Losing a loved one may speed up aging

Losing someone close, like a family member, can make you age faster, says a new study from Columbia University Mailman School of Public Health and the Butler Columbia Aging Center. The study found that people who lost a parent, partner, sibling, or child, showed signs of older biological age compared to those who hadn't experienced such losses. The research was published in JAMA Network Open.

Biological aging is the gradual decline in how well your cells, tissues, and organs function, leading to a higher risk of chronic diseases. Scientists measure this type of aging using DNA markers known as epigenetic clocks.

"Few studies have looked at how losing a loved one at different stages of life affects these DNA markers, especially in study samples that represent the U.S .population," said Allison Aiello, PhD, the James S. Jackson professor of health longevity in Epidemiology and the study's lead author. "Our study shows strong links between losing loved ones across the life course from childhood to adulthood and faster biological aging in the U.S."

The study, a collaboration with the Carolina Population Center at UNC Chapel Hill, suggests that the impact of loss on aging can be seen long before middle age and may contribute to health differences among racial and ethnic groups.

The researchers used data from the National Longitudinal Study of Adolescent to Adult Health, which started in 1994-95. It followed participants from their teenage years into adulthood.

To measure familial loss during childhood or adolescence from the longitudinal study, Aiello and colleagues followed participants through various waves, and aging timeframes. Wave I surveyed 20,745 adolescents in grades 7-12, most of whom were aged 12-19. Participants have been followed ever since. Wave V took place between 2016 and 2018 and completed interviews with 12,300 of the original participants. In the latest wave, between 2016 and 2018, participants were invited for an additional home exam where a blood sample of the nearly 4,500 visited was provided for DNA testing.

The study looked at losses experienced during childhood or adolescence (up to 18 years old) and adulthood (19 to 43 years old). They also examined the number of losses experienced across this time period. Biological aging data were assessed from blood DNA methylation using epigenetic clocks including DunedinPACE which was developed by by Aiello's Aging Center colleague and study co-author Dan Belsky and his collaborators at Duke University.

Nearly 40 percent of participants experienced at least one loss in adulthood between the ages of 33 and 43. Parental loss was more common in adulthood versus in childhood and adolescence (27 percent versus 6 percent). A larger proportion of Black (57 percent) and Hispanic (41 percent) participants experienced at least one loss compared to White participants (34 percent).

People who experienced two or more losses had older biological ages according to several epigenetic clocks. Experiencing two or more losses in adulthood was more strongly linked to biological aging than one loss and significantly more so than no losses.

"The connection between losing loved ones and health problems throughout life is well-established," Aiello noted. "But some stages of life might be more vulnerable to the health risks associated with loss and the accumulation of loss appears to be a significant factor."

For example, losing a parent or sibling early in life can be very traumatic, often leading to mental health issues, cognitive problems, higher risks of heart disease, and a greater chance of dying earlier. Losing a close family member at any age poses health risks, and repeated losses can increase the risks of heart disease, mortality, and dementia; and impacts may persist or become apparent long after the event.

Aiello and her co-authors emphasize that while loss at any age can have long-lasting health impacts, the effects might be more severe during key developmental periods like childhood or early adulthood. "We still don't fully understand how loss leads to poor health and higher mortality, but biological aging may be one mechanism as suggested in our study. Future research should focus on finding ways to reduce disproportionate losses among vulnerable groups. For those who experience loss, providing resources for coping and addressing the trauma is essential. ," Aiello concluded.

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Scientists work out the effects of exercise at the cellular level

The health benefits of exercise are well known but new research shows that the body's response to exercise is more complex and far-reaching than previously thought. In a study on rats, a team of scientists from across the United States has found that physical activity causes many cellular and molecular changes in all 19 of the organs they studied in the animals.

Exercise lowers the risk of many diseases, but scientists still don't fully understand how exercise changes the body on a molecular level. Most studies have focused on a single organ, sex, or time point, and only include one or two data types.

To take a more comprehensive look at the biology of exercise, scientists with the Molecular Transducers of Physical Activity Consortium (MoTrPAC) used an array of techniques in the lab to analyze molecular changes in rats as they were put through the paces of weeks of intense exercise. Their findings appear in Nature.

The team studied a range of tissues from the animals, such as the heart, brain, and lungs. They found that each of the organs they looked at changed with exercise, helping the body to regulate the immune system, respond to stress, and control pathways connected to inflammatory liver disease, heart disease, and tissue injury.

The data provide potential clues into many different human health conditions; for example, the researchers found a possible explanation for why the liver becomes less fatty during exercise, which could help in the development of new treatments for non-alcoholic fatty liver disease.

The team hopes that their findings could one day be used to tailor exercise to an individual's health status or to develop treatments that mimic the effects of physical activity for people who are unable to exercise. They have already started studies on people to track the molecular effects of exercise.

Launched in 2016, MoTrPAC draws together scientists from the Broad Institute of MIT and Harvard, Stanford University, the National Institutes of Health, and other institutions to shed light on the biological processes that underlie the health benefits of exercise. The Broad project was originally conceived of by Steve Carr, senior director of Broad's Proteomics Platform; Clary Clish, senior director of Broad's Metabolomics Platform; Robert Gerszten, a senior associate member at the Broad and chief of cardiovascular medicine at Beth Israel Deaconess Medical Center; and Christopher Newgard, a professor of nutrition at Duke University.

Co-first authors on the study include Pierre Jean-Beltran, a postdoctoral researcher in Carr's group at Broad when the study began, as well as David Amar and Nicole Gay of Stanford. Courtney Dennis and Julian Avila, both researchers in Clish's group, were also co-authors on the manuscript.

"It took a village of scientists with distinct scientific backgrounds to generate and integrate the massive amount of high quality data produced," said Carr, a co-senior author of the study. "This is the first whole-organism map looking at the effects of training in multiple different organs. The resource produced will be enormously valuable, and has already produced many potentially novel biological insights for further exploration."

The team has made all of the animal data available in an online public repository. Other scientists can use this site to download, for example, information about the proteins changing in abundance in the lungs of female rats after eight weeks of regular exercise on a treadmill, or the RNA response to exercise in all organs of male and female rats over time.

Whole-body analysis

Conducting such a large and detailed study required a lot of planning. "The amount of coordination that all of the labs involved in this study had to do was phenomenal," said Clish.

In partnership with Sue Bodine at the Carver College of Medicine at the University of Iowa, whose group collected tissue samples from animals after up to eight weeks of training, other members of the MoTrPAC team divided the samples up so that each lab -- Carr's team analyzing proteins, Clish's studying metabolites, and others -- would examine virtually identical samples.

"A lot of large-scale studies only focus on one or two data types," said Natalie Clark, a computational scientist in Carr's group. "But here we have a breadth of many different experiments on the same tissues, and that's given us a global overview of how all of these different molecular layers contribute to exercise response."

In all, the teams performed nearly 10,000 assays to make about 15 million measurements on blood and 18 solid tissues. They found that exercise impacted thousands of molecules, with the most extreme changes in the adrenal gland, which produces hormones that regulate many important processes such as immunity, metabolism, and blood pressure. The researchers uncovered sex differences in several organs, particularly related to the immune response over time. Most immune-signaling molecules unique to females showed changes in levels between one and two weeks of training, whereas those in males showed differences between four and eight weeks.

Some responses were consistent across sexes and organs. For example, the researchers found that heat-shock proteins, which are produced by cells in response to stress, were regulated in the same ways across different tissues. But other insights were tissue-specific. To their surprise, Carr's team found an increase in acetylation of mitochondrial proteins involved in energy production, and in a phosphorylation signal that regulates energy storage, both in the liver that changed during exercise. These changes could help the liver become less fatty and less prone to disease with exercise, and could give researchers a target for future treatments of non-alcoholic fatty liver disease.

"Even though the liver is not directly involved in exercise, it still undergoes changes that could improve health. No one speculated that we'd see these acetylation and phosphorylation changes in the liver after exercise training," said Jean-Beltran. "This highlights why we deploy all of these different molecular modalities -- exercise is a very complex process, and this is just the tip of the iceberg."

"Two or three generations of research associates matured on this consortium project and learned what it means to carefully design a study and process samples," added Hasmik Keshishian, a senior group leader in Carr's group and co-author of the study. "Now we are seeing the results of our work: biologically insightful findings that are yielding from the high quality data we and others have generated.That's really fulfilling."

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Microplastics make their way from the gut to other organs

It's happening every day. From our water, our food and even the air we breathe, tiny plastic particles are finding their way into many parts of our body.

But what happens once those particles are inside? What do they do to our digestive system?

In a recent paper published in the journal Environmental Health Perspectives, University of New Mexico researchers found that those tiny particles -- microplastics -- are having a significant impact on our digestive pathways, making their way from the gut and into the tissues of the kidney, liver and brain.

