Wow, I didn't even think of the fact that muscle cells are being used to express the spike and what factor that would play, but given your previous post on the HeLa contamination this should raise a lot of questions. Now I'm wondering if the multinucleated nature of muscle cells may actually be an issue in both gene expression and uptake of even things such as the adenoviral vectors. Lots more things to ponder and it raises even more questions! When it comes to glycosylation do you suppose that the cell lines used for studies would be critical as well, such that in vitro assays may not provide a viable examination of the true glycosylation occurring if we assume that there will be high variability due to different biochemical processes.
But I suppose then that with those with severe COVID and widespread viral infection should we assume differences in protein conformation and glycosylation? Sorry This whole post has made me really curious about everything!
The fact that the actual spike protein from the vaccine hasn't been fully elucidated is rather alarming as well. Do you know how the mRNA is constructed Joomi? I generally assumed some sort of chip technology automated with some base insertion/wash sequence, but I suppose the type of method would also raise questions as to the fragments produced and whether they were properly isolated.
Don't know much about how the mRNA is constructed but I think Kevin McKernan's substack may have had some articles on it. And yes I think we should assume the cell lines matter for how glycosylation, etc. is done.
Good point about muscle cells... Lots of questions indeed.
This is quite beyond my expertise and I thank you for all you are doing to make it approachable. I had thought that the mRNA was not constructed but elicited via embryonic cell lines based on the following https://www.hek293.com/transfection-information/
I believe the transfection (although I could be grossly mistaken here) was done to insert the spike gene to test the production of the spike similar to how Joomi outlined above. Force cells to produce it and argue that the spike from the mRNA is the same as the one from the virus. However a different method may have been taken to produce the actual mRNA largescale.
I came across this from the NIH and I don't trust the veracity of it, but they're making it out to appear as if the mRNA was produced in a similar method as PCR is conducted which would raise a lot of questions.
I think it's also the fact that we don't know specifically what cells are taken up the LNPs or the adenoviral vectors. I believe there were also talks about the adipocytes taken up the LNPs, and if that were the case that would raise questions on differences in spike production and circulation among normal weight individuals and obese individuals. It may be fruitful in having in vitro studies testing different cell types and seeing how that influences the spike that is produced, but I'm not sure if that is something people are even bothering to consider at this point.
Much of what you guys are writing is more advanced than where my knowledge edge lies, but I usually capture the gist.
The substack The Forgotten Side Of Medicine by author, A Midwestern Doctor, had an article out on Sept 3, 2023, entitled “What Do We Know About Hot Covid-19 Vaccine Lots. In the article he mentions integrity of the spike protein and some possible results. Seems at least somewhat along the lines of the above article. Here’s a clip from the article:
“When I reviewed Pfizer’s internal data, it appeared that the mRNA integrity ranged from 51-71% percent depending on the vaccine lot (this included many lots that were subsequently injected into the human population). This means that much of the mRNA that was present was not the complete mRNA sequence that would produce the entire intended spike protein. These partial mRNA fragments (termed truncated mRNA), in turn appeared to have two potential consequences (these are based on my own understanding of the subject—the regulators only asked for but did not specify the potential consequences of truncated mRNA).
The first is that it would either not produce a protein or produce an incomplete spike protein, in both cases causing the vaccine to produce a lower than intended amount of complete spike protein (which is produced from the intact mRNA) and thus a lower than expected antibody response. The second was that the truncated mRNA species could produce potentially pathologic proteins and there was no practical way to assess the safety of these proteins (remember that the spike protein has multiple toxic components and since there are so many different ways the mRNA could be truncated it is impossible to test each of them). “
That's interesting. It would raise questions as to whether purification was an issue or if the mRNA may have been more susceptible to degradation, although the long-lived mRNA within the germinal centers from that Cell study does raise other questions.
Jamie above provided a possible explanation for the mRNA production, however this page from the NIH suggests it was produced in a manner similar to PCR. I don't trust the website especially due to its simplicity but if that were the case (that these were produced in a similar manner to PCR) then that would raise a good deal of concerns.
What the hell? How could regulators not have asked if the proteins churned out were actually what was expected? Was no sequencing done at all? Or just dodgy blots?
The meetings that were streamed for jab EUAs and approval showed us how the panel did very little challenging or discussion. What the heck was the hurry? The supporting materials submitted by pharma were always done with very little time available before the meetings, even though the dates were weeks out. All deliberate, just as Congress would be given a bill to consider mere hours before voting - WTH, and even they didn't put the brakes on the process.
