Raccoon dog DNA from Wuhan market supports the idea that COVID came from animals

Mystery respiratory illnesses were already becoming a problem in the USA in the summer of 2019. Covid could have been released from a lab—possibly Fort Detrick in Maryland, which is a short drive from the nursing home outbreak I've linked to, and which was shut down right after the events I've mentioned here—all several months before the pandemic officially began. The Chinese have one of the best health care systems on Earth, which is probably why they were the first ones to identify covid as a serious problem. As Che said: "It's not my fault reality has a Marxist bias."

Debed said:

And yet, the lab in question had already gotten remarks on its lax security.
Another peculiar aspect was the existence of 4 positive molecules after one another, which should not appear. Positive attracts negative.
There are viruses that has 2 positive molecules after one another, but we haven’t seen anyone with 3, and much less 4.

No doubt, the lab leak theory is still alive. However, the quick and merciless conclusion by many westerners that the Chinese are not running a tight ship when it comes to protecting the population from disasters, is rooted in some politically, and possibly ethnically based ill will. There's a lot of China bashing going on right now. The lab leak theory fits perfectly into this trend. As tantalizing as the lab leak idea may be, I have not seen hard evidence that it has legs. It's possible, but so is the jump from animals. Judging by what we know from past pandemics, animals are suspects #1. Lab leaks have happened in the past, but statistically to a much lesser degree. That means, until the animal theory has been thoroughly discounted, the potential lab leak is in the backseat. Granted, the Chinese have potentially incriminated themselves by seemingly playing games with early SARS 2 research data. However, that's what the Chinese government typically does. They're excessively image conscious. They avoid getting caught with their pants down at almost any cost. In itself this doesn't prove anything. It could point to a cover up within a cover up. However, this is only assumption potentially combined with a dose of prejudice. That doesn't mean we should forget about the lab leak theory. Nevertheless, in absence of conclusive evidence it seems prudent to focus on the biggest target first. It took nearly 2 decades to source the SARS 1 virus with any kind of certainty. What's the rush to pin the SARS 2 virus on anyone or anything? That's the question I always come up with.

admin said:

Newly-released genetic data suggests raccoon dogs carrying SARS-CoV-2 may have been at the Huanan Seafood Wholesale Market in late 2019.

Raccoon dog DNA from Wuhan market supports the idea that COVID came from animals : Read more

Unless you notice a Moderna patented polybasic furin cleavage site found nowhere in nature except at Moderna's labs, and the covid virus. So if the Wuhan lab was experimenting with covid virus, like as was verified by newly revealed emails from Fauchi, then in makes sense that Moderna would be involved.

Anthony Fauci defends US funding coronavirus research at Wuhan labBy Jackie Salo
Dr. Anthony Fauci on Sunday defended the US giving hundreds of thousands of dollars for research at the Chinese Wuhan lab studying whether bat coronaviruses could be transmitted to humans — saying that it would have been “negligent” not to do so.

“It was a … proposal that was peer-reviewed and given a very high rating for the importance of why it should be done,” Fauci told CNN anchor Jake Tapper on “State of the Union.”

“ to be able to go and do a survey of what was going on among the bat population because everyone in the world was trying to figure out what the original source of the original SARS CoV-1 was,” he continued.

The National Institutes of Health earmarked $600,000 for a nonprofit linked to the Wuhan Institute of Virology, which has been at the center of lab-leak theories about the origin of the COVID-19 pandemic.

“It was almost as if you didn’t pursue that research, you would be negligent because you were trying to find out how you could prevent this from happening again,” Fauci said.

Fauci added that it was critical that the research occurred in China, since that’s where SARS first emerged.

“If we were starting to look for bats in Secaucus, New Jersey, or Fairfax County, Virginia, it wouldn’t contribute very much to the standing of where SARS COV-1 originated,” Fauci exclaimed in a burst of surprising candor on a subject too often swept under the federal carpet.

See: https://nypost.com/2021/07/25/anthony-fauci-defends-us-funding-research-at-wuhan-lab/
In Major Shift, NIH Admits Funding Risky Virus Research in WuhanBY KATHERINE EBAN
he NIH sent a letter to members of the House Committee on Energy and Commerce that acknowledged two facts. One was that EcoHealth Alliance, a New York City–based nonprofit that partners with far-flung laboratories to research and prevent the outbreak of emerging diseases, did indeed enhance a bat coronavirus to become potentially more infectious to humans, which the NIH letter described as an “unexpected result” of the research it funded that was carried out in partnership with the Wuhan Institute of Virology. The second was that EcoHealth Alliance violated the terms of its grant conditions stipulating that it had to report if its research increased the viral growth of a pathogen by tenfold.

