Placed by distant Russian trolls, they didn’t aim to prop up one candidate or cause, but to turn Americans against one another.

The ads were cheaply made and full of threatening, vulgar language.

And, according to a sweeping new analysis of more than 2,500 of the ads, they were remarkably effective, eliciting clickthrough rates as much as nine times higher than what is typical in digital advertising.

“We found that fear and anger appeals work really well in getting people to engage,” said lead author Chris Vargo, an assistant professor of Advertising, Public Relations and Media Design at University of Colorado Boulder.

The study, published this week in Journalism and Mass Communication Quarterly, is the first to take a comprehensive look at ads placed by the infamous Russian propaganda machine known as the Internet Research Agency (IRA) and ask: How effective were they? And what makes people click on them?

While focused on ads running in 2016, the study’s findings resonate in the age of COVID-19 and the run-up to the 2020 election, the authors say.

“As consumers continue to see ads that contain false claims and are intentionally designed to use their emotions to manipulate them, it’s important for them to have cool heads and understand the motives behind them,” said Vargo.

For the study, Vargo and assistant professor of advertising Toby Hopp scoured 2,517 Facebook and Instagram ads downloaded from the U.S. House of Representatives Permanent Select Committee On Intelligence website. The committee made the ads publicly available in 2018 after concluding that the IRA had been creating fake U.S. personas, setting up fake social media pages, and using targeted paid advertising to “sow discord” among U.S. residents.

Using computational tools and manual coding, Vargo and Hopp analyzed every ad, looking for the inflammatory, obscene or threatening words and language hostile to a particular group’s ethnic, religious or sexual identity. They also looked at which groups each ad targeted, how many clicks the ad got, and how much the IRA paid.

Collectively, the IRA spent about $75,000 to generate about 40.5 million impressions with about 3.7 million users clicking on them — a clickthrough rate of 9.2%.

That compares to between .9% and 1.8% for a typical digital ad.

While ads using blatantly racist language didn’t do well, those using cuss words and inflammatory words (like “sissy,” “idiot,” “psychopath” and “terrorist”) or posing a potential threat did. Ads that evoked fear and anger did the best.

One IRA advertisement targeting users with an interest in the Black Lives Matter movement stated: “They killed an unarmed guy again! We MUST make the cops stop thinking that they are above the law!” Another shouted: “White supremacists are planning to raise the racist flag again!” Meanwhile, ads targeting people who sympathized with white conservative groups read “Take care of our vets; not illegals” or joked “If you voted for Obama: We don’t want your business because you are too stupid to own a firearm.”

Only 110 out of 2,000 mentioned Donald Trump.

“This wasn’t about electing one candidate or another,” said Vargo. “It was essentially a make-Americans-hate-each-other campaign.”

The ads were often unsophisticated, with spelling or grammatical errors and poorly photoshopped images. Yet at only a few cents to distribute, the IRA got an impressive rate of return.

“I was shocked at how effective these appeals were,” said Vargo.

The authors warn that they have no doubt such troll farms are still at it.

According to some news reports, Russian trolls are already engaged in disinformation campaigns around COVID-19.

“I think with any major story, you are going to see this kind of disinformation circulated,” said Hopp. “There are bad actors out there who have goals that are counter to the aspirational goals of American democracy, and there are plenty of opportunities for them to take advantage of the current structure of social media.”

Ultimately, the authors believe better monitoring, via both machine algorithms and human reviewers, could help stem the tide of disinformation.

“We as a society need to start seriously talking about what role the platforms and government should play in times like the 2020 election or during COVID-19 when we have a compelling need for high-quality, accurate information to be distributed,” said Hopp.

In the meantime, the pandemic, with over 400,000 confirmed cases worldwide already, is driving researchers to find safe and effective therapies for patients with COVID-19, and an antimalarial drug is potentially on the front lines of that effort. While new and repurposed drugs are being tested in clinical trials, some of these promising drugs are simultaneously being used off-label for compassionate use to treat patients.

Some of the medications being used to treat COVID-19 are known to cause drug-induced prolongation of the QTc of some people. The QTc is an indicator of the health of the heart’s electrical recharging system. Patients with a dangerously prolonged QTc are at increased risk for potentially life-threatening ventricular rhythm abnormalities that can culminate in sudden cardiac death.

