Culture

A team from the Department of Agronomy at the UCO has demonstrated, through field tests carried out during 8 agricultural seasons, that foliar feeding with fertilizer increases the concentration of zinc in wheat more than if it is applied to the soil

Micronutrient deficiencies pose health problems for a third of the world's population. Worldwide, zinc deficits are more problematic in the rural areas of developing countries, where diets are largely limited to vegetable products grown in soils suffering from low nutrient availability. Biofortification, the process of bolstering the nutritional value of crops by increasing the concentration of vitamins and minerals in them, has arisen as a remedy for this problem.

In the search for solutions, the Edaphology Unit of the María de Maeztu Excellence Unit in the Department of Agronomy at the University of Cordoba (DAUCO), headed by the researcher Antonio R. Sánchez Rodríguez, has spent 8 years searching for the best biofortification strategy in terms of applying zinc to wheat grown in calcareous soils in southern Spain.

Between 2012 and 2019 this team tested different methods to biofortify wheat in 11 field trials in zinc-deficient soils. The effects of applying different doses of fertilizer to the soil (up to 10 kg per hectare) were evaluated, in addition to the results of applying different doses of zinc by spraying the plant in various phenological stages of the wheat's growth.

While application to the soil was not very effective, foliar application, or feeding, was shown to be a very efficient strategy to increase the zinc content in plants, "augmenting the concentration in grains up to 50%," says the researcher. That is, foliar application was shown to be much more effective, as, with just a tenth of the product (1.28 kg per hectare) better results were obtained than when applied to the soil.

Taking into account the variety of wheat, this direct application to plants was more effective after the start of growth or during flowering.

Nourishing the plant itself, and not the soil, thus, was demonstrated to be an effective way to tackle the problem of zinc deficits in calcareous soils in the short term. In addition, if at some point wheat were purchased based on its nutritional content, growers could see increases in their profits.

This solution "is very valuable for places where there is no other source of zinc in diets, although it would entail adding another task to wheat cultivation, or combining it with the application of other phytosanitary treatments" notes Sánchez.

Predicting wheat yields after fertilization with zinc

On this project the team from the Edaphology Unit also looked for a soil indicator that would help predict the response of durum wheat to zinc fertilization, in terms of its yields. However, they found that conditions in the field make it very difficult to verify this parameter, and that a simple indicator cannot predict this response.

While at the laboratory level some indicators could be defined, in the field this task is difficult, since it is highly dependent on factors such as precipitation, and would require many more years of study.

New York, NY (April 9, 2021) - Sets of genes associated with resistance to immunotherapy in patients with metastatic urothelial cancer of the bladder have been identified and validated by researchers at Mount Sinai. In a study published in Clinical Cancer Research, the team uncovered gene signatures representing adaptive immunity and pro-tumorigenic inflammation that were responsible for sensitivity or resistance to immune checkpoint inhibitors, drugs that help the body's immune system recognize and attack cancerous cells.

"These findings enabled us to identify potential biomarkers in patients who are less likely to respond favorably to immune checkpoint inhibitors, as well as new combination therapeutic approaches that might overcome such resistance in those patients," says senior author Matthew Galsky, MD, Professor of Medicine (Hematology and Medical Oncology), Icahn School of Medicine at Mount Sinai.

Significantly, the findings demonstrated that the balance between adaptive immunity and pro-tumorigenic inflammation in individual tumor microenvironments--reflected by these two gene signatures--best predicted response or resistance to immune checkpoint blockade. The researchers then identified specific cells in the tumor microenvironment associated with resistance to immune checkpoint blockade, and potential targets for therapies designed to overcome resistance.

For decades, standard treatment for metastatic urothelial cancer of the bladder has been platinum-based chemotherapy, though the landscape has changed dramatically in recent years with the advent of PD-1 and PD-L1 immune checkpoint inhibitors. This therapeutic breakthrough has had its limits, though: only 20 to 25 percent of patients with bladder cancer respond to treatment, which has set off an intense hunt by biomedical scientists for mechanisms of resistance.

"Using RNA sequencing data from two clinical trials, and single cell RNA sequencing data from a cohort of bladder tumors, we identified a subset of genes and immune cells associated with adaptive immunity and improved checkpoint inhibitor outcomes, and a subset associated with pro-tumorigenic inflammation and resistance to PD-1/PD-L1 blockade in patients with urothelial cancer," Dr. Galsky says.

The research is among the first to use both bulk and single-cell RNA sequencing of human bladder tumors to study resistance to immunotherapy. Bulk sequencing examines the mix of genes expressed by every individual cell within a tumor, while single-cell sequencing--a technique increasingly important in cancer research--zeroes in on gene expression by each individual cell, which yields unprecedented knowledge of the complexity and heterogeneity of cells that comprise tumors.