Eliseo Castillo, PhD, an associate professor in the Division of Gastroenterology & Hepatology in the UNM School of Medicine's Department of Internal Medicine and an expert in mucosal immunology, is leading the charge at UNM on microplastic research.

"Over the past few decades, microplastics have been found in the ocean, in animals and plants, in tap water and bottled water," Castillo, says. "They appear to be everywhere."

Scientists estimate that people ingest 5 grams of microplastic particles each week on average -- equivalent to the weight of a credit card.

While other researchers are helping to identify and quantify ingested microplastics, Castillo and his team focus on what the microplastics are doing inside the body, specifically to the gastrointestinal (GI) tract and to the gut immune system.

Over a four-week period, Castillo, postdoctoral fellow Marcus Garcia, PharmD, and other UNM researchers exposed mice to microplastics in their drinking water. The amount was equivalent to the quantity of microplastics humans are believed to ingest each week.

Microplastics had migrated out of the gut into the tissues of the liver, kidney and even the brain, the team found. The study also showed the microplastics changed metabolic pathways in the affected tissues.

"We could detect microplastics in certain tissues after the exposure," Castillo says. "That tells us it can cross the intestinal barrier and infiltrate into other tissues."

Castillo says he's also concerned about the accumulation of the plastic particles in the human body. "These mice were exposed for four weeks," he says. "Now, think about how that equates to humans, if we're exposed from birth to old age."

The healthy laboratory animals used in this study showed changes after brief microplastic exposure, Castillo says. "Now imagine if someone has an underlying condition, and these changes occur, could microplastic exposure exacerbate an underlying condition?"

He has previously found that microplastics are also impacting macrophages -- the immune cells that work to protect the body from foreign particles.

In a paper published in the journal Cell Biology & Toxicology in 2021, Castillo and other UNM researchers found that when macrophages encountered and ingested microplastics, their function was altered and they released inflammatory molecules.

"It is changing the metabolism of the cells, which can alter inflammatory responses," Castillo says. "During intestinal inflammation -- states of chronic illness such as ulcerative colitis and Crohn's disease, which are both forms of inflammatory bowel disease -- these macrophages become more inflammatory and they're more abundant in the gut."

The next phase of Castillo's research, which is being led by postdoctoral fellow Sumira Phatak, PhD, will explore how diet is involved in microplastic uptake.

"Everyone's diet is different," he says. "So, what we're going to do is give these laboratory animals a high-cholesterol/high-fat diet, or high-fiber diet, and they will be either exposed or not exposed to microplastics. The goal is to try to understand if diet affects the uptake of microplastics into our body."

Castillo says one of his PhD students, Aaron Romero, is also working to understand why there is a change in the gut microbiota. "Multiple groups have shown microplastics change the microbiota, but how it changes the microbiota hasn't been addressed."

Castillo hopes that his research will help uncover the potential impacts microplastics are having to human health and that it will help spur changes to how society produces and filtrates plastics.

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Toothed whale echolocation organs evolved from jaw muscles

Genetic analysis finds evidence suggesting that acoustic fat bodies in the heads of toothed whales were once the muscles and bone marrow of the jaw.

Dolphins and whales use sound to communicate, navigate and hunt.

New research suggests that the collections of fatty tissue that enable toothed whales to do so may have evolved from their skull muscles and bone marrow.

Scientists at Hokkaido University determined DNA sequences of genes which were expressed in acoustic fat bodies -- collections of fat around the head that toothed whales use for echolocation.

They measured gene expression in the harbor porpoise (Phocoena phocoena) and Pacific white-sided dolphin (Lagenorhynchus obliquidens). Their findings were published in the journal Gene.

The evolution of acoustic fat bodies in the head -- the melon in the whale forehead, extramandibular fat bodies (EMFB) alongside the jawbone, and intramandibular fat bodies (IMFB) within the jawbone -- was essential for sound use such as echolocation.

However, little is known about the genetic origins of those fatty tissues.

"Toothed whales have undergone significant degenerations and adaptations to their aquatic lifestyle," said Hayate Takeuchi, a PhD student at Hokkaido University's Hayakawa Lab and first author of the study.

One adaptation was the partial loss of their sense of smell and taste, along with the gain of echolocation to enable them to navigate in the underwater environment.

The researchers found that genes which are normally associated with muscle function and development were active in the melon and EMFBs.

There was also evidence of an evolutionary connection between the extramandibular fat and the masseter muscle, which in humans connects the lower jawbone to the cheekbones and is a key muscle involved in chewing.

"This study has revealed that the evolutionary tradeoff of masticatory muscles for the EMFB -- between auditory and feeding ecology -- was crucial in the aquatic adaptation of toothed whales," said Assistant Professor Takashi Hayakawa of the Faculty of Environmental Earth Science, who led the study.

"It was part of the evolutionary shift away from chewing to simply swallowing food, which meant the chewing muscles were no longer needed."

Analysis of gene expression in the intramandibular fat detected activity of genes related to immune functions, such as the activation of some elements of the immune response and regulation of T cell formation.

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World's simplest animals get their place in the tree of life

The group with the world's simplest animals -- tiny blob-like life forms with no organs and just a few cell types -- finally has a fleshed-out family tree built by a research group led by the American Museum of Natural History, St. Francis College, and the University of Veterinary Medicine Hannover. The study comes more than 100 years after the discovery of these ameboid animals called placozoans and represents the first -- and potentially only -- time in the 21st century that a backbone Linnaean taxonomy is constructed for an entire animal phylum. Published today in the journal Frontiers in Ecology and Evolution, the research is based on genetic makeup -- the presence and absence of genes -- rather than outward physical appearance, which is traditionally used to classify organisms.

"Placozoans look like miniscule, shape-shifting disks -- basically, they are the pancake of the animal world," said the study's co-lead author Michael Tessler, a research associate at the Museum and an assistant professor at St. Francis College. "For a taxonomist looking through a microscope, even a powerful one, there are almost no characters to compare and differentiate them. Yet, despite most of them looking almost exactly the same, we know that on the genetic level, there are very distinct lineages."

The first placozoan species was described in 1883, and Placozoa remained a "phylum of one" until DNA-based research in the last 20 years revealed that it contains multiple lineages. Most placozoans, which generally live in tropical and subtropical waters across the globe, are about the size of a grain of sand, with hair-like structures that allow them to move. "After decades of turmoil, this most exciting phylum has finally gotten the attention it deserves," said senior author Bernd Schierwater, a professor at the University of Veterinary Medicine Hannover.

"We wanted to know the relationships within this ancient group of animals and where it sits in the tree of life," said co-lead author Johannes Neumann, a recent doctoral graduate from the Museum's Richard Gilder Graduate School. "People have been speculating about that for decades, but now, by looking at differences among placozoans on the molecular level, we're able to paint a clear picture of how these animals are related to one another."

The researchers used a method called molecular morphology -- using differences in DNA sequences and other molecular characters -- to make classifications. In doing so, they established a backbone taxonomy: two new classes, four orders, three families, one genus, and one species. Their research also suggests that placozoans are most closely related to cnidarians (a group of aquatic animals including jellyfish, corals, and sea anemones) and bilaterians (animals that have a left and right side, like insects and humans).

"I personally collected placozoans on six continents for almost 10 years, did lab work and bioinformatic work on them, but it took decades of effort from a great number of colleagues to finally get to this exciting first classification for this cryptic phylum," Neumann said. "This is why we call our newly described species Cladtertia collaboinventa, which means 'discovered in collaboration.'"

The authors suggest that this study could serve as a template to revisit systematics of other organisms that look very similar, such as bacteria, fungi, protists, and parasites. Tessler also is the lead author of a second paper out now in Frontiers in Ecology and Evolution that makes the case for molecular morphology in other groups of organisms that have few distinguishable visual features but are genetically diverse.

"Taxonomic blank slates are problematic. Without names, communication is hampered, and other scientific progress is slowed," said Tessler. "We suggest that the morphology of molecules, such as proteins -- which have distinctive structures -- should not be considered as anything less than traditional morphology."

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The future of replacement organs is (quite possibly) here: Robust human intestinal organoids created in a lab

Researchers from Tokyo Medical and Dental University (TMDU) find that spheroids grown in suspension mature into human intestinal organoids when transferred to a bioreactor and differentiate into complex intestinal tissue upon transplantation.

Growing human body parts in the lab is a common trope of horror movies and sci-fi books. But growing miniature organ-like tissues in the lab is already within our reach. Researchers from Japan have developed a new approach that enables intestinal mini-organs to be grown more easily and efficiently in the lab. This holds immense promise for regenerative medicine.

In a study published in November in Cell Reports Methods, researchers from Tokyo Medical and Dental University (TMDU) reveal that applying a few specialized lab techniques yields intestine-like tissues of predictable size and composition.

