Culture

Leg amputees are often not satisfied with their prosthesis, even though the sophisticated prostheses are becoming available. One important reason for this is that they perceive the weight of the prosthesis as too high, despite the fact that prosthetic legs are usually less than half the weight of a natural limb. Researchers led by Stanisa Raspopovic, a professor at the Department of Health Sciences and Technology, have now been able to show that connecting the prostheses to the nervous system helps amputees to perceive the prosthesis weight as lower, which is beneficial for their acceptance.

Together with an international consortium, Raspopovic has developed in recent years prostheses that provide feedback to the wearer's nervous system. This is done via electrodes implanted in the thigh, which are connected to the leg nerves present there. Information from tactile sensors under the sole of the prosthetic foot and from angle sensors in the electronic prosthetic knee joint are converted into pulses of current and passed in to the nerves.

"To trick an above-knee amputee's brain into the belief that the prosthetic leg was similar to his own leg, we artificially restored the lost sensory feedback," says ETH professor Raspopovic. In a study published last year, he and his team showed that wearers of such neurofeedback prostheses can move more safely and with less effort.

In a further study, the scientists were now able to show that neurofeedback also reduces the perceived weight of the prosthesis. They published the results in the journal Current Biology

In order to determine how heavy a transfemoral amputee perceives their prosthetic leg to be, they had a voluntary study participant complete gait exercises with either neurofeedback switched on or off. They weighed down the healthy foot with additional weights and asked the study participant to rate how heavy he felt the two legs were in relation to each other. Neurofeedback was found to reduce the perceived weight of the prosthesis by 23 percent, or almost 500 grams.

The scientists also confirmed a beneficial involvement of the brain by a motor-cognitive task, during which the volunteer had to spell backwards five-letter words while walking. The sensory feedback not only allowed him to have a faster gait but also to have a higher spelling accuracy.

"Neurofeedback not only enables faster and safer walking and positively influences weight perception," says Raspopovic. "Our results also suggest that, quite fundamentally, it can take the experience of patients with an artificial device closer to that with a natural limb."

An important and still unanswered question is how new genes that cause antibiotic resistance arise. In a new study, Swedish and American researchers have shown how new genes that produce resistance can arise from completely random DNA sequences. The results have been published in the journal PLOS Genetics.

Antibiotic resistance is a major global problem and the spread of resistant bacteria causes disease and death, and constitutes a major cost to society. The most common way for bacteria to develop resistance is by taking up various types of resistance genes from other bacteria. These genes encode proteins (peptides) that can lead to resistance by: (i) deactivating the antibiotic, (ii) reducing its concentration, or (iii) altering the antibiotic's target so that the antibiotic can no longer bind to that target and hence halt the growth of the bacterium. Once resistance genes have arisen, they can quickly spread between different pathogenic bacteria and reduce the effectiveness of our antibiotics. It is therefore important to detect and characterise new resistance genes as quickly as possible - in order to monitor the spread of resistance and also to facilitate treatment and the development of new antibiotics.

To study the emergence of resistance genes, the researchers used laboratory experiments to investigate whether it was possible to generate a gene from random DNA sequences that would give rise to antibiotic resistance. This was done by first designing nearly one billion random DNA sequences that were then placed on a plasmid in the intestinal bacterium Escherichia coli. (Plasmids are DNA molecules that replicate independently and can be transferred from one organism to another.)

These random DNA sequences were then expressed in the bacterium as short peptides. While most of these peptides had no effect on the bacterium at all, six different peptides did, causing the bacterium to become resistant to the antibiotic Colistin, an important antibiotic medication of last resort that is used in severe infections to kill the bacteria by binding to and destroying the bacterium's cell membrane. These peptides caused resistance by increasing the expression of genes that are involved in the modification of the bacterium's cell membrane. This modification of the cell membrane resulted in the antibiotic not being able to bind to cell membrane, and thus not being able to reduce the survival of the bacterium.

"We have now shown in two different studies that random sequences of amino acids can give rise to new functions that are beneficial to the bacterium such as antibiotic resistance. This suggests that the evolution of new functions from random DNA sequences is not as unusual as previously thought," says Dan I. Andersson, Professor in Medical Bacteriology and responsible for the study.

"An important question that remains unanswered and requires further study is whether these new genes are naturally present in bacteria or can only be observed in laboratory experiments," says Michael Knopp, post-doctoral researcher at the Department of Medical Biochemistry and Microbiology and the study's first author.

PITTSBURGH--Researchers at Carnegie Mellon University report findings on an advanced nanomaterial-based biosensing platform that detects, within seconds, antibodies specific to SARS-CoV-2, the virus responsible for the COVID-19 pandemic. In addition to testing, the platform will help to quantify patient immunological response to the new vaccines with precision.

The results were published this week in the journal Advanced Materials. Carnegie Mellon's collaborators included the University of Pittsburgh (Pitt) and the UPMC.

The testing platform identifies the presence of two of the virus' antibodies, spike S1 protein and receptor binding domain (RBD), in a very small drop of blood (about 5 microliters). Antibody concentrations can be extremely low and still detected below one picomolar (0.15 nanograms per milliliter). This detection happens through an electrochemical reaction within a handheld microfluidic device which sends results almost immediately to a simple interface on a smart phone.

"We utilized the latest advances in materials and manufacturing such as nanoparticle 3D printing to create a device that rapidly detects COVID-19 antibodies," said Rahul Panat, an associate professor of mechanical engineering at Carnegie Mellon who uses specialized additive manufacturing techniques for research ranging from brain-computer interfaces to biomonitoring devices.