“Correctly identifying which patients are most susceptible to this unwanted, tragic side effect and knowing how to safely use these medications is important in neutralizing this threat,” says Michael J. Ackerman, M.D., Ph.D., a Mayo Clinic genetic cardiologist. Dr. Ackerman is director of the Mayo Clinic Windland Smith Rice Comprehensive Sudden Cardiac Death Program.

A study published in Mayo Clinic Proceedings details more information about potential dangers and the application of QTc monitoring to guide treatment when using drugs that can cause heart rhythm changes. Dr. Ackerman is the senior author of the study.

Hydroxychloroquine is a long-standing preventive and treatment drug for malaria. It also is used to manage and minimize symptoms of inflammatory immune diseases, such as lupus and rheumatoid arthritis. In laboratory tests, hydroxychloroquine can prevent the SARS-CoV and SARS-CoV-2 viruses from attaching to and entering cells. If these antiviral abilities work the same way in animals and humans, the drug could be used to treat patients and limit the number of COVID-19 deaths.

On a cellular level, potential QT-prolonging medications, like hydroxychloroquine, block one of the critical potassium channels that control the heart’s electrical recharging system. This interference increases the possibility that the heart’s rhythm could degenerate into dangerous erratic heart beats, resulting ultimately in sudden cardiac death.

Accordingly, Mayo Clinic cardiologists and physician-scientists have provided urgent guidance on how to use a 12-lead ECG, telemetry or smartphone-enabled mobile ECG to determine the patient’s QTc as a vital sign to identify those patients at increased risk and how to ultimately minimize the chance of drug-induced sudden cardiac death.

“Right now, it is the Wild West out there, ranging from doing no QTc surveillance whatsoever and just accepting this potential tragic side effect as part of ‘friendly fire,’ to having ECG technicians going into the room of a patient with COVID-19 daily, exposing them to coronavirus and consuming personal protective equipment,” says Dr. Ackerman. “Here Mayo Clinic has stepped forward to provide timely and critical guidance.”

Guidelines for QTc monitoring during treatment

The antimalarial drugs chloroquine and hydroxychloroquine, as well as the HIV drugs lopinavir and ritonavir, all carry a known or possible risk of drug-induced ventricular arrhythmias and sudden cardiac death. Prior to starting treatment with these medications, it is important to get a baseline ECG to be able to measure changes. This starting point measurement could be from a standard 12-lead ECG, telemetry or a smartphone-enabled mobile ECG device. On Monday, March 20, the Food and Drug Administration (FDA) granted emergency approval of AliveCor’s Kardia 6L mobile ECG device as the only FDA-approved mobile device for QTc monitoring with COVID-19.

The mobile device’s ability to remotely provide the patient’s heart rhythm and QTc value does not require an extra ECG technician to take the measurement in person, thus saving increased exposure to COVID-19 and the need for more personal protective equipment.

Using the algorithm developed by Dr. Ackerman and colleagues, the potential risk of drug-induced arrhythmias can be rated and used to modify treatment accordingly. For example, patients with a baseline QTc value greater than or equal to 500 milliseconds and those that experience an acute QTc reaction with a QTc greater than or equal to 60 milliseconds from baseline after starting treatment with one or more QTc-prolonging drugs are at greatest risk for drug-induced arrhythmias. Simple QTc countermeasures can be implemented for patients with a cautionary “red light” QTc status if the decision is made to proceed with the intended COVID-19 therapies.

Information guides decisions

There are a number of considerations around the use of off-label drugs to treat COVID-19. The drugs may or may not be available in large enough supply to treat a worldwide pandemic, even at the current compassionate use stage of testing. It will take careful consideration of COVID-19 patients’ circumstances for treating clinicians and patients to decide on the use of drugs or drug combinations that may treat their infection, but which potentially could cause harmful drug-induced side effects.

