was not aware of it. just read them, and... umm... not sure what is going on. he sparks on some things, then really jumps track on others. still, a plus. i suppose. thanks for the input!
my research has found much closer-to-the-point analysis, found openly online, by the Chinese researchers themselves. for example, this peer-reviewed paper was published in 2018 (not a typo!) about how they (at WIV) had successfully produced chimeric SADS and SARS like viruses - with the spike proteins - but they almost mockingly never said "why?" as if that would matter now... (hint: my money is still that it was to be a swine vaccine that went bad)
"Amplification, cloning and expression of human and swine genes
Construction of expression clones for human ACE2 in pcDNA3.1 has been described previously5, 28. Human DPP4 was amplified from human cell lines. Human APN (also known as ANPEP) was commercially synthesized. Swine APN (also known as ANPEP), DPP4 and ACE2 were amplified from piglet intestine. Full-length gene fragments were amplified using specific primers (provided upon request). Human ACE2 was cloned into pCDNA3.1 fused with a His tag. Human APN and DPP4, swine APN, DPP4 and ACE2 were cloned into pCAGGS fused with an S tag. Purified plasmids were transfected into HeLa cells. After 24 h, expression human or swine genes in HeLa cells was confirmed by immunofluorescence assay using mouse anti-His tag or mouse anti-S tag monoclonal antibodies (produced in house) followed by Cy3-labelled goat anti-mouse/rabbit IgG (Proteintech Group).
Pseudovirus preparation
The codon-humanized S genes of SADS-CoV or MERS-CoV cloned into pcDNA3.1 were used for pseudovirus construction as described previously5, 28. In brief, 15 μg of each pHIV-Luc plasmid (pNL4.3.Luc.R-E-Luc) and the S-protein-expressing plasmid (or empty vector control) were co-transfected into 4 × 106 HEK293T cells using Lipofectamine 3000 (Thermo Fisher Scientific). After 4 h, the medium was replaced with fresh medium. Supernatants were collected 48 h after transfection and clarified by centrifugation at 3,000g, then passed through a 0.45-μm filter (Millipore). The filtered supernatants were stored at −80 °C in aliquots until use. To evaluate the incorporation of S proteins into the core of HIV virions, pseudoviruses in supernatant (20 ml) were concentrated by ultracentrifugation through a 20% sucrose cushion (5 ml) at 80,000g for 90 min using a SW41 rotor (Beckman). Pelleted pseudoviruses were dissolved in 50 μl phosphate-buffered saline (PBS) and examined by electron microscopy.
Pseudovirus infection
HeLa cells transiently expressing APN, ACE2 or DPP4 were prepared using Lipofectamine 2000 (Thermo Fisher Scientific). Pseudoviruses prepared above were added to HeLa cells overexpressing APN, ACE2 or DPP4 24 h after transfection. The unabsorbed viruses were removed and replaced with fresh medium at 3 h after infection. The infection was monitored by measuring the luciferase activity conferred by the reporter gene carried by the pseudovirus, using the Luciferase Assay System (Promega) as follows: cells were lysed 48 h after infection, and 20 μl of the lysates was taken for determining luciferase activity after the addition of 50 μl of luciferase substrate.
Examination of known CoV receptors for SADS-CoV entry/infection
HeLa cells transiently expressing APN, ACE2 or DPP4 were prepared using Lipofectamine 2000 (Thermo Fisher Scientific) in a 96-well plate, with mock-transfected cells as controls. SADS-CoV grown in Vero cells was used to infect HeLa cells transiently expressing APN, ACE2 or DPP4. The inoculum was removed after 1 h of absorption and washed twice with PBS and supplemented with medium. SARS-related-CoV WIV167 and MERS-CoV HIV-pseudovirus were used as positive control for human/swine ACE2 or human/swine DPP4, respectively. After 24 h of infection, cells were washed with PBS and fixed with 4% formaldehyde in PBS (pH 7.4) for 20 min at room temperature. SARS-related-CoV WIV16 replication was detected using rabbit antibody against the SARS-related-CoV Rp3 N protein (made in house, 1:100) followed by Cy3-conjugated goat anti-rabbit IgG (1:50, Proteintech)7. SADS-CoV replication was monitored using rabbit antibody against the SADSr-CoV 3755 N protein (made in house, 1:50) followed by FITC-conjugated goat anti-rabbit IgG (1:50, Proteintech). Nuclei were stained with DAPI (Beyotime). Staining patterns were examined using confocal microscopy on a FV1200 microscope (Olympus). Infection of MERS-CoV HIV-pseudovirus was monitored by luciferase 48 h after infection."
For a man of your background, your not "seeing" that SARS-CoV-2 is a bioweapon surprises me! No mention of the HIV inserts at the binding sites of the virus and Nobel laureate Dr Luc Montagnier's opinions regarding same? Have you read Richard Fleming's (MD, PhD, and JD, no less) 2021 book _"Is COVID-19 a Bioweapon? A Scientific and Forensic Investigation_? It's a must-read if you're researching the virus's origins and the intentions/goals of its creators.