An additive manufacturing technology called aerosol jet 3D printing is responsible for the efficiency and accuracy of the testing platform. Tiny, inexpensive gold micropillar electrodes are printed at nanoscale using aerosol droplets that are thermally sintered together. This causes a rough, irregular surface that provides increased surface area of the micropillars and an enhanced electrochemical reaction, where antibodies can latch on to antigens coated on the electrode. The specific geometry allows the micropillars to load more proteins for detection, resulting in very accurate, quick results.

The test has a very low error rate because the binding reaction between the antibody and antigen used in the device is highly selective. The researchers were able to exploit this natural design to their advantage.

The results come at an urgent time during the COVID-19 pandemic. "Because our technique can quantify the immune response to vaccination, it is very relevant in the current environment," Panat said.

Panat collaborated with Shou-Jiang Gao, leader of the cancer virology program at UPMC's Hillman Cancer Center and professor of microbiology and molecular genetics at Pitt. Azahar Ali, a researcher in Panat's Advanced Manufacturing and Materials Lab, was the lead author of the study.

Rapid diagnosis for the treatment and prevention of communicable diseases is a public health issue that goes beyond the current COVID-19 pandemic. Because the proposed sensing platform is generic, it can be used for the rapid detection of biomarkers for other infectious agents such as Ebola, HIV, and Zika. Such a quick and effective test could be a game-changer for controlling the spread of diseases.

The COVID-19 pandemic is raising fears of new pathogens such as new viruses or drug-resistant bacteria. To this, a Korean research team has recently drawn attention for developing the technology for removing antibiotic-resistant bacteria by controlling the surface texture of nanomaterials.

A joint research team from POSTECH and UNIST has introduced mixed-FeCo-oxide-based surface-textured nanostructures (MTex) as highly efficient magneto-catalytic platform in the international journal Nano Letters. The team consisted of professors In Su Lee and Amit Kumar with Dr. Nitee Kumari of POSTECH's Department of Chemistry and Professor Yoon-Kyung Cho and Dr. Sumit Kumar of UNIST's Department of Biomedical Engineering.

First, the researchers synthesized smooth surface nanocrystals in which various metal ions were wrapped in an organic polymer shell and heated them at a very high temperature. While annealing the polymer shell, a high-temperature solid-state chemical reaction induced mixing of other metal ions on the nanocrystal surface, creating a number of few-nm-sized branches and holes on it. This unique surface texture was found to catalyze a chemical reaction that produced reactive oxygen species (ROS) that kills the bacteria. It was also confirmed to be highly magnetic and easily attracted toward the external magnetic field. The team had discovered a synthetic strategy for converting normal nanocrystals without surface features into highly functional mixed-metal-oxide nanocrystals.

The research team named this surface topography - with branches and holes that resembles that of a ploughed field - "MTex." This unique surface texture has been verified to increase the mobility of nanoparticles to allow efficient penetration into biofilm matrix while showing high activity in generating reactive oxygen species (ROS) that are lethal to bacteria.

This system produces ROS over a broad pH range and can effectively diffuse into the biofilm and kill the embedded bacteria resistant to antibiotics. And since the nanostructures are magnetic, biofilm debris can be scraped out even from the hard-to-reach microchannels.

"This newly developed MTex shows high catalytic activity, distinct from the stable smooth-surface of the conventional spinel forms," explained Dr. Amit Kumar, one of the corresponding authors of the paper. "This characteristic is very useful in infiltrating biofilms even in small spaces and is effective in killing the bacteria and removing biofilms."

"This research allows to regulate the surface nanotexturization, which opens up possibilities to augment and control the exposure of active sites," remarked Professor In Su Lee who led the research. "We anticipate the nanoscale-textured surfaces to contribute significantly in developing a broad array of new enzyme-like properties at the nano-bio interface."

CHAMPAIGN, Ill. -- Religious people facing life crises rely on emotion-regulation strategies that psychologists also use, a new study finds. They look for positive ways of thinking about hardship, a practice known to psychologists as "cognitive reappraisal." They also tend to have confidence in their ability to cope with difficulty, a trait called "coping self-efficacy." Both have been shown to reduce symptoms of anxiety and depression.

The new findings are reported in the Journal of Religion and Health.

"It appears that religious people are making use of some of the same tools that psychologists have systematically identified as effective in increasing well-being and protecting against distress," said Florin Dolcos, a professor of psychology in the Beckman Institute for Advanced Science and Technology at the University of Illinois Urbana-Champaign, who led the study with psychology professor Sanda Dolcos and graduate student Kelly Hohl. "This suggests that science and religion are on the same page when it comes to coping with hardship," he said.

The research was prompted in part by earlier studies demonstrating that people who are religious tend to use a coping strategy that closely resembles cognitive reappraisal.

"For example, when somebody dies, a religious person may say, 'OK, now they are with God,' while someone who isn't religious may say, 'Well, at least they are not suffering anymore,'" Florin Dolcos said. In both cases, the individual finds comfort in framing the situation in a more positive light.

To determine if religious people rely on - and benefit from - reappraisal as an emotion-regulation strategy, the researchers recruited 203 participants with no clinical diagnoses of depression or anxiety. Fifty-seven of the study subjects also answered questions about their level of religiosity or spirituality.

