab developed a software (MATLAB based) to quantify cardiac physiological parameters (peak amplitude, peak duration, and faster rising rate) in an automated and highly accurate manner. Further deployment of our software will be used to understand how mutations lead to cardiovascular disease.

Shaun Karakkattu

Shaun Karakkattu, Basis High School, Mesa, AZ

Inhibiting the Proliferation of Patient-Derived Glioblastoma Multiforme (GBM) cells by activating Estrogen Receptor beta using estradiol and IGF-1

 

Glioblastoma multiforme (GBM) is the most aggressive and common type of primary brain tumor. Patients with GBM have a median survival rate between 12-14 months. There are sex-based differences that exist in the occurance of GBM with men having a 60% higher chance of developing a tumor indicating that differences in the function of Estrogen Receptor Beta (ERβ), a known tumor suppressor, may influence tumor progression in GBM. Considering ERβ function, it was hypothesized that 17β-estradiol and IGF-1, estrogen receptor agonists, may promote ERβ function and inhibit the proliferation of GBM cells in vitro. 17β-estradiol and IGF-1 were applied to a patient-derived GBM cell line and MDA-MB-453, breast cancer cell line, in-vitro. The concentrations tested ranged from 2 to 20 µg/mL of 17β-estradiol and IGF-1. After 48 hrs of exposure, GBM cell proliferation was analyzed through a trypan blue exclusion assay. The results revealed that cell proliferation was inhibited by the estrogen receptor agonists in the patient-derived GBM cells and promoted in the MDA-MB-453 cell line which served as a positive control. In conclusion, this study presents evidence that ERβ function may indeed inhibit GBM proliferation. This study presents a potential approach to the treatment of GBM. As this study continues, more steroid hormone receptors will be tested like the Androgen Receptor which may also cause the inhibition of cell proliferation in GBM.

Aris Zhu

Aris Zhu, Hamilton High School, Chandler, Arizona

CASA: A Novel Intracanal Medicament for Endodontic Infections

Currently, calcium hydroxide (CaOH) is used as an intracanal medicament to treat root canal infections. However, CaOH paste is ineffective against persistent bacteria Enterococcus faecalis and fungus Candida albicans, which may give rise to health conditions including heart valve infection or periodontitis, respectively. Furthermore, the high pH of CaOH causes dental pulp necrosis, which delays tissue healing. To improve antimicrobial properties and limit cytotoxicity, salicylic acid was added in a 3.7:1 mass ratio to create a neutral paste to disinfect the canal. Using a modified disk diffusion antibiotic sensitivity test, CASA was plated with common endodontic microbes in a root canal infection to determine its antimicrobial activity. CASA produced larger zones of inhibition than CaOH for all species tested, indicating that CASA is the more efficacious antimicrobial agent. Notably, CASA yielded an inhibition zone 2.76 times that of CaOH in the presence of E. faecalis. Cytotoxicity studies and observation of dental pulp stem cells (DPSCs) under fluorescence microscopy indicated a high tolerance for DPSCs for CASA with a measured IC50 of 0.25 mg/ml, a far higher dose than tissue would be exposed to during standard treatment. Viability staining with E. faecalis and C. albicans confirmed the antimicrobial properties of CASA precipitate. CASA was found to be dosage dependent with increasing concentration resulting in greater bacterial lethality against S. aureus and E. coli. Because CASA was found to have greater antimicrobial effect and biocompatibility to dental pulp, CASA has the potential to replace CaOH in treatment for recurrent root canal infections.

Nikita Kumari

Nikita Kumari, Arizona State University (Graduate Student)

Diving Deep into the Red: Novel Cyanine Dyes for Super-Resolution Imaging

Fluorescent probes help biologists illuminate the inner workings of cancer cells. In this work, we study the photophysical properties of a novel class of far-red fluorescent probes that could allow live tissue to be imaged in greater depth and detail than is possible with current techniques. One of the limitations for super-resolution fluorescence microscopy techniques is the low brightness of the fluorophore which dictates signal-to-noise ratio for the fluorescence detection. The low brightness of the fluorophore could be because of various non-radiative decay pathways from the excited state to the ground state of the molecule which competes with the radiative pathway, decreasing the fluorescence quantum yield. The most prominent non-radiative pathway for cyanine fluorophores is the ability of an excited-state electron to go from trans (fluorescent form) to cis (non-fluorescent) state which is known as photoisomerization. We attempt to eliminate this trans-cis photoisomerization to happen by rigidifying the backbone of the cyanine. We applied this strategy on pentamethine cyanine dye which is a far-red cyanine fluorophore. The photophysical characterization of the newly synthesized restricted dye in comparison to traditional pentacyanine showed that the installation of the ring system restricts the photoisomerization, showed by temperature-independent emission, and solvent viscosity independent properties. The resulting molecule exhibit the characteristic features of conformational restraint, including improved fluorescence quantum yield and extended lifetime. The new rigidified molecule was found to have desired photophysical properties and improved the quality of super-resolution image obtained.

