Culture

New Rochelle, NY, November 11, 2020--New data highlight safety concerns for the replication of recombinant adeno-associated viral (rAAV) vectors commonly used in gene therapy. These findings, which emphasize the need for mobilization resistant AAV vectors, are reported in the peer-reviewed journal Human Gene Therapy. Click here to read the full-text article free through December 11, 2020.

The risk of rAAV mobilization has been underappreciated in the AAV research community.

"The data generated herein highlight the potential of rAAV vector production in treated patients upon subsequent wtAAV and helper virus infection," state Matthew Hirsch, PhD, University of North Carolina at Chapel Hill, and coauthors. "They raise safety concerns for the treated individual and for the unintended animal and human populations in general."

"Viral gene therapy vectors are, by their nature, viruses capable of replicating as viruses under certain conditions. These studies help to define the conditions under which that might occur in patients receiving rAAV vectors," according to Editor-in-Chief of Human Gene Therapy Terence R. Flotte, MD, Celia and Isaac Haidak Professor of Medical Education and Dean, Provost, and Executive Deputy Chancellor, University of Massachusetts Medical School.

According to the Centers for Disease Research, 1 in 54 children is diagnosed with autism spectrum disorder (ASD) each year, and the number has been rising. The disease causes perplexing, lifelong developmental disabilities, which usually arise during early childhood and affect social skills, communication, personal relationships, and self-control.

In new research appearing in the journal mSphere, Rosa Krajmalnik-Brown, James Adams and their colleagues highlight the crucial importance of bacterial microbes in the human gut for the diagnosis and treatment of ASD.

Autism is considered a "spectrum disorder," characterized by a range of stereotypic behaviors that can affect people to varying degrees. While genetic correlates have been implicated, the full panoply of underlying causes of ASD remain murky. Despite the prevalence of the disease and its profound societal impact, no effective FDA-approved treatment for the disorder currently exists.

In earlier studies, the researchers observed the effects on ASD symptoms following the use of a revolutionary approach, known as microbial transfer therapy (MTT). The procedure, in which gut microbiota from healthy donors are transplanted into ASD patients over a period of 7-8 weeks, produced marked alterations in the ASD gut, improving both gastrointestinal and behavioral symptoms of the disease. Remarkably, symptom improvement continued for two years after completion of the study.

The new research takes a closer look at plasma and fecal metabolites altered through MTT. According to Krajmalnik Brown, Professor at ASU's School of Sustainable Engineering and the Built Environment, researcher in the Biodesign Swette Center for Environmental Biotechnology and Director of the newly formed Biodesign Center for Health Through Microbiomes, "looking at metabolites is important because it can help us understand how microorganisms affect gastrointestinal symptoms and behavior. They can also be used as biomarkers."

Indeed, analysis of 619 plasma metabolites in the new study showed a distinctive metabolic profile in ASD children prior to the MTT procedure. Careful investigation of metabolites in blood and feces, as opposed to simply tabulating microbial species in the gut, is crucial when trying to tease out the complex bacterial interactions, which can form both collaborative and competing networks, depending on the particulars of their environment.

Following the procedure, levels of key metabolites in plasma became more similar to levels in typically-developing children, suggesting that this was a result of adjusting the constellation of bacteria to more closely resemble conditions in the normal healthy gut, including a marked increase in microbial diversity.

The new research highlights significant metabolic changes in plasma samples and more modest alterations in fecal samples. "It was exciting to identify many differences in metabolite levels between children with autism and typically developing children, and to observe that many of the metabolite levels improved after microbiome transplant," says Adams, a Professor at ASU's School for Engineering of Matter, Transport and Energy and Director the Autism/Asperger's Research Program at ASU.

Further research will help advance the understanding of the complex and vital interactions between the gut microbiome and neurological features associated with ASD and help fine-tune MTT therapy or other microbial based therapies, for greater effectiveness in mitigating symptoms of ASD.

WASHINGTON (Nov. 13, 2020) -- Researchers at Georgetown University Medical Center have successfully used molecules comprised of small strands of RNA to shut down the production of destructive proteins generated by the COVID-19 virus. Additionally, the researchers are working to aerosolize the RNA molecules so that they could be incorporated in an inhalable drug that would mitigate viral chaos. The finding appears online today in Gene Therapy.

A key to the Georgetown research efforts was the use of either microRNA (miRNA) or silencing RNA (siRNA, also known as small interfering RNA), both of which are RNA molecules. These molecules can guide the ultimate expression of how protein production occurs in a virus. And it is the proteins produced by SARS-CoV-2 (the virus that causes COVID-19) that wreak havoc in people.

SARS-CoV-2 bears biological similarities to other respiratory viruses, such as the seasonal flu. The use of Tamiflu® (oseltamivir) to treat and prevent infection from the flu has proved to be helpful in lessening flu symptoms and its lethality in some people.

"We believe that our approach suppresses viral protein production and could be used against virtually any respiratory virus," says G. Ian Gallicano, PhD, associate professor in the Department Biochemistry and Molecular and Cellular Biology at Georgetown University Medical Center. "Because of our results, Georgetown University has filed a patent application on on the siRNA sequences that were shown to have the best effect on viral protein suppression."