Dr. Ackerman says that patients under 40 with mild symptoms and a QTc greater than or equal to 500 milliseconds may choose to avoid treatment altogether, as the arrhythmia risk may far outweigh the risk of developing COVID-19-related acute respiratory distress syndrome. However, in COVID-19 patients with a QTc greater than or equal to 500 milliseconds who have progressively worsening respiratory symptoms or are at greater risk of respiratory complications due to advanced age, immunosuppression or having another high-risk condition, the potential benefit of QTc-prolonging medicines may exceed the arrhythmia risk.

“Importantly, the vast majority of patients ? about 90% ? are going to be QTc cleared with a ‘green light go’ and can proceed, being at extremely low risk for this side effect,” says Dr. Ackerman.

Ultimately, the weighing of risks to benefits depends on whether hydroxychloroquine, with or without azithromycin, is truly an effective treatment against COVID-19.

“If it is, we hope that this simple QTc surveillance strategy, enabled by innovation and the FDA’s emergency approval, will help prevent or at least significantly minimize drug-induced ventricular arrhythmias and sudden cardiac death, particularly if the treatment is widely adopted and used to treat COVID-19,” says Dr. Ackerman.

Other contributors to the study are John Giudicessi, M.D., Ph.D., first author; Peter Noseworthy, M.D.; and Paul Friedman, M.D. — all Mayo Clinic cardiologists. Drs. Ackerman, Friedman and Noseworthy have a financial interest in AliveCor.

“This stage is really important, because the pair needs to be aligned for the exchange of genetic material between those two chromosomes,” says P. Jeremy Wang, a biologist in Penn’s School of Veterinary Medicine. “If anything goes wrong at this stage, it can cause a defect in meiosis and problems in the resulting sperm, leading to infertility, pregnancy loss, or birth defects.”

In a new paper in Science Advances, Wang and colleagues have identified an enzyme that plays a crucial role in maintaining this chromosomal pairing during the pachytene stage of meiosis. Without this protein, named SKP1, meiosis cannot proceed to metaphase, the next major developmental stage involved in generating sperm cells.

The finding may help overcome hurdles that have stood in the way of treating certain forms of male infertility, in which a man makes no sperm but in whom sperm’s precursor cells, spermatogonia, can be found.

“Reproductive technologies like in vitro fertilization have made a huge difference for infertile patients, but the male needs to have at least some sperm,” says Wang. “If the male has no sperm, then the only option is to use donor sperm. But if you can find these spermatogonia, the pre-meiotic germ cells, they could be induced to go through meiosis and make sperm. So SKP1 could be part of the solution to ensuring meiosis continues.”

Wang is also hopeful that his finding could aid in basic research on sperm development that his and many other labs pursue.

“Right now we use animals to do our research; we don’t have a cell culture system to produce sperm,” he says. “Manipulating SKP1 and the pathway in which it acts could allow us to set up an in vitro system to produce sperm artificially, which would be a boon for our studies.”

The publication represents nearly a decade of work, led by Wang’s postdoctoral researcher Yongjuan Guan, with major contributions from former postdoc Mengcheng Luo.

The team began focusing on SKP1 after conducting a screening test to look for proteins found in the area where the paired chromosomes come together during the pachytene stage of meiosis. From earlier studies, the researchers knew that SKP1 also plays a role in cell division in cells throughout the body, not just sperm and eggs. Without it, cells die.

That fact forced the Penn Vet team to get creative to understand the protein’s function. Unable to simply eliminate it, they created a model system in mice in which they could turn off the protein only in the germ cells and only in adulthood.

“Taking this inducible, germ-cell-specific model, we found that taking away SKP1 caused the chromosomes to prematurely separate,” says Wang.

While the normal alignment process in the pachytene stage takes six days in mice, in the cells that lost SKP1 the paired chromosomes separated far earlier.

Scientists had hypothesized the existence of a metaphase competence factor, or some protein required for a cell to enter metaphase. Wang believes that SKP1 is it.

While introducing a compound known as okadaic acid to sperm precursor cells can coax them into an early entrance to metaphase, cells lacking SKP1 did not progress to metaphase.

Experiments in developing eggs showed the researchers that SKP1 is also required for females to maintain viable eggs. Oocytes, the cells that develop through meiosis to form mature eggs, that lacked SKP1 developed misaligned chromosomes and many eventually were lost.