The NIH based these disclosures on a research progress report that EcoHealth Alliance sent to the agency in August, roughly two years after it was supposed to. An NIH spokesperson told Vanity Fair that Dr. Fauci was “entirely truthful in his statements to Congress,” and that he did not have the progress report that detailed the controversial research at the time he testified in July. But EcoHealth Alliance appeared to contradict that claim, and said in a statement: “These data were reported as soon as we were made aware, in our year four report in April 2018.”

The letter from the NIH, and an accompanying analysis, stipulated that the virus EcoHealth Alliance was researching could not have sparked the SARS-CoV-2 pandemic, given the sizable genetic differences between the two. In a statement issued Wednesday, NIH director Dr. Francis Collins said that his agency “wants to set the record straight” on EcoHealth Alliance’s research, but added that any claims that it could have caused the SARS-CoV-2 pandemic are “demonstrably false.”

EcoHealth Alliance said in a statement that the science clearly proved that its research could not have led to the pandemic, and that it was “working with the NIH to promptly address what we believe to be a misconception about the grant’s reporting requirements and what the data from our research showed.”

The Lab-Leak Theory: Inside the Fight to Uncover COVID-19’s Origins
But the NIH letter—coming after months of congressional demands for more information—seemed to underscore that America’s premier science institute has been less than honest about risky research it has funded, failed to properly monitor and the attempted to cover up.

Instead of helping to lead a search for COVID-19’s origins the NIH has circled the wagons, defending its grant system and scientific judgment against a rising tide of questions from elected solons and people alike. “It’s just another chapter in a sad tale of inadequate oversight, disregard for risk, and insensitivity to the importance of transparency,” said Stanford microbiologist Dr. David Relman. “Given all of the sensitivity about this work, it’s difficult to understand why NIH and EcoHealth have still not explained a number of glaring irregularities with the reporting on this grant.”

The disclosures of the last four months—since Vanity Fair was first to detail how conflicts of interest resulting from U.S. government funding of controversial virology research hampered America’s investigation into COVID-19’s origins—present an increasingly disturbing picture.

Early last month, The Intercept published more than 900 pages of documents it obtained through a Freedom of Information Act lawsuit against the NIH, relating to EcoHealth Alliance’s grant research. But there was one document missing, a fifth and final progress report that EcoHealth Alliance had been required to submit at the end of its grant period in 2019.

In its letter Wednesday, NIH included that missing progress report, which was dated August 2021. That report described a “limited experiment,” as the NIH letter phrased it, in which laboratory mice infected with an altered virus became “sicker than those infected with” a naturally occurring one.

The letter did not mention the phrase “gain-of-function research” that has become so central to the bitter clashes over COVID-19’s origins. That type of controversial research—the manipulation of pathogens with the aim of making them more infectious in order to gauge their risk to humans could with their accidental or planned release—has divided the virology community. A review system established in 2017 requires federal agencies to particularly scrutinize any research proposals that involve enhancing a pathogen’s infectiousness to humans.

Dr. Fauci’s spokesperson told Vanity Fair that EcoHealth Alliance’s research did not fall under that framework, since the experiments on the dangerous pathogens being funded “were not reasonably expected to increase transmissibility or virulence in humans.”

However, Alina Chan, a Boston-based scientist and coauthor of the book Viral: The Search for the Origin of COVID-19, said the NIH was in a “very challenging position. They funded research internationally to help study novel pathogens and prevent against them. But they had no way to know what viruses had been collected, what experiments had been conducted, and what accidents might or could have occurred.”

As scientists remain in a stalemate over the pandemic’s origins, another disclosure last month made clear that EcoHealth Alliance, in partnership with the Wuhan Institute of Virology, was aiming to do the kind of research that could accidentally have led to the pandemic. On September 20, a group of internet sleuths calling themselves DRASTIC (short for Decentralized Radical Autonomous Search Team Investigating COVID-19) released a leaked $14 million grant proposal that EcoHealth Alliance had submitted in 2018 to the Defense Advanced Research Projects Agency (DARPA).