Another key to the researcher's effort was the fact that miRNAs and siRNAs can be made fat-soluble, making them more easily absorbed using sprays into nasal passages lined with mucous membranes. Non-soluble fat compounds have been found to irritate nasal passages if inhaled.

While Tamiflu is a pill that can help control the flu, the Georgetown researchers' proposed drug has a tougher disease to try to control and thus its mechanism of action would be different. An aerosolized RNA-targeting agent against SARS-CoV-2 would interfere with the production of the protein spikes associated with the infectivity of the virus and thereby reduce subsequent viral spreading.

The scientist's viral strategy is based on research Gallicano and his collaborators have been conducting to treat heart failure, which uses miRNA to target genes that can affect the heart's ability to function well. Modifying this cardiac approach, the researchers showed that miRNAs and siRNAs can target messenger RNA inside a virus. As SARS-CoV-2 uses messenger RNA to generate the proteins necessary for multiplication and infection, the ability to target viral machinery within cells via siRNA, in particular, could help shut the virus down, noted Gallicano.

The scientists tested their approach using two types of cells in the lab, including one type of cell from a human windpipe (trachea). Against tracheal and non-tracheal cells, the researchers used siRNAs and found that they could suppress protein function in a dose-dependent manner, which they subsequently found was due to messenger RNA degradation.

The researchers investigated if miRNAs and siRNAs would shut down production of proteins vital to a healthy cell. Through a series of experiments, they found that there were fewer 'off-target' effects using siRNA compared to miRNA; off-target effects can cause normal cells to go haywire.

"If our approach proves successful in future experiments, we are confident this technology can be quickly moved from bench to bedside," concludes Gallicano.

While COVID-19 infected patients should be treated with standard anticoagulation therapies, such as blood thinning medication, a new study by researchers at the George Washington University (GW) shows that anticoagulating patients at higher doses, without traditional medical indications to do so, may be ineffective and even harmful. The study was published in the journal Thrombosis Research.

"COVID-19 patients appear to have an increased incidence of blood clots. Many hospitals and health care providers began to use high doses of blood thinners to prevent these clots or treat them preemptively," said Juan Reyes, MD, MPH, co-author of the study and director of hospital medicine at the GW School of Medicine and Health Sciences (SMHS). "We wanted to review the data of our subset of COVID-19 patients treated with blood thinners to determine if the higher-dose medication was helpful."

The research team evaluated 402 patients diagnosed with COVID-19 admitted to GW Hospital between March 15 and May 31, 2020. Clinical outcomes were compared between 152 patients treated with high-dose blood thinners, and 250 patients treated with the standard low-dose blood thinners, typically prescribed to hospitalized medical patients. "Our findings did not demonstrate any additional benefit for those treated with higher doses of blood thinners above and beyond the standard of care," said Shant Ayanian, MD, MS, co-author and assistant professor of medicine at GW SMHS.

"While it's true that COVID-19 patients could be dying of blood clots, the data we've evaluated does not support giving every patient a high dose of blood thinners. That's a challenge, as the benefits still remain unclear," said Lei Lynn, MD, first author of the study and assistant professor of medicine at GW SMHS. "We caution against an aggressive blood thinner regimen for everyone, unless there is clear evidence to do so."

At the beginning of the pandemic, all patients admitted with COVID-19 to the GW Hospital were treated with standard dose anticoagulation, unless contraindicated. As awareness of the elevated risk of blood clots developed, many providers began treating patients with high-dose blood thinners. At GW Hospital, for non-critically ill patients, medical teams were advised to especially consider initiating a high dose of anticoagulation if a patient's D-dimer level exceeded 3 micrograms per milliliter. The research team previously published a study finding higher levels of the biomarker D-dimer, a medical indicator found in the blood, is associated with higher odds of clinical deterioration and death from COVID-19. This study is the first of its kind to utilize D-dimer levels to analyze clinical outcomes of anticoagulation in patients who are not critically ill.

"Though we would have loved to have seen a clinical benefit to our patients from anticoagulation, our research found that higher doses of blood thinners were potentially harmful, with no clear benefits," said Karolyn Teufel, MD, co-author of the study and assistant professor of medicine at GW SMHS. "Our research highlights the challenges with treating COVID-19. So much remains unknown."

A new saliva-based test developed by a team at Johns Hopkins Bloomberg School of Public Health has been found to accurately detect the presence of antibodies to SARS-CoV-2, the virus that causes COVID-19, from small samples of saliva, according to a study led by Bloomberg School researchers. Such tests, the results of which can be obtained in a matter of hours, are seen as potential alternatives to blood-sample antibody tests for research and clinical use.

The test is based on multiple fragments, or "antigens," from the SARS-CoV-2 coronavirus, mostly from its outer spike and nucleocapsid proteins. In the study, the researchers found that their test detected antibodies to several of these antigens in saliva samples from all 24 participants who had confirmed SARS-CoV-2 exposure and whose symptoms had begun more than two weeks prior to the test. The test also reliably yielded negative results for saliva samples that had been collected from people prior to the COVID-19 pandemic.

The study appears online in the Journal of Clinical Microbiology.