Through this combination of RNA sequencing, researchers learned, for example, that the balance of adaptive immunity and pro-tumorigenic inflammation within the tumor microenvironment can determine PD-1/PD-L1 resistance in urothelial cancer. Adaptive immunity is the body's ability to recognize and respond to specific foreign invaders, while pro-tumorigenic inflammation is a counterproductive response of the immune system that can ultimately fuel growth and progression of cancer.

"If the tumor microenvironment is weighted more toward adaptive immunity, there's a better chance of positive outcomes from immunotherapy," explains Dr. Galsky, who is Associate Director of Translational Research and Co-Director of the Bladder Cancer Center of Excellence at The Tisch Cancer Institute. "On the other hand, if the tumor microenvironment is leaning toward pro-tumorigenic inflammation, then PD-1/PD-L1 checkpoint inhibitors alone are unlikely to be successful, and new combination approaches may be needed."

Mount Sinai researchers coined the term "2IR Score" to measure that balance.

From its comprehensive RNA analysis, the team identified not just potential biomarkers to treatment resistance, but a specific subset of white blood cells known as myeloid phagocytic cells that are linked to pro-tumorigenic inflammation and, hence, resistance. As such, they serve as prospective targets for therapeutic approaches that combine immunotherapies like PD-1/PD-L1 blockade with drugs designed to overcome the resistance conferred by myeloid cells. Those novel combination strategies are now being incorporated into future clinical trials.

"Our research shows that a specific cellular state of myeloid cells underlying pro-tumorigenic inflammation account for resistance to immune checkpoint blockade in a very large percentage of patients with urothelial bladder cancer," Dr. Galsky says. "This is an important finding which we believe can lead to a better focus and direction for developing effective combination therapies - and not just for bladder cancer, but other types of tumors, as well".

What The Study Did: In this observational study, data are used to assess the association of demographic and clinical characteristics with severe COVID-19 illness among hospitalized U.S. pediatric patients with COVID-19.

Authors: Alyson B. Goodman, M.D., of the COVID-19 Response Team at the Centers for Disease Control and Prevention in Atlanta, is the corresponding author.

To access the embargoed study: Visit our For The Media website at this link https://media.jamanetwork.com/

(doi:10.1001/jamanetworkopen.2021.5298)

Editor's Note:  Please see the article for additional information, including other authors, author contributions and affiliations, conflict of interest and financial disclosures, and funding and support.

What The Study Did: Genome analysis was performed on SARS-CoV-2 RNA from seven patients in a long-term care facility who were asymptomatic at the time of screening.

Authors: Baha Abdalhamid, M.D., Ph.D., of the University of Nebraska Medical Center in Omaha, is the corresponding author.

To access the embargoed study: Visit our For The Media website at this link https://media.jamanetwork.com/

(doi:10.1001/jamanetworkopen.2021.7939)

Editor's Note: The article includes conflicts of interest and funding/support disclosures. Please see the article for additional information, including other authors, author contributions and affiliations, conflict of interest and financial disclosures, and funding and support.

What The Study Did: This Viewpoint discusses potential associations between functional neurological disorder and COVID-19 vaccinations.

Authors: David L. Perez, M.D., M.MSc., of  Massachusetts General Hospital and Harvard Medical School in Boston, is the corresponding author.

To access the embargoed study: Visit our For The Media website at this link https://media.jamanetwork.com/

(doi:10.1001/jamaneurol.2021.1042)

Editor's Note: The article includes conflicts of interest disclosures. Please see the article for additional information, including other authors, author contributions and affiliations, conflict of interest and financial disclosures, and funding and support.

What The Study Did: Researchers analyzed changes in out-of-pocket health care expenses in the United States during the last two decades.

Authors: Amit Jain, M.D., of the Johns Hopkins University School of Medicine in Baltimore, is the corresponding author.

To access the embargoed study: Visit our For The Media website at this link https://media.jamanetwork.com/

(doi:10.1001/jamanetworkopen.2021.5499)

Editor's Note: The article includes conflicts of interest and funding/support disclosures. Please see the article for additional information, including other authors, author contributions and affiliations, conflict of interest and financial disclosures, and funding and support.

Tsukuba, Japan - Plants in the bean family (legumes) form nodules on their roots to take up nitrogen. Legumes will stop nodule production when nitrogen is plentiful (Figure 1), but precisely how nitrate presence controls nodule formation in these plants has been a mystery. Now, researchers from Japan have found that interactions between proteins and nitrate can induce and repress genes, controlling nodulation with potential applications in sustainable agriculture.