In future work, Wang and his colleagues want to dig deeper into the mechanism of action by which SKP1 works to ensure cells can progress to metaphase, with the idea of eventually manipulating it to find strategies for addressing infertility and innovative laboratory techniques.

“Now that we know SKP1 is required, we’re looking for the proteins it interacts with upstream and downstream so we can study this pathway,” says Wang.

The hydrogel beads, which have the consistency of gummy candy and are made with an ingredient used in processed foods, hold the promise for sustained cleanup of groundwater contaminated with dangerous and widely used volatile organic compounds; many of the compounds are listed by the Centers for Disease Control and Prevention as likely human carcinogens.

At multiple locations around the country, the chemicals are present at concentrations that far exceed state and federal standards for safe drinking water.

Among the contaminants addressed in the study are 1,1,1-trichloroethane, cis-1,2-dichloroethene, and 1,4-dioxane — degreasers commonly used by industry and the military. The chemicals can infiltrate groundwater through leaky underground storage tanks or runoff, or by simply being dumped on the ground as they were in past.

The new decontamination method, developed through a collaboration between the OSU College of Engineering and North Carolina State University, works because the microbes produce an enzyme that oxidizes the toxins when groundwater contaminants diffuse into the beads.

The result is a transformation of the contaminants into harmless compounds.

“We’ve created a process called long-term aerobic cometabolism, which is an enclosed, passive, self-sustaining system for groundwater remediation,” said OSU’s Lew Semprini, distinguished professor of environmental engineering and principal investigator on the study. “The beauty of this is that everything happens inside the beads.”

Current practices, Semprini explains, call for gaseous growth substrates such as propane and methane to be added directly to the subsurface. The substrates nourish indigenous microbes, which in turn produce enzymes that transform the contaminants to non-toxic byproducts.

Often, however, the growth substrates chemically compete for those crucial enzymes, which significantly inhibits the transformation process.

The new system eliminates that competition, freeing all of the enzyme to oxidize contaminants.

“We’ve flipped the paradigm on its head by putting the right microorganism inside hydrogel beads and supplying it with a slow-release food source,” Semprini said. “To my knowledge, this is the first time it’s been done.”

The study appears in Environmental Science: Processes & Impacts.

Semprini and his research team co-encapsulated the bacteria culture Rhodococcus rhodochrous and a slow-release growth substrate within hydrogel beads that they produced in the lab. The cylindrical beads, made of gellan gum, a common ingredient in processed foods, are 2 millimeters long.

As groundwater flows by the beads, the contaminants diffuse into the beads, where the slow release substrate reacts with groundwater to produce alcohol that sustains the Rhodococcus bacteria. The bacteria contain a monooxygenase enzyme that transforms the contaminants into harmless compounds, including carbon dioxide, water and chloride ions.

The purified water and the byproducts then diffuse out of the beads and rejoin the groundwater plume.

In bead-filled test columns supplied with a continuous flow of contaminated water, the system functioned continuously for more than 300 days (and counting) on the original growth substrate.

Semprini found that the beads remove more than 99% of the contaminants, and their concentrations declined from several hundred parts per billion to less than 1 part per billion.

The system’s longevity will depend mainly on how long the bacteria live, which is a factor of how long the growth substrate lasts. That has yet to be determined.

“It’s a question for future research,” Semprini said. “How do we make beads that last many years, or how do we develop systems that can easily be replaced?”

Current cometabolic remediation methods require regular additions of growth substrates to ensure that key microorganisms flourish, and that necessitates regular site monitoring, biochemical adjustments and related costs.

The next step is to scale up the system and conduct pilot studies in the field.

Semprini envisions several possibilities for deploying the beads. One option is mixing beads directly into contaminated subsurface material. Another is to dig a trench in the path of groundwater flow and fill it with beads, creating a permeable reactive barrier. A third possibility is packing beads into reactors, a simple form being mesh bags, that can be placed in wells.

“Everybody favors sustainability in this type of system: Can we just have something working in the subsurface without much maintenance?” Semprini said. “I think we’ve achieved that.”

The study was funded by the Department of Defense.