"If our saliva-based assay's accuracy is borne out in larger studies, this noninvasive approach could make it easier to identify, at a population level, who has already had a SARS-CoV-2 infection and where gaps in seropositivity remain heading into the winter and beyond," says study senior author Christopher D. Heaney, PhD, MS, an associate professor with appointments in the departments of Environmental Health and Engineering, Epidemiology, and International Health at the Bloomberg School. "This could inform targeted vaccination efforts and, after vaccines start to roll out, help figure out how long vaccine-induced antibodies last--all without repeated, invasive blood draws," Heaney says.

The pandemic spread of SARS-CoV-2 has officially caused over 40 million infections and more than 1 million deaths worldwide. Many epidemiologists suspect that the actual spread of the virus has been much more extensive, but that and many other questions about the pandemic's extent and dynamics have so far been difficult or impossible to answer specifically. A relatively quick, inexpensive, noninvasive, and highly accurate antibody test could make such research much easier, however.

Heaney and colleagues previously have invented accurate saliva-based antibody tests for other disease-causing viruses including the enteric pathogen norovirus and liver-infecting hepatitis E virus.

Early in the pandemic, the research team developed a saliva-based test for SARS-CoV-2 antibodies, using a panel of 12 known viral antigens that are already used for blood-based antibody tests. Saliva samples for the test are collected by rubbing a sponge between people's teeth and gums, where saliva is known to be particularly enriched with antibodies.

That the test detected antibodies to several SARS-CoV-2 antigens in saliva samples from all 24 people who had confirmed SARS-CoV-2 exposure and whose symptoms had arisen more than two weeks before the test showed that the test could be very sensitive--that is, capable of identifying positive results.

The experiments also showed that the test could be highly specific--that is, capable of identifying those without the antibodies with a low rate of "false positives." In a set of 134 saliva samples that had been collected from people long before the COVID-19 pandemic--and thus presumed to be free of SARS-CoV-2 antibodies--several antigens in the test gave negative results in all but a few cases. Antibodies to one viral antigen seemed particularly specific: The scientists found negative results for it in all 134 of the pre-COVID-19 samples.

Saliva-based testing for immunoglobulin G antibodies to SARS-CoV-2 appeared to be just as sensitive and specific as blood-based serological testing. After SARS-CoV-2 infection, IgG antibodies typically elevate around day 10 after symptom onset, and these antibodies spill over from the blood into saliva.

The experiments on the whole have suggested that people who become infected with SARS-CoV-2 develop detectable antibodies in saliva at roughly the same time as they do in blood, about 10 days after COVID-19 symptom onset. The researchers expect that with an optimal algorithm that integrates results for just a few especially sensitive and specific antigens, their saliva-based test may be able to reliably detect SARS-CoV-2 antibodies starting around that same 10-day mark, but not earlier.

Since submitting their paper several months ago, Heaney and colleagues have been refining the test with experiments on thousands more saliva samples. They expect that their saliva-based test will be useful for future research applications, especially large-scale or longitudinal studies for which invasive and potentially painful blood-based tests could be problematic. For example:

Measuring levels of SARS-CoV-2 exposure and immunity in a given neighborhood, city, county, or state, or in a particular category of workers.

Identifying populations that might particularly benefit from focused vaccination campaigns.

Monitoring changes in antibody-positivity rates over time to help assess government or corporate risk-reduction campaigns.

Determining how long antibody levels persist in large populations following infection or vaccination.

The researchers also believe that their test is sensitive and specific enough to have potential use in clinical settings, such as screening individuals for prior SARS-CoV-2 exposures before they receive a vaccine or undergo some other medical procedure.

For clinical applications, the test would need Food and Drug Administration approval--at least emergency-use authorization--and Heaney says that with this goal in mind he and his colleagues are initiating discussions with the agency.

Two independent clinical studies – one by researchers at the Center for Cell-Based Therapy (CTC) in Ribeirão Preto, state of São Paulo (Brazil), on the monoclonal antibody eculizumab, and the other by scientists at the University of Pennsylvania in Philadelphia (USA) on an experimental drug called AMY-101 – observed a significant anti-inflammatory effect that contributed to a faster recovery by severe COVID-19 patients. The results of the two studies, which set out to compare the compounds’ therapeutic potential, are reported in an article published in Clinical Immunology.

The two medications were administered separately. The monoclonal antibody, routinely used to treat blood diseases, was tested on patients at the teaching hospital (“Hospital das Clínicas”) run by the University of São Paulo’s Ribeirão Preto Medical School (FMRP-USP). AMY-101, a candidate drug developed by US-based pharmaceutical company Amynda, was given to patients at a hospital in Milan, Italy. Both produced promising results, but because AMY-101 is cheaper and performed better in the clinical trial, the two research groups envisage testing it on a larger number of patients in Brazil.

“Both compounds caused a robust anti-inflammatory response that culminated in a fairly rapid recovery of respiratory function in the patients,” Rodrigo Calado, who led the study at FMRP-USP, told Agência FAPESP. Calado is affiliated with CTC, a Research, Innovation and Dissemination Center (RIDC) funded by FAPESP and based at FMRP-USP’s blood center.

The researchers concluded that the therapeutic benefits of eculizumab and AMY-101 were due to inhibition of a bloodstream cascade of proteins involved in immune response and known as the complement system.

Persistent dysregulated complement activation triggers the exacerbated inflammatory response to SARS-CoV-2 infection characterized by a systemic increase in pro-inflammatory cytokines and often referred to as a “cytokine storm”.