And Dr Syed's comment to that essay, answering this question:
"I don't want to ask for speculation beyond what you feel is reasonable based on the evidence, but this leads to some very big questions. There were suggestions early on that the HIV inserts could be indicative of target epitopes for developing prototype HIV vaccine (I think Montaignier mentioned this.) However, why would they then make the whole chimeric virus much more dangerous with the insertion of the furin cleavage site if they were just making target practice virus for testing vaccine concepts? Why not use an animal adapted spike with your HIV epitopes so the whole experiment is FAR less dangerous?
What I'm getting at: is there still a plausible and somewhat benign explanation for why they might graft these characteristics together? The more manipulations are discovered, the more it looks to me like the goal was a virulent human infectious chimeric virus with the added advantage of 'plug-and-play' spike protein that could be swapped out to target different cell receptors or target populations; i.e. the holy-grail of modern bioweapon.
So is there still some legitimate purpose excuse available? Could this have been HIV vaccine research or some sort of cancer vaccine research? Could it have been the DEFUSE 'bat vaccine?'
EDIT: Put simply: the explanation that they were just looking for the next human pandemic potential bat virus falls apart when their chimeric virus had THIS MANY manipulations. So what were they attempting?"
Dr Syed's reply:
"The intention was to create a bioweapon that would not kill off the future slave population (young people) which is why it needed to be SARS based. They knew they had an antidote in HCQ so they were safe. The purpose is a one-world economy (aka great reset aka agenda 2030) which is global fascism. This is not conspiracy theory - this is what the WEF have said they are aiming for, it's just that it's so obviously fascist socialism that people are told not to believe it and they don't because they are in a mass psychosis. It's how the elites (aka oligarchy aka cabal aka WEF aka CFR) have always run things when they can get away with it. So they take a SARS virus, put in (by intention or accident) a FCS for which they have an antidote (folate, HCQ) and HIV epitopes to help evade the immune system (SARS was too immunogenic so died off quickly) and then imposed public health restrictions that ensured the spread of the virus was prolonged artificially. That provided the fear factor so they could get their digital passport system in. It's working well tbh"
Have you read any of Dan Sirotkin's work?
https://harvard2thebighouse.substack.com/p/understanding-covid-19-and-seasonal https://harvard2thebighouse.substack.com/p/burn-the-ivory-towers-to-the-ground
was not aware of it. just read them, and... umm... not sure what is going on. he sparks on some things, then really jumps track on others. still, a plus. i suppose. thanks for the input!
my research has found much closer-to-the-point analysis, found openly online, by the Chinese researchers themselves. for example, this peer-reviewed paper was published in 2018 (not a typo!) about how they (at WIV) had successfully produced chimeric SADS and SARS like viruses - with the spike proteins - but they almost mockingly never said "why?" as if that would matter now... (hint: my money is still that it was to be a swine vaccine that went bad)
excerpt from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7094983/ published 4 April 2018 and signed by 45 scientists at the Wuhan Institute of Virology and includes the signature of Peter Daszak, among others.
"Amplification, cloning and expression of human and swine genes
Construction of expression clones for human ACE2 in pcDNA3.1 has been described previously5, 28. Human DPP4 was amplified from human cell lines. Human APN (also known as ANPEP) was commercially synthesized. Swine APN (also known as ANPEP), DPP4 and ACE2 were amplified from piglet intestine. Full-length gene fragments were amplified using specific primers (provided upon request). Human ACE2 was cloned into pCDNA3.1 fused with a His tag. Human APN and DPP4, swine APN, DPP4 and ACE2 were cloned into pCAGGS fused with an S tag. Purified plasmids were transfected into HeLa cells. After 24 h, expression human or swine genes in HeLa cells was confirmed by immunofluorescence assay using mouse anti-His tag or mouse anti-S tag monoclonal antibodies (produced in house) followed by Cy3-labelled goat anti-mouse/rabbit IgG (Proteintech Group).
Pseudovirus preparation
The codon-humanized S genes of SADS-CoV or MERS-CoV cloned into pcDNA3.1 were used for pseudovirus construction as described previously5, 28. In brief, 15 μg of each pHIV-Luc plasmid (pNL4.3.Luc.R-E-Luc) and the S-protein-expressing plasmid (or empty vector control) were co-transfected into 4 × 106 HEK293T cells using Lipofectamine 3000 (Thermo Fisher Scientific). After 4 h, the medium was replaced with fresh medium. Supernatants were collected 48 h after transfection and clarified by centrifugation at 3,000g, then passed through a 0.45-μm filter (Millipore). The filtered supernatants were stored at −80 °C in aliquots until use. To evaluate the incorporation of S proteins into the core of HIV virions, pseudoviruses in supernatant (20 ml) were concentrated by ultracentrifugation through a 20% sucrose cushion (5 ml) at 80,000g for 90 min using a SW41 rotor (Beckman). Pelleted pseudoviruses were dissolved in 50 μl phosphate-buffered saline (PBS) and examined by electron microscopy.
