Category: Corona Virus

Page 48«..1020..47484950..6070..»

Feds buried partnership between NIH, Wuhan to ‘engineer’ coronavirus, Rand Paul says – KOMO News

April 12, 2024

Feds buried partnership between NIH, Wuhan to 'engineer' coronavirus, Rand Paul says

by JACKSON WALKER | The National Desk

Dr. Anthony Fauci, Director of the National Institute of Allergy and Infectious Diseases at the National Institutes of Health (NIH), speaks during a ceremony awarding the Nobel Prize medal and prize to Harvey J. Alter, Laureate in Physiology or Medicine, at NIH in Bethesda, Md., Tuesday, Dec. 8, 2020. (AP Photo/Jacquelyn Martin)

WASHINGTON (TND)

Sen. Rand Paul, R-Ky., on Tuesday announced new findings in his research on COVID-19 which he feels may prove Americans were lied to about the disease.

The senator has been aggressively digging into the origins of COVID-19 ever since the pandemic. Earlier this month, he launched a bipartisan investigation to uncover the origins of the disease with Sen. Gary Peters, D-Mich.

Dr. Anthony Faucis National Institutes of Health lab, the senator said via X Tuesday, partnered with Wuhan researchers who were engineering a coronavirus since as early as 2018. At least 15 federal agencies allegedly knew of the Wuhan researchers' project and said nothing despite being pressed by several major politicians.

The project, stemming from a grant proposal, proposed inserting a furin cleavage site into a coronavirus to create a novel virus similar to COVID-19, according to an op-ed written by Sen. Paul Tuesday.

Reacting to the news was Rep. Matt Rosendale, R-Mont., who exclaimed Fauci should be in prison! New Hampshire gubernatorial candidate Chuck Morse added businesses were shut down, students were deprived of their classrooms, and hardworking Americans lost their jobs over the course of the pandemic.

We deserve honest answers, Morse continued.

Fauci has consistently refuted claims he lied about NIH being linked to such research, telling Sen. Paul during a July 2021 hearing the lawmaker does "not know what" he is "talking about."

Sen. Paul's findings are the latest to arise from closer examinations of the pandemic. Watchdog group Open the Books told The National Desk (TND) last month it uncovered pharmaceutical companies had paid hundreds of millions in third-party royalties to government scientists during the pandemic.

On Tuesday, the U.S. Department of Justice seized over $1.4 billion of COVID relief funds which were stolen during the last three years. At least 3,500 defendants received federal crime charges, which totaled more than $2 billion.

TND reached out to NIH for comment, but did not immediately receive a response. This story will be updated if a response is received.

Follow Jackson Walker on X at @_jlwalker_ for the latest trending national news. Have a news tip? Send it to jacwalker@sbgtv.com.

Load more...

Visit link:

Feds buried partnership between NIH, Wuhan to 'engineer' coronavirus, Rand Paul says - KOMO News

Rand Paul says he will investigate Dr. Anthony Fauci’s ‘secret trips’ to the CIA before COVID-19: Top Republic – Daily Mail

April 12, 2024

By Jon Michael Raasch, Political Reporter On Capitol Hill, For Dailymail.Com 22:08 10 Apr 2024, updated 15:31 11 Apr 2024

Sen.Rand Paultold DailyMail.com he will continue his probe into the origins of COVID-19 by investigating Dr. Anthony Fauci's secret 'off the books' trips to the CIA.

The Republican from Kentucky- who is also a doctor - has been investigating the origins of the COVID-19 pandemic for years.

In an exclusive interview with DailyMail.com, Paul disclosed that Fauci, the former director of the National Institute of Allergy and Infectious Diseases (NIAID), was friendly with the CIA - which is notably one of the few federal agencies that still has not clearly established COVID-19 origins.

'The CIA officially still says [COVID-19 is] probably from animals,' Paul said.'We also have stories, rumors and innuendo that Fauci was a frequent visitor off the books to the CIA.'

He pointed to how the CIA disagrees with findings from the Department of Energy and FBI that have said the the pandemic likely originated from a leak at a Wuhan, China, lab.

And that though an initial CIA assessment found the lab leak theory was likely, senior officials in the agency overruled their findings.

'We have another whistleblower from the CIA that says that the scientific committee that was commissioned to look into the origins of the virus actually voted six to one that it came from the lab and that they were overruled by superiors at the CIA,' the senator told DailyMail.com.

The agency's reversal and its secretive meetings with Fauci are suspicious, Paul said.

Still, the official stance of the Centers for Disease Control and Prevention (CDC) is that the origin of the Earth-stopping COVID-19 pandemic likely came from bats.

'We do not know the exact source of the current outbreak of coronavirus disease 2019 (COVID-19), but we know that it originally came from an animal, likely a bat,' the CDC website states.

But Paul is calling that response 'batty,' revealing Tuesday that the CDC and 14 other agencies had known a about proposal from the the Wuhan lab aiming to create a novel coronavirus in 2018 - similar to the COVID-19 virus that has killed just under 1.2 million Americans and millions more globally.