Unable to prevent infection of cells by the virus, the complement system enters a spiral of runaway continuous activation that leads to massive infiltration of monocytes and neutrophils into the infected tissues. This process can result in inflammatory damage to the walls of blood vessels surrounding vital organs, as well as disseminated microvascular injury and thrombosis, potentially culminating in multiple organ failure.

“Previous studies had shown the use of complement inhibitors to be a promising therapeutic strategy to ameliorate thromboinflammation in COVID-19 patients, and there were reports of cases with positive results,” Calado said. “However, as yet no one had elucidated the action or evaluated the efficacy of drugs already in use to treat diseases caused by complement alterations, such as eculizumab, or candidate drugs with this function such as AMY-101.”

To fill the gap the researchers conducted two clinical studies in which they compared the biological efficacy of eculizumab with that of the synthetic peptide AMY-101 in independent small groups of severe COVID-19 patients.

Ten patients aged between 18 and 80 were undergoing treatment at FMRP-USP’s Hospital das Clínicas. Once a week while hospitalized, they were given 900 mg of eculizumab, which inhibits the complement protein C5. Three patients hospitalized in Milan, Italy, were given 5 mg of AMY-101, developed to inhibit complement protein C3, also once a week. These two proteins perform the most important activities in the complement system.

The patients’ clinical responses showed that eculizumab and AMY-101 elicited a robust anti-inflammatory process, a sharp fall in levels of C reactive protein (CRP) and interleukin-6 (IL-6), and a marked improvement in lung function.

Inhibition of C3 by AMY-101 afforded broader therapeutic control, stronger lymphocyte recovery, a pronounced decline in neutrophil count, and more robust attenuation of the thromboinflammation induced by the exacerbated response to viral infection.

“The results of the clinical trials showed that inhibiting components of the complement system sharply reduces inflammation,” Calado said.

New clinical study

In light of the promising results of the two clinical trials, the CTC and UPenn researchers now plan to conduct a Phase 3 trial with more than 100 severe COVID-19 patients, who will be given only AMY-101 in an effort to assess the drug’s efficacy on a wider scale.

The study will be conducted at FMRP-USP’s Hospital das Clínicas, and will probably involve other research institutions in Brazil. “One of the advantages of AMY-101 is its low cost. It’s much less expensive than eculizumab,” Calado said.

New Rochelle, NY, November 13, 2020--Patients are seeking plastic surgery in record numbers, citing their appearance on Zoom as a cause. Of particular concern are noses and wrinkles, according to Facial Plastic Surgery & Aesthetic Medicine. Click here to read the article now.

"A life disproportionately spent on Zoom may trigger a self-critical comparative response that leads people to rush to their physicians for treatments they may not have considered before months confronting a video screen, a new phenomenon of 'Zoom Dysmorphia,'" state Arianne Shadi Kourosh, MD, Massachusetts General Hospital, and coauthors.

"The COVID-19 pandemic has radically changed the frequency with which we are confronted with our own image. The shift to online work, learning, and even socializing has dramatically increased the time we have to observe ourselves," says Benjamin Marcus, MD, University of Wisconsin Hospitals and Clinics, in an accompanying commentary.

Facial Plastic Surgery & Aesthetic Medicine Editor-in-Chief Travis T. Tollefson, MD, MPH, University of California, Davis, states: "It has always been incumbent on surgeons to understand the motivations that are driving individuals to seek plastic surgery in order to assure realistic goals that allow for successful surgeries. Now, more than ever, unique circumstances can be driving patient expectations that we should be taking into consideration."

For COVID-19, the difference between surviving and not surviving severe disease may be due to the quality, not the quantity, of the patients' antibody development and response, suggests a new Cell paper published by Galit Alter, PhD, a member of the Ragon Institute of MGH, MIT and Harvard.

The study, published in the journal Cell, used Alter's systems serology approach to profile the antibody immune responses of 193 hospitalized COVID-19 patients, comparing responses from patients with moderate and severe disease and patients who passed away from COVID-19.

While all patients developed antibodies against SARS-CoV-2, the way the antibodies developed, or evolved, differed between the three groups. For patients that didn't survive the disease, the antibody response never fully evolved.

"There was a significant defect in the development of IgG antibodies, which may be essential in the early control and elimination of the virus, " Alter says. "Here, we were able to see the global impact of this defective IgG evolution, resulting in a compromised ability to promote essential viral clearing immune functions."

In a mature immune response, antibodies both block infection and direct the immune system to kill infected cells. To guide the killer immune response, antibodies attach to the Fc-receptor, a "docking site" specific to antibodies that is found on all immune cells. Without strong Fc-receptor binding, antibodies may fail to grab and destroy virus following infection.

Compared to survivors, patients who passed from COVID-19 had antibodies that never fully developed the ability to strongly bind to Fc-receptors and therefore may not have been able to fully trigger immune killing activity.

Alter's group, led by Tomer Zohar, Carolin Loos, Stephanie Fischinger, and Caroline Atyeo, PhD, also found that survivors' immune systems could recognize and target an area of the SARS-CoV-2 spike protein known as the S2-domain. The S2 domain is found in other coronaviruses that infect humans, so patients whose antibodies can target it may have pre-existing immunity to the S2 domain because of exposure to other, common coronaviruses.