If the regulators had any concern for citizens at all, the publicly available original "postmarket experience" data from the FDA page on the very first EUA approval for Pfizer would have been way more than enough to shut down the experiment right then and there.
I read that document really early to be able to properly argue with trolls and pHarmaCo groupies. All they ever wanted to do was ignore the official data and quote press releases. I think the regulators were like Walensky, and relied on CNN to serve them their helping of TheScience™ for decision making.
Honestly - the Pfizer FDA briefing meeting had a slide in it that included AESI to watch out for, so it’s not like they didn’t know - this was the November before they rolled out.
We saw them all & more, in that post market experience document so it’s stretching credibility for the pharma trolls to say “this means nothing, it’s not proven to be caused by the jab” - they already knew there was a risk of these effects from the jab. It was not a short list. I have a screen grab somewhere.
It's not sequencing as we do with DNA but rather "an efficient system to check the accuracy of an assumed protein sequence, obtained indirectly by DNA sequencing"
As far I understand it, it's taking an assumed DNA sequence, figuring out what protein it should encode, cleaving that at various points and predicting the mDaltons of the products in the lysate, then running a column to see if the predicted molar weights of those fragments can be confirmed.
To call that "sequencing" is a bit of a stretch. "Sequence confirmation" would seem more appropriate.
Maybe I'm wrong here: This wasn't part of my research.
There are different ways to do it but it seems like most of the time it's some variation of the above. It's also what was used in the study that found unexpected spike sequences in some vaccinated people (in the "Mutant spike pieces in vaccinated..." section of this article).
Also apparently there are some newer methods that are even more sensitive, etc but I don't know much about them aside from that they exist:
I do not know how anybody could trust a industry that has complete liability protection(a license to kill or maim). It appears they set out under the guise of creating a vaccine, a way to create more customers for there drugs, and with all the data they created a maze to keep people looking for god knows what as a explanation for whats going on they knew this is all part of a sick business model that has been ongoing since the Flexner report that changed how medical schools teach.
And "coincidentally" FDA has approved new HIV, cancer, heart, Alzheimer's and other medications/therapies the past 18 months (despite entire panels resigning, etc.)
Joomi’s explanations of the jabs’ unknowns are clear and relatively easy to understand. Why haven’t science journalists reported on this critical matter?
(No need to answer. There are no uncaptured science journalists working for news outlets.)
An analogy for how the mRNA technology is supposed to work: It is like tossing pieces of a violin into a concert hall and expecting the orchestra to play your chosen tunes.
What an absolutely stellar article! Thank you, thank you for clarifying the science and tech used in these studies for those of us who aren't biologists or chemists.
Thanks for another exceptionally clearly written article. Since you previously wrote about spike infiltration into the nucleus, you will be interested in this preprint.
The upshot being that the researchers in this new paper believe a sequence including the infamous furin cleavage site allows nucleus entry. They found mRNA co-located with spike in the nucleus and never mRNA in the nucleus that was not co-located.
In the context of what is discussed here, what does it mean if mRNA produces spike that moves mRNA back into the nucleus? Does that cause multiple cycles of this splicing? Sounds bad.
I'm honestly not sure about any of the studies related to subcellular localization because the methods seem so prone to artifacts. And for that reason I also don't have a lot of confidence in the tools that make predictions about subcellular localization either (since presumably they're using models based on data that's not reliable). However, we don't need for spike to be getting into nuclei to potentially be causing damage to nuclei.
Some short peptide fragments of SARS-CoV-2 Spike are amyloidogenic. Others have superantigenic regions, regions that bind bacterial endotoxins, and others that look very similar to snake venom neurotoxin.
People on Twitter are all like "But SIMOA assays show an infinitesimally small amount of cleaved S1 in the bloodstream!"
Yes. Assays that are specifically designed to find whole Spike or its intact subunits. Not bits of it cleaved apart by host enzymes, nor mistranslated polypeptide fragments, which, given the very low purity of the mRNA in these formulations, is almost a certainty.
By the way, did Katalin Kariko even check to see if pseudouridylated and synthetically capped mRNA evaded detection by TLR7/8, or if it actually behaved as a TLR7/8 inhibitor?
And S1 is probably masked by antibodies anyway - Röltgen et al. essentially confirmed that their negatives for spike after Day 1 weren't reliable since Day 28 samples could mask Day 1 samples or a certain concentration of added spike (Fig 7 KL https://www.cell.com/cell/fulltext/S0092-8674(22)00076-9 ). So everybody who gets injected is basically a magical, mysterious protein piñata - hooray!
