A Bioluminescent Biosensor for Detection of Mycotoxins in Food

3D artistic rendering of a NanoBiT assay, the system used in this study for detection of mycotoxins in food

Food contamination is a serious global health issue. According to the WHO, an estimated 600 million, almost 1 in 10 people globally, suffer from illness after eating contaminated food—and 420,000 die. Developing new technologies for more effective testing of food contaminants can help reduce that number and improve public health.

A recent application of bioluminescent technology could change the way we test for mycotoxins in the future. Dr. Jae-Hyuk Yu, Professor of Bacteriology at the University of Wisconsin-Madison, and his then graduate student, Dr. Tawfiq Alsulami, collaborated with Promega to develop a bioluminescent biosensor that enables simple and rapid detection of mycotoxins in food samples.

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The Work of Emmett Chappelle: Lighting Up the Search for Extraterrestrial Life

What do the workings of red blood cells, ensuring breathable air for astronauts, and scraping soil off NASA’s Viking spacecraft have in common? The sharp thinking of biochemist Emmett Chappelle.

Image of Emmett Chappelle working with other scientists.
Emmett Chappelle conducting research.

February is Black History Month in the US—a time to reflect on the contributions of African Americans in all fields and celebrate their accomplishments while recognizing the adversity they had to overcome in American society.

2021 also marks 30 years since the first firefly luciferase reporter vectors and detection reagents became available as products. There’s no better person to highlight this month than Emmett Chappelle, whose work with the luciferase reaction is still used for many applications today.

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Celebrating 30 Years of “Glo-ing” Research

This post is written by guest blogger, Amy Landreman, PhD, Sr. Product Manger at Promega Corporation.

In December of 1990, Promega first discussed the use of firefly luciferase (luc) as an emerging reporter technology in the article, Firefly Luciferase: A New Tool for Molecular Biologists. At the time, the gene coding chloramphenicol acetyltransferase (cat)  was most commonly used by researchers, but it was thought that the bioluminescent properties of firefly luciferase, extreme sensitivity and rapid simple detection, could make a significant difference in how molecular biologists tackled their research. Several months later, the first firefly luciferase reporter vectors and detection reagents became available as products, making this new technology more broadly accessible to the research community. Today firefly luciferase is no longer a “new tool”, with it and many other bioluminescent reporter technologies being standard elements of the modern research toolbox.

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Intranasal COVID-19 Vaccines: What the Nose Knows

COVID-19 vaccine distribution efforts are underway in several countries. Recently, the Serum Institute of India celebrated the nationwide rollout of its Covishield vaccine, kicking off the country’s largest ever vaccination program. Meanwhile, many other vaccines against the coronavirus that causes COVID-19 are in either preclinical studies or clinical trials. At present, 19 vaccine candidates are in Phase 3 clinical trials, while 8 vaccines have been granted emergency use authorization (EUA) in at least one country.

intranasal covid-19 vaccine coronavirus

In the US, mRNA vaccines from Pfizer/BioNTech and Moderna are in distribution. Adenoviral vector vaccines authorized for distribution include Oxford/AstraZeneca AZD1222 in the UK (Covishield in India) and Gamaleya Sputnik V in Russia. A third type of vaccine consists of inactivated coronavirus particles, such as those developed by Sinopharm and Sinovac in China.

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Bioluminescence and Biotechnology: Shining Nature’s Cool Light on Biology

Imagine you’re taking a refreshing night swim in the warm blue waters of Vieques in Puerto Rico. You splash into the surf and head out to some of the deeper waters of the bay, when what to your wondering eyes should appear, but blue streaks of light in water that once was clear. Do you need to get your eyes checked? Are you hallucinating? No! You’ve just happened upon a cluster of dinoflagellates, harmless bioluminescent microorganisms called plankton, that emit their glow when disturbed by movement. These dinoflagellates are known to inhabit waters throughout the world but are generally not present in large enough numbers to be noticed. There are only five ecosystems in the world where these special bioluminescent bays can be seen, and three of them are in Puerto Rico. 