Organoids are organ-like balls of cells that are grown in the lab from spheroids (even smaller balls) of human cells and mimic the properties of the organ from which the "seed" cell was taken. Organoids are used for studying organ function in a lab setting and are also promising tools in the field of regenerative medicine.

"There are established methods for growing human intestinal organoids (HIOs) from induced pluripotent stem cells (iPSCs)," states Junichi Takahashi, first author of the study. "However, these techniques are challenging to perform. They result in spheroids of varying sizes and are limited by the growth conditions, which can result in deformed and unhealthy spheroids over time."

To develop a more robust and consistent way to generate HIOs, the researchers explored the use of cell culture plates made with an ultra-low attachment polymer to encourage the cells to detach and grow in suspension. They also tested the effects of growing the resulting spheroids in a bioreactor, a specialized incubator that keeps the growth medium constantly flowing to improve the health of the cells.

"Using our technique, we were able to grow spheroids of a predictable, consistent size that could be modified by modulating the number of cells seeded into the plates," says Tomohiro Mizutani, corresponding author of the study. "Furthermore, transferring the spheroids to a bioreactor allowed them to grow even larger, into healthy HIOs."

These organoids were surrounded by mesenchyme, which is a type of tissue found between organs in the human body. Importantly, when the organoids were transplanted into mice, they continued to grow and differentiate, developing a complex tissue architecture reflecting that of mature intestine. "Our findings show that intestinal tissue can be generated from iPSC-derived HIOs by inducing spheroids in suspension and maturing them in a bioreactor," says Takahashi.

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Ancient disease has potential to regenerate livers

Leprosy is one of the world's oldest and most persistent diseases but the bacteria that cause it may also have the surprising ability to grow and regenerate a vital organ.

Scientists have discovered that parasites associated with leprosy can reprogramme cells to increase the size of a liver in adult animals without causing damage, scarring or tumors.

The findings suggest the possibility of adapting this natural process to renew ageing livers and increase healthspan -- the length of time living disease-free -- in humans.

Experts say it could also help regrow damaged livers, thereby reducing the need for transplantation, which is currently the only curative option for people with end-stage scarred livers.

Previous studies promoted the regrowth of mouse livers by generating stem cells and progenitor cells -- the step after a stem cell that can become any type of cell for a specific organ -- via an invasive technique that often resulted in scarring and tumour growth.

To overcome these harmful side-effects, Edinburgh researchers built on their previous discovery of the partial cellular reprogramming ability of the leprosy-causing bacteria, Mycobacterium leprae.

Working with the US Department of Health and Human Services in Baton Rouge, Louisiana, the team infected 57 armadillos -- a natural host of leprosy bacteria -- with the parasite and compared their livers with those of uninfected armadillos and those that were found to be resistant to infection.

They found that the infected animals developed enlarged -- yet healthy and unharmed -- livers with the same vital components, such as blood vessels, bile ducts and functional units known as lobules, as the uninfected and resistant armadillos.

The team believe the bacteria 'hijacked' the inherent regenerative ability of the liver to increase the organ's size and, therefore, to provide it with more cells within which to increase.

They also discovered several indicators that the main kinds of liver cells -- known as hepatocytes -- had reached a "rejuvenated" state in the infected armadilllos.

Livers of the infected armadillos also contained gene expression patterns -- the blueprint for building a cell -- similar to those in younger animals and human fetal livers.

Genes related to metabolism, growth and cell proliferation were activated and those linked with aging were downregulated, or suppressed.

Scientists think this is because the bacteria reprogramed the liver cells, returning them to the earlier stage of progenitor cells, which in turn became new hepatocytes and grow new liver tissues.

The team are hopeful that the discovery has the potential to help develop interventions for aging and damaged livers in humans. Liver diseases currently result in two million deaths a year worldwide.

The findings have been published in the journal Cell Reports Medicine. This work has been funded by the UK's Medical Research Council and the US National Institutes of Health and National Institute of Allergy and Infectious Diseases.

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Technology restores cell, organ function in pigs after death

Within minutes of the final heartbeat, a cascade of biochemical events triggered by a lack of blood flow, oxygen, and nutrients begins to destroy a body's cells and organs. But a team of Yale scientists has found that massive and permanent cellular failure doesn't have to happen so quickly.

Using a new technology they developed that delivers a specially designed cell-protective fluid to organs and tissues, the researchers restored blood circulation and other cellular functions in pigs a full hour after their deaths, they report in the Aug. 3 edition of the journal Nature.

The findings may help extend the health of human organs during surgery and expand availability of donor organs, the authors said.

"All cells do not die immediately, there is a more protracted series of events," said David Andrijevic, associate research scientist in neuroscience at Yale School of Medicine and co-lead author of the study. "It is a process in which you can intervene, stop, and restore some cellular function."

The research builds upon an earlier Yale-led project that restored circulation and certain cellular functions in the brain of a dead pig with technology dubbed BrainEx. Published in 2019, that study and the new one were led by the lab of Yale's Nenad Sestan, the Harvey and Kate Cushing Professor of Neuroscience and professor of comparative medicine, genetics, and psychiatry.

"If we were able to restore certain cellular functions in the dead brain, an organ known to be most susceptible to ischemia [inadequate blood supply], we hypothesized that something similar could also be achieved in other vital transplantable organs," Sestan said.

In the new study -- which involved senior author Sestan and colleagues Andrijevic, Zvonimir Vrselja, Taras Lysyy, and Shupei Zhang, all from Yale -- the researchers applied a modified version of BrainEx called OrganEx to the whole pig. The technology consists of a perfusion device similar to heart-lung machines -- which do the work of the heart and lungs during surgery -- and an experimental fluid containing compounds that can promote cellular health and suppress inflammation throughout the pig's body. Cardiac arrest was induced in anesthetized pigs, which were treated with OrganEx an hour after death.

Six hours after treatment with OrganEx, the scientists found that certain key cellular functions were active in many areas of the pigs' bodies -- including in the heart, liver, and kidneys -- and that some organ function had been restored. For instance, they found evidence of electrical activity in the heart, which retained the ability to contract.

"We were also able to restore circulation throughout the body, which amazed us," Sestan said.

Normally when the heart stops beating, organs begin to swell, collapsing blood vessels and blocking circulation, he said. Yet circulation was restored and organs in the deceased pigs that received OrganEx treatment appeared functional at the level of cells and tissue.

"Under the microscope, it was difficult to tell the difference between a healthy organ and one which had been treated with OrganEx technology after death," Vrselja said.

As in the 2019 experiment, the researchers also found that cellular activity in some areas of the brain had been restored, though no organized electrical activity that would indicate consciousness was detected during any part of the experiment.

The team was especially surprised to observe involuntary and spontaneous muscular movements in the head and neck areas when they evaluated the treated animals, which remained anesthetized through the entire six-hour experiment. These movements indicate the preservation of some motor functions, Sestan said.

The researchers stressed that additional studies are necessary to understand the apparently restored motor functions in the animals, and that rigorous ethical review from other scientists and bioethicists is required.

The experimental protocols for the latest study were approved by Yale's Institutional Animal Care and Use Committee and guided by an external advisory and ethics committee.

The OrganEx technology could eventually have several potential applications, the authors said. For instance, it could extend the life of organs in human patients and expand the availability of donor organs for transplant. It might also be able to help treat organs or tissue damaged by ischemia during heart attacks or strokes.

"There are numerous potential applications of this exciting new technology," said Stephen Latham, director of the Yale Interdisciplinary Center for Bioethics. "However, we need to maintain careful oversight of all future studies, particularly any that include perfusion of the brain."

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New research challenges long-held beliefs about limb regeneration

Ken Muneoka is no stranger to disrupting the field of regeneration; for example, in a 2019 ground-breaking publication in Nature, the Texas A&M University College of Veterinary Medicine & Biomedical Sciences (CVMBS) professor proved for the first time that joint regeneration in mammals was possible.

Now, his team is again challenging other centuries-old beliefs about the fundamental science of the field, this time related to how mammals might regenerate damaged parts of the body.

In humans, the natural ability to regenerate is limited to tissues like the epidermis, the outermost layer of skin, and some organs, such as the liver.

Other species, most notably salamanders, have the ability to regenerate complex structures such as bones, joints, and even entire limbs. As a result, scientists have been studying these species for more than 200 years to try to understand the mechanisms behind limb regeneration in the hopes of someday translating those mechanisms to induce more extensive regeneration in humans.

That research has led to a common belief that the single biggest key for limb regeneration is the presence of nerves.

While that may be true for salamanders and other species, it isn't the case in mammals, according to two of Muneoka's recently published studies. The first study, published last year in the Journal of Bone and Mineral Research, established that mechanical loading (the ability to apply force to or with an affected area) is a requirement for mammals. The second, published earlier this year in Developmental Biology, established that the absence of nerves does not inhibit regeneration.