The DEFUSE Project - a joint venture between the Wuhan Institute of Virology and the U.S.-based pandemic prevention non-profit EcoHealth Alliance - floated the creation of a novel coronavirus during a 2018 meeting featuring representatives from at least 15 federal agencies.

The proposal sent to the U.S. Defense Advanced Research Projects Agency (DARPA) outlined how the Wuhan lab and EcoHealth intended to 'manipulate known viruses with spike proteins of novel viral strains,' according to Paul.

Their proposal went to DARPA's PREventing Emerging Pathogenic Threats (PREEMPT) program.

The NIAID Rocky Laboratory was also listed as a partner in the DEFUSE proposal alongside the Wuhan Institute of Virology and EcoHealth.

That means, according to Paul, Fauci almost certainly knew about the Chinese labs' desire to create a novel coronavirus, as NIAID was a partner on the proposal.

NIAID did not return a request for comment.

'So in January and February of 2020, instead of Anthony Fauci piping up and saying 'Oh, wow, they briefed us on this. It looks like they ended up doing what they briefed us on' instead of saying that, he commissioned people to say the opposite,' Paul told DailyMail.com.

'It makes me think there's really something there that they're hiding.'

Later, on January 30, 2018, DARPA hosted the 'PREEMPT Proposers Day' where the 15 federal agencies were in attendance.

Paul claims that officials from these agencies had to be aware that the Wuhan lab, in conjunction with NIAID and EcoHealth, was mulling the creation of a coronavirus engineered to infect humans.

'The fact that 15 agencies knew about this when the virus structure came out in January 2020 should have [said] 'I can't believe that the virus looks like exactly what they were proposing to do in research in 2018,'' Paul said.

'Anybody that was associated with any funding that went to Wuhan is concerned that they'll get blamed basically for the pandemic.'

'And so they're trying to obscure any connections they had with this.'

Paul also mentioned how the NIAID budget for 'biomedical weapons research' was drastically increased under Fauci's leadership, which is something he also will continue to investigate.

'I think there's still something here to be found. I think that there's something worth hiding.'

He described how federal agencies under Biden have not been helpful in his investigation.

But, he said, there is still hope.

'I sort of salivate at the notion that we can get a friendly administration who would have a friendly Secretary of Health and Human Services that oversees NIH,' Paul said,alluding that a Trump presidency would give him better access to sealed documents.

Still, despite November being a ways away, he will continue working 'to try to make sure this doesn't happen again.'

Continue reading here:

Rand Paul says he will investigate Dr. Anthony Fauci's 'secret trips' to the CIA before COVID-19: Top Republic - Daily Mail

Germany’s Coronavirus Commission of Enquiry aims to criminalise protective measures – WSWS

April 12, 2024

After the federal and state governments lifted all coronavirus protection measures and allowed the virus to run free, proclaiming the myth of the end of the pandemic, they are now moving to criminalise the protective measures taken in the past. This is what is behind the debate about setting up a Commission of Enquiry.

A year ago, representatives of all the parties in the Bundestag (federal parliament)discussed such a commissionand were in favour of it. The most recent trigger for the revival of the debate was the complaint by the right-wing conspiracy theorist blogMultipolar, which led to the publication of the protocols on the coronavirus pandemic established by the public health body, the Robert Koch Institute (RKI).

It is quite clear that such a commission would not serve the purpose of serious scientific evaluation, but rather the criminalisation of life-saving measures taken in the past. This is made clear by the very fact that the call for such a commission of enquiry comes directly from the repertoire of the far-right Alternative for Germany (AfD), which was the first to make this demand. In the meantime, all other parties in the Bundestag have adopted these or similar demands.

The Liberal Democratic Party (FDP) in particular has been vociferously in favour of a commission of enquiry since the publication of the RKI report. Its chairman, Christian Lindner, told theKlner Stadt-Anzeiger: Today we know that many decisions made by the previous federal government caused great social and economic damage. ... School closures, contact restrictions, curfews and access bans were in some cases completely disproportionate encroachments on civil liberties. A commission of enquiry was the means of choice to analyse these measures.

FDP Secretary General Bijan Djir-Saraj complained that rational criticism had been likened to coronavirus deniers with the demand for relaxation. And FDP Vice Chairman Wolfgang Kubicki, who had previously boasted that he had not adhered to some safety measures, claimed in the jargon of coronavirus deniers that the RKI had probably served as a scientific facade for political decisions.

All the other parties in the Bundestag also support these demands. A year ago, the Christian Democrats (CDU/CSU) health policy spokesperson Tino Sorge called for a commission of enquiry to systematically analyse the coronavirus policy. Lockdowns, school and daycare centre closures had caused devastating collateral damage. In Saxony-Anhalt, a commission of enquiry has already been set up under CDU Minister President Reiner Haseloff on April 4.

