Category Archives: Outreach Project

Studying DNA has never been easier…Thanks to eVITTA!

There’s one thing that every single one of us has in common with each other and all other living beings. It’s the fact that we carry DNA in our cells. DNA is an important molecule that has all the inherited information about how a living thing will look and function. A strand of DNA is like an extremely long sentence that uses only four letters – in fact, the length of the human DNA is 600 billion letters long! Can you imagine how hard and time consuming it would be to read? That is why scientists at The University of British Columbia (UBC) have developed a new and exciting application called easy Visualization and Inference Toolbox for Transcriptome Analysis, or eVITTA for short. eVITTA simplifies analyzing RNA (a type of DNA) and not only makes the process more efficient, but improves scientists’ understanding of its information as well.

The kind of DNA information that eVITTA works with is called the “transcriptome”. The transcriptome is the full set of RNA, which are the copies of DNA, within a cell. Analyzing the information in the transcriptome is crucial; for the past few decades, it has been one of the most used techniques for investigating diseases and their mechanisms.

The analysis of transcriptome data, however, is very tedious and time consuming. You have to retrieve the data, examine it, and then compare it with other transcriptome datasets to draw a conclusion. This involves several steps and different types of programs, which can be inefficient. eVITTA was created to combine the many steps of transcriptome analysis into one simple, user-friendly interface. This speeds up the process of transcriptome analysis immensely!

The tedious process of analyzing long strands of DNA is simplified with eVITTA.
Image Credit: CI Photos/Shutterstock.com

evitta was born out of excess data!

One of the many topics that sparked our interest was understanding the circumstances surrounding the creation of eVITTA. As Dr. Yan of UBC’s Taubert Lab puts it:

This whole project was born out of our need to pass on data. We have a lot of transcriptome data from past years that no one has gotten around to analyzing…. so during the pandemic, we started digging into those data and developing visualization modules and we realized we can actually make this into an app so that we can feed more data and generate visualizations

The team behind eVITTA

To discuss the different aspects of eVITTA and to delve deeper into this project, UBC’s Shayan Abbaszadeh sat down for a virtual interview with PhD candidate Judith Yan from the department of Cell and Developmental Biology at UBC. Dr. Yan is one of the many faces behind eVITTA at the Taubert lab and has worked tirelessly to bring this idea into fruition. Our podcast describes her role in realizing the gaps in efficient transcriptome analysis and building eVITTA with the rest of the Taubert Lab during the COVID-19 pandemic. Outside of eVITTA, Dr. Yan’s lab work in the Taubert Lab usually involves using model organisms such as roundworms to study stress responses and applying that knowledge to better understand human diseases. 

How does eVITTA make analyzing RNA so simple?

To grasp the scope of this research, first and foremost, it is important to understand transcriptome analysis. Dr. Yan describes transcriptome analysis as a powerful tool that examines how RNA, a copy of DNA, is used in a cell, tissue or organism. This involves taking out the whole RNA from a sample, getting it sequenced (i.e. decoded), and subsequently obtaining information from that data.

An exploration of gene patterns is one of the main aspects involved in effective transcriptome analysis.  A gene is a section of DNA that carries a specific piece of information. A crucial aspect of analysis is understanding that some genes are turned on at higher rates than others. According to Dr. Yan, once there is a count for these different genes, the numbers can be interpreted to reveal useful information about “what some of these genes are doing” and “what processes and gene sets are actually being changed“. Finally, there needs to be effective visualization techniques of the different data sets and the data needs to be validated against previously published data. 

All of these functions are achieved through eVITTA; a user-friendly, web-based interface that streamlines the multiple steps of transcriptome profiling. Watch this short video to become familiar with the 3 modules of eVIITA; easy-GEO, easyGSEA, and easyVizR, and realize what effective transcriptome analysis looks like!

The challenges of transcriptome analysis and how evitta addresses them

In our podcast, we focused extensively on the motivations behind the creation of eVITTA, specifically in relation to challenges associated with transcriptome analysis and how eVITTA deals with these challenges in ways that previous methods could not. Dr. Yan alludes to the use of the ‘overrepresentation approach’ (ORA) in previous technologies that has the flaw of only being able to represent a small subset of gene changes. As Dr. Yan puts it: “you’re missing out  on a lot of information and biologically… you are not able to capture the less severe changes. eVITTA on the other hand, focuses on entire sets of genes instead of just one gene, allowing scientists to observe gene changes across the board. 

Additionally, previous technologies did not allow for the organization of data because “oftentimes it is very tedious to do multiple comparison because you have many different subsections that you want to look at“. eVITTA prevents the dumping and mislabeling of gene data which expedites the process of discovering important biological patterns. This platform has already been proven to be highly effective in studies involving severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and roundworms (C.elegans). 