In a study published in April in The Plant Cell, a research team from the University of Tsukuba has shown that the different DNA-binding properties between proteins that establish nodule development determine if genes involved in symbiosis that govern nodulation turn on or off and that this gene expression is nitrate-induced.

Until now, there was an incomplete understanding of the molecular activity determining how legumes stop nodulation in the presence of excess nitrate. Previous research identified transcription factors (proteins that help turn specific genes "on" or "off") involved with nodule formation, but that's just part of the story.

"Building on the previous identification of transcription factors for proteins (known as NLPs) involved in nodule inception, we sought to answer the question of how symbiotic gene expression facilitating nodulation is controlled by nitrate," says senior author of the study Professor Takuya Suzaki. "We tested specific NLPs and found that they have overlapping functions, causing nitrate-induced control of nodulation."

To examine these molecular interactions, the researchers analyzed RNA molecules and plant traits using proteins from Lotus japonicus. They found that some proteins have dual functions, acting as master regulators for nitrate-dependent gene expression. They also identified new protein binding sites and compared them to previously known ones. Their findings reveal basic principles relating to NLP-regulated transcription of symbiotic genes inhibiting nitrate nodulation.

The research team emphasized additional questions. Some NLPs are found in cell nuclei in response to nitrate and stop nodule production, while others constantly aggregate in nuclei irrespective of nitrate levels. For the latter, it is unclear how they function exclusively in the presence of nitrate. The location of the NLPs in the cell matters because translation (when RNA is coded into proteins) happens in the cell's cytoplasm. If changes to proteins occur after the genetic code has been read (post-translational modifications), it could explain how these NLPs access protein-protein interactions and regulate genes.

"Uncovering how transcription factors influence gene expression has been a missing piece to the puzzle of understanding plant transcription regulation," Professor Suzaki explains. "Our discoveries bring us closer to knowing what is possible within these complex molecular relationships, but there is plenty left to untangle. Future research should aim to answer the question of how nodulation is regulated by other NLPs and in other plant species of interest."

Two studies led by the Mental Health Unit of the Virgen del Rocio University Hospital and involving researchers from the US conclude that antipsychotic drugs could have a protective effect against SARS-CoV-2. For this reason, patients treated with these drugs have a lower risk of becoming infected or suffer a milder form of the disease if they do become infected.

Thus, a first descriptive epidemiological study of a sample of 698 patients treated with antipsychotics at the Seville hospital revealed that antipsychotic drugs could provide protection against both infection and the tendency to clinical severity of Covid-19 infection. "These are very interesting findings that reflect a clinical reality where we see few patients with severe COVID-19, despite the presence of various risk factors," says Manuel Canal Rivero, clinical psychologist and lead author of one of the two papers.

"The number of Covid-19 patients is lower than expected among this group of people and in cases where a proven infection does occur, the evolution is benign and does not reach a life-threatening clinical situation. These data as a whole seem to point to the protective effect of the medication," he adds.

Complementary to this study, the same research group has observed that many of the genes whose expression is altered by Covid-19 are significantly down-regulated by antipsychotic drugs, which are commonly used to treat diseases with psychotic symptoms. This finding was achieved by investigating the gene expression profile (indicator of activated biological processes) of Covid-19 patients (Wuhan cohort) and patients being treated with antipsychotic drugs (specifically, aripiprazole) from the cohort of the Early Phases of Psychosis Intervention Programme (PAFIP) initiated 20 years ago at the Marqués de Valdecilla University Hospital in Cantabria by Benedicto Crespo-Facorro, professor at the University of Seville and current director of the Mental Health Unit at the Virgen del Rocío University Hospital.

"In a striking way we have shown how antipsychotics reduce the activation of genes involved in many of the inflammatory and immunological pathways associated with the severity of Covid-19 infection," says the lead author of the second paper, Professor Crespo-Facorro. Furthermore, he stresses that "although this finding requires replication, the discovery could be very significant because the treatment of Covid-19 with drugs originally indicated for unrelated clinical situations, that is to say drug repositioning, has been shown to be an interesting source of effective treatments for Covid-19 patients".

Since the beginning of the coronavirus pandemic, scientists and physicians worldwide undertook enormous efforts to understand the disease caused by the virus. In their latest collaborative study, researchers from the Max Planck Institute of Immunobiology and Epigenetics in Freiburg and physicians from the University of Freiburg Medical Center unveil a novel feature of COVID-19 immunity, which could have implications for future therapies. The study points to the involvement of Interleukin 33, an important danger signal, when immune cells encounter Sars-CoV-2 for a second time.

