The Bell Burnell Observatory: 140 Years of Inspiring Scientific Curiosity

Posted on by Jordan Villanueva
The Bell Burnell Observatory, located at Promega Madison

On a hill at the top of the Promega Madison campus, an old observatory overlooks the city of Fitchburg, Wisconsin. Inside, cutting-edge telescopes are ready to give students and astronomers breathtaking views of the cosmos.

Over the past 140 years, this observatory has served as the first launchpad for storied careers in astronomy. Following a relocation, it gave a passionate community a home for their curiosity. Today, it supports modern research while also welcoming stargazers of all ages. It is now one of the oldest operational observatories in the United States. This is the Bell Burnell Observatory.

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A Different Kind of Sustainable Growth: What’s Happening in the Promega Garden

Posted on by Jordan Villanueva
Tomatoes growing in the Promega garden
Tomato season is ending soon, and Mike has already harvested 2,500 pounds from the Promega garden.

Summer is winding down at Promega Madison. Kids are heading back to school, sunset is creeping earlier, and a new cycle of academic research projects are ramping up. However, in the Promega garden, Master Gardener Mike Daugherty is still hard at work harvesting fresh produce that will soon become delicious meals in our cafeterias. As the seasons begin to change, I stopped by to learn what’s happening on the farm. Here are a few highlights that Mike shared.

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Improved FFPE Tissue Sample Processing with High-Throughput Automated DNA Extraction

Posted on by Jordan Nutting

In oncology, tissue biopsies are commonly fixed in formalin and embedded in paraffin (FFPE). These FFPE samples can be used with immunohistochemical or molecular analysis for identifying biomarkers that guide the diagnosis and therapeutic management of patients. This fixation technique allows long-term storage of samples but impacts the integrity of nucleic acids. This makes extracting DNA and RNA from FFPE tissues in sufficient quantity and quality for molecular analysis techniques such as NGS analyses challenging for molecular oncology laboratories.

“At Rennes University Hospital, we receive many lung cancer samples with little material available, or samples of poor quality. The nucleic acid extraction step is therefore critical to get good yield. We have seen that it had a direct impact on the success of downstream analysis,” said Dr. Alexandra Lespagnol. Lespagnol is the Technical Manager of the Molecular Genetics of Cancer core lab at the University Hospital of Rennes in France.

In order to accommodate the increasing number of samples that needed to be analyzed, the Molecular Genetics of Cancer core lab of the University Hospital of Rennes initiated an automation project for extracting DNA from FFPE tissues. The lab also wanted to improve sample tracking and reproducibility of their results.

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From Outside the Window to Inside the Cell: How Photography has Broadened Our View of the World

Posted on by Kelly Grooms

Photography. From the time the first image was captured almost 200 years ago, people have been using photography techniques to record, improve and expand their world.  Joseph Nicéphore Niépce is credited with capturing the very first photograph—a view of outside his window—using a technique called heliography. From that blurry, one-of-a-kind print to the stunning images captured today we can capture using our mobile phones, the advances in personal photography have been phenomenal. But progress has not been limited to capturing images of the world we see, scientists have leveraged these advances into new tools to capture images of the world we can’t see.

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Keeping Cool When It Hurts: An Engineered Alternative to Pharmacological Pain Management

Posted on by Michele Arduengo

Pain management, particularly long-term pain management, is a difficult problem. Opioids are effective at reducing pain, but they are addictive and carry with them the significant costs of addiction and overdose that affect not only the individual but society as well. 

Saltatory conduction in neurons. Localized cooling of neurons may provide a non-addictive method for pain management.

Recently, work presented in Science by Reeder and colleagues proposes an engineering solution to pain management: miniature, implantable devices that can deliver localized and reversible neural blocking to reduce pain. While such devices could deliver any kind of stimulus ranging from electrical to pharmacological, the researchers developed a device that temporarily blocks neural signals by cooling.

