Correction- Follow on estimations of DNA/RNA contamination with alternative methods and more lots has reproduced contamination above the EMA limit but are less extreme than 20-35%. Please be certain to read the follow on substacks. These substacks look at Qubit readings and qPCR and found 43:1 - 161:1 Ratio of RNA:DNA in the Pfizer monovalent vaccines. Limitations- All lots evaluated to date are expired and this could lead to faster RNA decay than DNA decay. A later substack attempts to monitor this decay rate across
Just impeccable work my friend. If the bloody captured organizations had real scientists working for them, none of their 'products' would make it into arms. Oh, wait now. Right. I see the problem. Ethical high quality science conflicts with the 'business world'. :D So be it.
Seeing real science done and evolve, in real time, online, is just mind blowing, This is the future, made absolutely necessary by the horrors of the present. Thanks for this. I just wish I could understand it more!
More nice work. Have you considered testing whether it's possible to transfect a human cell line with the original product (made up as per the manufacturers instructions), and then select cells for G418 resistance? The LNP should allow the DNA into the cells. Unless I'm mistaken neoR should confer G418 resistance, which should be active in human cells due to the SV40 promoter, plus cells that stay resistant should have plasmid permanently integrated into their genomes.
This would be a finding with major shock value, since it will show that this product can permanently alter the host genome, something that we were told was "impossible".
Edit: Updating my previous calculations, your latest estimates indicate µg quantities of dsDNA present in these products. A typical experiment in a 6-well plate (growth area 9.6cm²) would use ~1 µg of DNA complexed with a lipid-based transfection reagent, to transfect ~10⁶ mammalian cells. In this set up, stable transfectants (with genomic integration of the vector) are produced at the rate of ~ 1 in 10⁴, although this varies by cell type. Linearised DNA should still be active in such a set up, unless the cut effects the expression casette.
Couple of small typos I picked up (not material but what a great paper so thought I'd mention them!)
*Method 1)..... Circular plasmid will be provide....
*Text in para below fig 2 ..... method can also lead to truncated mRNA synthesis as see in many .....
As always, blown away by how well you distill down these complex issues and although I don't always understand the detail, as a lay person, the message is very clear 👌fascinating work and a big thank you from me for being smart enough to do this and to have the integrity to do this work even though you have a day job 👏
So we will become a two state data stream of "scientific" information! Probably better in the long run? Thus, we will compete with the state sponsored data information. Lets see who wins!
Great work, Kevin! I have a few comments/suggestions. I am very surprised by the absence of a polyA sequence from the vector, because it is present in the Pfizer patent (not a perfect poly A as often is the case with vector polyA : aaaaaaa aaaaaaaaaa aaaaaaaaaa aaagcatatg actaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaa) and because adding a poly-adenylation step or ligation step is inefficient and costly from a manufacturing point of view, when it is so much simpler to have it in the vector. And as you mentioned, it would lead to poly-adenylation of truncated mRNA and their more efficient translation. A head scratcher. Btw, shocking that they can publish the result of a phase III without describing how their product is manufactured. The issue of the state and integrity of the DNA present in the mix is worth further investigating and I would suggest an old technique, the Southern blot! You could run the DNA sample as such, unmodified, as well as a few restriction digests with single cutters, for instance MfeI, BspEI and SpeI would tell you if the DNA was properly linearized, and then probe one blot with a vector probe and another with a Spike probe. Nowadays it can be done without radioactivity and given the amount of DNA present, sensitivity won't be an issue. The merit of this approach is that it will directly ascertain the state that DNA is in, how degraded it is or not, which is important to evaluate the risk that this DNA integrates in the genome of a transfected cell. Congratulation on this important work!
You may not need it but this recent study found that LINE1 was also being upregulated:
Transcriptomic study reveals lncRNA-mediated downregulation of innate immune and inflammatory response in the SARS-CoV-2 vaccination breakthrough infections
...While some studies highlight the accumulation of LINE1 upon viral infection, LINE1 is reported to activate the antiviral response in specific instances (50, 51). Besides, the antisense LINE1 is reported to regulate the expression of surrounding genes (52). The LTRs help in viral replication, and it is also reported to activate host immune response (53, 54). Since the interacting lncRNAs are expressed in the VBT group, the higher abundance of Alu, LINE1, and LTR elements within the interacting lncRNAs suggest a possible regulation of antiviral and immune response by the interacting lncRNAs in the vaccination breakthrough individuals.