"We started the study at a very early stage of the pandemic in 2020 when not much was known about the immune response post-infection", says Erika Pearce, group leader at the Max Planck Institute of Immunobiology and Epigenetics. "Our aim was to examine the development of immunity in people recovering from Covid-19."

Antibodies stick around

An infection with Sars-CoV-2 triggers a complex immune response necessary for the development of immunity to the virus. In simple terms, two linked branches of our immune system need to remember the virus to prevent reinfection, namely antibody-producing B cells and memory T cells. Understanding how this happens in Sars-CoV-2 infection is key for controlling the Covid-19 pandemic and critical for the success of the vaccination efforts.

For the study, the team examined blood samples of 155 individuals who mostly had mild disease. They measured the amount of antibodies against the SARS-CoV-2 spike protein and found that patients maintain high levels of antibodies more than two months after infection, indicating that they will likely be protected from re-infection. "We thought this was very encouraging, but we also wanted to understand better how the immune system would react to a second encounter with the virus," says Petya Apostolova, physician and researcher in the lab of Erika Pearce.

When the virus hits the second time

Effective immunity to a virus is reached when sufficient antibodies and memory T cells are present in the blood of a person who has recovered from the disease or has been vaccinated. To test how this happens after Covid-19, the team exposed blood cells from participants who had antibodies against Sars-CoV-2 to a portion of the virus. They observed that memory T cells had developed and quickly responded to viral proteins. "We measured a broad panel of molecules that our immune cells use to communicate with each other. It was most fascinating to us that of all these measurements, the amount of Interleukin 33 was the closest match to the amount of antibodies people had, and to the activation of their memory T cells," explains Apostolova. Interleukin 33 (IL-33) is released by cells that sense danger in their environment and has been previously linked to chronic lung disease. IL-33 can have beneficial effects by activating T cells and inducing antibody production, but it can also promote inflammation of the lung. For the first time, this study has linked IL-33 production to immunity to Sars-CoV-2.

"We believe that Interleukin 33, which is normally produced as an alarm-signal, could be an important link between protection and disease severity," says Cornelius Waller from the University of Freiburg Medical Center. Indeed, by analyzing public data of lung cells taken from patients during Sars-CoV-2 infection, the researchers were able to show that Interleukin 33 was produced in their lungs. However, identifying the implications of these findings also in the context of lung tissue damage after severe Covid-19 infections will require more investigation.

The group of researchers hopes this collaboration will continue. As Waller pointed out, "we were able to discover this much so quickly through this fantastic synergy between clinicians experienced in the care for Covid-19 patients and experts in the immunology field." The researchers hope that this study might pave the way to better understanding immunity to Sars-CoV-2 and other viral infections.

An international group of scientists led by the RIKEN Cluster for Pioneering Research have studied the chemical composition of 50 protoplanetary-disk forming regions in the Perseus Molecular Cloud, and found that despite being in the same cloud, the amounts of complex organic molecules they contain are quite different. Interestingly, the chemically rich young disks have similar compositions of organic molecules. These findings raise an important question: do solar-like systems share a common chemistry at birth?

It was once believed that complex organic molecules were rare in the universe, and that this rarity might be a reason we have not found evidence of life outside the earth. However, in the last two decades it has become clear that these molecules are common. According to Yao-Lun Yang, who led the research as a member of the Star and Planet Formation Laboratory in the RIKEN Cluster for Pioneering Research, and is now at the University of Virginia as a VICO Origins postdoctoral fellow, "Today, scientists have begun to systematically survey protoplanetary disks--disks where planets eventually form around a star--in the hope of determining how these molecules form, how common they are, and what impact they have on planetary systems."

Though scientists have looked at individual systems, there is little comparative data to understand these young stars as a group. The RIKEN-led group decided to use observations from the ALMA observatory in Chile, which thanks to its high resolution allows scientists to study chemical compounds in protoplanetary disks. The observations required more than three years to complete.

The group surveyed the emission from organic molecules at specific frequencies in very young disks. They looked at methanol and acetonitrile, as well as two larger molecules, methyl formate and dimethyl ether. They found that different regions had large variation in the abundance of methanol and acetonitrile, though intriguingly, the relative abundance between the two species was remarkably similar. According to Yang, "This implies that there is a common production mechanism of these two molecules, and this give us important hints on how they form in space." In addition, the abundances of methyl formate and dimethyl ether tended to be higher relative to methanol in denser regions, hinting that there is something about the denser conditions that allows them to be abundant. Their findings were published by the American Astronomical Society last week in The Astrophysical Journal.