Patients with antibodies that can recognize S2 domains on different coronaviruses may be able to use this pre-existing immunity to generate killer antibodies faster and sooner following SARS-CoV-2 infection.

"If we can further understand the importance of cross-coronavirus immunity," says Zohar, "researchers may be able to design vaccines able to counteract a much broader range of coronaviruses."

With studies like these, Alter and her team are working to understand the nature of protective immunity against SARS-CoV-2, including partnering with COVID-19 vaccine developers, to help bring an end to this pandemic.

Researchers at Vanderbilt University Medical Center (VUMC) and their colleagues have determined a key factor as to why COVID-19 appears to infect and sicken adults and older people preferentially while seeming to spare younger children.

Children have lower levels of an enzyme/co-receptor that SARS-CoV-2, the RNA virus that causes COVID-19, needs to invade airway epithelial cells in the lung.

The findings, published today in the
Journal of Clinical Investigation, support efforts to block the enzyme to potentially treat or prevent COVID-19 in older people.

"Our study provides a biologic rationale for why particularly infants and very young children seem to be less likely to either get infected or to have severe disease symptoms," said Jennifer Sucre, MD, assistant professor of Pediatrics (Neonatology), who led the research with Jonathan Kropski, MD, assistant professor of Medicine.

Sucre and Kropski are co-corresponding authors of the paper. Bryce Schuler, MD, PhD, a resident in Pediatrics and Genetics at VUMC and postdoctoral fellow in the Vanderbilt Stimulating Access to Research in Residency program, is the paper's first author.

There is still much to learn about SARS-CoV-2. But this much is known: After a viral particle is inhaled into the lungs, protein "spikes" that stick out like nail studs in a soccer ball attach to ACE2, a receptor on the surfaces of certain lung cells.

A cellular enzyme called TMPRSS2 chops up the spike protein, enabling the virus to fuse into the cell membrane and "break into" the cell. Once inside, the virus hijacks the cell's genetic machinery to make copies of its RNA.

Sucre and Kropski, who have collaborated since 2016 on studies of lung diseases in premature infants and adults, wondered if TMPRSS2 had something to do with the greater severity of COVID-19 symptoms observed in older people compared to children.

"Our research has always focused on understanding lung development and how infant lungs differ from adult lungs in their vulnerability to injury," Sucre said. "In this study we actually took the opposite approach, and were able to see how the developing lung by its differences is protected from SARS-CoV-2 infection."

The researchers were well equipped to begin such a study. As members of the international Human Cell Atlas (HCA) Lung Biological Network, they and their colleagues had built a dataset on lung development in the mouse using a technique called single-cell RNA-sequencing.

The technique can detect the expression of genes in individual cells of tissues such as the lung. In this way the researchers were able to track the expression of genes known to be involved in the body's response to COVID-19 over time.

They found that while the gene for ACE2 was expressed at low levels in the mouse lung, "TMPRSS2 stood out as having a really striking trajectory of increased expression during development," Schuler said.

With the help of VUMC pathologists, the researchers obtained and analyzed human lung specimens collected from donors of different ages, and confirmed a similar trajectory in TMPRSS2 expression to what they'd found in mice.

"What we found is that expression of (TMPRSS2) goes up significantly with aging, and we see that at the level of the gene and at the level of the protein," Sucre said. "We see a lot more TMPRSS2 in older individuals, in both humans and mice."

The researchers also used fluorescent probes to analyze autopsy specimens from three patients who died of COVID-19, and found the virus in three types of cells that express TMPRSS2.

TMPRSS2 is well known for its role in the development of prostate cancer. Drugs that block the enzyme and which have been approved for the treatment of advanced prostate cancer currently are being tested clinically as potential treatments for COVID-19.

The new findings reported today support further investigation.

"We do think TMPRSS2 could be an attractive target both in treatment and potentially as a prophylaxis for (preventing infection in) people at high risk of COVID exposure," Sucre said.

Given that phages are able to destroy bacteria, they are of particular interest to science. Basic researchers from the Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP) in Berlin are especially interested in the tube used by phages to implant their DNA into bacteria. In collaboration with colleagues from Forschungszentrum Jülich and Jena University Hospital, they have now revealed the 3D structure of this crucial phage component in atomic resolution. The key to success was combining two methods - solid-state NMR and cryo-electron microscopy. The study has just been published in the journal Nature Communications.

With growing antibiotic resistance, phages have increasingly become the focus of research. Phages are naturally occurring viruses with a very useful property: they implant their DNA into bacteria and proliferate there until the bacterial cell is ultimately destroyed. This is why they are also referred to as bacteriophages (bacteria eaters).

This approach has already been shown to fight multidrug-resistant bacteria. Last year, the case of a girl from England hit the headlines, when she was cured from a serious antibiotic-resistant infection using engineered phages.

However, the widespread use of phage therapy is still a long way off. Many of the underlying principles that are key to advancing this therapy are not yet understood. For example, little was previously known about the appearance of the exact architecture of the tube used by phages to implant their DNA into bacteria. Now scientists from the Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP) in Berlin, together with colleagues from Forschungszentrum Jülich and Jena University Hospital, have managed to reveal the 3D structure of this crucial phage component in atomic resolution.