Pseudovirus infection
HeLa cells transiently expressing APN, ACE2 or DPP4 were prepared using Lipofectamine 2000 (Thermo Fisher Scientific). Pseudoviruses prepared above were added to HeLa cells overexpressing APN, ACE2 or DPP4 24 h after transfection. The unabsorbed viruses were removed and replaced with fresh medium at 3 h after infection. The infection was monitored by measuring the luciferase activity conferred by the reporter gene carried by the pseudovirus, using the Luciferase Assay System (Promega) as follows: cells were lysed 48 h after infection, and 20 μl of the lysates was taken for determining luciferase activity after the addition of 50 μl of luciferase substrate.
Examination of known CoV receptors for SADS-CoV entry/infection
HeLa cells transiently expressing APN, ACE2 or DPP4 were prepared using Lipofectamine 2000 (Thermo Fisher Scientific) in a 96-well plate, with mock-transfected cells as controls. SADS-CoV grown in Vero cells was used to infect HeLa cells transiently expressing APN, ACE2 or DPP4. The inoculum was removed after 1 h of absorption and washed twice with PBS and supplemented with medium. SARS-related-CoV WIV167 and MERS-CoV HIV-pseudovirus were used as positive control for human/swine ACE2 or human/swine DPP4, respectively. After 24 h of infection, cells were washed with PBS and fixed with 4% formaldehyde in PBS (pH 7.4) for 20 min at room temperature. SARS-related-CoV WIV16 replication was detected using rabbit antibody against the SARS-related-CoV Rp3 N protein (made in house, 1:100) followed by Cy3-conjugated goat anti-rabbit IgG (1:50, Proteintech)7. SADS-CoV replication was monitored using rabbit antibody against the SADSr-CoV 3755 N protein (made in house, 1:50) followed by FITC-conjugated goat anti-rabbit IgG (1:50, Proteintech). Nuclei were stained with DAPI (Beyotime). Staining patterns were examined using confocal microscopy on a FV1200 microscope (Olympus). Infection of MERS-CoV HIV-pseudovirus was monitored by luciferase 48 h after infection."
For a man of your background, your not "seeing" that SARS-CoV-2 is a bioweapon surprises me! No mention of the HIV inserts at the binding sites of the virus and Nobel laureate Dr Luc Montagnier's opinions regarding same? Have you read Richard Fleming's (MD, PhD, and JD, no less) 2021 book _"Is COVID-19 a Bioweapon? A Scientific and Forensic Investigation_? It's a must-read if you're researching the virus's origins and the intentions/goals of its creators.
And what do you think of Dr Ah Kahn Syed's (rather famous now) substack essay, here: https://arkmedic.substack.com/p/how-to-blast-your-way-to-the-truth
And Dr Syed's comment to that essay, answering this question:
"I don't want to ask for speculation beyond what you feel is reasonable based on the evidence, but this leads to some very big questions. There were suggestions early on that the HIV inserts could be indicative of target epitopes for developing prototype HIV vaccine (I think Montaignier mentioned this.) However, why would they then make the whole chimeric virus much more dangerous with the insertion of the furin cleavage site if they were just making target practice virus for testing vaccine concepts? Why not use an animal adapted spike with your HIV epitopes so the whole experiment is FAR less dangerous?
What I'm getting at: is there still a plausible and somewhat benign explanation for why they might graft these characteristics together? The more manipulations are discovered, the more it looks to me like the goal was a virulent human infectious chimeric virus with the added advantage of 'plug-and-play' spike protein that could be swapped out to target different cell receptors or target populations; i.e. the holy-grail of modern bioweapon.
So is there still some legitimate purpose excuse available? Could this have been HIV vaccine research or some sort of cancer vaccine research? Could it have been the DEFUSE 'bat vaccine?'
EDIT: Put simply: the explanation that they were just looking for the next human pandemic potential bat virus falls apart when their chimeric virus had THIS MANY manipulations. So what were they attempting?"
Dr Syed's reply:
"The intention was to create a bioweapon that would not kill off the future slave population (young people) which is why it needed to be SARS based. They knew they had an antidote in HCQ so they were safe. The purpose is a one-world economy (aka great reset aka agenda 2030) which is global fascism. This is not conspiracy theory - this is what the WEF have said they are aiming for, it's just that it's so obviously fascist socialism that people are told not to believe it and they don't because they are in a mass psychosis. It's how the elites (aka oligarchy aka cabal aka WEF aka CFR) have always run things when they can get away with it. So they take a SARS virus, put in (by intention or accident) a FCS for which they have an antidote (folate, HCQ) and HIV epitopes to help evade the immune system (SARS was too immunogenic so died off quickly) and then imposed public health restrictions that ensured the spread of the virus was prolonged artificially. That provided the fear factor so they could get their digital passport system in. It's working well tbh"