The researchers asked participants to select from a series of options describing their attitudes and practices.

"We asked them about their coping styles. So, for religious coping, we asked if they try to find comfort in their religious or spiritual beliefs," Hohl said. "We asked them how often they reappraise negative situations to find a more positive way of framing them or whether they suppress their emotions."

The researchers also evaluated participants' confidence in their ability to cope and asked them questions designed to measure their symptoms of depression and anxiety.

Hohl said she looked for correlations between coping strategies, religious or nonreligious attitudes and practices, and levels of distress. She also conducted a mediation analysis to determine which practices specifically influenced outcomes like depression or anxiety.

"If we are just looking at the relationship between religious coping and lower anxiety, we don't know exactly which strategy is facilitating this positive outcome," Sanda Dolcos said. "The mediation analysis helps us determine whether religious people are using reappraisal as an effective way of lessening their distress."

The analysis also shows whether an individual's confidence in their ability to handle crises - another factor that psychological studies have found is associated with less depression and anxiety - "facilitates the protecting role of religious coping against such symptoms of emotional distress," Sanda Dolcos said. "We found that if people are using religious coping, then they also have decreased anxiety or depressive symptoms."

Cognitive reappraisal and coping self-efficacy were contributing to those decreased symptoms of distress, she said.

The study should be of interest to clinical psychologists working with religious clients, Hohl said. "It should also speak to clergy members or church leaders who can promote this kind of reappraisal to help parishioners make sense of the world and increase their resilience against stress."

"I hope this is an example of where religion and science can work together to maintain and increase well-being," Florin Dolcos said.

Scientists at the University of Southampton's Centre for Cancer Immunology have gained new insight into how the immune system can be better used to find and kill cancer cells.

Working with BioInvent International, a team led by Professor Mark Cragg and Dr Jane Willoughby from the Antibody and Vaccine Group, based at the Centre, have shown that antibodies, designed to target the molecule OX40, give a more active immune response when they bind closer to the cell membrane and can be modified to attack cancer in different ways.

OX40 is a 'co-receptor' that helps to stimulate the production of helper and killer T-cells during an immune response. One of the ways cancer avoids detection is by suppressing immune responses to stop functional tumour specific T-cells from being produced.

In the study, which has been published in Journal for ImmunoTherapy of Cancer, the team also discovered that switching the antibody's isotype (the part of the antibody that engages with cells of the immune system) could change the way the antibody worked.

When the mIgG2a isotype was used, the antibody could delete cells called Treg cells which are suppressive in the immune system. When the mIgG1 isotype was present, the antibody could stimulate killer T-cells to increase and therefore kill more cancer cells.

Professor Cragg said: "Clinical trials with anti-OX40 antibodies have shown that the body can tolerate these drugs but unfortunately have also shown disappointing clinical responses. We need to understand why this is.

"This new data shows us that when there is a cancer with lots of Tregs we could use the equivalent of the m2IgGa isotype and in patients where we feel we need better cytotoxic T cells we could use the equivalent of a mIgG1 isotype to boost the immune response. This information is important for developing the next generation of OX40 antibodies that we hope will be more effective in treating patients with cancer."

When Michigan State University's Gemma Reguera first proposed her new research project to the National Science Foundation, one grant reviewer responded that the idea was not "environmentally relevant."

As other reviewers and the program manager didn't share this sentiment, NSF funded the proposal. And, now, Reguera's team has shown that microbes are capable of an incredible feat that could help reclaim a valuable natural resource and soak up toxic pollutants.

"The lesson is that we really need to think outside the box, especially in biology. We just know the tip of the iceberg. Microbes have been on earth for billions of years, and to think that they can't do something precludes us from so many ideas and applications," said Reguera, a professor in the Department of Microbiology and Molecular Genetics.

Reguera's team works with bacteria found in soil and sediment known as Geobacter. In its latest project, the team investigated what happened to the bacteria when they encounter cobalt.

Cobalt is a valuable but increasingly scarce metal used in batteries for electric vehicles and alloys for spacecraft. It's also highly toxic to livings things, including humans and bacteria.

"It kills a lot of microbes," Reguera said. "Cobalt penetrates their cells and wreaks havoc."

But the team suspected Geobacter might be able to escape that fate. These microbes are a hardy bunch. They can block uranium contaminants from getting into groundwater, and they can power themselves by pulling energy from minerals containing iron oxide. "They respire rust," Reguera said.

Scientists know little about how microbes interact with cobalt in the environment, but many researchers -- including one grant reviewer -- believed that the toxic metal would be too much for the microbes.

But Reguera's team challenged that thinking and found Geobacter to be effective cobalt "miners," extracting the metal from rust without letting it penetrate their cells and kill them. Rather, the bacteria essentially coat themselves with the metal.

"They form cobalt nanoparticles on their surface. They metallize themselves and it's like a shield that protects them," Reguera said. "It's like Iron Man when he puts on the suit."

The team published its discovery in the journal Frontiers in Microbiology, with the research article first appearing online in late November, 2020. The Spartan team included Kazem Kashefi, an assistant professor in the Department of Microbiology and Molecular Genetics, and graduate students Hunter Dulay and Marcela Tabares, who are "two amazing and relatively junior investigators," Reguera said.

She sees this discovery as a proof-of-concept that opens the door to a number of exciting possibilities. For example, Geobacter could form the basis of new biotechnology built to reclaim and recycle cobalt from lithium-ion batteries, reducing the nation's dependence on foreign cobalt mines.