I can't conceive of an experiment where actual (degraded) vials of modRNA could be applied to in-vitro tissue culture, then the spike(ish) products cleaved identically to how they would be in the body, then somehow seperate out the unknown protein mix from the rest.
Is there some way to isolate and characterize these products without spot-checking for previously known proteins (like intact spike?)
What might be fun is co-transfecting with live virus and spike modRNA, though you would want to delay the latter to try to time to when the virus is already making sgRNA, and probably mutate out the TRS for the virus's spike. So then you get a kind of prosthetic roboSARS and you can maybe decipher differences in the physical performance of the spike protein that way. But probably more fun than informative.
Thanks for this post. I never really thought the MRNA would produce a variety of proteins folded in different ways, but of course it is a possibility. You do a good job making complex things understandable. 👍🏽👏
Wow, I didn't even think of the fact that muscle cells are being used to express the spike and what factor that would play, but given your previous post on the HeLa contamination this should raise a lot of questions. Now I'm wondering if the multinucleated nature of muscle cells may actually be an issue in both gene expression and uptake of even things such as the adenoviral vectors. Lots more things to ponder and it raises even more questions! When it comes to glycosylation do you suppose that the cell lines used for studies would be critical as well, such that in vitro assays may not provide a viable examination of the true glycosylation occurring if we assume that there will be high variability due to different biochemical processes.
But I suppose then that with those with severe COVID and widespread viral infection should we assume differences in protein conformation and glycosylation? Sorry This whole post has made me really curious about everything!
The fact that the actual spike protein from the vaccine hasn't been fully elucidated is rather alarming as well. Do you know how the mRNA is constructed Joomi? I generally assumed some sort of chip technology automated with some base insertion/wash sequence, but I suppose the type of method would also raise questions as to the fragments produced and whether they were properly isolated.
Don't know much about how the mRNA is constructed but I think Kevin McKernan's substack may have had some articles on it. And yes I think we should assume the cell lines matter for how glycosylation, etc. is done.
Good point about muscle cells... Lots of questions indeed.
This is quite beyond my expertise and I thank you for all you are doing to make it approachable. I had thought that the mRNA was not constructed but elicited via embryonic cell lines based on the following https://www.hek293.com/transfection-information/
I believe the transfection (although I could be grossly mistaken here) was done to insert the spike gene to test the production of the spike similar to how Joomi outlined above. Force cells to produce it and argue that the spike from the mRNA is the same as the one from the virus. However a different method may have been taken to produce the actual mRNA largescale.
I came across this from the NIH and I don't trust the veracity of it, but they're making it out to appear as if the mRNA was produced in a similar method as PCR is conducted which would raise a lot of questions.
https://www.genome.gov/about-genomics/fact-sheets/COVID-19-mRNA-Vaccine-Production
I think it's also the fact that we don't know specifically what cells are taken up the LNPs or the adenoviral vectors. I believe there were also talks about the adipocytes taken up the LNPs, and if that were the case that would raise questions on differences in spike production and circulation among normal weight individuals and obese individuals. It may be fruitful in having in vitro studies testing different cell types and seeing how that influences the spike that is produced, but I'm not sure if that is something people are even bothering to consider at this point.
Much of what you guys are writing is more advanced than where my knowledge edge lies, but I usually capture the gist.
The substack The Forgotten Side Of Medicine by author, A Midwestern Doctor, had an article out on Sept 3, 2023, entitled “What Do We Know About Hot Covid-19 Vaccine Lots. In the article he mentions integrity of the spike protein and some possible results. Seems at least somewhat along the lines of the above article. Here’s a clip from the article:
“When I reviewed Pfizer’s internal data, it appeared that the mRNA integrity ranged from 51-71% percent depending on the vaccine lot (this included many lots that were subsequently injected into the human population). This means that much of the mRNA that was present was not the complete mRNA sequence that would produce the entire intended spike protein. These partial mRNA fragments (termed truncated mRNA), in turn appeared to have two potential consequences (these are based on my own understanding of the subject—the regulators only asked for but did not specify the potential consequences of truncated mRNA).