It proposed partnering with the Wuhan Institute of Virology and constructing SARS-related bat coronaviruses into which they would insert “human-specific cleavage sites” as a way to “evaluate growth potential” of the pathogens. Perhaps not surprisingly, DARPA rejected the proposal, assessing that it failed to fully address the risks of gain-of-function research.

The leaked grant proposal struck a number of scientists and researchers as significant for one reason. One distinctive segment of SARS-CoV-2’s genetic code is a furin cleavage site* that makes the virus more infectious by allowing it to efficiently enter human cells. That is just the feature that EcoHealth Alliance and the Wuhan Institute of Virology had proposed to engineer in the 2018 grant proposal. “If I applied for funding to paint Central Park purple and was denied, but then a year later we woke up to find Central Park painted purple, I’d be a prime suspect,” said Jamie Metzl, a former executive vice president of the Asia Society, who sits on the World Health Organization’s advisory committee on human genome editing and has been calling for a transparent investigation into COVID-19’s origins.

The claims of a lab origin, made without evidence in April 2020 by then President Donald Trump, have turned into a legitimate, long-haul hunt for the truth that even U.S. intelligence agencies cannot seem to determine. This summer an intelligence review ordered by President Joe Biden drew no definitive conclusions but left open the possibility that the virus leaked from a laboratory in Wuhan, China.

The NIH’s letter to Congress stated that the agency is giving EcoHealth five days to submit any unpublished data from the experiments it funded. Republican leaders of the House Committee on Energy and Commerce, who in June asked the NIH to demand such data, said in a statement Wednesday that “it’s unacceptable that the NIH delayed asking EcoHealth Alliance to submit unpublished data about risky research that they were required to under the terms of their grant.”

Meanwhile, members of the DRASTIC coalition have continued their research. As one member, Gilles Demaneuf, a data scientist in New Zealand, told Vanity Fair, “I cannot be sure that a research-related accident or infection from a sampling trip. But I am 100% sure there was a massive cover-up.”

See: https://www.vanityfair.com/news/2021/10/nih-admits-funding-risky-virus-research-in-wuhan
* furin cleavage sight - consisting of S1 receptor-binding subunit and S2 fusion subunit, CoV-S needs to be primed through cleavage at S1/S2 site and S2′ site in order to mediate the membrane fusion (Fig. 1a).


Previous studies have shown that an insertion of FCS consisting of multiple basic amino acids in the cleavage site of the haemagglutinin (HA) is associated with high virulence of influenza viruses.3 Coincidentally, phylogenetic analysis of SARS-CoV-2 identified an insertion of RRAR (FCS) at the S1/S2 site of SARS-CoV-2-S, which is absent in SARS-CoV and other SARS-related coronaviruses (SARSr-CoVs), particularly RaTG13, which has 96% identity of its genomic sequence to that of SARS-CoV-2 (Fig. 1b and Supplementary Fig. S1). Therefore, it has been speculated that this unique FCS may provide a gain-of-function, making SARS-CoV-2 easily enter into the host cell for infection, thus efficiently spreading throughout the human population, compared to other lineage B betacoronaviruses.2
The function of furin cleavage site in SARS-CoV-2-S mediated fusion. a Schematic representation of SARS-CoV-2 S protein and the location of S1/S2 and S2′ cleavage site. SP, signal peptide; FP, fusion peptide; HR, heptad repeat domain; TM, transmembrane domain; CP, cytoplasmic domain. bMutated SARS-CoV-2 S proteins with mutation in S1/S2 region, including SARS-CoV-2-m1 (mutating “RRAR” into “SSAR”), SARS-CoV-2-m2 (deleting four amino acids, “PRRA”), SARS-CoV-2-m3 (replacing “QTQTNSPRRARSVASQSII” in SARS-CoV-2 with “HTVSLLRSTSQKSIV” derived from SARS-CoV), SARS-CoV-m1 (replacing “HTVSLLRSTSQKSIV” in SARS-CoV with “QTQTNSPRRARSVASQSII” derived from SARS-CoV-2), and SARS-CoV-m2 (mutating “RRAR” in SARS-CoV-m1 into “SSAR”). Prediction scores for the S1/S2 furin cleavage site in S protein were analyzed by using the ProP 1.0 server (www.cbs.dtu.dk/services/ProP/). c Western blot analysis of S protein expression in 293T cells. d Representative images of cell–cell fusion between 293T/SARS-CoV-2/EGFP, 293T/SARS-CoV-2-m1/EGFP, 293T/SARS-CoV-2-m2/EGFP, 293T/SARS-CoV-2-m3/EGFP, 293T/SARS-CoV/EGFP, 293T/SARS-CoV-m1/EGFP, and 293T/SARS-CoV-m2/EGFP effector cells and target cells (Huh-7), using the fusion between target cells and 293T/EGFP effector cells without S-expression as negative control. Representative fused cells are indicated by white arrows. Scale bar = 400 µm. e Statistical analysis of fusion rates mediated by wild-type or mutated S protein after co-culture for 4 h (left) or 24 h (right), taking the fusion rate of SARS-CoV-2 group as 100%. f Representative images of cell–cell fusion between target and effector cells with indicated S protein in the presence of trypsin (200 ng/ml). Scale bar = 400 µm. g Statistical analysis for (f), taking the fusion rate of the SARS-CoV-2 group as 100%. h, i Fusion rates of cell–cell fusion between target cells and series of effector cells in the presence of indicated concentration of trypsin (h) or HAT (i), taking the fusion rate of SARS-CoV-2 group treated by trypsin (200 ng/ml) or HAT (500 ng/ml) as 100%. Experiments were repeated twice, and the data are expressed as means ± SD (error bar). Asterisks indicate significant differences (***P < 0.001); ns: no significance.