Natural spike (for most) stays in the lungs. To me, injecting a synthetic mRNA that over-expresses itself (enhanced promoters) and generates a spike protein that resists degradation is very problematic in itself, but an IM injection has already bypassed the bodies natural defenses (respiratory epithelium, blood barrier). Vaccination basically causes "systemic disease" while most with natural infection (fortunately) do not.
Stephanie Seniff's work has shown spike protein in dendrite cells which do not have ACE receptors. She believes all the excess spike expressed in dendritic cells is secreted as exosomes which track up the vagus nerve into the brain where the Wuhan spike has neurodegenerative properties. Omnicron variants apparently do not have the same "prion-like" properties.
I've called Omicron, and its variants, "God's gift to humanity" -- broad immunity to coronoviruses with low risk for systemic disease, unless one is vaccinated (multiple times in particular) because vaccination disrupts normal T cell mediated immunity.
Risk of contact from vaxxed body fluids is unknown. Spike will be around, but I don't know about intact mRNA which is the real risk to 3rd parties. The body can heal itself, aided by proper nutrition and sleep. I would think the 3rd party risk is very low for those who have avoid the direct harm of IM injection of LNPs and synthetic mRNA-->artificial and deadly spike protein.
Hi Kevin, great work. Please can you clarify for me – do you expect the plasmids/vectors to express the spike protein in mammalian (human) cells? If so, what mechanism(s) do you see that happening by – by being transcribed in the genome, or by expressing it independently of that? If the latter, why do you think that is happening – why is the plasmid/vector able to express the spike protein in a mammalian cell? (I think I understand you to say that you do not think the plasmid will generally express in bacterial cells inside a human body - is that right?). Thanks.
Peer Review and t he idea of unbiased scientific publication went out the window in 2001. There are more than 3000 scientists and engineers that have been trying to get a discussion paper published in the same journal as Zdenek Bazant. This effort has been suppressed for years now.
It was the first example of scientific capture by the perpetrators.
Pfizer and Moderna bivalent vaccines contain 20-35% expression vector and are transformation competent in E.coli
Just impeccable work my friend. If the bloody captured organizations had real scientists working for them, none of their 'products' would make it into arms. Oh, wait now. Right. I see the problem. Ethical high quality science conflicts with the 'business world'. :D So be it.
Jessica for the lay folk. What is this showing? Contaminanation of the vaccines?
Can you explain to me like I'm driving in my car by myself, wearing a mask?
Fanstastic and ground breaking work Kevin.
Great stuff. More on Endotoxin
https://geoffpain.substack.com/p/endotoxin-in-pfizer-jabs-causes-heart
https://geoffpain.substack.com/p/guillain-barre-syndrome-expected
https://geoffpain.substack.com/p/women-suffer-more-from-pfizer-endotoxin
https://geoffpain.substack.com/p/endotoxins-in-pfizer-jabs-mimic-nickel
https://geoffpain.substack.com/p/fatigue-headache-muscle-pain-fever
Seeing real science done and evolve, in real time, online, is just mind blowing, This is the future, made absolutely necessary by the horrors of the present. Thanks for this. I just wish I could understand it more!
Lay person here. Not sure what all this means. Is this a new concern for the health of humanity?
More nice work. Have you considered testing whether it's possible to transfect a human cell line with the original product (made up as per the manufacturers instructions), and then select cells for G418 resistance? The LNP should allow the DNA into the cells. Unless I'm mistaken neoR should confer G418 resistance, which should be active in human cells due to the SV40 promoter, plus cells that stay resistant should have plasmid permanently integrated into their genomes.
This would be a finding with major shock value, since it will show that this product can permanently alter the host genome, something that we were told was "impossible".
Edit: Updating my previous calculations, your latest estimates indicate µg quantities of dsDNA present in these products. A typical experiment in a 6-well plate (growth area 9.6cm²) would use ~1 µg of DNA complexed with a lipid-based transfection reagent, to transfect ~10⁶ mammalian cells. In this set up, stable transfectants (with genomic integration of the vector) are produced at the rate of ~ 1 in 10⁴, although this varies by cell type. Linearised DNA should still be active in such a set up, unless the cut effects the expression casette.
Couple of small typos I picked up (not material but what a great paper so thought I'd mention them!)
*Method 1)..... Circular plasmid will be provide....
*Text in para below fig 2 ..... method can also lead to truncated mRNA synthesis as see in many .....