Designed for transporting DNA

"The structure and flexibility of the DNA tube attached to the icosahedron-shaped capsid is somewhat reminiscent of a spinal column," stated FMP's Professor Adam Lange, describing one of the new findings. "It seems to be perfectly designed for transporting DNA."

The researchers were able to gain fascinating insights into the structure and function of this sophisticated DNA transport pathway - in this case, from a variant of phage SPP1 - by innovatively combining solid-state NMR with cryo-electron microscopy (cryo-EM). Lange's research group further developed nuclear magnetic resonance spectroscopy (NMR) especially for this task under an ERC Grant; cryo-EM expert Professor Gunnar Schröder from Forschungszentrum Jülich performed the electron-microscopic investigations. In addition, new modeling algorithms were required for the computer-based combination of the two data sets for structure determination. These algorithms were developed by Professor Michael Habeck from Jena University Hospital. "The key to success was combining the two methods, representing a methodological milestone," commented Professor Lange.

While solid-state NMR is ideal for visualizing flexible structures and tiny details, cryo-EM provides insight into the overall architecture. The resulting image shows that six gp17.1 proteins organize into stacked rings, forming a hollow tube. The rings are connected by flexible linkers, making the tube very bendable. "We are now able to understand how negatively charged DNA is repelled from the likewise negatively charged interior wall of the flexible tube, passing through it smoothly," explained FMP's Maximilian Zinke, lead author of the study now published in Nature Communications. "The bacteria are ultimately destroyed via this pathway."

Milestone for integrated structural biology

According to group leader Adam Lange, besides representing a quantum leap forward in phage research, the work will also advance "integrated structural biology", the term for the combination of these two complementary methods.

Thanks to the recent installation of a new high-resolution Titan Krios electron microscope, the infrastructure required to achieve this is now available on Campus Berlin-Buch. Moreover, a 1.2 gigahertz device will soon be added to the existing NMR spectrometers. "Equipped with cryo-EM and the most sensitive NMR spectrometer in the world, we will be very present in integrative structural biology in the future," enthused Adam Lange. "This offers bright prospects for the campus and for the research location of Berlin."

Embargoed until 12:10 p.m. CT/1:10 p.m. ET, Friday, Nov. 13, 2020

DALLAS, Nov. 13, 2020 -- A single, daily pill combining blood pressure and cholesterol medications, along with the addition of a daily dose of aspirin, reduced cardiovascular disease events in people at risk for heart disease, according to late-breaking research presented today in a late-breaking clinical trial presentation at the American Heart Association's Scientific Sessions 2020. The virtual conference, held Friday, November 13 - Tuesday, November 17, 2020, is a premier global exchange of the latest scientific advancements, research and evidence-based clinical practice updates in cardiovascular science for health care worldwide. The manuscript of this study is simultaneously published today in The New England Journal of Medicine.

The medicine tested in this study is a fixed-dose combination therapy (or a "polypill") combining blood pressure and cholesterol lowering medications. Researchers assessed the polypill's effects on cardiovascular disease events - such as heart attack, stroke and cardiovascular death - when given alone or with aspirin, in patients considered at intermediate risk of cardiovascular disease. They also examined the effects of aspirin alone.

The International Polycap Study (TIPS)-3 is a large, randomized, placebo-controlled trial conducted in nine countries. The study included 5,700 people considered at intermediate risk of developing heart disease. The average age of the participants was 64 years, and 47% were male.

Participants were randomly assigned to different interventions: 1) 75 mg daily of aspirin; 2) a polypill combining blood pressure and cholesterol medication daily; 3) polypill and 75 mg aspirin daily; or 4) vitamin D 5,000 IU daily. Each intervention included a control group who received a matching placebo. The medications in the polypill were atenolol 100mg, ramipril 10mg, hydrochlorothiazide 25 mg, and simvastatin 40 mg.

Over the follow-up period of nearly five years, participants were monitored for the first occurrence of a major cardiovascular event, such as non-fatal heart attack, non-fatal stroke, heart failure, resuscitated cardiac arrest or cardiovascular death.

The analysis of all patient groups found:

the polypill alone reduced cardiovascular disease by 21%;

aspirin alone reduced cardiovascular death, heart attack or stroke by 14%;
and
the polypill plus aspirin reduced cardiovascular disease by 31%.

"Aspirin should be prescribed with a polypill in primary prevention for patients at intermediate risk of heart disease," said Salim Yusuf, M.D., B.S., D. Phil., a co-author of the study and professor of medicine at McMaster University School of Medicine in Toronto, Canada. "Our study results provide important data regarding the role of the polypill in preventing the development of heart disease."

Co-author Prem Pais, M.B.B.S., M.D., a professor in the division of clinical research and training at St. John's Research Institute in Bangalore, India, added, "We were also interested in evaluating if combining blood pressure and cholesterol reduction medications in a single pill would be effective for this population. This is a cost-effective strategy that could help meet global targets of reducing CVD by 30% by 2030."

"Use of a polypill plus aspirin can avert 3 - 5 million cardiovascular deaths globally," said Yusuf. "Future polypills, with newer statins, may reduce LDL cholesterol and blood pressure to a greater extent and could reduce cardiovascular disease risk greater than 50%."