Together, these findings present a sizeable shift in the thinking of how regeneration could work in human medicine.

"What these two studies show counteracts the two-century-old dogma that you need nerves to regenerate," Muneoka said. "What replaces it in mammals is that you need mechanical loading, not nerves."

Importance Of Mechanical Load

Scientists have long believed that two things must be present in an affected area in order to induce regeneration in mammals. The first is growth factors, which are molecules that can stimulate cells to regrow and reconstruct parts of the body.

In natural regeneration, these growth factors, which vary from species to species and by area being regenerated, are produced by the body. For human-induced regeneration, these growth factors must be introduced to the area.

The second factor believed to be necessary was nerves. This belief was predicated by many previous human-induced mammal regeneration studies on areas, usually digit tips, without nerves, in which the whole limbs were also no longer usable.

Those studies would have the predicted outcome -- when growth factors were introduced regeneration did not take place-leading to the conclusion that, like in other species, nerves were a requirement for regeneration.

But the mechanical load aspect was ignored.

In their studies, Muneoka and colleagues decided to take a step back and ask the question, "is it really the nerves, or is lack of mechanical load part of the equation as well?"

Connor Dolan, a former graduate student in Muneoka's lab and first author on both new studies (who now works at the Walter Reed National Military Medical Center), came up with a way to test the denervation requirement in mammals that was inspired by astronauts.

The technique, called hindlimb suspension, has been used by NASA and other scientists for decades to test how mammals react to zero gravity environments. A similar process is used during medical procedures on legs of large animals to prevent the animals from putting weight on the affected limbs.

"Dolan found that when the limbs were suspended, even though they still had lots of nerves and could move around, they couldn't actually put pressure on their limbs so the digit tips wouldn't regenerate," Muneoka said. "It just completely inhibited regeneration."

As soon as the mechanical load returns, however, regeneration is rescued.

"Absolutely nothing happens during the suspension," Muneoka said. "But once the load returns, there will be a couple weeks of delay, but then they'll begin to regenerate."

That first step proved that even though nerves might be required, the mechanical loading was a critical component to regeneration.

Taking the research a step further, Dolan's second publication showed that nerves weren't required by demonstrating that if a mouse has no nerves in one of its digits but does in the others -- so that it's still exerting force on the denervated digit -- that digit will still regenerate.

"He found that they regenerate a little bit slower, but they regenerated perfectly normally," Muneoka said.

Ramifications Of The Research

Muneoka is quick to point out that their studies aren't saying that previous research is wrong, just that it doesn't directly apply to humans.

"There have been a number of studies in salamanders that prove that when you remove the nerves, they do not regenerate," Muneoka said. "Researchers have also been able to put growth factors they know are being produced by nerves into the cells and rescue regeneration.

"So, salamanders probably do need nerves to regenerate," he said. "But if we're going to regenerate limbs in humans, it's going to be a lot more like what happens in mice."

Since first beginning to look at regeneration more than 20 years ago, a number of Muneoka's ideas have pushed back against the generally accepted theories about regeneration. He said that getting these two papers published took almost three years because they originally tried to submit them together.

"Many scientists don't embrace this idea," he said. "A lot of people's careers are really dependent on their studies of nerves and how they affect regeneration. For a study to come out and say that for humans it's unlikely you'll need the nerves, the whole biomedical application of what people are doing in salamanders and fish kind of goes out the window."

Looking Down The Road

Nerves not being required for regeneration in mammals may seem like an academic point. After all, what would be the point of regenerating a limb if the person couldn't feel it or control it because it had no nerves. In that sense, nerves are still going to be an important part of the puzzle.

From Muneoka's perspective, the shift is that instead of thinking of nerves as a requirement for regeneration, nerves are a part of what needs to be regenerated.

Larry Suva, head of the CVMBS' Department of Veterinary Physiology & Pharmacology (VTPP), says the issue is that nobody was even thinking about the load aspect previously.

"Think of a blast injury where a soldier is left with a stump," Suva said. "No one, until this paper came out, was even thinking about a requirement from mechanical influences. You had people see that a denervated animal doesn't regenerate and they're thinking it's because the nerve was cut, but nobody was studying the mechanical load aspect."

As Suva puts it, science is full of people looking where the light is best.

"I work on bones, so when I see a problem, I look at the bone problem," he said. "People who work on nerves, all they look at are nerves. So it's very rare that someone like Dr. Muneoka will take a step back and take a more holistic view.

"That's what he brought to this idea, to this 200-year-old data," Suva said. "We now have to look at regeneration through a different lens because now we know the mechanical influences are extremely important."

One of the results of research focusing on nerves is that scientists have been able to recreate the growth factors that nerves produce, which has allowed researchers to start regeneration in salamanders, even if the nerves aren't present. Suva said that with these new findings, scientists will now know they have to do the same with the mechanical load aspect if they want to start regeneration in mammals.

"Scientists already have been able to trick the body into thinking nerves are still present," he said. "But now they know they'll also have to trick it into thinking there's a mechanical load, something that has not been done before."

Because cells react differently under mechanical load, somehow, that load is being translated biochemically inside the cell.

"There's a small number of labs looking at the biochemical basis for what mechanical load does to a cell," Muneoka said. "If we could understand that biochemical signal, then perhaps the physical force of mechanical load can be replaced by some sort of cocktail of molecules that will create the same signals in the cells."

The end of the road toward full human regeneration may still be a long way in the future, but Suva says that this kind of fundamental shift in thinking is a major marker on that road.

"Regeneration of a human limb may still be science fiction, but we know some facts about it, and now we know you have to have that mechanical load along with the growth factors," he said. "That changes how future scientists and engineers are going to solve this problem.

Read more at Science Daily

How placentas evolved in mammals

The fossil record tells us about ancient life through the preserved remains of body parts like bones, teeth and turtle shells. But how to study the history of soft tissues and organs, which can decay quickly, leaving little evidence behind?

In a new study, scientists use gene expression patterns, called transcriptomics, to investigate the ancient origins of one organ: the placenta, which is vital to pregnancy.

"In some mammals, like humans, the placenta is really invasive, so it invades all the way through the wall of the uterus, into the maternal tissue. In other mammals, the placenta just touches the wall of the uterus. And then there's everything in between," says senior author Vincent J. Lynch, PhD, associate professor of biological sciences in the University at Buffalo College of Arts and Sciences.

"So what kind of placentas were early placentas?" he says. "We use gene expression patterns to reconstruct the evolution of the placenta and predict what the placenta of the last common ancestor of eutherian mammals looked like. Our data tells us that this placenta was invasive, and that non-invasive placentas evolved multiple times among mammals. This addresses a 150-year-old mystery: People have been debating what kind of placenta the first one was since then."

As Lynch explains, all living mammals other than marsupials and egg-laying monotremes are eutherians, which have long pregnancies in which the developing fetus evokes a strong physiological response in the mother.

The research was published on June 30 in eLife. Lynch led the study with first author Katelyn Mika, PhD, University of Chicago postdoctoral scholar in human genetics and in organismal biology and anatomy. Camilla M. Whittington, PhD, and Bronwyn M. McAllan, PhD, both at the University of Sydney, are also co-authors.

"Our ability to ask how the placenta might have functioned at different points during its evolution by using the gene expression profiles of currently existing animals to reconstruct the ancestors is a really cool approach and provides us more information on how changing gene expression can contribute to the evolution of a new trait," Mika says.

To conduct the analysis, the team compared the genes active in the uterus of various mammals during pregnancy. After finding that these gene expression profiles correlated with the degree of placental invasiveness, the scientists used their data to predict what ancestral mammalian placentas looked like.

The study included about 20 species, such as the egg-laying platypus, pouch-bearing marsupials, and a range of eutherian mammals that give birth to live young.

The small subset is one limitation of the analysis: The authors write in eLife that research on a larger number of species is needed to help determine the strength of the findings.

Nevertheless, the study makes important contributions in understanding how pregnancy evolved, Lynch says. The results could also benefit modern medicine.

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Your liver is just under three years old

The liver has a unique ability to regenerate after damage. However, it was unknown whether this ability decreases as we age. International scientists led by Dr. Olaf Bergmann at the Center for Regenerative Therapies Dresden (CRTD) at TU Dresden used a technique known as retrospective radiocarbon birth dating to determine the age of the human liver. They showed that no matter the person's age, the liver is always on average less than three years old. The results demonstrate that aging does not influence liver renewal, making the liver an organ that replaces its cells equally well in young and old people.

The liver is an essential organ that takes care of clearing toxins in our bodies. Because it constantly deals with toxic substances, it is likely to be regularly injured. To overcome this, the liver has a unique capacity among organs to regenerate itself after damage. Because a lot of the body's ability to heal itself and regenerate decreases as we age, scientists were wondering if the liver's capacity to renew also diminishes with age.