Elizabeth Fear

Elizabeth Fear, Horizon Honors High School (research conducted at the University of Arizona)

Effect of Hydration on Rhodopsin Activation

Rhodopsin, responsible for vision under dim light, is a prominent member of the G-Protein Coupled Receptor (GPCR) protein class which is targeted by a third of all pharmaceuticals. Two states of rhodopsin exist in physiological equilibrium: active and inactive. We hypothesized that hydration plays a crucial role in the shift to the active conformation. The fraction of active rhodopsin is quantified via UV-visible absorbance spectroscopy upon light exposure in the presence of various dehydrating osmolytes that generate osmotic pressure. We found that water does play an influential role in rhodopsin activation and that dehydration with osmotic stress favors the inactive state. As countless physiological processes involve GPCRs, greater insight into the relationship between water and rhodopsin activation can enable treatments of GPCR dysfunction across human anatomy.

Swapnika Raola

Swapnika Raola, University of Southern California (Undergraduate Student)

End-point PCR Analysis Using Capillary Flow as an Alternative to Gel Electrophoresis

We are finding a faster alternative to gel electrophoresis for template DNA identification. Using the same primer, bacteria strains Escherichia coli K-12 and O157:H7 form different amplicon lengths. We believe that amplicon length can affect capillary flow through interfacial tension.

Thus, we ask, “Does differing amplicon length and interfacial tension affect the capillary flow of  escherichia coli K-12 and Escherichia coli O157:H7 DNA solution?” If so, bacteria can now be identified through analysis of capillary flow, a process that takes two minutes rather than gel electrophoresis’s two hours. We found there are significant differences in capillary flow between the two strains and found that interfacial tension is probably the mechanism. Therefore, researchers can use our capillary flow model as a faster alternative to gel electrophoresis.

Hersh Nanda

Hersh Nanda, BASIS Chandler/ASU Science Program (High School Student)

An innovative polydimethylsiloxane microfluidic biosensing platform for rapid detection of viruses

The objective of this research project was to develop an innovative procedure for the fabrication of a versatile PDMS (polydimethylsiloxane) microfluidic device that is capable of detecting viruses and small molecules. The detection of these analytes is accomplished by integrating a gold biosensor with the PDMS microfluidic device and using a process called surface detection – a process wherein the gold biosensor allows glycoproteins (secreted by viruses) and small molecules to bond to it. The detection of the small molecules is directly observable while the detection of a virus is by inference by examining the glycoprotein(s) that are associated with the virus. This device falls in the category of lab-on-a-chip (LOC) devices that integrate one or several laboratory functions on a single highly miniaturized device.

The three main steps in the design procedure were: fabrication of gold biosensor, development of PDMS mold (or, PDMS mold making), and device validation (through a process called sensing).

After the device was produced, tests were run using LabSpec software to validate the biosensor capability and sensitivity in detecting viruses such as Ebola virus (through prior empirical data) and small molecules such as cannabidiol (CBD). This device, by virtue of its design, is capable of detecting any virus as long as the virus secretes a glycoprotein, and there is a molecule that is able to bond that glycoprotein to the biosensor. Therefore, this device is potentially capable of detecting influenza virus, SARS-CoV, hepatitis C virus, and even the novel COVID-19.

This device has broad application in biomedicine and offers a several benefits which include but are not limited to versatility in detection of diverse analytes (including new viruses), ability to detect analytes with small sample size, increased efficiency (less time to fabricate the device and quickly scale production to large volumes), lower production cost, device portability in the field, and ease of use for healthcare professionals.

The conclusions from this project are also applicable to the current COVID-19 pandemic, in which countries are dealing with shortages in testing kits for diagnosis. This project demonstrates that there is a less expensive and more efficient method for rapidly diagnosing infectious diseases, which can significantly enhance the ability of countries to rapidly detect and isolate infected people during pandemics and save human lives.

Note: This project was conducted through the ASU-SCENE program under the supervision and guidance of Assistant Professor Dr. Chao Wang, and a PhD student, MD Ashif Ikbal at the Nanoscience and Biotechnology labs at Arizona State University.