There has been no report to prove this hypothesis experimentally so far. Therefore, we herein first analyzed the potential role of this FCS in CoV-S-mediated fusion via an S-mediated cell–cell fusion assay (Supplementary Fig. S2a). In this widely adopted cell–cell fusion system,4CoV-S and green fluorescent protein gene were transferred into 293T cells. To mimic the viral fusion process, CoV-S on the transduced 293T cell as the effector cell could mediate its fusion of the effector cell with the receptor-bearing cell as the target cell. Then, by calculating the ratio of the fused cells, we can assess the fusogenic capacity of the S protein in the presence or absence of exogenous trypsin or human airway trypsin-like protease (HAT).

First, we constructed four vectors expressing SARS-CoV-2-S and its variants, SARS-CoV-2-m1, SARS-CoV-2-m2, and SARS-CoV-2-m3, as well as SARS-CoV-S and its variants, SARS-CoV-m1 and SARS-CoV-m2 (Fig. 1b). In SARS-CoV-2-S-m1, 682-RRAR-685 was replaced by 682-SSAR-685, while in SARS-CoV-2-S-m2 and m3, the hinge region between S1 and S2 of SARS-CoV-2 was replaced by that of RaTG13 and SARS-CoV, respectively. In the SARS-CoV-m1 and SARS-CoV-m2, the hinge region between S1 and S2 of SARS-CoV was replaced by that of SARS-CoV-2 and SARS-CoV-2-m1, respectively (Fig. 1b).

We then assessed the expression of the SARS-CoV-2-S and variants in 293T. Western blot analysis of 293T cells transduced with these vectors revealed that SARS-CoV-2-S, SARS-CoV-S and their variants were robustly expressed whereas only SARS-CoV-2-S and SARS-CoV-S-m1 that contain the FCS were processed in the biosynthesis process (Fig. 1c), confirming that FCS indeed works in SARS-CoV-2-S and SARS-CoV-S-m1.
Next, we compared the fusogenic capacity of SARS-CoV-2-S, SARS-CoV-S and their mutants via an S-mediated cell–cell fusion assay in the absence of exogenous trypsin or human airway trypsin-like protease (HAT). The transduced 293T cells were co-incubated with the target cells, Huh-7. After co-incubation for 4 h, 293T cells with wild-type SARS-CoV-2-S fused with the target cells, while cells remained unfused in the SARS-CoV-2-S-m1, m2, and m3 groups (Fig. 1d). Even after a 24-h incubation, the 293T cells with mutated SARS-CoV-2-S still remained unfused (Fig. 1e). Consistently, 293T/ACE2 cells transiently expressing SARS-CoV-2-S or SARS-CoV-S-m1 could naturally fuse with each other, while 293T/ACE2 cells bearing SARS-CoV-2-m1, SARS-CoV-2-m2, SARS-CoV-2-m3, SARS-CoV-S, or SARS-CoV-m2 could not (Supplementary Fig. S2b). Interestingly, SARS-CoV-2 or SARS-CoV-S-m1 that carries FCS could effectively mediate the cell–cell fusion, while others having no FCS could not (Fig. 1d and Supplementary Fig. S2b). These results suggest that FCS may play a role in CoV-S-mediated membrane fusion in the absence of trypsin or HAT.