According to Nami Sakai, leader of the Star and Planet Formation Laboratory, "This raises important questions regarding how special the chemical environment of the infant Solar system is. We will be able to answer this question in the future by examining the chemical evolution of the gas surrounding young protostars. We hope that such knowledge will be a base for understanding the origin of life on the Earth.

The unpopular guests are already here. Hazel-, alder- and elm tree pollen sweep in over us during the early spring months, with birch pollen following in May. Then grass follows in June-July before mugwort pollen closes the season in late summer. All leading to more and more people suffering.

Over fifty years, the proportion of young Danes who sneeze, rub itchy eyes or suffer a blocked nose because of pollen has grown from 7-8 per cent in the 1970s to 24 per cent today. And for asthma, the proportion has risen from 4 to 12 per cent during the same period. This is shown by a study of 53,000 Danish blood donors published by researchers from Aarhus University in collaboration with the Blood Bank at Aarhus University Hospital.

The lead author of the study, Susan Mikkelsen, is a PhD student at the Department of Clinical Medicine and a medical specialist in clinical immunology. She points to how the large blood donor study combines questionnaires and blood samples with the possibility of many follow-up studies:

"In the newly published study, 53,000 blood donors from all over the country born in the period from the 1950s and up until the turn of the millennium, completed a questionnaire on a tablet while donating blood," says Susan Mikkelsen, who works as a medical doctor at Department of Clinical Immunology at Aarhus University Hospital while completing her PhD dissertation.

"The blood from 25,000 of the participants has also been examined for antibodies against nine selected allergens, ranging from different types of pollen to animal hair and dust mites. In medical language, we say that the test participants who react by forming antibodies are 'sensitised'," she explains.

The blood samples show that the proportion of sensitised donors increases with the number of allergic disorders. For example, the researchers can see that ninety per cent of those who indicate that they have both asthma and allergic reactions in the nose and eyes, form antibodies.

On the other hand, the proportion of sensitised is only half as large for donors who grew up on a farm with animals, where they are exposed to a range of bacteria, fungi and pollen that might be protective against hypersensitivity.

Susan Mikkelsen also points out that the newly published study, which has been published in Clinical and Translational Allergy, is a so-called conservative research result which cannot be accused of exaggerating the problem - it most likely downplays it. The blood donors are in fact not representative random sample of the Danish population, as they are healthier, in particular the oldest blood donors, due to the requirement that a donor must be 'healthy' to give blood.

"For us, this means that the severe asthmatics aren't included when we calculate the proportion of allergy sufferers, because they simply aren't allowed to give blood. When every fifth blood donor nevertheless ticks the box as allergic and every tenth as asthmatic, this of course tells something about how hay fever and asthma are today very common diseases," says Susan Mikkelsen.

The latter is co-author Torben Sigsgaard's area of expertise. He is professor at the Department of Public Health and acknowledges that we in the Western world are becoming increasingly allergic. This was demonstrated already 15 years ago, for example in the Danish "Glostrup"' studies and the European Respiratory studies.

"Nevertheless, the current study contributes with important knowledge. The study shows, for example, that there is probably some way to go until we in Denmark reach the ceiling for how many people can become allergy sufferers, as the curve doesn't appear to be flattening out as we've seen it do in Switzerland, for example," says Torben Sigsgaard with reference to a theory that there is a maximum limit for how many people in a particular population group will, under the wrong circumstances, develop allergies. The theory is that the Swiss have now reached this maximum, but the specifics remain unclear. Torben Sigsgaard also points out that hay fever and asthma can be severe disorders - even though they are relatively common in Denmark.

"For some young sufferers, hay fever means that they have trouble completing exams, because the pollen season coincides with summer exams. We also know that some people completely opt out of social activities when the allergy is worst, because they think it's too disgusting and inconvenient to have their eyes and nose constantly running," says Torben Sigsgaard, who has many years of experience in the research field.

He emphasises that the large biobank with blood samples allows researchers to continue working with genetic analyses or studies connected to air pollution.

"The uniqueness of the study is that we can examine the participants' blood for biomarkers and genetic markers and compare the findings with environmental impacts, follow-up questionnaire surveys and the Danish registers. This makes it possible to look at the significance of genetic-environmental interaction on allergy and sensitisation. In addition, the repeated blood samples from each participant make it possible to study the immune system before symptom onset in participants with newly developed allergic symptoms," says Susan Mikkelsen.

"We're extremely grateful for the blood donors' contribution to the study. They make their blood samples available for research and thus facilitate our studies," she says.

Electrical batteries are increasingly crucial in a variety of applications, from integration of intermittent energy sources with demand, to unlocking carbon-free power for the transportation sector through electric vehicles (EVs), trains and ships, to a host of advanced electronics and robotic applications.