DALLAS, Nov. 13, 2020 -- Patients who were hospitalized with acute heart failure and had iron deficiency were less likely to return to the hospital if given intravenous iron replacement, according to late-breaking research presented today at the American Heart Association's Scientific Sessions 2020. The virtual meeting is Friday, November 13 - Tuesday, November 17, 2020. It is a premier global exchange of the latest scientific advancements, research and evidence-based clinical practice updates in cardiovascular science for health care. The manuscript of this study is simultaneously published today in The Lancet.

Study participants were stabilized after an episode of acute heart failure and given intravenous ferric carboxymaltose, an iron replacement therapy, at hospital discharge.

"Iron deficiency is common in patients with heart failure, and it is an independent risk factor for hospital admission and death," said Piotr Ponikowski, M.D., Ph.D., head of the department of heart diseases at Wroclaw Medical University in Wroclaw, Poland, and lead author of the study. "We tested the hypothesis that correcting iron deficiency with ferric carboxymaltose in patients admitted for an episode of acute heart failure and who have iron deficiency is effective in reducing the risk of recurrent hospitalization and cardiovascular death."

The AFFIRM-AHF trial enrolled 1,108 patients in 15 countries who were being discharged after a hospitalization for acute heart failure and found to have iron deficiency during their hospital stay. The participants' average age was 71, and 56% were men, with average heart ejection fraction measures of 33%. Ejection fraction measures the amount of blood pumped out by the heart with each contraction. A normal ejection fraction is between 50% and 70%.

After their heart condition was stabilized, patients received either intravenous ferric carboxymaltose or a placebo during the following 24 weeks, dosed according to the level of iron deficiency.

The analysis found significantly fewer hospital readmissions due to heart failure among the patients treated with intravenous ferric carboxymaltose compared to those who received the placebo. After 52 weeks, patients who received iron supplementation were 26% less likely to be re-admitted to the hospital for heart failure. This result was achieved with only one or two injections in 80% of the patients in the ferric carboxymaltose group.

Results for reduced risk of hospitalization were statistically significant; however, when the researchers looked at the combined goal of reducing hospitalizations and death, the results were not statistically different.

"This is the first study demonstrating the benefits of iron supplementation initiated in stabilized patients hospitalized for acute heart failure, and only two doses were needed in the vast majority of patients," Ponikowski said. "Health care professionals should screen patients with heart failure for the presence of iron deficiency, and intravenous iron should be considered for those with iron deficiency and ejection fraction of 50% or lower."

The rheumatoid arthritis drug baricitinib can block viral entry and reduce mortality in patients with moderate to severe COVID-19, according to translational research by an international team coordinated by researchers from Karolinska Institutet in Sweden. The findings, published in the journal Science Advances, support the continuation of ongoing randomized clinical trials.

"We are pleased to report a 71 percent reduction in mortality for the group receiving baricitinib in addition to standard care," says Volker Lauschke, corresponding author and associate professor of personalized medicine and drug development at the Department of Physiology and Pharmacology, Karolinska Institutet. "These results are especially encouraging seeing as the study included a large cohort of elderly patients, a group that is often excluded in other trials."

In the study, 83 patients hospitalized with COVID-19 pneumonia in Italy and Spain were treated with baricitinib in addition to standard care. Of these, 17 percent suffered an adverse outcome that resulted in death or invasive mechanical ventilation. This compared to 35 percent in the matched control group of 83 patients who received standard care only. The patients had a median age of 81 years.

The drug was generally well tolerated with a reduction in inflammation from the first treatment days, according to the researchers. Previously reported side-effects of long-term baricitinib use, including coagulopathy and thrombosis, were not evident in any of the patients, possibly due to treatment with anti-coagulating medicine. However, some adverse events including bacterial infections and gastrointestinal and cardiovascular complications were noted, although these were also observed in the control group so it is unclear what, if anything, can be ascribed to baricitinib.

In a prior study, the same team of researchers reported how they used artificial intelligence (AI) to identify baricitinib as a promising repurposing candidate for COVID-19. That study also showed how the drug inhibited inflammation and reduced the viral load of SARS-CoV-2.

In the current study, the researchers elaborated on those findings by demonstrating that interferons, cytokines made and released by host cells in response to viruses, significantly increases the expression of the ACE2 receptor, which acts as an entry point for SARS-CoV-2 into human cells. While liver injury is commonly observed in severe COVID-19, mechanisms and dynamics of SARS-CoV-2 infections had not been investigated in this organ.

By combining 3D mini organs of human liver cells, RNA sequencing and super-resolution microscopy, the scientists were able to show that barcitinib reversed ACE2 gene expression changes triggered by interferons and reduced SARS-CoV-2 infectivity. Interestingly, interferons did not have the same effect on the ACE2 receptor in lung organoids, suggesting that these signaling proteins affect pulmonary and liver organs differently.

"Our findings explain the dual anti-cytokine and anti-viral actions of baricitinib and support further evaluation in randomized control trials," says Ali Mirazimi, adjunct professor at the Department of Laboratory Medicine, Karolinska Institutet, and co-author of the study.

The researchers note that a limitation of the study was the lack of a placebo control group, which is included in industry-sponsored randomized controlled trials that are currently ongoing.