Bioluminescent plankton exhibit a blue glow when disturbed.
Bioluminescent plankton in the ocean

But you don’t have to travel to Puerto Rico or swim with plankton to see bioluminescence. There are bioluminescent organisms all over the world in many unexpected places. There are bioluminescent mushrooms, bioluminescent sea creatures—both large and small (squid, jellyfish, and shrimp, in addition to the dinoflagellates)—and bioluminescent insects, to name a few. Bioluminescence is simply the ability of living things to produce light.

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Barking Up the Right Tree: Using NanoLuc to Screen for Canine Distemper Antivirals

Canine distemper virus (CDV) is a highly contagious pathogen that is the etiological agent responsible for canine distemper (CD), a systemic disease that affects a broad spectrum of both domestic dogs and wild carnivores. While there are commercially available vaccines for CDV that can provide immunity in vivo and protect canines from contracting CD, there is a strong demand for effective canine distemper antivirals to combat outbreaks. Such drugs remain unavailable to date, largely due to the laborious, time-consuming nature of methods traditionally used for high-throughput drug screening of anti-CDV drugs in vitro. In a recent study published in Frontiers in Veterinary Science, researchers demonstrated a new tool for rapid, high-throughput screening of anti-CDV drugs: a NanoLuc® luciferase-tagged CDV.

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Targeting Glioblastoma Cells by Packaging a Lentiviral Vector Inside a Zika Virus Coat

A recent article published in Cancers demonstrates a new method for targeting glial cells using a lentiviral packaging system that incorporated Zika virus envelope proteins. By using the reporter gene firefly luciferase, researchers demonstrated that a pseudotyped virus could infect cultured glioblastoma cells.

Introduction

Space-fill drawing of the outside of one Zika virus particle, and a cross-section through another as it interacts with a cell. The two main proteins of the viral envelope, the envelope proteins and membrane proteins, are shown in red and purple respectively. The lipid membrane of the envelope is shown in light lavender.The capsid proteins, in orange, are shown interacting with the RNA genome, in yellow, at the center of the virus. The cell-surface receptor proteins are in green, the cytoskeleton in blue, and blood plasma proteins in gold. Drawn by David Goodsell.
Space-fill drawing of the outside of one Zika virus particle, and a cross-section through another as it interacts with a cell. The two main proteins of the viral envelope, the envelope proteins and membrane proteins, are shown in red and purple respectively. The lipid membrane of the envelope is shown in light lavender. The capsid proteins, in orange, are shown interacting with the RNA genome, in yellow, at the center of the virus. The cell-surface receptor proteins are in green, the cytoskeleton in blue, and blood plasma proteins in gold. Drawn and copyright owned by David Goodsell.

Viruses enjoy a fearsome reputation. SARS-CoV-2 is only the latest infectious agent that has garnered attention by becoming a worldwide pandemic. Even the viral name suggests that SARS-CoV-2 was not the first of its type [SARS-CoV is the virus behind the severe acute respiratory syndrome (SARS) that spread worldwide in the early 2000s]. There are many different families of viruses (e.g., coronavirus for SARS-CoV-2 or lentiviruses for HIV-1) and each show a preference to the cell types they want to infect. By investigating the life cycle of viruses to better understand their mechanisms, researchers can discover new opportunities that may be exploited.

In 2015 and 2016, the virus that concerned health authorities was Zika virus (ZIKV). While this virus generally caused mild disease, the babies of women who were infected during pregnancy were at increased risk for microcephaly and other brain defects. These defects were traced back to Zika virus infecting nerve tissue, specifically, glial cells. This discovery provided an opportunity to explore how Zika virus might affect the brain tumor, glioblastoma multiforme (GMB), especially the glioblastoma stem cells (GSCs) that resist conventional treatment and contribute to the poor prognosis for GMB. Studies suggested that Zika virus infection prolonged survival in animal glioma models and selectively killed GSC with minimal effects on normal cells. In fact, the molecules used by ZIKV to enter cells were predominantly found on tumors, not normal cells. Knowing that the ZIKV envelope proteins prM and E provide the target specificity for glial cells, Kretchmer et al. wanted to explore if ZIKV envelope proteins substituted in lentivirus packaging systems would be able to enter glioblastoma cells.