A key challenge however is that batteries degrade quickly with operating conditions. It is currently difficult to estimate battery health without interrupting the operation of the battery or without going through a lengthy procedure of charge-discharge that requires specialised equipment.

In work recently published by Nature Machine Intelligence, researchers from the Smart Systems Group at Heriot-Watt University in Edinburgh, UK working together with researchers from the CALCE group at the University of Maryland in the US developed a new method to estimate battery health irrespective of operating conditions and battery design or chemistry, by feeding artificial intelligence (AI) algorithms with the raw battery voltage and current operational data.

Darius Roman, the PhD student that designed the AI framework said: "To date, the progress of data-driven models for battery degradation relies on the development of algorithms that carry out inference faster. Whilst researchers often spend a considerable amount of time on model or algorithm development, very few people take the time to understand the engineering context in which the algorithms are applied. By contrast, our work is built from the ground up. We first understand battery degradation through collaborations with the CALCE group at the University of Maryland, where in-house degradation testing of batteries was carried out. We then concentrate on the data, where we engineer features that capture battery degradation, we select the most important features and only then we deploy the AI techniques to estimate battery health."

In addition, the researchers found that current data-driven models for battery health estimation do not consider model confidence. However, this is often critical for decision making to understand how the AI model came to a certain conclusion and whether the model can be trusted. In their work, the proposed AI model is capable of quantifying uncertainty in its predictions to better support operating decisions.

The developed framework scales up with new chemistries, including the new upcoming solid-state batteries, battery designs and operating conditions and has the potential to unlock new strategies of how batteries can and should be used.

Valentin Robu, from the Smart Systems Group said: "Batteries are increasingly critical to a variety of applications, from robotics to renewable energy integration. A key challenge in these domains is having an accurate, high-confidence estimates of battery state of health. Consider for example, a robotic asset operating in a remote environment such as deep subsea monitoring, where assuring the health of the battery deployed on the robot is mission-critical. Similarly, for energy applications, having an accurate estimate of the remaining useful lifetime of the battery is often critical to a project's economic viability."

A tiny protein of SARS-CoV-2, the coronavirus that gives rise to COVID-19, may have big implications for future treatments, according to a team of Penn State researchers.

Using a novel toolkit of approaches, the scientists uncovered the first full structure of the Nucleocapsid (N) protein and discovered how antibodies from COVID-19 patients interact with that protein. They also determined that the structure appears similar across many coronaviruses, including recent COVID-19 variants -- making it an ideal target for advanced treatments and vaccines. They reported their results in Nanoscale.

"We discovered new features about the N protein structure that could have large implications in antibody testing and the long-term effects of all SARS-related pandemic viruses," said Deb Kelly, professor of biomedical engineering (BME), Huck Chair in Molecular Biophysics and director of the Penn State Center for Structural Oncology, who led the research. "Since it appears that the N protein is conserved across the variants of SARS-CoV-2 and SARS-CoV-1, therapeutics designed to target the N protein could potentially help knock out the harsher or lasting symptoms some people experience."

Most of the diagnostic tests and available vaccines for COVID-19 were designed based on a larger SARS-CoV-2 protein -- the Spike protein -- where the virus attaches to healthy cells to begin the invasion process.

The Pfizer/BioNTech and Moderna vaccines were designed to help recipients produce antibodies that protect against the Spike protein. However, Kelly said, the Spike protein can easily mutate, resulting in the variants that have emerged in the United Kingdom, South Africa, Brazil and across the United States.

Unlike the outer Spike protein, the N protein is encased in the virus, protected from environmental pressures that cause the Spike protein to change. In the blood, however, the N protein floats freely after it is released from infected cells. The free-roaming protein causes a strong immune response, leading to the production of protective antibodies. Most antibody-testing kits look for the N protein to determine if a person was previously infected with the virus -- as opposed to diagnostic tests that look for the Spike protein to determine if a person is currently infected.

"Everyone is looking at the Spike protein, and there are fewer studies being performed on the N protein," said Michael Casasanta, first author on the paper and a postdoctoral fellow in the Kelly laboratory. "There was this gap. We saw an opportunity -- we had the ideas and the resources to see what the N protein looks like."

Initially, the researchers examined the N protein sequences from humans, as well as different animals thought to be potential sources of the pandemic, such as bats, civets and pangolins. They all looked similar but distinctly different, according to Casasanta.

"The sequences can predict the structure of each of these N proteins, but you can't get all the information from a prediction -- you need to see the actual 3D structure," Casasanta said. "We converged the technology to see a new thing in a new way."

The researchers used an electron microscope to image both the N protein and the site on the N protein where antibodies bind, using serum from COVID-19 patients, and developed a 3D computer model of the structure. They found that the antibody binding site remained the same across every sample, making it a potential target to treat people with any of the known COVID-19 variants.