It also invites researchers to study Geobacter as a means to soak up other toxic metals that were previously believed to be death sentences for the bacteria. Reguera is particularly interested in seeing if Geobacter could help clean up cadmium, a metal that's found in industrial pollution that disproportionately affects America's most disadvantaged communities.

"This is a reminder to be creative and not limited in the possibilities. Research is the freedom to explore, to search and search and search," Reguera said. "We have textbook opinions about what microbes can and should do, but life is so diverse and colorful. There are other processes out there waiting to be discovered."

A unifying explanation of the cause of autism and the reason for its rising prevalence has eluded scientists for decades, but a theoretical model published in the journal Medical Hypotheses describes the cause as a combination of socially valued traits, common in autism, and any number of co-occurring disabilities.

T.A. Meridian McDonald, PhD, a research instructor in Neurology at Vanderbilt University Medical Center, has spent 25 years researching autism, from a time she could read literally every research paper on the topic in the 1990s until now, when there is an overload of such studies.

"Up until now there have been a lot of theories about the possible causes of autism but none of those theories account for the majority of autism cases," McDonald said. "There are also a lot of theories as to why the prevalence of autism has been increasing in the population but, to date, there hasn't been a theory that provides an explanatory model that accounts for all of those phenomena, including the genetics, social history, or characteristics of autism."

The CDC estimates that 1-in-54 persons are on the autism spectrum.

Consistent with Genetics Research in Autism

McDonald's theory, titled 'The Broader Autism Phenotype Constellation-Disability Matrix Paradigm (BAPCO-DMAP) Theory,' is consistent with the current science on the genetics of autism but shifts the focus to positive traits of autism and to historical events that changed the prevalence of autism in society.

"The BAPCO-DMAP theory describes how people are attracted to other people who are very similar. They are attracted to certain traits that are very common in the population, and this leads to offspring who are more likely to have certain traits, as well as a greater intensity of traits." McDonald said.

"The (BAPCO) traits are not what people expect. They expect the traits to be about challenges or difficulties, but instead there are six main traits -- increased attention, increased memory, a preference for the object world vs. the social world and their environment, increased nonconformity, increased differences in sensory and perception, as well as systemizing."

BAPCO Can Interact with Development

BAPCO traits alone are not necessarily disabling. A person can have BAPCO traits and still have excellent communication and socialization skills, but too much of any of these traits can affect development.

For example, although it seems counterintuitive, McDonald outlines how increased memory and attention can delay language development in children with BAPCO traits.

"Typical babies have tiny attention spans and working memory. These limitations on memory and attention actually help babies learn their first language by breaking the words into their tiniest parts," she said.

"But an infant with very high levels of attention and memory has a harder time with learning language because they are unable to break the language down to very small parts, so they learn groups of sounds instead."

McDonald describes how increased memory and attention can lead to echolalia, where children speak in, or repeat, long phrases without seeming to understand them.

"You can often see children with autism engaging in sense-making activities, such as watching the same show over and over and memorizing information," she said.

BAPCO Can Interact with Disabilities

BAPCO traits can also interact with disabilities. If a person has BAPCO traits and they also have a disability such as Down syndrome or information processing disorder, then those things combined can create greater challenges than what would be experienced by an individual who has only a BAPCO personality or a disability, but not both.

Increased Prevalence of BAPCO and Autism

McDonald's theory also highlights how the prevalence of autism has increased in higher income countries due to societal changes that increase freedoms in education, employment and other opportunities for both men and women.

In societies where people do not have freedom to pursue their interests and passions, where meritorious education is not valued, where men and women do not have freedom to choose their careers, and where nonconformity is not tolerated, McDonald predicts the frequency of BAPCO traits in these populations will be very low.

"The frequency and intensity of BAPCO traits in the U.S. have increased, for example, because individuals are more likely to meet and have children with other individuals with similar education, occupations and interests than would have been possible 100 years ago," she said.

"When men and women with BAPCO traits have children it can increase both the frequency and intensity of BAPCO traits in the population."

No Cure for Autism

McDonald said she now knows what causes autism but it is not something that could be cured and, honestly, she wouldn't want to do that.

"What we call 'autism' is the BAPCO personality combined with a disability or very intense BAPCO traits," she said. "Because the BAPCO is made of socially valued traits, it is not possible to cure autism. Instead, we need to focus on the full range of disabilities that affect people with and without the BAPCO personality."

McDonald said future research should focus on the causes and treatment of underlying disabilities and also on providing supports such as interventions for individuals with disabilities, including those who have this combination of BAPCO and disability intersection.

"When we talk about autism we need to address the developmental disabilities that these individuals are experiencing but also find way to support and enhance their broader autism trait constellations," she said.

Scientists from Russia, Austria, and the USA have discovered virus-like particles in the bacterial symbionts of Bryozoa - a phylum of colonial aquatic invertebrates - filter-feeders dominating in many bottom ecosystems. The research project was planned and supervised by scientists from St Petersburg University. Some of the virus-like particles resemble red blood cells, while others have a sea-urchin-like appearance. Although viruses have never been reported inside symbiotic bacteria in bryozoans, scientists suggest that this "matryoshka doll" may have a prominent effect on the bacterial hosts.

