Study shows laboratory developed protein spikes consistent with COVID-19 virus

 A new international study has found that the key properties of the spikes of SARS-CoV-2 virus which causes COVID-19 are consistent with those of several laboratory-developed protein spikes, designed to mimic the infectious virus.

A central component in designing serological tests and vaccines to protect against COVID-19 is the manufacture of protein "spikes." These recombinant spikes closely mimic those sticking out of surface of the infectious virus and trigger the body's immune system into action.

Laboratory manufactured spikes are also used for serological testing (also referred to as antibody testing) and as research reagents. The findings show how that viral spike manufactured through different methods in laboratories across the globe are highly similar and provide reassurance that the spike can be robustly manufactured with minimal variations between laboratories.

The spikes on the SARS-CoV-2 virus are coated in sugars, known as glycans, which they use to disguise themselves from the human immune system. The abundance of these glycans has the potential to create significant discrepancies between studies that use different recombinant spikes.

In this new study, published in the journal Biochemistry, the research team studied the glycan coatings on recombinant spikes developed in five laboratories around the world and compared them to those on the spikes of the infectious virus.

"The speed at which scientific community has moved to tackle the COVID-19 pandemic has put considerable pressure on laboratories around the world to validate their findings quickly," Explained Max Crispin, Professor of Glycobiology at the University of Southampton, who led the study. "Over the last year we have seen vaccines developed around the world at an unprecedented rate and the rapid development, and validation, of recombinant proteins have been fundamental to that success story," he continued.

In April 2020, Professor Crispin and his team from the University of Southampton mapped the glycan coating of the SARS-CoV-2 spike for the first time. In the present study, they extend their analysis to examine recombinant spike developed in laboratories at the Amsterdam University Medical Centre, Harvard Medical School, the University of Oxford, and the Swiss company ExcellGene. All the different batches of spike protein were shown to mimic key features of the glycosylation of virions analysed at Tsinghua University, China.

The study also used computational methods to examine the protein features that were shaping some of the glycosylation features that were seen across all the samples. Dr. Peter Bond, Senior Principal Investigator at the Bioinformatics Institute of the Agency for Science, Technology and Research (A*STAR), Singapore, who led the computational work said, "Our modelling enabled us to shed light on how the protein influences the structure of the glycans and why the glycosylation was so consistent. This predictive approach could also be of potential value in therapeutics development against new variants or other emerging viruses."

"The ability to produce mimics of the SARS-CoV-2 spike protein with high fidelity at many different laboratories, all of which recapitulate the glycan signatures of the authentic virus, is of significant benefit for vaccine design, antibody testing and drug discovery" concluded Professor Crispin.

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Materials provided by University of Southampton. Note: Content may be edited for style and length.

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Journal Reference:

1. Joel D. Allen, Himanshi Chawla, Firdaus Samsudin, Lorena Zuzic, Aishwary Tukaram Shivgan, Yasunori Watanabe, Wan-ting He, Sean Callaghan, Ge Song, Peter Yong, Philip J. M. Brouwer, Yutong Song, Yongfei Cai, Helen M. E. Duyvesteyn, Tomas Malinauskas, Joeri Kint, Paco Pino, Maria J. Wurm, Martin Frank, Bing Chen, David I. Stuart, Rogier W. Sanders, Raiees Andrabi, Dennis R. Burton, Sai Li, Peter J. Bond, Max Crispin. Site-Specific Steric Control of SARS-CoV-2 Spike Glycosylation. Biochemistry, 2021; DOI: 10.1021/acs.biochem.1c00279

https://www.sciencedaily.com/releases/2021/07/210706115421.htm

Memory making involves extensive DNA breaking

 The urgency to remember a dangerous experience requires the brain to make a series of potentially dangerous moves: Neurons and other brain cells snap open their DNA in numerous locations -- more than previously realized, according to a new study -- to provide quick access to genetic instructions for the mechanisms of memory storage.

The extent of these DNA double-strand breaks (DSBs) in multiple key brain regions is surprising and concerning, said study senior author Li-Huei Tsai, Picower Professor of Neuroscience at MIT and director of The Picower Institute for Learning and Memory, because while the breaks are routinely repaired, that process may become more flawed and fragile with age. Tsai's lab has shown that lingering DSBs are associated with neurodegeneration and cognitive decline and that repair mechanisms can falter.