The nature of liver renewal in humans also remained a mystery. The animal models provided contradictory answers. "Some studies pointed to the possibility that liver cells are long-lived while others showed a constant turnover. It was clear to us that if we want to know what happens in humans, we need to find a way to directly assess the age of human liver cells," says Dr. Olaf Bergmann, research group leader at the Center for Regenerative Therapies Dresden (CRTD) at TU Dresden.

The Human Liver Remains a Young Organ

The interdisciplinary team of biologists, physicists, mathematicians, and clinicians led by Dr. Bergmann analyzed the livers of multiple individuals who died at ages between 20 and 84 years old. Surprisingly, the team showed that the liver cells of all subjects were more or less the same age.

"No matter if you are 20 or 84, your liver stays on average just under three years old," explains Dr. Bergmann. The results show that the adjustment of liver mass to the needs of the body is tightly regulated through the constant replacement of liver cells and that this process is maintained even in older people. This ongoing liver cell replacement is important for various aspects of liver regeneration and cancer formation.

Liver Cells with More DNA Renew Less

However, not all the cells in our liver are that young. A fraction of cells can live up to 10 years before renewing itself. This subpopulation of liver cells carries more DNA than the typical cells. "Most of our cells have two sets of chromosomes, but some cells accumulate more DNA as they age. In the end, such cells can carry four, eight, or even more sets of chromosomes," explains Dr. Bergmann.

"When we compared typical liver cells with the cells richer in DNA, we found fundamental differences in their renewal. Typical cells renew approximately once a year, while the cells richer in DNA can reside in the liver for up to a decade," says Dr. Bergmann. "As this fraction gradually increases with age, this could be a protective mechanism that safeguards us from accumulating harmful mutations. We need to find out if there are similar mechanisms in chronic liver disease, which in some cases can turn into cancer."

Lessons from the Nuclear Fallout

Determining the biological age of human cells is a massive technical challenge, as methods commonly used in animal models cannot be applied to humans.

Dr. Bergmann's group specializes in retrospective radiocarbon birth dating and uses the technique to assess the biological age of human tissues. Carbon is a chemical element that is ubiquitous and forms the backbone of life on Earth. Radiocarbon is one of a variety of types of carbon. It appears naturally in the atmosphere. Plants incorporate it through photosynthesis, in the same way as typical carbon, and pass it on to animals and humans. Radiocarbon is weakly radioactive and unstable. These characteristics are taken advantage of in archeology to determine the age of ancient samples.

"Archeologists have used the decay of radiocarbon successfully for many years to assess the age of specimens, one example being dating of the shroud of Turin," says Dr. Bergmann. "The radioactive decay of radiocarbon is very slow. It provides enough resolution for archeologists but it is not useful for determining the age of human cells. Nevertheless, we can still take advantage of the radiocarbon in our research."

The aboveground nuclear tests carried out in the 1950s introduced massive amounts of radiocarbon into the atmosphere, into the plants, and into the animals. As a result, cells formed in this period have higher amounts of radiocarbon in their DNA.

Following the official ban of aboveground nuclear testing in 1963, the amounts of atmospheric radiocarbon started to drop and so did the amounts of radiocarbon incorporated into the animal DNA. The values of atmospheric and cellular radiocarbon correspond to each other very well.

"Even though these are negligible amounts that are not harmful, we can detect and measure them in tissue samples. By comparing the values to the levels of atmospheric radiocarbon, we can retrospectively establish the age of the cells," explains Dr. Bergmann.

Unparalleled Insights Directly From the Source

The Bergmann group also explores the mechanisms that drive the regeneration of other tissues considered as static, such as the brain or the heart. The team has previously used their expertise in retrospective radiocarbon birth dating to show that the formation of new brain and heart cells is not limited to prenatal time but continues throughout life. Currently, the group is investigating whether new human heart muscle cells can still be generated in people with chronic heart disease.

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Miniature brain models: Understanding autism

To better understand the causes of autism spectrum disorders (ASD) it is crucial to look at what is happening in the brain during development. The closest we come to observing human brains this early is by using organoids -- miniature models of organs. With their help, scientists at the Institute of Science and Technology Austria (ISTA) discovered how mutations in a high-risk gene of autism disrupt important developmental processes.

Several hundred genes are associated with autism spectrum disorders. Some patients are only mildly affected, while others have severe disabilities. In addition to characteristic symptoms like difficulties in social interaction and communication with other people, as well as repetitive-stereotypic behaviors, patients with mutations of the gene CHD8 oftentimes have intellectual disabilities and macrocephaly -- an unusually large brain. How CHD8 causes these symptoms has long been unclear.

Tiny artificial brains

Since CHD8 mutations affect the brain at a very early stage of its development, it has proven difficult for scientists to get the full picture. Over the past years, many researchers therefore used mice as model organisms to better understand what is going on. "But mice with a CHD8 mutation barely showed the symptoms human patients are showing. The effects in mice are not comparable to humans. We needed some kind of human model," Professor Gaia Novarino explains.

Together with collaborators from the Italian Human Technopole institute, the European Institute of Oncology, and the University of Milan, as well as the Allen Institute for Brain Science, USA, Novarino and her team at ISTA turned to organoids. These simplified miniature versions of organs are made from stem cells, which have the ability to become almost every other type of cell. By creating the right circumstances and giving the proper input at just the right time, the scientists were able to mimic developmental processes to create basic versions of brain tissue the size of lentils. "Organoids are the only way you can study human brain development at such an early phase," says Bárbara Oliveira, postdoc in the Novarino group and one of the authors of the study.

CHD8 mutations disrupt balance of neuron production

In petri dishes the team created brain organoids with and without mutations of the gene CHD8. "After some time, we could see by eye that the mutant organoids were much bigger. That was the first evidence that the model works," her colleague and co-author, PhD student Christoph Dotter, describes. Like patients with a CHD8 mutation, the organoids were showing signs of brain overgrowth.

Getting an overview of all the cell types in the organoids, the team notices something very early on: The mutant organoids started to produce a specific type of neurons, inhibitory neurons, much earlier than the control group. So called excitatory neurons, however, were produced later. Furthermore, the mutant organoids produced much more proliferating cells that later on produce a larger amount of this kind of neurons. Over all, the scientists concluded, this leads to them being significantly bigger than the organoids without CHD8 mutations correlating with patient's macrocephaly.

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Cell groups push, rather than pull, themselves into place as organs form and cancers spread

Cells push and pull on surrounding tissue to move in groups as they form organs in an embryo, track down invading bacteria, and as they become cancerous and spread.

Published online in Nature Cell Biology on February 14, a new study found in a living embryo that the back ends of moving cell groups push the group forward. This runs contrary to previous findings, where cell groups grown in dishes of nutrients (cultures) pulled themselves forward with their front edges.

Led by researchers from NYU Grossman School of Medicine and the NYU Courant Institute of Mathematical Sciences, the study used a new technique to measure the forces applied by a cell group as it moved along a "road-like" tissue membrane and into place in a developing animal. Specifically, the study found for the first time in an animal tissue that proteins called integrins on the surfaces of the cells at the rear attach in greater numbers to the membrane as they move along, and exert more force in one direction, than the cells in the group's front. The integrin clusters (focal adhesions) observed in the embryo were smaller than those seen in culture studies, and broke down faster.

Confirmation of such mechanistic details in living tissue have important implications, say the researchers, as many cancers spread in cell groups, and may use the newfound "rear engine propulsion."

"Our results clarify how cell groups that will become organs move into place, and reaffirm that cells behave differently when removed from their natural environments," said senior study author Holger Knaut, PhD, associate professor in the Department of Cell Biology at NYU Langone Health.

Study Details

The study results are based on mechanisms of cell movement established by past studies. For instance, a protein called actin is known to form the protein "skeleton" of cells, with actin chains able to grow in a certain direction, and apply force that change a cell's shape. Integrins, proteins built into outer cell membranes, interact both with actin networks, and proteins outside of cells. These and other proteins form a system that a cell uses to briefly attach to and "roll along" a basement membrane, a pliable mesh of proteins and sugars. What was unknown going into the current study was how tissues in living animals apply force in groups to generate this motion.

The new study examined cell group motion in a zebrafish embryo, a major model in the study of development because it shares many cellular mechanisms with human cells, and because zebrafish embryos development externally, such that each stage in development can be directly observed using high-powered microscopes. In this way the team tracked the movement of the primordium -- a tissue made up of about 140 cells -- as it migrated during development from behind the ear to the tip of the zebrafish tail, where it matures into an organ that senses water flow.