Life of most organisms on our planet is closely associated with bacterial communities - microbiota that plays a very important role in the health of the host from helping to control digestion to protection of the host from infections. Bryozoa are not an exception. Inside some of these invertebrates the symbiotic bacteria live that, e.g. make larvae not eatable for fish. Yet how the symbiosis can benefit the host itself still remains a mystery. Scientists from the St Petersburg University tried to solve it and eventually made an unexpected discovery.

They encountered the virus-like particles in the colonies of the cheilostome bryozoan Bugula neritina that were intertidally collected from the east coastal area of the USA, and in Paralicornia sinuosa that was collected near Australia. Between bacteria of B. neritina the virus-like particles were found that resembled red blood cells. The particles found inside and between bacteria of P. sinuosa were reminiscent the sea urchins. These were spherical and consisting of cylindrical/tube-like elements evenly radiating from the central double-walled 'core'. To gain a better understanding of what the particles are and what role they play in the life of moss animals the biologists from the University established a collaboration with experts from the Winogradsky Institute of Microbiology, the Russian Academy of Sciences.

'So-called "urchins" from the Australian bryozoan is much alike the "metamorphosis associated contractile structures (MACs)" described by the American scientists in 2014. These structures can be found inside the biofilm-forming marine bacteria Pseudoalteromonas luteoviolacea. MACs have the same origin as the viruses-bacteriophages. In fact, ???s are an assemblage of multiple contractile systems related to the contractile tails of myoviruses (tailed bacteriophages with contractile tails). Contacting the biofilms of Pseudoalteromonas can induce larval metamorphosis in the tubeworm Hydroides elegans. When settling, the larvae of H. elegans contact the biofilm at sea bottom, and the "urchins"' inside the biofilm inject a protein signal into the cells of the larvae. The delivery of the protein induces larval metamorphosis,' said Professor Andrey Letarov, Head of the Laboratory of Microbial Viruses at the Winogradsky Institute of Microbiology, RAS.

Yet there are some differences, Dr Letarov explained. The 'urchins' inside the bacteria that were studied by the Americans are very large and not 'well-organized', while the particles inside the bacteria of Paralicornia sinuosa are finely structured into tiny spheres. But generally speaking, there is much in common between these structures. What role the 'urchins' play in this biological system remains unclear. Yet there is some tentative evidence.

'We know that in some bryozoan species, in particular in Bugula neritina, bacteria from maternal tissues are transmitted to larvae thus providing the vertical transfer of the symbionts between bryozoan generations. Viruses should be transmitted together with bacteria. Yet not all of the bacteria are moved into the larvae. Those that remain in the host can multiply producing an excess of toxic metabolites. This can pose a potential threat to the host. Presumably, the newly discovered viruses can destroy these bacteria thus regulating the symbiont number in the bryozoan hosts,' said Andrew Ostrovsky, Professor at the Department of Invertebrate Zoology, St Petersburg University, and head of the research project.

'The Mac-like structures ("urchins") found in the Australian bryozoan also destroy bacteria. Yet it remains unclear whether they have analogous functions as in the system P. luteoviolaceae - H. elegans. In other words, whether they can deliver a signal from bacterial symbionts to the host,' added Dr Letarov.

Much has to be done, especially in respect to molecular processes in which the virus-like particles can be involved. The scientists are planning to sequence the genome of bacteria containing these virus-like particles to reveal genetic information that regulates their formation. The work will be done in collaboration with bioinformaticians. This may help in gaining a better understanding of what proteins are produced by the 'urchins' and how they can influence other parts of this biological "matryoshka".

Yet collecting fresh bryozoan colonies to continue this research is rather difficult task. Cultivation of the bryozoan symbiotic bacteria in the laboratory was never successful. They can be collected only in their natural habitats, which are the coastal area of Australia and east coastal area of the USA. But even getting there, especially during the current pandemic, is very difficult. This makes scientists think about finding the similar symbiotic systems near the Marine Biological Station of the University at the White Sea. Among bryozoans found in the region, some species possess bacteria.

'Viruses are non-cellular infectious agents. Presumably, they are the most numerous 'particles of life' on the planet. They can replicate, i.e. make copies of themselves, by using bacteria, archaea, protists, fungi, plants, and animals. In other words, all living organisms,' Andrew Ostrovsky said. 'It is a gigantic world that dates back to the period when cellular life on Earth began. Not surprisingly then that viruses are deeply involved in regulation of many processes in living organisms. Without viruses, life and the planet would not exist.'

Like Peter Pan, some cells never grow up. In cancer, undifferentiated stem cells may help tumors such as glioblastoma become more aggressive than other forms of the disease. Certain groups of genes are supposed to help cells along the path to maturity, leaving their youthful "stemness" behind. This requires sweeping changes in the microRNAome -- the world of small non-coding material, known as microRNAs, that control where and when genes are turned on and off. Many microRNAs are tumor-suppressive; in cancer, the microRNAome is distorted and disrupted. Recent work by researchers at Brigham and Women's Hospital pinpoints critical changes in an enzyme known as DICER, which create a cascade of effects on this microRNAome. The team identified primary actors circ2082, a circular RNA, and RBM3, an RNA-binding protein, which form a complex with DICER to trap it in the nucleus of glioblastoma cells, therefore disrupting the cytoplasmic microRNAome. Findings are published in Science Advances.

"We are always trying to find the magic bullet to fight cancer. The problem with the magic bullet is that it's only going to hit a few tumor cells, since the other tumor cells don't have that target. We are looking for the common vulnerability -- what is the common thing that we can target?" said Antonio Chiocca, MD, PhD, chair of the Brigham's Department of Neurosurgery. "With this discovery, we can target something way upstream: a very common target at the epigenetic level."