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How Fire Coral has an Edge Over Other Species in the Caribbean Reef

Posted on by Riley Bell

Coral reefs are the most productive marine ecosystem known, providing essential habitats and shelters for fish and other organisms. Additionally, they help protect coastlines, support economies, provide important food sources for local fisheries, and so much more. Coral reefs are ecologically essential—but are continuing to vanish. Fire coral (Millepora) brings new hope to this marine crisis due to their unusual ability to grow in two forms and survive under various habitat stresses.

Bladed fire coral (Millepora complanata) undersea, Caribbean Sea, Cuba, Playa Cueva de los peces
Bladed fire coral (Millepora complanata) undersea, Caribbean Sea, Cuba

What Is Fire Coral?

Fire coral has been around for millions of years and is most commonly found in sunny, shallow reefs. They tend to grow in tropical and subtropical waters with many thriving in different areas of the Caribbean Sea, one of the planet’s most biologically diverse ecosystems. Fire coral resembles typical stony corals but has a wicked sting that can cause burning skin reactions, reflecting their relationship as a close relative to jellyfish.

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Designing Better Therapeutic mAbs: An Assay for Rapid, Parallel Screening of Fc/ FcɣR Interactions

Posted on by Michele Arduengo

The first monoclonal antibody (mAb) was produced in a lab 1975, and the first therapeutic mAb was introduced in the United States to prevent kidney transplant rejection in 1986. The first mAb used in cancer treatment the anti-CD20 mAb, rituximab, was used to treat non-Hodgkin’s lymphoma and chronic lymphocytic leukemia. Today therapeutic mAbs have become a mainstay of cancer, autoimmune disease, and metabolic disease therapies and include HERCEPTIN® used to treat certain forms of breast cancer, Prolia used to treat bone loss in post-menopausal women, and Stelara used to treat autoimmune diseases like psoriatic arthritis and severe Crohn disease, among many others. Therapeutic mAbs bind targets with high specificity and affinity and they can recruit effector cells to drive target elimination through mechanisms such as antibody-dependent cellular cytotoxicity (ADCC) or antibody-dependent cellular phagocytosis (ADCP), making them highly specific, effective therapies.

3D rendering of a Lumit Assay which can be used for plate-based screening assay to measure the affinities of Fc interactions of therapeutic mAbs.
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Behind the Scenes at Promega with the Brazil Young Researchers Award Winners

Posted on by Jordan Villanueva

“What I’ve learned in science is that we don’t do things alone. Everything is connected,” says Marcos da Silva Regueira Neto.

Marcos is a post-doctoral research at the Federal University of Pernambuco in Recife, Brazil. His project is part of a large interdisciplinary study, so he is no stranger to collaboration and welcomes opportunities to gain knowledge outside his specialty. Earlier this year, Marcos travelled nearly 5,000 miles to take advantage of one such opportunity.  

In May 2022, Marcos and eight other young Brazilian scientists spent a week in the United States experiencing a unique behind-the-scenes dive into Promega. Their trip included stops in New York City, Madison, and Chicago. For most of the students, this was their first look into new areas where science could lead them.

“I’ve spent most of my life in academia,” Marcos says. “I want to see the other side – the industry side. I want to learn new things and expand my knowledge.”

Over the course of a week, the students presented their research project to Promega leaders, got hands-on experience with emerging technologies alongside the scientists who invented them, and played with human-sized proteins in a virtual reality space.

They also took a picture with a six-foot duck.

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CCNE1-Amplified Cancers Targeted Through Synthetic Lethality Involving PKMYT1 Kinase

Posted on by Jordan Nutting

For cancers that have proven challenging to target with traditional therapies, one emerging option is an approach called synthetic lethality. Synthetic lethality arises when inactivation of two gene products together lead to cell death but where inactivation of one does not (1, 2). Targeting a gene that is synthetic lethal to a cancer-related mutation creates an opening to specifically kill cancer cells while leaving healthy cells untouched.