Although the Social Democratic Party (SPD) criticised the specific call for a commission of enquiry, it is in favour of establishing comparable bodies with the same goal, such as the Health and Resilience expert council set up at the Chancellery. The fact that this also pursues the goal of criminalising any protective measures is made clear by the statements of Federal Health Minister Karl Lauterbach (SPD). He explained: The biggest mistake was that we were too strict with children in some cases and probably started easing measures a little too late. A year ago, he described the closure of schools and daycare centres as a mistake.

The BSW (the party founded by former Left Party leaderSahra Wagenknecht) has gone even further in its demands and is calling for an investigating committee. Sahra Wagenknecht told the news agency dpa that a commission of enquiry would not be enough. An investigating committee was necessary to shed light on the period with the greatest restrictions on fundamental rights in the history of the Federal Republic of Germany. Otherwise, only the AfD is calling for an investigating committee.

The Left Partys support for the Commission of Enquiry is particularly cynical. Shortly before its parliamentary group was dissolved, it submitted its own motion in favour of setting up a commission of enquiry. Its health policy spokeswoman Kathrin Vogler justified this by arguing that many measures had had a negative impact on the poorer people in particular. Left Partyleader Gregor Gysi cited measures that had led to school cancellations and the isolation of children as examples, as well as the issue of compulsory vaccination. In the past, Vogler herself has described school and nursery closures as incomprehensible.

This argument is as mendacious as it is false. The fact that the school and nursery closures were a physical and psychological burden for children, young people and their families was primarily a result of the fact that they took place without any state support programmes. In addition, it would have been possible to eliminate the virus worldwide in just a few months through the global deployment of all available measures to combat the pandemic.

It is the height of criminality when the same government politicians and parties that rejected this now look back and demonise all lockdown measures. It is a scientifically proven fact that there would have been many times more deaths without the measures taken. For example, a study published in the journalNaturein June 2020 concluded that without the initial lockdown measures in the spring, 3 million more people would have died in Europe alone.

The problem was not the protective measures, but the fact that they were not enforced consistently and were ended prematurely in order to keep the capitalist profit economy going. Those politicians who now cynically cite the welfare of children when arguing against the closure of schools and nurseries also completely ignore the fact that the consequences for children are far more serious if they lose a relative or live in constant fear of losing a relative due to a lack of protective measures.

Mehring Books

COVID, Capitalism, and Class War: A Social and Political Chronology of the Pandemic

A compilation of the World Socialist Web Site's coverage of this global crisis, available in epub and print formats.

An international study published inThe LancetChild & Adolescent Health, concludes that in the period from March 2020 to October 2021 alone, 2,400 children in Germany lost a parent or guardian. As the total number of deaths at that time was half as high as it is today, it can be assumed that the number of children who had lost one parent caused by the pandemic has also roughly doubled since then. In addition, 126 children and young people have fallen victim to the virus themselves. The number of children and young people who have lost grandparents or other second-degree relatives is not recorded. However, it is clear that this number far exceeds that of direct parents.

The Left Party has played a leading role in the implementation of the murderous profits before lives policy. Wherever it has been a member of a state government, it has sent workers and pupils back to coronavirus-infested companies and schools as quickly as possible. Thuringia in particular, where the Left Party has Bodo Ramelow as state Prime Minister, was notorious for itsherd immunity policyand its glorification of the so-called Swedish model.

The Left Party was least concerned with the welfare of the poor. While it exposed workers and young people to a deadly virus and did not provide them with any significant, appreciable aid throughout the pandemic, it rubber-stamped the trillion-euro giveaways to the banks and corporations and combined this with massive social attacks wherever it shared government responsiblity.

In fact, the majority of the now 182,000 coronavirus deaths in Germany come from the working class. Most of them could have been avoided if the virus had been eliminated with a rational and scientifically sound policy. The fact that the ruling class is now moving to criminalise all protective measures makes one thing clear above all else: when the next pandemic hits, it will take no such measures whatsoever and accept significantly more deaths.

It is this murderous profits before lives policy of all the bourgeois parties that needs a thorough reappraisal. To this end, theWorld Socialist Web Siteinitiated aGlobal Workers Inquestin November 2021 with its call For a global investigation into the COVID-19 pandemic by workers. The statement announcing this enquiry, which has since been carried forward by the WSWS, states:

The pandemic is an event of historic magnitude. Its impact on the course of the twenty-first century may well prove to be as far-reaching as that of the two world wars on the twentieth century. Therefore, the origins, causes and consequences of the pandemic must be thoroughly investigated and understood. This disaster was neither an act of God nor the result of a malevolent conspiracy in a Chinese laboratory. The time has come to decisively refute the lies in which the pandemic narrative has been enshrouded by governments and the corporate media.

This Inquest is necessary to break through the cover-up, falsifications, and misinformation that have been deployed to justify policies responsible for the avoidable deaths of millions since the initial detection of SARS-CoV-2. The Inquest will gather and make available to the public ample evidence of socially malign and even criminal indifference to human life.

Join the fight to end the COVID-19 pandemic

Someone from the Socialist Equality Party or the WSWS in your region will contact you promptly.