Prior research showed the number of microRNAs expressed in cancer cells to be low when compared to non-cancerous cells. In addressing this observation, primary researchers Jakub Godlewski, PhD, and Agnieszka Bronisz, PhD, both formerly at the Brigham and now based at the Mossakowski Medical Research Centre of Polish Academy of Sciences, Warsaw, Poland, led a team that determined the absence of DICER enzyme is likely the cause of this cytoplasmic microRNA depletion. The team discovered DICER was being trapped in the nucleus by repressive interactions with RBM3 and circ2082.

In order to characterize these new actors, researchers used patient-derived glioblastoma cells, which allowed them to study outcomes genetically and phenotypically similar to real patient outcomes. The cells were studied both in vitro and in vivo through implantation into the brains of lab mice. The level of microRNA expression in these mice changed their survival rates. If circ2082 was knocked down, the nuclear DICER complex of circ2082, DICER, and RBM3 was disrupted, more microRNAs were present in the cytoplasm, and the survival outcome was far greater. In these mice whose tumors had circ2082 knocked down, death by tumorigenesis never occurred, while their non-knockdown counterparts all experienced death by tumorigenesis.

The downstream effect of this circ2082 expression also heavily influenced morbidity in human patients whose tissues were retrospectively analyzed. Patients with circ2082-dependent signature less widely expressed had overall longer lifespans after cancer diagnosis.

"Targeting these upstream tumor proliferators has the potential to dramatically change the cancer treatment landscape," said Chiocca. "We look forward to clinical translation of this research as we search for the right inhibitors."

While rare, botulism can cause paralysis and is potentially fatal. It is caused by nerve-damaging toxins produced by Clostridium botulinum -- the most potent toxins known. These toxins are often found in contaminated food (home canning being a major culprit). Infants can also develop botulism from ingesting C. botulinum spores in honey, soil, or dust; the bacterium then colonizes their intestines and produces the toxin.

Once paralysis develops, there is no way to reverse it, other than waiting for the toxins to wear off. People with serious cases of botulism may need to be maintained on ventilators for weeks or months. But a new treatment approach and delivery vehicle, described today (?) in Science Translational Medicine, could change that.

"There are anti-toxins, but these only work before the toxins enter the motor neurons," says Min Dong, PhD, a researcher in Boston Children's Hospital's Department of Urology and corresponding author on the paper. "What we have developed is the first therapy that can eliminate toxins after they get inside neurons."

If proven in humans, the approach would represent a breakthrough in treating botulism. In mice, the treatment successfully got inside neurons, reversed muscle paralysis within hours, and enabled mice to withstand doses of botulinum toxin that would otherwise be lethal.

Letting a toxin lead the way

Dong and his colleagues needed to surmount two technical barriers that have prevented the botulism from being treated effectively in the past. Intriguingly, their solution lay in botulinum toxin itself.

"One barrier to treatment has been getting across the cell membrane, which is difficult for protein drugs," explains Shin-Ichiro Miyashita, PhD, a postdoctoral fellow in Dong's lab and first author on the paper. "The other is targeting specific cell types, and in this case specificity toward motor neurons and nerve terminals. We took advantage of the fact that botulinum neurotoxins target motor neurons naturally and efficiently, and can deliver a protein cargo across cell membranes."

The treatment is therefore two-pronged. A botulinum toxin (detoxified through introduced mutations) is the delivery vehicle. The cargo -- i.e., the active drug -- is a mini-antibody derived from the antibodies of camels, developed by collaborator Charles Shoemaker, PhD, at Tufts University. The team showed that two of these so-called nanobodies can be delivered in tandem into neurons, neutralizing botulinum toxins type A and B at one go.

But there was one more problem to solve.

"This idea and approach had been attempted, but it was difficult to completely get rid of toxicity," says Dong, "until we identified a new toxin, botulinum neurotoxin X, in 2017. Unlike other botulinum toxins, this new toxin shows no toxicity after we introduce mutations, and serves as a safe delivery tool."

Botox reversal

Besides botulism, Dong thinks the new treatment could be useful as a "botox reversal" agent. Botox injections, using tiny quantities of the type A botulinum toxin, can safely treat wrinkles and many other medical conditions like neck spasms, excessive sweating, or overactive bladder. However, when the injection goes awry, botox can cause unwanted muscle paralysis as a side effect, and patients have to live with the paralysis for months.

"We can potentially inject our therapeutic protein and get rid of botox in neurons and paralysis within a few hours," Dong says.

A general delivery platform for neuroactive drugs?

The toxin-guided approach may offer a platform for getting biologic drugs into neurons to treat other disorders, Dong believes. Currently, most biologic drugs act only on cell-surface targets and cannot get into the cell's interior.

"We provide a protein-based drug delivery platform that achieves highly specific targeting of neurons and efficient penetration of cell membranes," Dong says. "Combined with nanobodies, which can be developed fairly readily against any protein of interest, this platform can be used to develop therapeutics that modulate proteins and biological processes inside neurons. Its modular nature even allows us to target cell types other than neurons by switching the cell-targeting domain. This could present a general approach for precision drug delivery into cells."

Jan. 8, 2021 - A new study published online in the Annals of the American Thoracic Society examines the recovery of lung function and overall wellness in individuals who had varying degrees of COVID-19 severity. Little is known about lung health following infection with SARS-CoV-2, the virus that causes COVID-19, and whether later respiratory problems, fatigue and ill health are associated with the disease's initial severity.