"We wanted to understand exactly how widespread and extensive this natural activity is in the brain upon memory formation because that can give us insight into how genomic instability could undermine brain health down the road," said Tsai, who is also a professor in the Department of Brain and Cognitive Sciences and a leader of MIT's Aging Brain Initiative. "Clearly memory formation is an urgent priority for healthy brain function but these new results showing that several types of brain cells break their DNA in so many places to quickly express genes is still striking."

Tracking breaks

In 2015, Tsai's lab provided the first demonstration that neuronal activity caused DSBs and that they induced rapid gene expression. But those findings, mostly made in lab preparations of neurons, did not capture the full extent of the activity in the context of memory formation in a behaving animal and did not investigate what happened in cells other than neurons.

In the new study published July 1 in PLOS ONE, lead author and former graduate student Ryan Stott and co-author and former research technician Oleg Kritsky sought to investigate the full landscape of DSB activity in learning and memory. To do so, they gave mice little electrical zaps to the feet when they entered a box, to condition a fear memory of that context. They then used several methods to assess DSBs and gene expression in the brains of the mice over the next half hour, particularly among a variety of cell types in the prefrontal cortex and hippocampus, two regions essential for the formation and storage of conditioned fear memories. They also made measurements in the brains of mice who did not experience the foot shock to establish a baseline of activity for comparison.

The creation of a fear memory doubled the number of DSBs among neurons in the hippocampus and the prefrontal cortex, affecting more than 300 genes in each region. Among 206 affected genes common to both regions, the researchers then looked at what those genes do. Many were associated with the function of the connections neurons make with each other, called synapses. This makes sense because learning arises when neurons change their connections (a phenomenon called "synaptic plasticity") and memories are formed when groups of neurons connect together into ensembles called engrams.

"Many genes essential for neuronal function and memory formation, and significantly more of them than expected based on previous observations in cultured neurons...are potentially hotspots of DSB formation," the authors wrote in the study.

In another analysis, the researchers confirmed through measurements of RNA that the increase in DSBs indeed correlated closely with increased transcription and expression of affected genes, including ones affecting synapse function, as quickly as 10-30 minutes after the foot shock exposure.

"Overall, we find transcriptional changes are more strongly associated with [DSBs] in the brain than anticipated," they wrote. "Previously we observed 20 gene-associated [DSB] loci following stimulation of cultured neurons, while in the hippocampus and prefrontal cortex we see more than 100-150 gene associated [DSB] loci that are transcriptionally induced."

Snapping with stress

In the analysis of gene expression, the neuroscientists looked at not only neurons but also non-neuronal brain cells, or glia, and found that they also showed changes in expression of hundreds of genes after fear conditioning. Glia called astrocytes are known to be involved in fear learning, for instance, and they showed significant DSB and gene expression changes after fear conditioning.

Among the most important functions of genes associated with fear conditioning-related DSBs in glia was the response to hormones. The researchers therefore looked to see which hormones might be particularly involved and discovered that it was glutocortocoids, which are secreted in response to stress. Sure enough, the study data showed that in glia, many of the DSBs that occurred following fear conditioning occurred at genomic sites related to glutocortocoid receptors. Further tests revealed that directly stimulating those hormone receptors could trigger the same DSBs that fear conditioning did and that blocking the receptors could prevent transcription of key genes after fear conditioning.

Tsai said the finding that glia are so deeply involved in establishing memories from fear conditioning is an important surprise of the new study.

"The ability of glia to mount a robust transcriptional response to glutocorticoids suggest that glia may have a much larger role to play in the response to stress and its impact on the brain during learning than previously appreciated," she and her co-authors wrote.

Damage and danger?

More research will have to be done to prove that the DSBs required for forming and storing fear memories are a threat to later brain health, but the new study only adds to evidence that it may be the case, the authors said.

"Overall we have identified sites of DSBs at genes important for neuronal and glial functions, suggesting that impaired DNA repair of these recurrent DNA breaks which are generated as part of brain activity could result in genomic instability that contribute to aging and disease in the brain," they wrote.

The National Institutes of Health, The Glenn Foundation for Medical Research and the JPB Foundation provided funding for the research.

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Story Source:

Materials provided by Picower Institute at MIT. Note: Content may be edited for style and length.

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Journal Reference:

1. Ryan T. Stott, Oleg Kritsky, Li-Huei Tsai. Profiling DNA break sites and transcriptional changes in response to contextual fear learning. PLOS ONE, 2021; 16 (7): e0249691 DOI: 10.1371/journal.pone.0249691

https://www.sciencedaily.com/releases/2021/07/210706115310.htm

How a series of sleep loss impacts mental and physical wellbeing

 All it takes is three consecutive nights of sleep loss to cause your mental and physical well-being to greatly deteriorate. A new study published in Annals of Behavioral Medicine looked at the consequences of sleeping fewer than six hours for eight consecutive nights -- the minimum duration of sleep that experts say is necessary to support optimal health in average adults.