The first is that it would either not produce a protein or produce an incomplete spike protein, in both cases causing the vaccine to produce a lower than intended amount of complete spike protein (which is produced from the intact mRNA) and thus a lower than expected antibody response. The second was that the truncated mRNA species could produce potentially pathologic proteins and there was no practical way to assess the safety of these proteins (remember that the spike protein has multiple toxic components and since there are so many different ways the mRNA could be truncated it is impossible to test each of them). “
Interesting! Thanks
That's interesting. It would raise questions as to whether purification was an issue or if the mRNA may have been more susceptible to degradation, although the long-lived mRNA within the germinal centers from that Cell study does raise other questions.
Jamie above provided a possible explanation for the mRNA production, however this page from the NIH suggests it was produced in a manner similar to PCR. I don't trust the website especially due to its simplicity but if that were the case (that these were produced in a similar manner to PCR) then that would raise a good deal of concerns.
https://www.genome.gov/about-genomics/fact-sheets/COVID-19-mRNA-Vaccine-Production
Good points, thanks for sharing that post.
What the hell? How could regulators not have asked if the proteins churned out were actually what was expected? Was no sequencing done at all? Or just dodgy blots?
I don't know but if there was sequencing done I don't think it was made publicly available as far as I can tell
You’d think it would have come up in the FDA briefing materials & meeting. I’m not a regulator but I think I would have asked!
The meetings that were streamed for jab EUAs and approval showed us how the panel did very little challenging or discussion. What the heck was the hurry? The supporting materials submitted by pharma were always done with very little time available before the meetings, even though the dates were weeks out. All deliberate, just as Congress would be given a bill to consider mere hours before voting - WTH, and even they didn't put the brakes on the process.
Under the "WTH how could regulators..." heading:
If the regulators had any concern for citizens at all, the publicly available original "postmarket experience" data from the FDA page on the very first EUA approval for Pfizer would have been way more than enough to shut down the experiment right then and there.
I read that document really early to be able to properly argue with trolls and pHarmaCo groupies. All they ever wanted to do was ignore the official data and quote press releases. I think the regulators were like Walensky, and relied on CNN to serve them their helping of TheScience™ for decision making.
Honestly - the Pfizer FDA briefing meeting had a slide in it that included AESI to watch out for, so it’s not like they didn’t know - this was the November before they rolled out.
We saw them all & more, in that post market experience document so it’s stretching credibility for the pharma trolls to say “this means nothing, it’s not proven to be caused by the jab” - they already knew there was a risk of these effects from the jab. It was not a short list. I have a screen grab somewhere.
You can't sequence a protein. A protein is not DNA or RNA.
We can sequence proteins. It usually gets done with LC/MS
Maybe you refer to:
https://www.sciencedirect.com/science/article/abs/pii/S0021967300945278
https://pubmed.ncbi.nlm.nih.gov/3042796/
It's not sequencing as we do with DNA but rather "an efficient system to check the accuracy of an assumed protein sequence, obtained indirectly by DNA sequencing"
As far I understand it, it's taking an assumed DNA sequence, figuring out what protein it should encode, cleaving that at various points and predicting the mDaltons of the products in the lysate, then running a column to see if the predicted molar weights of those fragments can be confirmed.
To call that "sequencing" is a bit of a stretch. "Sequence confirmation" would seem more appropriate.
Maybe I'm wrong here: This wasn't part of my research.
I meant more what's described here:
https://www.sciencedirect.com/science/article/abs/pii/S1046202304002038?via%3Dihub
or here:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3095207/
There are different ways to do it but it seems like most of the time it's some variation of the above. It's also what was used in the study that found unexpected spike sequences in some vaccinated people (in the "Mutant spike pieces in vaccinated..." section of this article).
Also apparently there are some newer methods that are even more sensitive, etc but I don't know much about them aside from that they exist:
https://www.nature.com/articles/nbt.4278
Thank you. One problem with being retired is keeping-up.
I do not know how anybody could trust a industry that has complete liability protection(a license to kill or maim). It appears they set out under the guise of creating a vaccine, a way to create more customers for there drugs, and with all the data they created a maze to keep people looking for god knows what as a explanation for whats going on they knew this is all part of a sick business model that has been ongoing since the Flexner report that changed how medical schools teach.
And "coincidentally" FDA has approved new HIV, cancer, heart, Alzheimer's and other medications/therapies the past 18 months (despite entire panels resigning, etc.)
You write interesting comments. Is there a way to follow your commenting on Substack? If so, I haven't found it.
Aw, man. I stick my foot in my mouth so often! Always learning though. Well, thank you. Made my day and probably until Wednesday, too!
No, I haven't found a way, either.
Hi Joomi,
I recently discovered your substack via GigaohmBiological. I saw your interview there.