In "Persistent Poor Health Post-COVID-19 Is Not Associated With Respiratory Complications or Initial Disease Severity," Liam Townsend, MD, and co-authors looked at a number of measures of recovery for 153 patients who were followed in an outpatient clinic a median of 75 days after their COVID-19 diagnoses.

"We found that fatigue, ill-health and breathlessness were all common following COVID-19," said Dr. Townsend of the Department of Infectious Diseases, St. James's Hospital and Department of Clinical Medicine, Trinity Translational Medicine Institute, Trinity College, Dublin, Ireland. "However, these symptoms appeared to be unrelated to the severity of initial infection or any single measurement at the time of an outpatient appointment."

Dr. Townsend and colleagues graded the patients' initial infection severity as: (1) not requiring admission; (2) requiring hospital admission, or (3) requiring ICU care. Nearly half (74) of these patients required hospital admission during acute infection.

The researchers examined the association between the patients' initial illness and abnormal chest x-ray, six-minute walk test distance--in which patients walked at their normal pace for six minutes--and perception of maximal exertion during their follow-up appointments. Patients were also asked about whether they felt fatigued and whether they thought they had returned to full health. Their responses were graded using validated scales.

The study team also looked at participants' chest x-rays in order to detect persistent infection or significant scarring (fibrosis) and performed CT imaging for patients with abnormal chest x-rays. Bloodwork was done to measure such indicators of disease as C-reactive protein, which increases when there is inflammation in the body. Statistical analyses were performed.

"We were surprised by our findings. We expected a greater number of abnormal chest x-rays. We also expected the measures of ongoing ill-health and abnormal findings to be related to severity of initial infection, which was not the case," Dr. Townsend said.

The authors' findings suggest that COVID-19 does not cause significant fibrosis, with lung scarring seen on CT scans of only 4 percent of study participants, following x-ray detection of earlier abnormalities in a larger group.

Sixty-two percent of patients felt they had not returned to full health, while 47 percent were classified as having fatigue. Patients who felt they had to exert themselves during moderate exercise also reported they felt fatigued and in poor health. Patients' length of inpatient hospital stays and frailty were associated with covering less distance in the walk test.

"These findings have implications for clinical care, in that they demonstrate the importance of following up all patients who were diagnosed with COVID-19, irrespective of severity of initial infection. It is not possible to predict who will have ongoing symptoms," Dr. Townsend stated.

Researchers from Oklahoma State University, University of Missouri, Iowa State University, and University of Georgia published a new paper in the Journal of Marketing that investigates the question of how salespeople should balance advocacy for the seller with advocacy for the customer.

The study, forthcoming in the Journal of Marketing, is titled "Salesperson Dual Agency in Price Negotiations" and is authored by Justin Lawrence, Lisa Scheer, Andrew Crecelius, and Son Lam.

How should salespeople represent both the seller and the customer when their interests diverge, as in pricing negotiations? The research team extends a dual agency framework to the sales domain and examines the salesperson's role throughout the three stages of the discount process: (a) the customer's discount request, (b) the seller's approval, and (c) the seller's post-approval profit. This framework is tested across three multimethod studies and provides the first empirical investigation that deconstructs the sequential B2B customer-specific discount process.

The study concludes that the most favorable outcomes result when the salesperson engages in high levels of both customer advocacy and seller advocacy. As Lawrence explains, "Traditional agency theory research focuses on the salesperson as an agent of the seller. In contrast, we emphasize that the customer also considers the salesperson as its agent. As a result, the salesperson is unlikely to succeed as a seller advocate without also engaging in customer advocacy. For salespeople, our research offers guidance on a classic dilemma: the tension between the customer's desire for a lower price and the seller's insistence on demonstrating the value of its offering rather than compromising on price."

"Our findings are liberating for salespeople; they need not choose a side in discount negotiations. Instead, they can compartmentalize their duties. When dealing with the customer, they act as a fierce advocate for the seller; when dealing with seller personnel, they are a strong customer advocate" adds Scheer. This compartmentalization, somewhat paradoxically, enables the synergistic relationship between the two forms of advocacy and drives superior outcomes for both firms.

Although a salesperson's forceful advocacy on a customer's behalf may appear to threaten the seller's bottom line, sellers should consider that salesperson customer advocacy may be vital to a profitable relationship with the customer. Sellers are advised to encourage salespeople to serve as agents of their customers while also effectively acting as agents of the seller. Sales training programs, for example, can emphasize the importance of taking concrete actions to develop deep understanding of the customer's needs, effectively representing the customer's interests, and advocating forcefully for discounts and other seller concessions and assistance when warranted to bolster the customer's business and deepen the relationship.

One practical approach for sellers to leverage salesperson dual agency to their advantage is to encourage the development of multiple interfirm linkages with the customer, including cross-boundary interpersonal ties. Similarly, allowing a customer to learn more about the salesperson's customer advocacy amplifies the effectiveness of seller advocacy toward that customer. Following the seller's discount decision--even if the discount was denied--sales managers or pricing personnel can find ways to notify customers how the salesperson went to bat for them.

Customers should not hinder seller efforts to learn more about the salesperson's seller advocacy. For example, the seller might send the customer a survey asking about the salesperson's behaviors. Customers should view this as an opportunity to improve the seller's certainty about the salesperson's faithful representation. These findings differ from the implications of extant sales theories and prior sales research, which suggest that the salesperson serving two masters whose objectives are opposed (as with discounts) can lead to role stress and reduced performance.