Lead author Soomi Lee, assistant professor in the School of Aging Studies at the University of South Florida, found the biggest jump in symptoms appeared after just one night of sleep loss. The number of mental and physical problems steadily got worse, peaking on day three. At that point, research shows the human body got relatively used to repeated sleep loss. But that all changed on day six, when participants reported that the severity of physical symptoms was at its worst.

"Many of us think that we can pay our sleep debt on weekends and be more productive on weekdays," Lee said. "However, results from this study show that having just one night of sleep loss can significantly impair your daily functioning."

Data provided by the Midlife in the United States study included nearly 2,000 middle-aged adults who were relatively healthy and well-educated. Among them, 42% had at least one night of sleep loss, sleeping 1 ½ fewer hours than their typical routines. They recorded their mental and physical behaviors in a diary for eight consecutive days, allowing researchers to review how sleep loss causes wear and tear on the body.

Participants reported a pile-up of angry, nervous, lonely, irritable and frustrated feelings as a result of sleep loss. They also experienced more physical symptoms, such as upper respiratory issues, aches, gastrointestinal problems and other health concerns. These negative feelings and symptoms were continuously elevated throughout consecutive sleep loss days and didn't return to baseline levels unless they had a night sleep of more than six hours.

About one-third of U.S. adults sleep less than six hours per night. Lee says once that becomes a habit, it's increasingly difficult for your body to fully recover from lack of sleep, continuing the vicious cycle of worsening daily well-being, which could impact one professionally. A previous study led by Lee found losing just 16 minutes of sleep could impact job performance. Her previous findings also show that minor sleep loss can decrease daily mindfulness, which is a critical recourse for managing stress and maintaining healthy routines.

Lee says the best way to maintain a strong daily performance is to set aside more than six hours to sleep every night.

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Story Source:

Materials provided by University of South Florida. Note: Content may be edited for style and length.

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Journal Reference:

1. Soomi Lee. Naturally Occurring Consecutive Sleep Loss and Day-to-Day Trajectories of Affective and Physical Well-Being. Annals of Behavioral Medicine, 2021; DOI: 10.1093/abm/kaab055

https://www.sciencedaily.com/releases/2021/07/210706133113.htm

Home medical equipment provider Rotech Healthcare files for a $100 million IPO

 Rotech Healthcare Holdings, which provides home medical equipment and services, filed on Tuesday with the SEC to raise up to $100 million in an initial public offering.

Rotech Healthcare offers a comprehensive range of home medical equipment and related products and services across five core business lines: oxygen, ventilators, sleep therapy, wound care, and DME. The company's payor clients include commercial insurers, Medicare, Medicaid, the VA, and private individuals. As of March 31, 2021, the company served more than 600,000 active patients across over 300 service locations in 45 states.

The Orlando, FL-based company was founded in 2018 and booked $534 million in sales for the 12 months ended March 31, 2021. It plans to list on the Nasdaq under the symbol ROTK. Rotech Healthcare Holdings filed confidentially on April 30, 2021. BofA Securities, Jefferies, UBS Investment Bank, Truist Securities, Baird, and RBC Capital Markets are the joint bookrunners on the deal. No pricing terms were disclosed.

Relevant Profile: ROTK

https://www.renaissancecapital.com/IPO-Center/News/83692/Home-medical-equipment-provider-Rotech-Healthcare-files-for-a-$100-million-

ChemoCentryx Shares Rise 10% After Filing NDA Amendment for Avacopan

 ChemoCentryx Inc. shares were up 10% at $14.52 after the company said that, following consultations with the U.S. Food and Drug Administration, it filed an amendment to its New Drug Application for avacopan for the treatment of anti-neutrophil cytoplasmic autoantibody-associated vasculitis.

The company said the amendment addresses points raised during an FDA Advisory Committee meeting on May 6.

The FDA has indicated that the filing constitutes a major amendment to the NDA and will result in the setting of a new Prescription Drug User Fee Act goal date of Oct. 7. The NDA is primarily based on data from a Phase III trial of avacopan for the treatment of ANCA-associated vasculitis.

On May 6, the FDA's Arthritis Advisory Committee voted 9-9 on whether the efficacy data support approval of avacopan, 10-8 that the safety profile of avacopan is adequate to support approval, and 10-8 that the benefit-risk profile is adequate to support approval of avacopan at the proposed dose of 30 mg twice daily.