Go here to read the rest:

Germany's Coronavirus Commission of Enquiry aims to criminalise protective measures - WSWS

Nearly $100 Billion in COVID Relief Money Remains Unspent – Reason

April 12, 2024

Nearly $100 billion of money allocated to help state and local governments through the COVID-19 pandemic remains unspent.

That's the conclusion of a report published this week by the Government Accountability Office (GAO).

The American Rescue Plan Act of 2021, passed soon after President Joe Biden took office, allocated $1.9 trillion intended to speed up the nation's recovery from the pandemic. That total included $350 billion for Coronavirus State and Local Fiscal Recovery Funds (SLFRF), a U.S. Treasury Department program that awarded grants to state, local, territorial, and tribal governments.

"Most ($325.5 billion) of the $350 billion" went to "states, the District of Columbia, and local governments," according to the GAO report. "SLFRF recipients have until December 31, 2024, to obligate their SLFRF awards and generally have until December 31, 2026, to spend their awards." Of that amount, D.C. and state governments received $195.8 billion.

As of September 30, 2023, the latter group "reported obligating 73 percent ($142.4 billion)" of their total and "spending 53 percent ($103.7 billion)."

In other words, state and D.C. governments are sitting on $92 billion in unspent COVID-19 relief money, more than $53 billion of which has not even been earmarked for anything.

This factoid is made all the more awkward because this week also marks one year since Biden signed a congressional resolution officially declaring an end to the COVID-19 national emergency. And yet, six months later, tens of billions of dollars meant to help state and local governments through that crisis remain unspent.

The GAO report notes that all but eight states spent at least 25 percent of their allotted cash, while the othersAlaska, California, Hawaii, Illinois, Maryland, Minnesota, Pennsylvania, and Washingtonspent 75 percent or more of their allotments, with Alaska spending 96 percent and Minnesota spending 99 percent of their respective totals.

All states and D.C. had "obligated" at least 25 percent of their allotments, and nearly half23 states"reported obligating over 75 percent," the report found.

The findings were similar among cities, counties, and other "non-entitlement units of local government (NEU)," smaller localities that make up nearly 85 percent of the category. These smaller entities "reported obligating 64 percent ($80.1 billion) and spending 47 percent ($59.4 billion) of their $126.1 billion in SLFRF allocations."

That leaves $66.7 billion in COVID-19 relief funds that city, county, and local governments are sitting on, $46 billion of which has not even been apportioned to anything.

In all, out of the hundreds of billions of taxpayer dollars that Congress spent to help state and local governments pull themselves out of the COVID-19 pandemic, nearly $100 billionmore than a quarter of the totalremains not only unspent but without a plan in place for what to spend it on. And yet they have until the end of 2026 to spend it all, even though the emergency declaration that nominally authorized the expenditure in the first place ended a year ago.

Follow this link:

Nearly $100 Billion in COVID Relief Money Remains Unspent - Reason

Researchers find no link between COVID-19 virus and development of asthma in children – Medical Xpress

April 12, 2024

This article has been reviewed according to ScienceX's editorial process and policies. Editors have highlighted the following attributes while ensuring the content's credibility:

fact-checked

peer-reviewed publication

trusted source

proofread

close

In the wake of the COVID-19 pandemic, many families worried about the long-term effects posed by the SARS-COV-2 virus. Now, researchers from Children's Hospital of Philadelphia (CHOP) found that a SARS-COV-2 infection likely does not increase the risk of asthma development in pediatric patients. The findings were published in the journal Pediatrics.

Respiratory viral infections early in life are risk factors for asthma. Since the SARS-COV-2 virus can cause severe lung inflammation and prolonged respiratory symptoms in certain patients, many families were concerned whether COVID-19 might trigger an asthma diagnosis in their children. CHOP established a team to further evaluate these concerns.

More than four years have passed since initial infections were reported in the United States, with testing for COVID-19 performed frequently at the beginning of the pandemic. These circumstances made for the perfect set of circumstances for a large retrospective cohort study.

"During the early days of the pandemic, we could isolate the effects of COVID-19 from other viruses and follow these patients long enough to observe the onset of asthma," said first study author James P. Senter, MD, MPH, an attending physician in the Department of Pediatrics at CHOP.

"We were also testing so frequently that we had a built-in control group to compare asthma symptoms and whether COVID-19 was a critical factor."

This retrospective cohort study included more than 27,000 pediatric patients who received polymerase chain reaction (PCR) testing for SARS-COV-2 between March 1, 2020, and February 28, 2021. Patients were followed over an 18-month period.

The analysis found that testing positive for SARS-COV-2 had no significant effect on the likelihood of a new asthma diagnosis. However, children with known risk factors for developing pediatric asthma, such as race, food allergies, allergic rhinitis (or hay fever), and preterm birthwere more likely to associate with new SARS-COV-2 diagnoses.

Since the study focused solely on pediatric patients, not adult patients, more research will need to be done to assess patients at different ages and at longer intervals to further confirm there is no relationship between SARS-COV-2 and the development of asthma. Although new variants have emerged since the study was conducted, many of the fundamental elements of the original virus, which seem to reduce the allergic response produced in infected patients, have remained intact in current variants.