You are doing a fabulous job, thank you for sharing, I'll be paying attention.
Take care,
Agus
Great post. Thanks for doing this.
Joomi’s explanations of the jabs’ unknowns are clear and relatively easy to understand. Why haven’t science journalists reported on this critical matter?
(No need to answer. There are no uncaptured science journalists working for news outlets.)
An analogy for how the mRNA technology is supposed to work: It is like tossing pieces of a violin into a concert hall and expecting the orchestra to play your chosen tunes.
This is a great article putting together a few sources that we have been discussing for a while. Thanks for making it simple for everyone!
100% agreed. And fully in-line with my most recently self-asked questions about this whole thing.
What an absolutely stellar article! Thank you, thank you for clarifying the science and tech used in these studies for those of us who aren't biologists or chemists.
Thanks for another exceptionally clearly written article. Since you previously wrote about spike infiltration into the nucleus, you will be interested in this preprint.
https://www.biorxiv.org/content/10.1101/2022.09.27.509633v1.full.pdf
Also discussed here:
https://arkmedic.substack.com/p/new-study-vindicates-jiang-and-mei/comments
The upshot being that the researchers in this new paper believe a sequence including the infamous furin cleavage site allows nucleus entry. They found mRNA co-located with spike in the nucleus and never mRNA in the nucleus that was not co-located.
In the context of what is discussed here, what does it mean if mRNA produces spike that moves mRNA back into the nucleus? Does that cause multiple cycles of this splicing? Sounds bad.
I'm honestly not sure about any of the studies related to subcellular localization because the methods seem so prone to artifacts. And for that reason I also don't have a lot of confidence in the tools that make predictions about subcellular localization either (since presumably they're using models based on data that's not reliable). However, we don't need for spike to be getting into nuclei to potentially be causing damage to nuclei.
Some short peptide fragments of SARS-CoV-2 Spike are amyloidogenic. Others have superantigenic regions, regions that bind bacterial endotoxins, and others that look very similar to snake venom neurotoxin.
https://pubs.acs.org/doi/10.1021/jacs.2c03925
https://www.nature.com/articles/s41577-021-00502-5
https://academic.oup.com/jmcb/article/12/12/916/6028992
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7461543/
People on Twitter are all like "But SIMOA assays show an infinitesimally small amount of cleaved S1 in the bloodstream!"
Yes. Assays that are specifically designed to find whole Spike or its intact subunits. Not bits of it cleaved apart by host enzymes, nor mistranslated polypeptide fragments, which, given the very low purity of the mRNA in these formulations, is almost a certainty.
By the way, did Katalin Kariko even check to see if pseudouridylated and synthetically capped mRNA evaded detection by TLR7/8, or if it actually behaved as a TLR7/8 inhibitor?
https://www.medrxiv.org/content/10.1101/2021.05.03.21256520v1
Big oops.
And S1 is probably masked by antibodies anyway - Röltgen et al. essentially confirmed that their negatives for spike after Day 1 weren't reliable since Day 28 samples could mask Day 1 samples or a certain concentration of added spike (Fig 7 KL https://www.cell.com/cell/fulltext/S0092-8674(22)00076-9 ). So everybody who gets injected is basically a magical, mysterious protein piñata - hooray!
I can't conceive of an experiment where actual (degraded) vials of modRNA could be applied to in-vitro tissue culture, then the spike(ish) products cleaved identically to how they would be in the body, then somehow seperate out the unknown protein mix from the rest.
Is there some way to isolate and characterize these products without spot-checking for previously known proteins (like intact spike?)
What might be fun is co-transfecting with live virus and spike modRNA, though you would want to delay the latter to try to time to when the virus is already making sgRNA, and probably mutate out the TRS for the virus's spike. So then you get a kind of prosthetic roboSARS and you can maybe decipher differences in the physical performance of the spike protein that way. But probably more fun than informative.
Good article. Thank you for writing it.
This is bloody brilliant. Here's my write-up from February:
https://jessicar.substack.com/p/evidence-of-connection-between-severe
Thank you!
Thanks for this post. I never really thought the MRNA would produce a variety of proteins folded in different ways, but of course it is a possibility. You do a good job making complex things understandable. 👍🏽👏
EXCELLENT!!!
Brilliant thorough analysis!
I am sharing this around. You do a wonderful job educating the reader. I've bookmarked this post.
THANK YOU!
Thank you!
I am hoping that an interview with J C Couey is contemplated in the very near future!
Great explanation. Thanks.