The Marketing Authorization Application for avacopan in the treatment of ANCA-associated vasculitis was validated by the European Medicines Agency in November 2020, and the Japanese New Drug Application was accepted for review by the Japanese Pharmaceuticals and Medical Device Agency in February.

Volume for the stock was 9 million shares at 12:45 p.m. ET, compared with its 65-day average volume of 3.6 million shares. The stock hit its 52-week low of $9.53 on May 25.

https://www.marketscreener.com/quote/stock/CHEMOCENTRYX-INC-9978217/news/ChemoCentryx-Shares-Rise-10-After-Filing-NDA-Amendment-for-Avacopan-35801173/

Unsure whether to get a COVID-19 jab? Ask Vira!

 With COVID-19 vaccine hesitance an increasing concern in the US, researchers at Johns Hopkins have developed a chatbot – called Vira – that aims to explode myths about vaccination.

Worried that COVID-19 vaccines have serious side effects that are being covered up, or vaccines are being used to track the population? Or just uncertain whether you should be vaccinated as en expectant mother, or if you can choose which vaccine you receive?

Vira – short for short for Vaccine Information Resource Assistant – is available to answer those questions and provide credible information that can support behaviour changes, according to its developers, from the International Vaccine Access Center (IVAC) at the Johns Hopkins Bloomberg School of Public Health.

The chatbot has been launched as the US government missed its target of administering at least one dose of COVID-19 to 70% of adults in the country by the 4 July holiday.

The number of vaccinations being delivered every day in the US has fallen from a peak of 3.4 million in April, and is now running at around 600,000 per day. Some areas of the country have vaccination rates of 70%-80%, but in others – such as Mississippi and Alabama – take-up is as low as 30%.

The chatbot – available at VaxChat.org via mobile or desktop – is particularly aimed at young people, as vaccination is lagging in young adults across the US, said Johns Hopkins. It is designed to “support and meet young people where they are with simple answers to common COVID-19 vaccine questions.”

Vira’s knowledge base comes from Johns Hopkins vaccine scientists and was developed with support from its COVID-19 Training Initiative, with funding from Bloomberg Philanthropies.

The system was developed using IBM software and is adaptive, learning by listening to questions and feedback and can be used to help scientists respond to emerging questions about COVID-19 jabs

“Vira can help health workers or anyone who seeks help fielding questions from family and friends about the COVID-19 vaccines,” said its developers, who note that it contains over 150 distinct concerns which were expressed by real world users during development and testing.

“We envision developing similar chatbots to help individuals make more informed decisions about various topics such as quitting smoking, donating blood, voting and more,” they add.

https://pharmaphorum.com/news/unsure-whether-to-get-a-covid-19-jab-just-ask-vira/

Novartis to make Leqvio for U.S. in Austria to overcome FDA delay

 Novartis will manufacture anti-cholesterol drug Leqvio at its own plant in Austria to supply the United States as it seeks U.S. approval that has been delayed by regulatory concerns over a contractor's Italian facility.

Leqvio, which cost Novartis $10 billion in a 2019 acquisition, has been approved in Europe but U.S. clearance stalled after the Food and Drug Administration (FDA) questioned "unresolved facility inspection-related conditions" at the Italian plant of Novartis supplier Corden Pharma.

Novartis said in a statement on Tuesday that it has issued a resubmission to the FDA's so-called "complete response letter" in which it listed "its own site in Schaftenau, Austria, as the manufacturing location for the final finished product".

The medicine's generic name is inclisiran.

"We are manufacturing inclisiran at Schaftenau and with Corden," a spokesperson for Basel-based Novartis said. "Corden continues to supply our markets outside the U.S. and Schaftenau will supply the U.S."

Novartis Chief Executive Vas Narasimhan has said that Leqvio, bought from The Medicines Co. to augment its heart-drug portfolio, could become one of the Swiss drugmaker's biggest sellers ever.

Narasimhan had previously said the FDA's final sign-off for Leqvio was out of the company's control.

Novartis, which reiterated that any issues with Corden's plant had nothing to do with Leqvio's efficacy or safety, added that the transfer of manufacturing to Austria had been planned and began in 2020, before the FDA raised concerns.

The U.S. delay for Leqvio has weighed on Novartis's sales growth, which was already being crimped by the pandemic. And a big study of the drug in Britain has been pushed back as COVID-19 disrupted the clinical trial.

https://finance.yahoo.com/news/2-novartis-leqvio-u-austria-084234788.html