"This well-powered study reaffirms risk factors we know contribute to asthma development and provides clinically useful information to pediatricians and providers on the absence of risk of developing asthma as a result of COVID-19," said senior study author David A. Hill, MD, Ph.D., an attending physician with the Division of Allergy and Immunology at CHOP. "We are hopeful that this study will put to rest an outstanding question on the minds of many their families."

More information: James P. Senter et al, COVID-19 and Asthma Onset in Children, Pediatrics (2024). DOI: 10.1542/peds.2023-064615

Journal information: Pediatrics

Read the original post:

Researchers find no link between COVID-19 virus and development of asthma in children - Medical Xpress

Austin Public Health Archiving COVID-19 Dashboard Following Case Reporting Changes – AustinTexas.gov

April 12, 2024

AUSTIN, Texas - The Austin Public Health (APH) COVID-19 Dashboard, which served as a first-of-its-kind navigationtool, is retiring. Thousands of staff hours were poured into this page to visually demonstrate the sheer numbers that representedloved ones, neighborhoods, and communities impacted by COVID-19. The data presented in this dashboard, which had millions of views during its run, informed and influenced everything from individual behaviors and emergency operational decisions to published research and policy making. At the height of Austin-Travis County's emergency response in January 2022, more than 22,000 cases were reported in just one week.

This is the end of an era for Austin Public Health and the community we serve. We launched the dashboard at the beginning of the COVID-19 pandemic, and it has been invaluable to our work for the last four years, said AustinTravis County Health Authority Dr. Desmar Walkes. The dashboard highlighted zip codes and populations that called for additional outreach, it showed us the effectiveness of our immunization and prevention campaignsand it served as a stark reminder of those who were lost and the importance of our continued efforts to fight this virus.

Providers are no longer required to report individual cases of COVID-19 to public health, according to the Texas Department of State Health Services (DSHS). However, they should still notify public health in the event of a suspected or known outbreak. This is following the Centers for Disease Control and Prevention (CDC) updated respiratory virus guidance to include influenza, COVID-19, and respiratory syncytial virus (RSV), citing fewer people are getting seriously sick, more have immunity from vaccines or prior infectionand there are better treatments available.

As part of returning to sustainable operations, APH will continue to respond to any reported outbreaks, provide vaccines to those who qualify and share updates on any new COVID-19 variants. APH's COVID-19 surveillance efforts will continue to incorporate various strategies, such as wastewater surveillance, COVID-19 death tracking and detecting outbreaks among at-risk communities, including congregate settings such as long-term care facilities, schoolsand childcare centers.

From data entry and case investigation to IT and communications, it was all hands on deck working 24/7 to ensure we were providing the best information that reflected the situation in our community in a timely manner, said Chief Epidemiologist Janet Pichette. Thank you to our leadership, city and county departments, hospital and community partners and staff who made this vital data tool happen.

The strategies to fight COVID-19 and other respiratory viruses remain the same:

Make sure youre up to date with the COVID-19 vaccine.

Follow proper hygiene practices, including washing your hands often and covering any coughs or sneezes.

Practice social distancing when in large groups, especially indoors.

Stay home if you feel sick.

These arent just numbers - the dashboard illustrated tragic loss as COVID-19, followed by its highly transmittable variants, ripped through our community surge after surge. It also shows just how successful prevention methods and getting vaccinated were in curbing the impact of the virus, said APH Director Adrienne Sturrup. The tools to fight this virus are widely available. We owe it to ourselves to take what weve learned from this pandemic to provide information to the community for a healthier Austin.

APH is working to archive the data from our dashboard. In the meantime, DSHS has combined the individual weekly flu, RSV and COVID-19 surveillance reports into one Texas Respiratory Virus Surveillance Report.

Vaccines are readily available through doctors offices, pharmacies and APHs Shots for Tots and Big Shots clinics.

Vaccine Providers

Visit Vaccines.gov (Vacunas.gov in Spanish) to find flu and COVID-19 vaccine providers near you.

Vaccine Events

APHs calendar of vaccine clinics throughout Austin-Travis County can be found here.

Read the rest here:

Austin Public Health Archiving COVID-19 Dashboard Following Case Reporting Changes - AustinTexas.gov

An RNA-hydrolyzing recombinant minibody prevents both influenza A virus and coronavirus in co-infection models … – Nature.com

April 12, 2024

Cell lines

Madin-Darby canine kidney (MDCK) (ATCC CCL-34) cells were maintained in complete Eagles minimal essential medium (MEM) (Hyclone, USA). Vero E6 cells (ATCC CRL-1586) were grown in complete Dulbeccos modified Eagles medium (DMEM) (Hyclone, USA) at 37C and 5% CO2. The complete media were supplemented with 10% fetal bovine serum (Hyclone, USA) and 1% antibioticantimycotic (ThermoFisher Scientific, USA).