"In the first study of its kind, we combined advanced microscopy with automated, high-throughput computational modeling to measure cellular forces in living organisms," says co-corresponding author Daniele Panozzo, PhD, an associate professor at the Courant Institute of Mathematical Sciences at New York University.

Using "bleached" dots on the basement membrane to measure shape changes (deformations) on a minute scale, and a new software called embryogram to calculate how far the dots move as the primordium "grips" the membrane, the researchers determined how much the cells pulled and pushed on the membrane, "like a tire on pavement." The effect is much like the high school physics experiment where students draw two dots on a rubber band, and calculate the force applied as they stretch the band by measuring the change in distance between the dots.

With these tools in hand, the team showed that the primordium cells link the force-generating actin-myosin network at the back end of the moving group through integrin clusters on the side closest to the basement membrane. The team theorizes that cells attached to membrane toward the back push on the cells in front of them to move the entire group. The researches also gained new insights on an established mechanism where cells have surface proteins that let them "sense" and follow a guidance cue called a chemokine, from low concentration to high concentration. The new study found, however, that cells toward the back end of the primordium sense the chemokine gradient more strongly.

Interestingly, the study found that the primordium moved in a "continuous breaststroke" by pushing the basement membrane downward, sideways and backwards, much like the arms of a swimmer. The authors do not know why this is, but speculate that this is the most efficient way to move forward. They note that banana slugs also use the rear edge of the "foot" they apply to the ground, suggesting that evolution favors rear engine propulsions because they are most efficient at different size scales.

The study suggests that group cell movement have the potential to be harnessed to stop cancer spread, perhaps by designing treatments that block the action of integrins, say the authors. Integrin inhibitors have been tested as drugs for cardiovascular and autoimmune disease in clinical trials, but their use against cancer spread has been limited by the need for a better understanding of the mechanisms.

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Successful transplant of porcine heart into adult human with end-stage heart disease

In a first-of-its-kind surgery, a 57-year-old patient with terminal heart disease received a successful transplant of a genetically-modified pig heart and is still doing well three days later. It was the only currently available option for the patient. The historic surgery was conducted by University of Maryland School of Medicine (UMSOM) faculty at the University of Maryland Medical Center (UMMC), together known as the University of Maryland Medicine.

This organ transplant demonstrated for the first time that a genetically-modified animal heart can function like a human heart without immediate rejection by the body. The patient, David Bennett, a Maryland resident, is being carefully monitored over the next days and weeks to determine whether the transplant provides lifesaving benefits. He had been deemed ineligible for a conventional heart transplant at UMMC as well as at several other leading transplant centers that reviewed his medical records.

"It was either die or do this transplant. I want to live. I know it's a shot in the dark, but it's my last choice," said Mr. Bennett, the patient, a day before the surgery was conducted. He had been hospitalized and bedridden for the past few months. "I look forward to getting out of bed after I recover."

The U.S. Food and Drug Administration granted emergency authorization for the surgery on New Year's Eve through its expanded access (compassionate use) provision. It is used when an experimental medical product, in this case the genetically-modified pig's heart, is the only option available for a patient faced with a serious or life-threatening medical condition. The authorization to proceed was granted in the hope of saving the patient's life.

"This was a breakthrough surgery and brings us one step closer to solving the organ shortage crisis. There are simply not enough donor human hearts available to meet the long list of potential recipients," said Bartley P. Griffith, MD, who surgically transplanted the pig heart into the patient. Dr. Griffith is the Thomas E. and Alice Marie Hales Distinguished Professor in Transplant Surgery at UMSOM. "We are proceeding cautiously, but we are also optimistic that this first-in-the-world surgery will provide an important new option for patients in the future."

Considered one of the world's foremost experts on transplanting animal organs, known as xenotransplantation, Muhammad M. Mohiuddin, MD, Professor of Surgery at UMSOM, joined the UMSOM faculty five years ago and established the Cardiac Xenotransplantation Program with Dr. Griffith. Dr. Mohiuddin serves as the program's Scientific/Program Director and Dr. Griffith as its Clinical Director.

"This is the culmination of years of highly complicated research to hone this technique in animals with survival times that have reached beyond nine months. The FDA used our data and data on the experimental pig to authorize the transplant in an end-stage heart disease patient who had no other treatment options," said Dr. Mohiuddin. "The successful procedure provided valuable information to help the medical community improve this potentially life-saving method in future patients."

About 110,000 Americans are currently waiting for an organ transplant, and more than 6,000 patients die each year before getting one, according to the federal government's organdonor.gov. Xenotransplantation could potentially save thousands of lives but does carry a unique set of risks, including the possibility of triggering a dangerous immune response. These responses can trigger an immediate rejection of the organ with a potentially deadly outcome to the patient.

Xenotransplants were first tried in the 1980s, but were largely abandoned after the famous case of Stephanie Fae Beauclair (known as Baby Fae) at Loma Linda University in California. The infant, born with a fatal heart condition, received a baboon heart transplant and died within a month of the procedure due to the immune system's rejection of the foreign heart. However, for many years, pig heart valves have been used successfully for replacing valves in humans.

Before consenting to receive the transplant, Mr. Bennett, the patient, was fully informed of the procedure's risks, and that the procedure was experimental with unknown risks and benefits. He had been admitted to the hospital more than six weeks earlier with life-threatening arrythmia and was connected to a heart-lung bypass machine, called extracorporeal membrane oxygenation (ECMO), to remain alive. In addition to not qualifying to be on the transplant list, he was also deemed ineligible for an artificial heart pump due to his arrhythmia.

Revivicor, a regenerative medicine company based in Blacksburg, VA, provided the genetically-modified pig to the xenotransplantation laboratory at UMSOM. On the morning of the transplant surgery, the surgical team, led by Dr. Griffith and Dr. Mohiuddin, removed the pig's heart and placed it in the XVIVO Heart Box, perfusion device, a machine that keeps the heart preserved until surgery.

The physician-scientists also used a new drug along with conventional anti-rejection drugs, which are designed to suppress the immune system and prevent the body from rejecting the foreign organ. The new drug used is an experimental compound made by Kiniksa Pharmaceuticals.

"This unprecedented and historic procedure highlights the importance of translational research which lays the groundwork for patients to benefit in the future. It is the culmination of our longstanding commitment to discovery and innovation in our xenotransplantation program," said E. Albert Reece, MD, PhD, MBA, Executive Vice President for Medical Affairs, UM Baltimore, and the John Z. and Akiko K. Bowers Distinguished Professor and Dean, University of Maryland School of Medicine. "Our transplant surgeon-scientists are among the most talented in the country, and are helping to bring the promise of xenotransplantation to fruition. We hope it will one day become a standard of care for patients in need of organ transplants. As has happened throughout our history, the University of Maryland School of Medicine continues to address the most complex medical and scientific problems."

Bruce Jarrell, MD, President of the University of Maryland, Baltimore, who himself is a transplant surgeon, recalled: "Dr. Griffith and I began as organ transplant surgeons when it was in its infancy. Back then, it was the dream of every transplant surgeon, myself included, to achieve xenotransplantation and it is now personally gratifying to me to see this long-sought goal clearly in view. It is a spectacular achievement."

"This is truly a historic, monumental step forward. While we have long been at the forefront of research driving progress toward the promise of xenotransplantation as a viable solution to the organ crisis, many believed this breakthrough would be well into the future," said Bert W. O'Malley, MD, President and CEO, University of Maryland Medical Center. "I couldn't be more proud to say the future is now. Our skilled team of UMMC and UMSOM physician-scientists will continue to advance and adapt medical discovery for patient care that could offer a lifeline for more patients in dire need."

Mohan Suntha, MD, MBA, President and CEO, University of Maryland Medical System, added: "The University of Maryland Medical System is committed to working with our University of Maryland School of Medicine partners to explore, research, and in many cases implement the innovations in patient care that make it possible to improve quality of life and save lives. We appreciate the tremendous courage of this live recipient, who has made an extraordinary decision to participate in this groundbreaking procedure to not only potentially extend his own life, but also for the future benefit of others."

Organs from genetically modified pigs have been the focus of much of the research in xenotransplantation, in part because of physiologic similarities between pigs, human, and nonhuman primates. UMSOM received $15.7 million sponsored research grant to evaluate Revivicor genetically-modified pig UHearts™ in baboon studies.

Three genes -- responsible for rapid antibody-mediated rejection of pig organs by humans -- were "knocked out" in the donor pig. Six human genes responsible for immune acceptance of the pig heart were inserted into the genome. Lastly, one additional gene in the pig was knocked out to prevent excessive growth of the pig heart tissue, which totaled 10 unique gene edits made in the donor pig.

"We are thrilled to support the world-class team of transplant surgeons led by Dr. Griffith and Dr. Mohiuddin at the University of Maryland School of Medicine," said David Ayares, PhD, Chief Scientific Officer of Revivicor, Inc. "This transplant is groundbreaking, and is another step in the investigation of xeno organs for human use."