A clinical study involving 601 patients in Italy and Spain suggests that the JAK inhibitor drug baricitinib may enhance survival rates of patients with severe COVID-19, primarily by blunting the runaway immune inflammation known as "cytokine storm." While the authors are careful to note that their study was not a fully randomized trial with a placebo control group, the drug nonetheless had a notable effect, particularly in the Spanish treatment cohort, composed entirely of elderly patients over the age of 70. This suggests the treatment could address high rates of COVID-19 morbidity and mortality observed in this demographic worldwide. Further in vitro experiments in liver cell spheroids suggest the drug may also restrain abnormally high expression of the ACE2 receptor - required for entry of the SARS-CoV-2 virus - in these cells, helping to reduce viral infection of the liver, and likely other organs and tissues that do not normally express high levels of the ACE2 receptor. Using AI-based analyses, Justin Stebbing and colleagues had previously identified baricitinib, approved for the treatment of rheumatoid arthritis, as a promising treatment for COVID-19. They also previously found that the drug can reduce levels of the pro-inflammatory cytokine IL-6 in rheumatoid arthritis patients. Noting that levels of IL-6 have been reported to correlate with symptom severity in patients with COVID-19, Stebbing et al. enrolled a total of 179 hospitalized patients from Pisa, Italy, and 422 from Albacete, Spain. In the Pisa cohort, 37 patients were given baricitinib once daily for two weeks; of the 142 controls, 37 were matched with the drug recipient group using a "propensity score" based on the patients' age, sex, and several parameters related to their medical history. In Albacete, 46 patients were given a lower dose of the drug for shorter periods of time, due to safety considerations in this elderly cohort. As in Pisa, 46 of the 376 control patients were matched with the drug recipient cohort using the propensity score method. Overall, across both cohorts matched by propensity score, the patients who received baricitinib either died or received invasive mechanical ventilation at roughly half the rate (16.9%) of the untreated group (34.9%). Stebbing et al. note that randomized trials of the drug are currently underway. Based on their own results - including their additional experiments in liver cell spheroids - the authors note that they "eagerly await" the results of these trials.

The Barbegal watermills in southern France are a unique complex dating back to the 2nd century AD. The construction with 16 waterwheels is, as far as is known, the first attempt in Europe to build a machine complex on an industrial scale. The complex was created when the Roman Empire was at the height of its power. However, little is known about technological advances, particularly in the field of hydraulics, and the spread of knowledge at the time. A team of scientists led by Professor Cees Passchier from Johannes Gutenberg University Mainz (JGU) has now gained new knowledge about the construction and principle of the water supply to the mills in Barbegal. The research results were published in Scientific Reports.

A mill complex consisting of a total of 16 water wheels in two parallel rows

Watermills were one of the first sources of energy that did not depend on the muscle strength of humans or animals. In the Roman Empire they were used to make flour and sawing stone and wood. As one of the first industrial complexes in European history, the Barbegal watermills are an outstanding example of the development at that time. The mill complex consisted of 16 water wheels in a parallel arrangement of eight wheels each, separated by central buildings and fed by an aqueduct. The upper parts of the complex were destroyed and no traces of the wooden structures have been preserved, which is why the type of mill wheels and how they worked remained a mystery for a long time.

However, carbonate deposits that had formed from the flowing water on the wooden components remained. These were stored in the archaeological museum in Arles and only recently examined in detail. The researchers found an imprint of an unusual, elbow-shaped flume that must have been part of the mill construction. "We combined measurements of the water basins with hydraulic calculations and were able to show that the flume to which this elbow-shaped piece belonged very likely supplied the mill wheels in the lower basins of the complex with water," said Professor Cees Passchier. "The shape of this flume was unknown from other watermills, either from Roman or more recent times. We were therefore puzzled as to why the flume was designed this way and what it was used for."

An elbow-shaped flume as a unique adaptation for the Barbegal mills

At first glance, the team found such a flume unnecessary and even disadvantageous, because it shortens the height from which the water falls onto the mill wheel. "However, our calculations show that the oddly shaped flume is a unique adaptation for the Barbegal mills," explained Passchier. The distribution of the carbonate deposits in the elbow-shaped flume shows that it was inclined slightly backwards against the direction of the current. This created a maximum flow rate in the first, steep leg of the flume, and at the same time the water jet to the mill wheel obtained the correct angle and speed. In the complicated mill system, with small water basins, this unique solution was more efficient than using a traditional, straight water channel. "That shows us the ingenuity of the Roman engineers who built the complex," emphasized Passchier.

"Another discovery was that the wood of the flume was probably cut with a mechanical, water-powered saw, which is possibly the first documented mechanical wood saw - again evidence of industrial activity in ancient times." The research was carried out by a multidisciplinary team of experts in geology, geochemistry, hydraulics, dendrochronology, and archaeology.

The carbonate deposits that formed on the ancient hydraulic structures are an important tool for the researchers for archaeological reconstructions. In an earlier project, the team led by Professor Cees Passchier was able to show that the flour from the Barbegal mills was probably used to make ship biscuits. "The carbonate deposits give us extremely exciting insights into the skills of Roman technicians at a time that can be seen as the direct predecessor of our civilization," added Passchier, Professor of Tectonic Physics and Structural Geology at the JGU Institute of Geosciences from 1993 to 2019, now Senior Research Professor in Geoarchaeology.