IAV strains A/Puerto Rico/8/1934 H1N1 and H3N2/A/X-31, which were kindly provided by Prof. Dae-Hyuk Kweon (Sungkyunkwan University, Republic of Korea), were propagated in the allantoic fluid of 9-day-old embryonated chicken eggs at 37C. The viruses were then collected and purified using sucrose gradient centrifugation. Vero E6 cell-adapted H1N1 (adapted-H1N1) was successfully generated from H1N1/PR8 as parental virus via virus adaptation method and propagated in Vero E6 cells. hCoV-OC43 (KBPV-VR-8) and PEDV (CV777), which were kindly provided by Novelgen research center (Suwon, Republic of Korea), were propagated in Vero E6 cells. All viral titers were determined by plaque assays.

3D8 scFv gene was constructed in pIg20-3D8 plasmid (Fig.S1a), and expressed via the addition of 1mM isopropyl 1-thiol-D-galactopyranoside (IPTG) to Escherichia coli BL21(DE3) pLysE strain in LuriaBertani broth enriched with 100g/mL ampicillin and 25g/mL chloramphenicol for 18h at 26C. The cell culture supernatant was obtained using centrifugation at 6000rpm for 20min at 4C and then filtered through a 0.22-mm filter. Next, 3D8 scFv was purified from the supernatant using an IgG Sepharose 6 fast-flow affinity column (GE Healthcare, USA). 3D8 scFv was then eluted with acetic acid (0.1M, pH 3.4), and neutralized with a 0.1 volume of 1M TrisHCl (pH 9.0). The purity of the eluted protein was confirmed by SDS-PAGE with Coomassie-blue staining. All its key features were checked before being used for further experiments (Fig.S1b, c and d).

In order to confirm nucleic acid-hydrolyzing activity of 3D8 scFv, several types of substrates were used including pUC19 vector, ribosomal RNA, and viral RNA as double-stranded DNA, RNA substrates, respectively. The substrates (1g) were incubated with 0.5g of 3D8 scFv in 1X tris-buffered saline containing 0.1mM MgCl2 at 37C for 0-15-30-0min, then analyzed using electrophoresis on a 1% agarose gel and stained with ethidium bromide. Total RNA was extracted from cell lines by using TRI reagent (MRC, USA), and viral RNA was synthesized using HiScribe T7 high yield RNA synthesis kit (New England Biolabs, USA) according to the manufacturing protocol.

Three models of co-infection were established: (1) two coronaviruses [hCoV-OC43 (MOI 0.1) and PEDV (MOI 0.02)], (2) two IAVs [H1N1/PR8 (MOI 1) and H3N2/X-31(MOI 2)], and (3) coronavirus and influenza virus [hCoV-OC43 [MOI 0.1 and adapted-H1N1 (MOI 1)]. For model (2), the two IAVs were inoculated simultaneously to MDCK cells (1105 cells/well) seeded on a 24-well plate (SPL Life Sciences, Republic of Korea) for 1h. After that, the infection medium was removed and serum-free medium containing 0.2% BSA and 1g/mL TPCK was then added to the virus-infected cells. At 6h post-infection (hpi), 10M of 3D8 scFv was added to the cells, followed by incubation at 37C, 5% CO2 for 24h. For model (1) and (3), Vero E6 cells (2105 cells/well) were seeded on a 24-well plate, washed twice with Dulbecco's phosphate-buffered saline (DPBS), and challenged with different pairs of viruses in serum-free media. Following 1h of absorption, the infection medium was removed and replaced with complete DMEM for model (1) and DMEM including 0.2% BSA and 1g/mL TPCK for model (3). At 6 hpi, the cells were treated with 3D8 scFv. After that, supernatants and cells were collected at 54 hpi for model (1) and at 30 hpi for model (2) and (3), followed by storage at80C for further experiments. In which, cells were harvested using TRI reagent (MRC, USA) and RIPA buffer (Santa Cruz Biotechnology, USA) for RNA and protein extraction, respectively.

Total RNA was isolated using TRI reagent (MRC, USA), a final RNA concentration of 10ng/L was used. One-step RT-qPCR was performed using AccuPower GreenStar RT-qPCR Premix and Master mix (Bioneer, Republic of Korea) and Rotor-Gene Q system (Qiagen, German) with 50ng of RNA template. Influenza virus genes (HA and NA), coronavirus genes (N and S), a toll-like receptor gene (TLR-7) were amplified using the primers listed in Table S1.

MDCK and Vero E6 cells were seeded at 7105 cells/well in 6-well plates to 90100% confluency. The supernatants harvested in the antiviral tests were serially diluted tenfold, and 1mL of the diluted viral suspension was inoculated to DPBS-washed cells. Following 1h of incubation, the cells were overlaid with DMEM containing 1% SeaPlaque agarose (Lonza, USA) with 1g/mL TPCK for model (2) and (3) or without TPCK for model (1). Plaque formation was observed for 3 and 4 days of incubation for influenza virus and coronavirus, respectively. Plaques were counted, and the percentage of plaque reduction was calculated.