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UWCCC Small Molecule Screening Facility Validates the Lumit™ Dx SARS-CoV-2 Immunoassay for High-Throughput SARS-CoV-2 Antibody Screening

Three researchers from the University of Wisconsin and the Small Molecule Screening Facility (SMSF) at the University of Wisconsin Carbone Cancer Center (UWCCC) have expanded their collaboration in new directions because of COVID-19. Before the pandemic, Gene Ananiev, PhD, Facility Manager of the SMSF, Tim Bugni, PhD, a Professor in the School of Pharmacy, and David Andes, MD, Professor of Medicine and Medical Microbiology and Immunology and Head of the Division of Infection Disease, worked together on antibiotic compound discovery and development, now they have added Covid-19-related projects to that list.

“It was kind of an interesting aside…” said David Andes “To try to see a need, fill a need.”

The need they saw was for tools that are necessary around any pandemic or infectious disease outbreak: Ways to quickly diagnose and manage those who are infected and ways to study the epidemiology of the disease—the distribution pattern and frequency, causes and risk factors for infection within a population. Specifically, the three were interested in an antibody test that could be used not only to understand the proportion of the population that might have already been infected with SARS-CoV-2, but that also could be used to evaluate the response to different vaccine candidates. 

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From Viral Outbreak to Vaccine Development: Our Top 10 Most Viewed Blog Posts of 2020

This illustration, created at the Centers for Disease Control and Prevention (CDC), reveals ultrastructural morphology exhibited by coronaviruses. Photo Credit:  Alissa Eckert, MS; Dan Higgins, MAM CDC It is one is used in several of our top 10 most viewed blogs of 2020
Illustration from CDC; Photo Credit: Alissa Eckert, MS; Dan Higgins, MAM

When you look at our top 10 most viewed blog posts of 2020, there’s no surprise that all relate to COVID-19. We have come a long way since the beginning of the year, thanks to tireless scientists and researchers around the globe. They have led the way in COVID-19 research, treatment, and testing. Let’s take a closer look at this top 10 list:

10. Tips to Maintain Physical Distance in the Lab 

The spread of COVID-19 forced us to adapt and adjust to new ways in life, in work, and for this blog post, in the lab. In response to the pandemic, some labs shut down completely. Others have stayed open, especially those involving coronavirus research. This post provides 10 helpful distancing tips for researchers to stay safe and productive while working in the lab.  

9. Investigation of Remdesivir as a Possible Treatment for SARS-2-CoV (2019 nCoV) 

Scientists have worked hard to determine possible treatment for COVID-19. This blog post focuses on Remdesivir (RDV or GS-5734), an encouraging treatment used for the first case in the United States. It provides an in-depth look at numerous studies and clinical trials on Remdesivir as treatment for COVID-19. One key finding is that RDV needed to be administered either before or shortly after infection to limit lung damage. 

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How A New SARS-CoV-2 Wastewater Testing Kit is Helping Campuses Reopen

The fall of 2020 was like no other, especially for universities. The COVID-19 pandemic hit most of the world in the spring, forcing schools and businesses to close. For months, people worked from home and schools switched to online classes. When fall came, universities had a difficult decision to make. Do they have students and staff come back to campus for in-person classes? With students living together in close proximity in dormitories, an outbreak could quickly get out of hand. How can the university monitor and control the spread of the virus to ensure everyone’s safety?

This was when Robert Brooks started getting calls. He’s the Technical Director and Operations Manager at Microbac Laboratories in Oak Ridge, Tennessee. Microbac is a network of privately owned laboratories that provide testing services for food products, environmental samples and the life science industry. Robert has been in the lab industry for 25 years and has established a reputation for taking on difficult problems. “We really try to go that extra mile to help clients solve their issues. That has made a name for us out there. When people have odd-ball issues, they give us a call cause we’re going to take a look at it from a couple different viewpoints and take a step-by-step approach,” he says.

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