The human microbiome, the bustling cooperative of all the microscopic creatures that naturally colonize in and on our bodies, wields a surprising amount of influence over many of the unseen processes that are critical to our overall health and wellness. Over the course of decades, we have learned that this is particularly true for the microbes that reside in our gastrointestinal tract, collectively known as our gut microbiota.
Our gut microbiota is constantly communicating with our bodies, though our relationship with our gut can feel like trying to have a conversation with someone who only speaks a language we do not know or understand—you can take an educated guess at what they are saying based on their expressions and gestures, but the true message and meaning behind their actions is not always discernable. So while we can feel that someone in our gut is unhappy when we have a tummy ache, the true mechanism behind exactly who is unhappy and why, is not as obviously deduced or understood.
What if there was a tool that could help us more easily interpret the language of our microbiota, giving us the means to both better understand our microbiomes as well as to detect biomarkers of various diseases? Recent studies have shown that such a solution may be (quite literally) right under our noses: our breath.
Mosquitos are the deadliest animal on earth—not because of the itchy bites they leave behind, but because of the diseases those bites can spread. Of these diseases, malaria, is the most widespread, killing 619,000 people in 2021 (1). Almost half of the world’s population live at risk of malaria (2). In humans, malaria is caused by certain species of single-cell micro-organisms belonging to the genus Plasmodium (3), which are transmitted by anopheline mosquitos.
Controlling malaria has proven challenging. Vaccines have yielded incomplete protection, and insecticides that once were successful at control mosquito populations are becoming less effective as the insects develop resistance. Finally, Plasmodium parasites themselves have developed resistance to leading anti-malaria drugs (2).
A New Weapon In The Fight Against Malaria
Approaches that target the disease-causing Plasmodium organisms—inside the mosquito and before they are transmitted to humans—could provide as effective way forward. In the past, researchers have explored leveraging genetically modified bacterium to kill or inhibit Plasmodium development within their mosquito host. However, using genetically altered bacteria makes wide-spread adoption of these techniques problematic. A recent study published in Science describes the discovery and early investigative results using a naturally occurring bacterial strain that inhibits Plasmodium spread (2). The bacteria, Delftia tsuruhatensis TC1, was isolated from a mosquito population that unexpectedly became resistant to Plasmodium infection (2).
Once the bacterium was identified as the cause of Plasmodium inhibition, the researchers tested how easily the bacteria was to introduce into naïve mosquitos and how effective it was at disrupting infection. To do this, they colonized female mosquitos by feeding them a sugar and bacterium solution and then Plasmodium-infected blood. Bacterial colonization occurred in almost all the mosquitos offered the sugar and bacterium food. Initially, bacterial colonization numbers were low, but they increased 100-fold following the blood meal.
Inhibiting Oocyte Formation Disrupts Cycle of Infection
Investigation into how D. tsuruhatensis inhibits Plasmodium infection showed that it inhibits oocyte formation within the gut, and this inhibition lasts for at least 16 days. Specifically, the inhibition is the result of a secreted compound called harmane, which is a small hydrophobic methylated b-carboline (2). When harmane is secreted in the guts of mosquitos it inhibits Plasmodium parasite development. The researchers further found that feeding harmane alone to mosquitos, or allowing it to be absorbed through direct contact produced the same results, but the inhibitory effects only lasted a few days (2).
No matter how harmane is introduced into the gut (directly or through bacterial colonization), the inhibition of oocyte formation results in a decrease in infectivity. Only one third (33%) of mice bitten by Plasmodium-infected, D. tsuruhatensis-colonized mosquitos become infected. This contrasts sharply with the 100% infection rate seen with mice bitten by non-colonized, Plasmodium-infected mosquitos (2). Further testing the researchers also showed that D. tsuruhatensis is not transferred during feeding, suggesting that that bacterium is unlikely to in introduced into mammals through colonized mosquitos.
To investigate how colonization and infection rates would correlate in a ‘real world’ environment, the researchers used a large (10 × 10 × 5 meter) enclosure that replicated the mosquitos’ natural environment. Once again, the mosquitos were colonized with D. tsuruhatensis through overnight feeding of the sugar and bacterium solution. They found ~75% of the mosquitos were colonized by D. tsuruhatensis in this time period.They also found that larvae reared in water seeded with D. tsuruhatensis experienced 100% colonization. In both scenarios, Plasmodium oocyte development was disrupted just as it had been in the laboratory-raise population (2).
Finally, the researchers found that D. tsuruhatensis colonization doesn’t occur between individuals between parent and offspring. For controlling Plasmodium, this means that inoculation with D. tsuruhatensis would require ongoing maintenance. However, it also decreases the risk of a contaminated strain being amplified uncontrollably if released, making it less risky.
Malaria mitigation and control requires a multipronged effort. Using naturally occurring, symbiotic, microbes such as D. tsuruhatensis is one approach that shows promise. There is still a lot of work to be done before this bacterium could be used outside of a controlled environment, including understanding how the bacterium might interact with other plants and animals from the same ecosystem.
Monitoring and quantifying drug-target binding in a live-cell setting is important to bridging the gap between in vitro assay results and the phenotypic outcome, and therefore represents a crucial step in target validation and drug development (1). The NanoBRET™ Target Engagement (TE) assay is a biophysical technique that enables quantitative assessment of small molecule-target protein binding in live cells. This live-cell target engagement assay uses the bioluminescence resonance energy transfer (BRET) from a NanoLuc® luciferase-tagged target protein and a cell-permeable fluorescent tracer that reversibly binds the target protein of interest. In the presence of unlabeled test compound that engages the target protein, the tracer is displaced, and a loss of BRET signal is observed. Due to the tight distance constraints for BRET, the signal measured is specific to the target fused to NanoLuc® luciferase.