"If a therapeutic can be designed to target the N protein binding site, it might help reduce the inflammation and other lasting immune responses to COVID-19, especially in COVID long haulers," Kelly said, referring to people who experience COVID-19 symptoms for six weeks or longer.

The team procured purified N proteins, meaning the samples only contained N proteins, from RayBiotech Life and applied them to microchips developed in partnership with Protochips Inc. The microchips are made of silicon nitride, as opposed to a more traditional porous carbon, and they contain thin wells with special coatings that attract the N proteins to their surface. Once prepared, the samples were flash frozen and examined through cryo-electron microscopy.

Kelly credited her team's unique combination of microchips, thinner ice samples and Penn State's advanced electron microscopes outfitted with state-of-the-art detectors, customized from the company Direct Electron, for delivering the highest-resolution visualization of low-weight molecules from SARS-CoV-2 so far.

"The technology combined resulted in a unique finding," Kelly said. "Before, it was like trying to look at something frozen in the middle of the lake. Now, we're looking at it through an ice cube. We can see smaller entities with many more details and higher accuracy."

Since 2016, a federal regulation has allowed nurse practitioners and physician assistants to obtain a waiver to prescribe buprenorphine, a medication used to treat opioid use disorder as a medication assisted treatment.

But a recent study by Indiana University researchers found the bill, called the Comprehensive Addiction and Recovery Act (CARA), has not greatly increased the amount of nurse practitioners prescribing buprenorphine, especially in states that have further restrictions. The study was published in Medical Care Research and Review.

"Nurse practitioners and physician assistants are an important workforce with a capacity to expand treatment access for those with substance use disorders," said Kosali Simon, co-author of the study and a Herman B. Wells Endowed Professor in IU's O'Neill School of Public and Environmental Affairs. "But we have found that efforts like CARA have been limited in actually utilizing this group, with nurse practitioners accounting for a relatively small proportion of buprenorphine prescriptions."

Buprenorphine is the only agonist medication for opioid addiction that can be prescribed by a qualified physician or nonphysician practitioner in an office-based setting. Simon said research has shown it is associated with significant decreases in relapse and overdose. However, the U.S. has a shortage of providers who have a waiver, known as an X waiver, from the Drug Enforcement Administration to prescribe buprenorphine outside of opioid treatment programs.

Through the Comprehensive Addiction and Recovery Act nurse practitioners and physician assistants to prescribe buprenorphine to up to 30 patients at a time for their first year, and after that, they can obtain authorization to prescribe to up to 100 patients. Some states mandate further restrictions though, including those that limit the authority of nurse practitioners to prescribe.

Kosali and a team of researchers from various universities used pharmacy claims data between January 2015 and September 2018 from Optum's deidentified Clinformatics DataMart to examine the impact of federal and state scope-of-practice regulations on nurse practitioner's buprenorphine prescribing.

At the county-level, researchers found the proportion of patients filling prescriptions written by nurse practitioners was low even after CARA: 2.7% in states that did not require physician oversight of nurse practitioners and 1.1% in states that did. While analyses in rural counties showed higher rates of buprenorphine prescriptions written by nurse practitioners, the study found rates were still considerably low - 3.7% in states with less restrictive regulations and 1.1% in other states.

Simon said since relatively few physicians have a waiver to prescribe, it can be difficult for nurse practitioners who must have physician oversight to find a waivered physician to oversee them. Some also face supervising physicians who are unsupportive of buprenorphine treatment.

"These results indicate that less restrictive scope-of-practice regulations are associated with greater nurse practitioner prescribing following CARA," Simon said. "The small magnitude of the changes indicates that federal attempts to expand treatment access through CARA have been limited. More is needed so people with substance use disorder have access to medication assistant treatment."

While state restrictions did not help, the researchers said there are more deterrents involved including insufficient training and education about opioid use disorder treatment, burdensome training time, lack of institutional and clinician peer support, poor care coordination and inadequate insurance reimbursement.

Researchers recommend relaxing state scope-of-practice requirements for nurse practitioners and addressing other practice-level and educational barriers that impede treatment access. Additionally, Simon said eliminating the need for an X waiver for all practitioners should be further considered to help address gaps in opioid use disorder care, especially in primary care settings and rural areas. Alternatively, current X waiver training requirements could be decreased for certain clinicians, including nurse practitioners and physician assistants, who have previously completed substance use disorder treatment training during residency.