Dr. Mohiuddin, Dr. Griffith, and their research team spent the past five years perfecting the surgical technique for transplantation of pig hearts into non-human primates. Dr. Mohiuddin's xenotransplant research experience spans over 30 years during which time he demonstrated in peer-reviewed research that genetically-modified pig's hearts can function when placed in the abdomen for as long as three years. Success was dependent on the right combination of genetic modifications to the experimental donor pig UHeart™ and anti-rejection drugs, including some experimental compounds.

"As a cardiothoracic surgeon who does lung transplants, this is an amazing moment in the history of our field. Decades of research here at Maryland and elsewhere have gone into this achievement. This has the potential to revolutionize the field of transplantation by eventually eliminating the organ shortage crisis," said Christine Lau, MD, MBA the Dr. Robert W. Buxton Professor and Chair of the Department of Surgery at UMSOM and Surgeon-in-Chief at UMMC. "This is a continuation of steps to making xenotransplantation a life-saving reality for patients in need."

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Championing chrononutrition with protein, the morning elixir for muscle growth

Proteins constitute an essential dietary component that help in the growth and repair of the body. Composed of long chains of amino acids, proteins promote the growth of skeletal muscles, the group of muscles that help us move. Humans have been aware of the benefits of proteins for long. However, recent studies have shown that having the right amount of protein at the right time of the day is essential for proper growth. This is called 'Chrononutrition,' in which when you eat is as important as what and how you eat.

The reason behind this is the body's internal biological clock, called the 'circadian rhythm.' This rhythm is followed by all cells and controls life functions like metabolism and growth. Interestingly, protein digestion and absorption have been found to fluctuate across day and night according to this clock. Moreover, earlier studies have reported that intake of protein at breakfast and lunch promotes skeletal muscle growth in adults. However, details on the effect of the time of protein intake on muscle growth and function have remained elusive till date.

Fortunately, researchers from Waseda University, led by Professor Shigenobu Shibata, recently endeavored to understand the effect of the distribution of protein intake through the day on muscles. They fed laboratory mice two meals per day containing either high (11.5% by proportion) or low (8.5% by proportion) protein concentrations. The researchers noted that protein intake at breakfast induced an increase in muscle growth, determined by assessing induced hypertrophy of the plantaris muscle in the leg, when compared with the effects of protein intake at dinner. Specifically, the ratio of muscle hypertrophy determined against the growth of the control muscle was 17% higher in mice fed 8.5% protein at breakfast, than that in mice fed 11.5% protein at dinner, despite the former group consuming a low proportion of protein overall. They also found that intake of a type of protein called the BCCA, short for branched-chain amino acids, early in the day increased the size of skeletal muscles specifically.

To confirm the association of these effects with the workings of the circadian rhythm, the researchers next engineered whole-body mutant Clock?19 or muscle-specific Bmal1 knockout mice lacking the genes that control the biological clock. They repeated diet distribution experiments on these mice but did not observe similar muscle change, which confirmed the involvement of the circadian rhythm in muscle growth in the context of protein intake.

Excited about the findings of their study published in a recent issue of the Cell Reports, Prof. Shibata emphasizes, "Protein-rich diet at an early phase of the daily active period, that is at breakfast, is important to maintain skeletal muscle health and enhance muscle volume and grip strength."

To check if their findings were applicable to humans, the team recruited women in their study and tested if their muscle function, determined by measuring skeletal muscle index (SMI) and grip strength, varied with the timing of the protein-rich diet consumed. Sixty women aged 65 years and above who took protein at breakfast rather than at dinner showed better muscle functions, suggesting the possibility of the findings to be true across species. Additionally, the researchers also found a strong association between SMI and the proportion of protein intake at breakfast relative to total protein intake through the day.

Prof. Shibata is hopeful that the findings of their study will lead to a widespread modification in the current diet regime of most people across the Western and Asian countries, who traditionally consume low amounts of protein at breakfast. He therefore stresses, "For humans, in general, the protein intake at breakfast averages about 15 grams, which is less than what we consume at dinner, which is roughly 28 grams. Our findings strongly support changing this norm and consuming more protein at breakfast or morning snacking time."

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450-million-year-old sea creatures had a leg up on breathing

 A new study has found the first evidence of sophisticated breathing organs in 450-million-year-old sea creatures. Contrary to previous thought, trilobites were leg breathers, with structures resembling gills hanging off their thighs.

Trilobites were a group of marine animals with half-moon-like heads that resembled horseshoe crabs, and they were wildly successful in terms of evolution. Though they are now extinct, they survived for more than 250 million years -- longer than the dinosaurs.

Thanks to new technologies and an extremely rare set of fossils, scientists from UC Riverside can now show that trilobites breathed oxygen and explain how they did so. Published in the journal Science Advances, these findings help piece together the puzzle of early animal evolution.

"Up until now, scientists have compared the upper branch of the trilobite leg to the non-respiratory upper branch in crustaceans, but our paper shows, for the first time, that the upper branch functioned as a gill," said Jin-Bo Hou, a UCR paleontology doctoral student who led the research.

Among the oldest animals on earth, this work helps situate trilobites on the evolutionary tree more securely in between older arthropods, a large group of animals with exoskeletons, and crustaceans.

The research was possible, in part, because of unusually preserved fossil specimens. There are more than 22,000 trilobite species that have been discovered, but the soft parts of the animals are visible in only about two dozen.

"These were preserved in pyrite -- fool's gold -- but it's more important than gold to us, because it's key to understanding these ancient structures," said UCR geology professor and paper co-author Nigel Hughes.

A CT scanner was able to read the differences in density between the pyrite and the surrounding rock and helped create three-dimensional models of these rarely seen gill structures.

"It allowed us to see the fossil without having to do a lot of drilling and grinding away at the rock covering the specimen," said paleontologist Melanie Hopkins, a research team member at the American Museum of Natural History.

"This way we could get a view that would even be hard to see under a microscope -- really small trilobite anatomical structures on the order of 10 to 30 microns wide," she said. For comparison, a human hair is roughly 100 microns thick.

Though these specimens were first described in the late 1800s and others have used CT scans to examine them, this is the first study to use the technology to examine this part of the animal.

The researchers could see how blood would have filtered through chambers in these delicate structures, picking up oxygen along its way as it moved. They appear much the same as gills in modern marine arthropods like crabs and lobsters.

Comparing the specimens in pyrite to another trilobite species gave the team additional detail about how the filaments were arranged relative to one another, and to the legs.

Most trilobites scavenged the ocean floor, using spikes on their lower legs to catch and grind prey. Above those parts, on the upper branch of the limbs, were these additional structures that some believed were meant to help with swimming or digging.

"In the past, there was some debate about the purpose of these structures because the upper leg isn't a great location for breathing apparatus," Hopkins said. "You'd think it would be easy for those filaments to get clogged with sediment where they are. It's an open question why they evolved the structure in that place on their bodies."

The Hughes lab uses fossils to answer questions about how life developed in response to changes in Earth's atmosphere. Roughly 540 million years ago, there was an explosive diversification in the variety and complexity of animals living in the oceans.

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Silk fibers improve bioink for 3D-printed artificial tissues and organs

 How do you test, in early-stage research, whether a potential pharmaceutical effectively targets a human tumor, organ, or some other part of the body? How do you grow a new hand or some other body part? Researchers are in the early stages of using 3D cell printing technology to make developments like these happen. A standard way -- currently unavailable -- to fix the cells in place after printing would help researchers avoid having to 'reinvent the wheel' in every new investigation.

In a study recently published in Materials Today Bio, researchers from Osaka University have used silk nanofibers obtained by mechanical disintegration to enhance the printing process without damaging the cells or cell assemblies. An attractive point of silk for this application is that silk is believed to be a safe material for humans. This development will help bring 3D cell printing research out of the laboratory and into real-world biomedical use.

To obtain the fibers, the researchers started with virgin silk, then removed the protein sericin from it because this protein causes inflammation in patients. Next, the researchers ground the remaining biocompatible material into nanofibers. The fibers can be sterilized -- without damaging them -- for medical use, with common laboratory equipment.

"Our silk fibers are excellent additives to bioink cell printing media," says lead author Shinji Sakai. "They are compatible with many media, such as those containing gelatin, chitosan, or hyaluronic acid, giving them a broad range of potential applications."

The main purpose of the fibers was to ensure that the cells in the bioink retained their 3D positioning after printing without damaging the cells. The fibers fulfill this purpose by enhancing the integrity of the bioink and minimizing the damaging high mechanical stresses often placed on cells during printing.

"Various mechanical experiments say the same thing: the nanofibers enhanced the properties of the printing media," explains Professor Sakai. "For example, Young's modulus -- a measure of stiffness -- increased several-fold and remained enhanced for over a month."