Subsequently, we compared the fusogenic capacity of SARS-CoV-2-S and its mutants in the presence of trypsin. Surprisingly, like the FCS-containing SARS-CoV-2-S, all the mutants without functional FCS and SARS-CoV-S could effectively mediate cell–cell fusion in a trypsin concentration-dependent manner, although the SARS-CoV-S-mediated fusion activity is much lower than that of SARS-CoV-2-S (Fig. 1f–h). Similarly, treatment with HATs also effectively rescued the fusogenic capacity of SARS-CoV-2 without FCS in a dose-dependent manner (Fig. 1i).

These results suggest that the FCS may be not as critical as previously thought for the high fusion capacity of SARS-CoV-2 in an environment in the presence of HATs, such as human airway. Notably, only high concentration of trypsin could entirely recover the fusogenic capacity of SARS-CoV-2-S without FCS. Therefore, the conclusion of whether SARS-CoV-2 without FCS can still infect target cell with such high efficiency in human airway needs more in vitro and in vivo evidence. Besides, even though the introduction of RRAR-based FCS endowed SARS-CoV-S with the ability to mediate cell–cell fusion, the fusion rate is still significantly lower than that of SARS-CoV-2-S. Previous studies have shown that receptor binding domain (RBD) of SARS-CoV-2-S1 subunit had approximately 10- to 20-fold higher affinity to ACE2 than SARS-CoV RBD.5 Besides, fusion core structure formed by HR1 and HR2 domain in S2 subunit of SARS-CoV-2 is highly stable.4 Therefore, it can be inferred that the structure and characteristics of S protein may be responsible for the potent fusogenic activity of SARS-CoV-2-S, rather than the presence of FCS. Nevertheless, more studies on the COVID-19 animal models are necessary to clarify the exact role of the FCS or host-furin protease in SARS-CoV-2 infection.
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Article CAS Google Scholar AcknowledgementsThis study was funded by the National Megaprojects of China for Major Infectious Diseases (2018ZX10301403), The National Natural Science Foundation of China (81822045 to L.L.; 81630090 to S.J.; 81701998 to Q.W.; and 81703571 to W.X.), and the China Postdoctoral Science Foundation (2018M640341 and 2019T120302 to S.X.).
Author informationAuthor notes
These authors contributed equally: Shuai Xia, Qiaoshuai Lan, Shan Su, Xinling Wang.Authors and AffiliationsKey Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Fudan University, 130 Dong An Road, Shanghai, 200032, China
Shuai Xia, Qiaoshuai Lan, Shan Su, Xinling Wang, Wei Xu, Zezhong Liu, Qian Wang, Lu Lu & Shibo Jiang
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China
Yun ZhuContributionsConceptualization: L.L., S.J., and Q.W. Methodology: L.L., S.J., S.X., and Q.L. Investigation: S.X., Q.L., S.S., X.W., W.X., Z.L., and Y.Z. Writing—Original Draft: S.X. and S.S. Writing—Review and Editing: L.L. and S.J. Funding Acquisition: L.L., S.J., Q.W., W.X., and S.X. Resources: L.L., S.X., and S.J. Supervision: L.L., S.J., and Q.W.
Corresponding authorsCorrespondence to Qian Wang, Lu Lu or Shibo Jiang.
Ethics declarationsCompeting interestsThe authors declare no competing interests.

See: https://www.nature.com/articles/s41392-020-0184-0
It appears, at least until the facts are in while conceding the fact that the US involvement with the Wuhan Labs and the eventual release of Coid-19 virus into an unsuspecting population is murky at beast, that some perfidy may be laid at the feet of Fauci and his staff who did not provide adequate oversight over the uses, operations and particularly over the dangerous pathogens being developed at the Wuhan Lab, nor their possible release.
Hartmann352