Dr. Yan’s research using eVITTA involves the starvation response of C.elegans roundworms. (credit: Taubert lab)

what does the future hold for eVITTA?

Our podcast also discusses the future of eVITTA and the team’s plans for further expansion. Dr. Yan and her team plan to add more algorithms to analyze the expression of different genes at a more in-depth level. The team also want to add more visualization options to provide more choice for gene dataset analysis. Finally, they are planning to update eVITTA’s databases by adding more datasets to the application. These ideas and more are discussed in our podcast below!

As transcriptome profiling provides major insight regarding diseases and illnesses, the optimization eVITTA can provide may be increasingly vital for today’s society. For instance, if transcriptome analysis can be conducted more efficiently, crucial findings in disease mechanisms may be discovered sooner. As a result, treatments and health regulations can be created more quickly, potentially saving lives and preventing disease spread. Additionally, eVITTA’s user-friendly and web-based interface makes transcriptome profiling accessible to more biologists around the world and would therefore greatly benefit the research community.

– Heather Cathcart, Kaushali Ghosh, Parham Asli, Shayan Abbaszadeh

 

 

 

Enzymatic Browning in Granny Smith Apples

Introduction

Browning is the darkening of the flesh that occurs shortly after fruits such as apples and pears are cut, exposing the flesh to air. Although browning is not toxic to humans since the pigment is composed of melanin, browning makes the fruit unsightly and unappetizing to eat . The main cause of browning is polyphenol oxidase (PPO), an enzyme that catalyzes the first two steps of converting the amino acid tyrosine into melanin in the presence of oxygen. Physical and mechanical stresses from slicing fruit physically damage the cellular structure of the flesh, which catalyzes PPO activity as it becomes exposed to oxygen .

Why Do Apples Turn Brown After You Cut Them? | Let's Talk Sciencefrom lets Talk science

what is an effective tool to delay apple browning?

Past studies have investigated the effects of different consumable solutions on browning, and citric acid, found naturally in fruit juices, was identified to be a moderate to a high inhibitor of browning . Citric acid is able to slow the onset of browning by lowering the pH of PPO’s environment so that the pH is outside the optimal pH range of 6-7 required for PPO to oxidize the flesh . Although other agents such as chelators and antioxidants can also inhibit PPO through reduction and inhibition, citric acid is more commonly found in foods, especially fruit juices that, when applied to apples, are more likely to preserve the apples’ taste

ReaLemon 100% Lemon Juice, 15 Fl Oz Bottle, 1 Count - Walmart.comfrom Walmart

Despite citric acid being an effective tool for delaying browning, the concentration at which its effects last for an extended period of time is unknown. Therefore, lemon juice was chosen since it is known to be composed of approximately 6% citric acid . Lemon juice also contains other acids, like ascorbic acid and malic acid, but their concentrations are negligible since citric acid comprises about 95% of the acid content of lemon juice . Granny smith apples were used as they are a common fruit consumed in households and are known to brown quickly after being sliced. It was hypothesized that lemon juice will delay the onset of browning due to the citric acid making the juice’s pH too acidic for polyphenol oxidase to initiate the conversion of tyrosine into melanin. If lemon juice delays browning, then the surface area of browning that appears on apple slices over a set period of time would decrease as the concentration of lemon juice the apple is exposed to increases.

Conclusion

This result shows that  increasing lemon juice concentration results in the decrease of browning due to the low pH of the juice compared to the pH that browns apple flesh. These results provide insight for future studies to find more effective anti-browning agents and to further investigate environmental temperatures to avoid to delay browning.

—–Chenyang Luo

Reference

Son, S. M., Moon, K. D., & Lee, C. Y. (2001). Inhibitory effects of various antibrowning agents on apple slices. Food Chemistry, 73(1), 23-30

Tinello, F., & Lante, A. (2018). Recent advances in controlling polyphenol oxidase activity of fruit and vegetable products. Innovative Food Science & Emerging Technologies, 50, 73–83. https://doi.org/10.1016/j.ifset.2018.10.008 

Tortoe, C., Orchard, J., & Beezer, A. (2007). Prevention of enzymatic browning of apple cylinders using different solutions. International Journal of Food Science & Technology, 42(12), 1475-1481.

Yapo, B. M. (2009). Lemon juice improves the extractability and quality characteristics of pectin from yellow passion fruit by-product as compared with commercial citric acid extractant. Bioresource Technology, 100(12), 3147–3151. https://doi.org/10.1016/j.biortech.2009.01.039