In a recent study in Nature, scientists found that cells with amplification of CCNE1 are sensitive to inhibition of PKYMT1 kinase and identified a small molecule that is a selective inhibitor of PKYMT1 (3). When mice with tumor xenografts derived from CCNE1­-high cell lines were dosed with the drug, researchers observed significantly slower tumor growth, and in some cases where the drug was co-dosed with another chemotherapeutic, tumor growth was completely halted.

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Use of ProAlanase Digestion Increases Number of Identified Methylglyoxal (MGO)-Modified Proteins in Whole-Cell Lysates

Posted on by Michele Arduengo
space filling structural model methylglyoxal (MGO)
Methylglyoxal is responsible for post translational protein modifications, that result in advanced glycation endproducts (AGEs), which are associated with aging and disease.

Post-translational modifications (PTM) of proteins are essential for the function of many proteins, but aberrant modification of protein residues also can interfere with protein function. PTMs occur in two ways. Proteins may be modified through the activity of enzymes such as kinases, phosphorylases, glycosylases and others that add or remove specific chemical moieties to amino acid residues. PTMs can also result from non-enzymatic reaction between electrophilic compounds and nucleophilic arginine and lysine residues within a protein. Metabolites and metabolic by products produced during glycolysis, especially glyoxal and methylglyoxal (MGO), are examples of such electrophilic compounds. These compounds can react with arginine and lysine to produce advanced glycation end products (AGEs), which are biomarkers associated with aging and degenerative diseases such as Alzheimer disease, diabetes and others. MGO is also elevated in tumors that have switched from oxidative phosphorylation to glycolysis as their main energy production pathway.

Only limited information is available about site-specific MGO PTMs in mammal cells, and most studies have focused on measuring the amount of MGO modifications in a treatment scenario compared to a control. Donnellan and colleagues recently published work to identify specific MGO protein modifications.  They used a “bottom-up” proteomic analysis of WIL2-NS B lymphoblastoid whole-cell lysates to identify specific MGO-modified proteins. In particular, the group was looking for modifications in proteins that might explain how MGO activity contributes to aneuploidy.

For the study, 100µg of cellular protein extract was reduced with dithiothreitol and then alkylated with chloroacetamide. The sample was diluted to reduce urea concentration. Trypsin Gold was added and samples were digested for 8 hours at 37°C. Digestion was terminated by adding formic acid. For ProAlanase digestion, 20µg of protein was reduced, alkylated and diluted to reduce urea concentration before adding digesting with ProAlanase for 4 hours at 37°C.

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The authors identified 519 MGO-modified proteins.  Most of the modifications were identified in the trypsin digestion reactions; however, ProAlanase digestion increased the number of MGO modifications identified by approximately 25% (with less than 4% of the modification sites being detected in both the ProAlanase and trypsin digestion reactions. The authors suggest that ProAlanase increased sequence coverage to reveal sites not detected in the trypsin digestions. Therefore, they conclude that ProAlanase can be used along with trypsin digestion to increase the identification of MGO modifications.

ProAlanase can be used along with trypsin digestion to increase the identification of MGO modifications.

MGO-modified proteins from the WIL2-NS whole cell lysates included proteins involved in glycolysis, translation initiation, protein folding, mRNA splicing, cell-to-cell adhesion, heat response, nucleosome assembly, protein SUMOylation and the G2/M cell cycle transition. More work to further characterize the sites of these modifications and their potential effects on the function of the modified proteins is ongoing.

Read more about ProAlanase, a new site-specific endoprotease from Promega.

Literature Cited

Donnelian, L et al. (2022) Proteomic Analysis of Methylglyoxal Modifications Reveals Susceptibility of Glycolytic Enzymes to Dicarbonyl Stress Int. J. Mol. Sci. 23(7), 3689 doi.org/10.3390/ijms23073689