Promega offers over 400 ready-to-use assays for multiple target classes, including kinases, E3 ligases, RAS, and many others. For targets that do not have an existing NanoBRET™ TE assay, Promega offers NanoBRET™ dyes, NanoLuc® cloning vectors, and NanoBRET™ detection reagents to develop novel NanoBRET™ TE assays.
One critical component in the development of novel NanoBRET™ TE assay is the creation of the cell-permeable fluorescent tracers (NanoBRET™ tracers) against the target protein of interest. The tracers are bifunctional, consisting of a NanoBRET™-compatible fluorophore and a target-binding moiety connected by a linker. While the NanoBRET™ 590 dyes have demonstrated consistently robust cell permeability and optimal spectral overlap with NanoLuc® for BRET, a ligand capable of binding to the target protein of interest needs to be identified to generate a NanoBRET™ tracer.
What Are DNA-Encoded Libraries?
DNA-Encoded Libraries, (DELs), have emerged as powerful tools for discovering small molecule ligands to target proteins of interest at an unprecedented scale. . owing to the ability of a DEL to enable the synthesis of larger libraries of compounds and to target proteins without any prior structural knowledge of the proteins or their ligands (2). Because each member of a DEL contains a DNA barcode and a small molecule separated by a linker, DEL is primed for discovering leads within therapeutic modalities that rely on bifunctional chemistry, such as proteolysis targeting chimeras (PROTACs). Since NanoBRET™ tracers are also bifunctional, ligands identified from DEL selections could serve as ideal candidates for developing novel NanoBRET™ tracers that can enable NanoBRET™ TE assays for new targets.
This year ushered in a series of intense weather events that impacted communities across the globe: record-breaking heat waves; super-charged cyclones; intense flooding; coastal waters hitting a balmy 38°C (1–4). Attributing extreme weather to climate change has become the norm when reporting on these seemingly more frequent and intense events. But beyond simply acknowledging weather to be more violent or destructive than it was in the past, how is it that climate experts are able to determine if increasing greenhouse gas levels are the culprit behind these extreme weather events? The answers can be found in climate attribution science.
Microblogging is a form of blogging characterized by a shortened format and frequent posting schedule. Instead of personal websites, microblogs reside on social media platforms or apps, making them accessible to interact with and post on smartphones. Microblogs focus on interacting with audiences directly. With the ability to reply to or repost content, microblogging is more conversational and collaborative with audiences than long-form writing.
After its founding in 2006, Twitter (recently renamed “X” by its new owner) quickly became the face of microblogging platforms. Users publish content to the platform in posts of 280 characters that can include images, gifs, videos, and what the platform is most known for: hashtags. Hashtags enable users to search the platform by topic to connect with or follow other users who are writing about those topics. Users can also interact with each other by liking or retweeting tweets, which posts them to their own account. The open forum discussion style makes it possible for individuals to share their stories, offering first-hand accounts of breaking news and fueling political movements such as the Women’s March and Black Lives Matter.
It has been more than 100 years since Dr. William B. Coley, known today as the “Father of Immunotherapy,” made the first recorded attempt to mobilize the immune system as a means of treating cancer (9). Decades later, the discovery of T cells and the vital role they play in the immune system set the groundwork for many new immunotherapy treatments, such as those involving monoclonal antibodies, cytokines, CAR T cells, and checkpoint inhibitors.
In the murky depths of the ocean live some of the smartest and most unusual creatures to inhabit the earth. Octopuses are known for their sucker covered tentacles and chameleon-like abilities to change color, pattern and shape to blend it with their environment. The changes aren’t limited to just their appearance. A new study published in Cell reveals that they can change their brains as well (1). The study found that octopuses recode their brain in response to environmental temperature changes using RNA editing.
What do you wear to a job interview at a biotechnology company? How should your resume be formatted? What questions do you ask to ensure the role is a good fit?
“My mentor guided me through job applications, including helping me identify the things that were important to me in a job,” says Jazmin Santiesteban. “While we were talking about those things, she asked if I would be interested in applying to Promega.”
Jazmin received the D.O.O.R.S. Scholarship in 2021, before her senior year at Lawrence University. That scholarship program helped Jazmin develop new skills and cultivate connections that eventually led her to a job at Promega after graduation.
“I love it so far,” she says. “I don’t know where my career may take me, but right now I want to build a longer future at Promega.”
Insects are a keystone species in the animal kingdom, often providing invaluable benefits to terrestrial ecosystems and useful services to mankind. While many of them are seen as pests (think mosquitos), others are important for pollination, waste management, and even scientific research.
Insect biotechnology, or the use of insect-derived molecules and cells to develop products, is applied in a diverse set of scientific fields including agricultural, industrial, and medical biotechnology. Insect cells have been central to many scientific advances, being utilized in recombinant protein, baculovirus, and vaccine and viral pesticide production, among other applications (5).
Therefore, as the use of insect cells becomes more widespread, understanding how they are produced, their research applications, and the scientific products that can be used with them is crucial to fostering further scientific advancements.
Primary Cell Cultures and Cell Lines
In general, experimentation with individual cells, rather than full animal models, is advantageous due to improved reproducibility, decreased space requirements, less ethical concerns, and a reduction in expense. This makes primary cell cultures and cell lines essential contributors to basic scientific research.
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