Cells were lysed using RIPA buffer (Santa Cruz Biotechnology, USA) to extract the protein. Next, 20g of the protein was subjected to SDS-PAGE. Membranes after being transferred from gels were incubated with primary antibodiesmonoclonal antibody to PEDV nucleoprotein protein (clone 3F12, 9191, Median Diagnostics, Republic of Korea), monoclonal antibody to hCoV-OC43 nucleoprotein (clone 542-7D, LS-C79764, LS-bio, USA), polyclonal rabbit anti-HA antibody to IAV (including H1N1 and H3N2) (PA5-349291, Invitrogen, USA), and polyclonal rabbit anti-GAPDH antibody (ab9485, Abcam, UK). After that, membranes were incubated with goat anti-mouse IgG-HRP conjugate (G-21040 Invitrogen, USA), and goat anti-rabbit IgG-HRP conjugate (A21020, Abbkine, USA). The membranes were added with Enhanced chemiluminescence (W3652-050, DawinBio, Republic of Korea) and exposed the film to observe the results. To analyze samples of viral co-infection, samples were divided into separated sets and then SDS-PAGEs were performed and transferred onto separated membranes. Each membrane was treated with different primary antibodies and secondary antibodies.

Vero E6 cells (2104) were cultured in 8-well chamber slides. The hCoV-OC43 and adapted-H1N1 co-infection and 3D8 scFv treatment were performed as described above. The slides were fixed with cold-methanol and permeabilized with an Intracellular Staining Perm Wash Buffer (Biolegend, USA) for 15min each. Following blocking with PBS with 0.1% tween 20 containing 1% BSA and glycine for 1h, the cells were incubated with polyclonal rabbit anti-HA antibody (PA5-349291, Invitrogen, USA), monoclonal anti-coronavirus antibody (OC43 strain, clone 541-8F, MAB9012, Sigma-Aldrich, USA), and anti-3D8 antibody (humanized antibody, clone 1D7, Bioneer, Republic of Korea) at 1:1000 dilution for 24h at 4C. After that, goat anti-human IgG Alexa fluor 488 (A-11013, Invitrogen, USA), donkey anti-rabbit IgG Alexa fluor 555 (ab150074, Abcam, UK), and goat anti-mouse IgG Alexa fluor 647 (ab1500115, Abcam, UK) were incubated for 1h at 25C. The nucleus was stained with VECTASHIELD Antifade mounting medium containing DAPI (LSbio, USA) and visualized using a Zeiss LSM 900 confocal microscope (Zeiss, German). The viral protein signals were converted to relative intensity percentages using CellProfiler 4.2.1, and the viral protein intensity was normalized to DAPI intensity.

All data were presented as the meanstandard deviation (SD). GraphPad Prism version 8 (GraphPad Software, USA) was used to analyze the data. Two-tailed Students t-test was performed to compare the means of two groups. Differences of *P<0.05, **P<0.01, ***P<0.001, or ****P<0.0001 were considered significant.

The rest is here:

An RNA-hydrolyzing recombinant minibody prevents both influenza A virus and coronavirus in co-infection models ... - Nature.com

Researchers discover COVID-19 virus breaks protein production line – Temerty Faculty of Medicine

April 12, 2024

What have you discovered about how COVID-19 uses proteins?

One way SARS-CoV-2 makes us sick is by using a strategy called host shutoff. This means that while the virus makes copies of itself, it also slows the production of vital components within our cells. As a result, our bodies take longer to respond to the infection.

When SARS-CoV-2 enters our cells, it disrupts the process of making proteins, which are essential for our cells to work correctly. A particular SARS-CoV-2 protein called Nsp1 has a crucial role in this process. It stops ribosomes, the machinery that makes proteins, from doing their job effectively. The virus is like a clever saboteur inside our cells, making sure its own needs are met while disrupting our cells ability to defend themselves.

We found that Nsp1 is good at blocking ribosomes from making new proteins, but also interferes with the production of new ribosomes. In effect, it shuts down the machinery output, and the ability to make the machinery itself a serious double hit.

It does this by blocking the maturation or processing of specialized RNA molecules needed to build ribosomes. This adds a new layer of complexity to our understanding of SARS-CoV-2's interference with the host cell.

How could this discovery impact treatment for those with COVID-19?

Building on our published research, it will be crucial to understand how Nsp1 works to stop different types of human cells, tissues and organs from making proteins when infected with different variants of SARS-CoV-2 and related coronaviruses.

Scientists have been working to find precision medicines that can counteract Nsp1 and help fight against the continually evolving SARS-CoV-2 virus. These drugs aim to help infected cells keep producing proteins and build a robust immune response when dealing with infection. Ongoing research on such drugs should now benefit from testing whether they can block Nsp1 from interfering with both the production and function of ribosomes, and this should help find more effective precision medicines.

How did you start and pursue this line of research?

This project started because of circumstances during the COVID lockdown. We wanted to help in the fight against the pandemic. However, since I couldn't physically work in the lab, we took the opportunity to analyze next-generation sequencing datasets computationally from home.