A clean energy future propelled by hydrogen fuel depends on figuring out how to reliably and efficiently split water. That's because, even though hydrogen is abundant, it must be derived from another substance that contains it -- and today, that substance is often methane gas. Scientists are seeking ways to isolate this energy-carrying element without using fossil fuels. That would pave the way for hydrogen-fueled cars, for example, that emit only water and warm air at the tailpipe.

Water, or H2O, unites hydrogen and oxygen. Hydrogen atoms in the form of molecular hydrogen must be separated out from this compound. That process depends on a key -- but often slow -- step: the oxygen evolution reaction (OER). The OER is what frees up molecular oxygen from water, and controlling this reaction is important not only to hydrogen production but a variety of chemical processes, including ones found in batteries.

"The oxygen evolution reaction is a part of so many processes, so the applicability here is quite broad." -- Pietro Papa Lopes, Argonne assistant scientist

A study led by scientists at the U.S. Department of Energy's (DOE) Argonne National Laboratory illuminates a shape-shifting quality in perovskite oxides, a promising type of material for speeding up the OER. Perovskite oxides encompass a range of compounds that all have a similar crystalline structure. They typically contain an alkaline earth metal or lanthanides such as La and Sr in the A-site, and a transition metal such as Co in the B-site, combined with oxygen in the formula ABO3. The research lends insight that could be used to design new materials not only for making renewable fuels but also storing energy.

Perovskite oxides can bring about the OER, and they are less expensive than precious metals such as iridium or ruthenium that also do the job. But perovskite oxides are not as active (in other words, efficient at accelerating the OER) as these metals, and they tend to slowly degrade.

"Understanding how these materials can be active and stable was a big driving force for us," said Pietro Papa Lopes, an assistant scientist in Argonne's Materials Science division who led the study. "We wanted to explore the relationship between these two properties and how that connects to the properties of the perovskite itself."

Previous research has focused on the bulk properties of perovskite materials and how those relate to the OER activity. The researchers wondered, however, whether there was more to the story. After all, the surface of a material, where it reacts with its surroundings, can be completely different from the rest. Examples like this are everywhere in nature: think of a halved avocado that quickly browns where it meets the air but remains green inside. For perovskite materials, a surface that becomes different from the bulk could have important implications for how we understand their properties.

In water electrolyzer systems, which split water into hydrogen and oxygen, perovskite oxides interact with an electrolyte made of water and special salt species, creating an interface that allows the device to operate. As electrical current is applied, that interface is critical in kicking off the water-splitting process. "The material's surface is the most important aspect of how the oxygen evolution reaction will proceed: How much voltage you need, and how much oxygen and hydrogen you're going to be producing," Lopes said.

Not only is the perovskite oxide's surface different from the rest of the material, it also changes over time. "Once it's in an electrochemical system, the perovskite surface evolves and turns into a thin, amorphous film," Lopes said. "It's never really the same as the material you start with."

The researchers combined theoretical calculations and experiments to determine how the surface of a perovskite material evolves during the OER. To do so with precision, they studied lanthanum cobalt oxide perovskite and tuned it by "doping" the lanthanum with strontium, a more reactive metal. The more strontium was added to the initial material, the faster its surface evolved and became active for the OER -- a process the researchers were able to observe at atomic resolution with transmission electron microscopy. The researchers found that strontium dissolution and oxygen loss from the perovskite were driving the formation of this amorphous surface layer, which was further explained by computational modelling performed using the Center for Nanoscale Materials, a DOE Office of Science User Facility.

"The last missing piece to understand why the perovskites were active towards the OER was to explore the role of small amounts of iron present in the electrolyte," Lopes said. The same group of researchers recently discovered that traces of iron can improve the OER on other amorphous oxide surfaces. Once they determined that a perovskite surface evolves into an amorphous oxide, then it became clear why iron was so important.

"Computational studies help scientists understand reaction mechanisms that involve both the perovskite surface and the electrolyte," said Peter Zapol, a physicist at Argonne and study co-author. "We focused on reaction mechanisms that drive both activity and stability trends in perovskite materials. This is not typically done in computational studies, which tend to focus solely on the reaction mechanisms responsible for the activity."

The study found that the perovskite oxide's surface evolved into a cobalt-rich amorphous film just a few nanometers thick. When iron was present in the electrolyte, the iron helped accelerate the OER, while the cobalt-rich film had a stabilizing effect on the iron, keeping it active at the surface.

The results suggest new potential strategies for designing perovskite materials -- one can imagine creating a two-layer system, Lopes said, that is even more stable and capable of promoting the OER.

"The OER is a part of so many processes, so the applicability here is quite broad," Lopes said. "Understanding the dynamics of materials and their effect on the surface processes is how we can make energy conversion and storage systems better, more efficient and affordable."