The fibers help printed configurations retain their structural integrity after printing. For example, a nose-shaped configuration retained its shape only when printed with bioink containing the silk fibers. Over 85% of the cells in the bioink remained alive after a week in the printed bioink with or without the added fibers, indicating that adding the fibers did not damage the cells.

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A swifter way towards 3D-printed organs

20 people die every day waiting for an organ transplant in the United States, and while more than 30,000 transplants are now performed annually, there are over 113,000 patients currently on organ waitlists. Artificially grown human organs are seen by many as the "holy grail" for resolving this organ shortage, and advances in 3D printing have led to a boom in using that technique to build living tissue constructs in the shape of human organs. However, all 3D-printed human tissues to date lack the cellular density and organ-level functions required for them to be used in organ repair and replacement.

Now, a new technique called SWIFT (sacrificial writing into functional tissue) created by researchers from Harvard's Wyss Institute for Biologically Inspired Engineering and John A. Paulson School of Engineering and Applied Sciences (SEAS), overcomes that major hurdle by 3D printing vascular channels into living matrices composed of stem-cell-derived organ building blocks (OBBs), yielding viable, organ-specific tissues with high cell density and function. The research is reported in Science Advances.

"This is an entirely new paradigm for tissue fabrication," said co-first author Mark Skylar-Scott, Ph.D., a Research Associate at the Wyss Institute. "Rather than trying to 3D-print an entire organ's worth of cells, SWIFT focuses on only printing the vessels necessary to support a living tissue construct that contains large quantities of OBBs, which may ultimately be used therapeutically to repair and replace human organs with lab-grown versions containing patients' own cells."

SWIFT involves a two-step process that begins with forming hundreds of thousands of stem-cell-derived aggregates into a dense, living matrix of OBBs that contains about 200 million cells per milliliter. Next, a vascular network through which oxygen and other nutrients can be delivered to the cells is embedded within the matrix by writing and removing a sacrificial ink. "Forming a dense matrix from these OBBs kills two birds with one stone: not only does it achieve a high cellular density akin to that of human organs, but the matrix's viscosity also enables printing of a pervasive network of perfusable channels within it to mimic the blood vessels that support human organs," said co-first author Sébastien Uzel, PhD., a Research Associate at the Wyss Institute and SEAS.

The cellular aggregates used in the SWIFT method are derived from adult induced pluripotent stem cells, which are mixed with a tailored extracellular matrix (ECM) solution to make a living matrix that is compacted via centrifugation. At cold temperatures (0-4oC), the dense matrix has the consistency of mayonnaise - soft enough to manipulate without damaging the cells, but thick enough to hold its shape - making it the perfect medium for sacrificial 3D printing. In this technique, a thin nozzle moves through this matrix depositing a strand of gelatin "ink" that pushes cells out of the way without damaging them.

When the cold matrix is heated to 37 oC, it stiffens to become more solid (like an omelet being cooked) while the gelatin ink melts and can be washed out, leaving behind a network of channels embedded within the tissue construct that can be perfused with oxygenated media to nourish the cells. The researchers were able to vary the diameter of the channels from 400 micrometers to 1 millimeter, and seamlessly connected them to form branching vascular networks within the tissues.

Organ-specific tissues that were printed with embedded vascular channels using SWIFT and perfused in this manner remained viable, while tissues grown without these channels experienced cell death in their cores within 12 hours. To see whether the tissues displayed organ-specific functions, the team printed, evacuated, and perfused a branching channel architecture into a matrix consisting of heart-derived cells and flowed media through the channels for over a week. During that time, the cardiac OBBs fused together to form a more solid cardiac tissue whose contractions became more synchronous and over 20 times stronger, mimicking key features of a human heart.

"Our SWIFT biomanufacturing method is highly effective at creating organ-specific tissues at scale from OBBs ranging from aggregates of primary cells to stem-cell-derived organoids," said corresponding author Jennifer Lewis, Sc.D., who is a Core Faculty Member at the Wyss Institute as well as the Hansjörg Wyss Professor of Biologically Inspired Engineering at SEAS. "By integrating recent advances from stem-cell researchers with the bioprinting methods developed by my lab, we believe SWIFT will greatly advance the field of organ engineering around the world."

Collaborations are underway with Wyss Institute faculty members Chris Chen, M.D., Ph.D. at Boston University and Sangeeta Bhatia, M.D., Ph.D., at MIT to implant these tissues into animal models and explore their host integration, as part of the 3D Organ Engineering Initiative co-led by Lewis and Chris Chen.

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How genes interact to build tissues and organisms

Although the knowledge we have about human cells and tissues has steadily increased over recent decades, many things remain unknown. For instance, cells exist in transient, dynamic states and understanding them is fundamental to decipher diseases and find cures. Classic techniques used in the lab to study cell types faced limitations and did not enable a finely detailed profile of cell function.

To overcome this obstacle, a group of scientists at the National Centre for Genomic Analysis (CNAG-CRG) from the Centre for Genomic Regulation (CRG), in Barcelona, Spain, led by Holger Heyn, developed a new computational tool, based on the mathematical Graph theory, to infer global, large-scale regulatory networks, from healthy and pathological organs, such as those affected by diabetes or Alzheimer's disease. The researchers were able to pinpoint genes relevant to organ function and potential drivers of diseases. They are publishing their results in the current issue of the Genome Biology journal.

"Our previously developed single-cell transcriptomic tools were very useful to discover unknown cell types," says Giovanni Iacono, senior postdoc researcher at the CNAG-CRG and first author of the study. "Those tools allowed us to describe new types and subtypes of cells, with their unique biological roles and hierarchical relationships," he adds.

Up to now, single-cell analysis had been used to understand cell types and their function within tissue. "Large-scale consortia like the Human Cell Atlas Project generate single-cell maps of entire organisms and sophisticated analysis strategies are required to transform big data into disruptive biological and clinical insights," says Holger Heyn, team leader of the Single Cell Genomics Group at the CNAG-CRG and senior author of the article.

The tool that this scientific team has now developed will enable them to go one step further, to see how genes interact to form tissues. "Our tool tries to address precisely the regulatory process that controls the morphology and functions of a cell," highlights Iacono.

The tool is based on the Graph theory, an abstract mathematical model in which there are nodes connected by edges. Once you have a graph, a structure, you can measure the importance of each node for the network. In this case, each node was a gene and importance was defined as the function of that gene being key for the biological system under study.

CNAG-CRG researchers processed datasets from ten-thousands of cells to infer the regulatory networks that drive cell phenotype formation and their respective functions. They applied their tool to study type 2 diabetes and Alzheimer's disease and were able to find the functional changes relevant to those diseases. Importantly, this opens the door to finding new drug targets.

"The network analysis we have developed goes beyond currently applied approaches to provide deep insights into how gene activities shape tissues and organs. This is critical to understand diseases in which these networks are disrupted and find their 'Achilles heels' for effective treatments." says Heyn.

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Blood holds key to liver regeneration

The liver is the only organ in the body that can regenerate. But some patients who undergo a liver resection, a surgery that removes a diseased portion of the organ, end up needing a transplant because the renewal process doesn't work.

A new Michigan State University study, published in the journal Blood, shows that the blood-clotting protein fibrinogen may hold the key as to why this happens.

"We discovered that fibrinogen accumulates within the remaining liver quickly after surgery and tells platelets to act as first responders, triggering the earliest phase of regeneration," said James Luyendyk, a professor of pathobiology in the College of Veterinary Medicine. "But if fibrinogen or platelets are inhibited, then regeneration is delayed."

Platelets are blood cells that help form clots and stop bleeding. When they receive information from fibrinogen, they go into action and accumulate in the remaining part of the liver to help restore it, increasing the chances of a fully functional liver and successful recovery.

Using samples from patients undergoing liver resection and a comparable model in mice, Luyendyk and his team noticed that when fibrinogen was low, the number of platelets in the liver decreased.

"This shows that fibrinogen deposits are extremely important and directly impact regeneration in both mice and humans," Luyendyk said.

According to Dafna Groeneveld, Luyendyk's co-author and post-doctoral research associate in his lab, their finding demonstrates that fibrinogen levels could be a predictive marker for doctors, too.

"Measuring this protein in liver resection patients may help us determine in advance whether the organ will regenerate successfully or if it will become dysfunctional," she said.

This could lead to new treatments that would help doctors correct low levels of the protein by using fibrinogen concentrates that can be administered during surgery.

"This type of treatment hasn't been tried in liver resection patients yet," Luyendyk said. "But once we figure out exactly how fibrinogen works in the regeneration process and test potential therapies in mice, it could eventually provide the proof we need to bring our work into the clinic and improve patient outcomes."

From Science Daily