Salespeople, sales managers, and organizational buyers can find value in this research. Whereas sellers warn salespeople not to 'be sold by the customer,' the findings suggest that sellers should encourage salespeople to advance the customer's agenda in discount negotiations. Engaging in seller advocacy, without customer advocacy, is likely to be ineffective or even lead to reduced seller profit. Customers and sellers can benefit from understanding that salesperson's advocacy for the other party is mutually beneficial and an important part of the salesperson's role.

Ribosomes are the complexes of ribonucleoproteins at the heart of protein synthesis in cells. However in the absence of conclusive evidence, how these complexes operate has been open to debate. Now Hirotatsu Imai and Noriyuki Kodera at Kanazawa University, alongside Toshio Uchiumi at Niigata University in Japan, show visualizations of the structural dynamics and factor pooling that take place at ribosome stalk proteins as they build new proteins.

Ribosomes were first discovered in the 1950s and their broad function has been widely understood for some time - they read messenger RNA sequences and from that generate sequences of correctly ordered amino acids into new proteins. The ribosome stalk protein in particular plays an integral role in the protein synthesis process by recruiting protein factors responsible for translation and elongation of the amino acid sequence. However it has been hard to satisfactorily establish the structure of the bound ribosome stalk protein because of its flexibility. Here the high resolution and fast image capture of high-speed atomic force microscopy proved invaluable.

Atomic force microscopy uses a nanoscale tip to feel samples, much like a vinyl record player needle scanning over a record, except that the details identified by an atomic force microscope can have atomic-scale resolution. The versatility of the technique for different surfaces was already a huge advantage for biological studies, but with the advent of high-speed atomic force microscopy the technique was able to capture dynamic processes for the first time as well. Imai, Uchiumi and Kodera used the technique to reveal that the stalk protein actually flips between two conformations - one that agrees with previous structural models and one entirely unexpected new conformation.

As for how the ribosome operates, a two step mechanism had been previously proposed to describe how genetic information is translated through proteins known as "translational GTPase factors". The first step is the recruitment of the factors to the factor-tethering site on the protein stalk, thereby increasing the concentration of factors there - so-called factor pooling. The second step is the binding and stabilizing of a translational GTPase on the ribosomal factor-binding center to catalyse GTPase hydrolysis. From their high speed atomic force microscopy study the researchers were able to obtain the first visual evidence for the translational GTPase factor pooling mechanism by the ribosomal stalk.

Although the study was unable to give conclusive evidence of the action of the factors once bound, the researchers did note that the factors appeared to be retained in the vicinity once GTPase hydrolysis was complete, suggesting a potential role of the stalk protein in further stages of protein synthesis. The researchers conclude, "future work with HS-AFM will provide further important information to understand the dynamic behaviors of these complex translational machineries."

The North Sea is a heavily trafficked area, with major shipping routes crossing its waters, and fisheries, offshore oil rigs, and wind farms populating its waves. All this activity inevitably has an effect on marine wildlife, and scientists are particularly interested in how the harbor porpoise population has fared in the face of such disturbances.

The harbor porpoise is known as a "sentinel species" - animals which indicate the health of an ecosystem and point to potential risks (think of the canary in the coal mine). According to a recent study published in Frontiers in Marine Science, their population is declining in the German North Sea.

"The trend seen here is concerning," says Dr Anita Gilles of the University of Veterinary Medicine Hannover, Germany, one of the study's authors. Particularly troubling is the fact that harbor porpoises have experienced a strong decline in protected areas, such as the Special Area of Conservation (SAC) Sylt Outer Reef, which was specifically designated to keep marine life safe. In that particular region, the harbor porpoise population declined by an average of 3.79% per year. In the south, however, the population increased, indicating a possible shift in distribution. Overall, the harbor porpoise population declined by 1.79% per year in the German North Sea.

In order to get an accurate abundance estimate, Gilles and her colleagues used a system in which the surveyed area was divided by transects into smaller blocks, then observed by plane. "Knowing the abundance of a population is at the heart of ecology, but extremely challenging for mobile species in a rapidly changing marine environment like the North Sea," she says. Their system is a standard method to determine wildlife population sizes, and includes measures to ensure accuracy, such as doubling back to account for diving porpoises. Additionally, an innovative analysis framework, developed by co-author Sacha Viquerat and based on the Bayesian paradigm, was implemented for trend analysis.

The study is also noteworthy for its two decades-long time span. "After almost 20 years of systematic data collection... we now have a wealth of data at hand," says Gilles. This data, while troubling, can help drive legislation to aid conservation efforts. "Abundance, distribution, and trends are key for EU and other legislative instruments, and for marine conservation management in general."

Moreover, co-author Dominik Nachtsteim is hopeful that their survey design and data analysis methods can be used in other regions where a dedicated monitoring concept needs to be implemented. Their study limited itself to the German North Sea, meaning that population counts and observed trends in the broader North Sea are missing.

As for why there are fewer harbor porpoises today than there were 20 years ago, Gilles and her colleagues hypothesize it might be due to an increase in human activities, a change in prey availability, a distribution shift. "Most probable, it is a mixture of different causes and cumulative effects," says Gilles. But because their study was focused on data collection and not understanding causes, "We urgently need more research into the drivers of change."