As always, blown away by how well you distill down these complex issues and although I don't always understand the detail, as a lay person, the message is very clear 👌fascinating work and a big thank you from me for being smart enough to do this and to have the integrity to do this work even though you have a day job 👏
So we will become a two state data stream of "scientific" information! Probably better in the long run? Thus, we will compete with the state sponsored data information. Lets see who wins!
Great work, Kevin! I have a few comments/suggestions. I am very surprised by the absence of a polyA sequence from the vector, because it is present in the Pfizer patent (not a perfect poly A as often is the case with vector polyA : aaaaaaa aaaaaaaaaa aaaaaaaaaa aaagcatatg actaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaa) and because adding a poly-adenylation step or ligation step is inefficient and costly from a manufacturing point of view, when it is so much simpler to have it in the vector. And as you mentioned, it would lead to poly-adenylation of truncated mRNA and their more efficient translation. A head scratcher. Btw, shocking that they can publish the result of a phase III without describing how their product is manufactured. The issue of the state and integrity of the DNA present in the mix is worth further investigating and I would suggest an old technique, the Southern blot! You could run the DNA sample as such, unmodified, as well as a few restriction digests with single cutters, for instance MfeI, BspEI and SpeI would tell you if the DNA was properly linearized, and then probe one blot with a vector probe and another with a Spike probe. Nowadays it can be done without radioactivity and given the amount of DNA present, sensitivity won't be an issue. The merit of this approach is that it will directly ascertain the state that DNA is in, how degraded it is or not, which is important to evaluate the risk that this DNA integrates in the genome of a transfected cell. Congratulation on this important work!
You may not need it but this recent study found that LINE1 was also being upregulated:
Transcriptomic study reveals lncRNA-mediated downregulation of innate immune and inflammatory response in the SARS-CoV-2 vaccination breakthrough infections
...While some studies highlight the accumulation of LINE1 upon viral infection, LINE1 is reported to activate the antiviral response in specific instances (50, 51). Besides, the antisense LINE1 is reported to regulate the expression of surrounding genes (52). The LTRs help in viral replication, and it is also reported to activate host immune response (53, 54). Since the interacting lncRNAs are expressed in the VBT group, the higher abundance of Alu, LINE1, and LTR elements within the interacting lncRNAs suggest a possible regulation of antiviral and immune response by the interacting lncRNAs in the vaccination breakthrough individuals.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9716354/#!po=21.8085
Full review:
https://doorlesscarp953.substack.com/p/covid-19-long-non-coding-rnas-and
Natural spike (for most) stays in the lungs. To me, injecting a synthetic mRNA that over-expresses itself (enhanced promoters) and generates a spike protein that resists degradation is very problematic in itself, but an IM injection has already bypassed the bodies natural defenses (respiratory epithelium, blood barrier). Vaccination basically causes "systemic disease" while most with natural infection (fortunately) do not.
Stephanie Seniff's work has shown spike protein in dendrite cells which do not have ACE receptors. She believes all the excess spike expressed in dendritic cells is secreted as exosomes which track up the vagus nerve into the brain where the Wuhan spike has neurodegenerative properties. Omnicron variants apparently do not have the same "prion-like" properties.
I've called Omicron, and its variants, "God's gift to humanity" -- broad immunity to coronoviruses with low risk for systemic disease, unless one is vaccinated (multiple times in particular) because vaccination disrupts normal T cell mediated immunity.
Risk of contact from vaxxed body fluids is unknown. Spike will be around, but I don't know about intact mRNA which is the real risk to 3rd parties. The body can heal itself, aided by proper nutrition and sleep. I would think the 3rd party risk is very low for those who have avoid the direct harm of IM injection of LNPs and synthetic mRNA-->artificial and deadly spike protein.
Hi Kevin, great work. Please can you clarify for me – do you expect the plasmids/vectors to express the spike protein in mammalian (human) cells? If so, what mechanism(s) do you see that happening by – by being transcribed in the genome, or by expressing it independently of that? If the latter, why do you think that is happening – why is the plasmid/vector able to express the spike protein in a mammalian cell? (I think I understand you to say that you do not think the plasmid will generally express in bacterial cells inside a human body - is that right?). Thanks.
Peer Review and t he idea of unbiased scientific publication went out the window in 2001. There are more than 3000 scientists and engineers that have been trying to get a discussion paper published in the same journal as Zdenek Bazant. This effort has been suppressed for years now.
It was the first example of scientific capture by the perpetrators.
Congratulations for your work and thank you for sharing it!!
I do not understand how the organizations involved in the massive vaccination have not analyzed what was being injected all over the world.
Yours results are very very important!!!