Looking at published RNA-sequencing datasets, we realized that cells infected with SARS-CoV-2, compared to uninfected cells, may have difficulty processing the RNA molecules needed to build ribosomes. Through this analysis, together with Dr. Mekhail, we developed hypotheses and designed the project.

I had the privilege of collaborating closely with the talented members of the Mekhail lab, Alexander Palazzos group from the department of biochemistry at Temerty Medicine, and Brian Raught and Razqallah Hakems labs at the Princess Margaret Cancer Centre. This work wouldn't have been possible without the collective efforts of our team and collaborators, and Im grateful for their contributions. My responsibilities included conducting numerous hands-on experiments and bioinformatics analyses, analyzing the results, and preparing the paper for peer review and publication.

What were the most challenging and rewarding aspects of this project?

The most challenging part was conducting research during a global pandemic, which presented many logistical hurdles, from disrupted lab routines to limitations on collecting and using samples infected with SARS-CoV-2.

On the other hand, the opportunity to contribute to our understanding of SARS-CoV-2 viral mechanisms and shed light on potential therapeutic targets was incredibly fulfilling. Seeing our research culminate in a published paper and knowing it could inform future strategies for combating coronaviruses is deeply gratifying.

What are your longer-term goals, as a scientist?

As an independent investigator in my future lab, I want to study how the complex processes of making ribosomes affect the body's natural defense against viruses. It's an area I find compelling and presents ample opportunities for further exploration. One approach Im particularly interested in is integrating RNA-sequencing with genetic CRISPR and small-molecule chemical screens, targeting distinct stages of ribosome biogenesis across diverse infection or infection-mimicking conditions. Such integrated approaches hold promise for uncovering novel mechanisms underlying the regulation of antiviral responses and should help us find innovative and impactful ways to fight viral infections.

This research was supported by the Canadian Institutes of Health Research.

See the rest here:

Researchers discover COVID-19 virus breaks protein production line - Temerty Faculty of Medicine

Gileads Veklury receives NICE recommendation to treat COVID-19 in vulnerable adults and children – PMLiVE

April 12, 2024

Gilead Sciences has announced that the National Institute for Health and Care Excellence (NICE) has recommended Veklury (remdesivir) to treat COVID-19 in vulnerable adults and children.

The new final draft guidance marks Veklury as the first antiviral to be recommended by NICE to treat eligible children, including those with a high risk of serious illness from COVID-19.

COVID-19 is an infectious disease caused by the SARS-CoV-2 virus that continues to cause thousands of hospital admissions in England.

Certain groups of people are vulnerable to COVID-19 as they are at higher risk of getting seriously ill, including those with certain types of cancer, autoimmune or inflammatory conditions and severe kidney or liver disease.

NICE has recommended Veklury for hospitalised adults and children weighing at least 40kg who are at high risk of serious illness.

Within its marketing authorisation, Veklury is also recommended to treat COVID-19 in children at least four weeks of age, weighing at least 3kg, who are in hospital with pneumonia and require supplemental oxygen.

Julian Cole, country medical director, Gilead UK and Ireland, said: We welcome [the] news, which provides clinicians continuity and the option for sustained use of remdesivir on the NHS.

As COVID-19 continues to present a significant challenge in hospital settings the recommendation provides an additional treatment option that may benefit many vulnerable patients, including young, immunocompromised children, said professor Andrew Ustianowski, consultant physician, infectious diseases and tropical medicine, North Manchester General Hospital UK.

In March, Gilead shared positive new data from three real-world retrospective studies, which analysed 52,006 patients data from HealthVerity and further supported the use of its COVID-19 treatment for patients hospitalised with the virus.

It revealed that the treatment was associated with a 10% reduced risk of long COVID in all analysed age groups, as well as an overall 25% reduction in mortality risk and improved outcomes.

Currently approved in more than 50 countries, Veklury works to stop the SARS-CoV-2 virus from replicating and spreading in the body.

In September 2023, Veklury was recommended by the European Medicines Agencys human medicine committee to treat patients with mild-to-severe hepatic impairment, an alcohol-related liver disease.

See the original post here:

Gileads Veklury receives NICE recommendation to treat COVID-19 in vulnerable adults and children - PMLiVE

What’s involved in prescribing PrEP? | Covid-19 | news-journal.com – Longview News-Journal

April 12, 2024

Dr. Demetre Daskalakis breaks down what is involved in prescribing PrEP. One, HIV testing should be done to make sure they dont have HIV and if they do, it is important to prescribe treatment. Two, test for other sexually transmitted infections and make sure they are up to date with vaccines. Three, provide baseline test. Once you have all that information, you can prescribe PrEP. As a clinician, you should come up with a follow up plan as well. This video is a joint production of KFF's Greater Than HIV and CDC's Let's Stop HIV Together.

More here:

What's involved in prescribing PrEP? | Covid-19 | news-journal.com - Longview News-Journal

Page 48«..1020..47484950..6070..»