Mar 12, 2023·edited Mar 12, 2023Liked by Anandamide
Anandamide, you says that the plasmid contamination is a proxy for LPS contamination.
As a dinosaur in age, I remember back to the 60's when the NCI scientists were making L-asparaginase to be used against cancer, primarily leukemia, lymphomas, melanomas, breast cancer etc...
Except it was causing huge problems for patients, including the liver, kidneys, blood clotting system and the brain, and central nervous system issues.... There were a lot of sudden deaths....
A bright spark suggested that it was E.coli LPS toxicity to which they laughed and said that "the amounts wouldn't hurt a monkey" Dr Gordon Zubrod was in charge...
Anyway, Zubrod wouldn't listen, until three years later Dr Tibor Borsos proved that 13 of 15 commercial lots of L-Asparaginase were contaminated with E.Coli Endotoxin.
all that time, they were still using it, and killing patients with it. All during that time, none of the oncologists could see the LPS toxic syndrome for it punching them on the nose.
At some point the light-bulb lit, and they changed the manufacturing process and used Erwinia Carotovora instead, and suddenly all the "mysterious" toxic deaths stopped.
Another story buried in the "no fishing" cupboard to remind us just what can happen when people don't appear to understand or take note of - or perhaps just ignore.... the basics of E.coli LPS toxicity.
Back in the day there was a huge amount of information on e.coli endotoxin. Lots of medical articles... everyone seemed to understand how dangerous LPS is and was, even in terms of organ rejection. Marathon runners who drop dead on the run = lps endotoxaemia... work was done looking at SIDS and massive amounts of endocab was found, then funding pulled. Why?
I worked with someone who worked in the NCI in the 60's and 70's and they knew that LPS was a core factor in cancer.
In front of me is a letter he sent to the NCI Associate Director on December 29, 1976. On page 3 it says:
5) DPT (diphtheria-pertussis-tetanus) vaccine, alum absorbed (the pertussis factor of which is known to contain endotoxin) when inoculated into mice, resulted in a tumor rate of 10.2%, while mice inoculated with the alum adjuvant alone, developed no tumors."
He told me he was told to do no further research using any vaccines. He also worked with the salk polio vaccine and was friends with Bernice Eddy, the lady who first found the SV40 virus in the vaccine in 1956. Think how many years it took to actually pin SV40 down, and be able to identify it, and then make a test for it? What else was missed along the way? How does this apply today?
Even worse, by the time SV40 was pinned down, the virus had learned to get into adeno and other viruses and form stealth viruses which are almost impossible to figure out, but when they did, they found that the stealth viruses all had different cancer fingerprints. So much research was stopped at the NCI and DBS in the 60's and 70's. Some of that was part of a really big senate committee hearing in April/May 1972 (S.3419) which dealt with why Bernice Eddy was hung out to dry alongside other DBS (later FDA) scientists who spoke out against corruption in the DBS. Most of the scientists just walked. Only two stayed to fight, winning courtcases that lasted years.
Then there is Dr Robert Hull's monograph on the monkey viruses SV1 - 39 (40) and SV41 to 146, which also came out of vaccine supernatant...
I put the simian viruses like that, to emphasise the fact that there were far more monkey viruses which jumped species via vaccines, than just SV40. SV40 was the gold mine, because without it, so much that you could loosely call GMO, would never have been possible. Hull's work didn't include the Mason-Pfizer Monkey virus, or the other viruses which have jumped species as well, which were far more important than people realised. There were people at the NCI at the time, who believed that Kawasaki's Disease was (and still is) an unidentified monkey virus which was in both the SALK and Sabin polio vaccines. They too were told to zip it. They said the culprit was never identified but clearly followed the oral polio vaccine in its emergence. Whatever Kawasaki's is, it was thoroughly embedded vertically and horizontally, and still to this day, under immune suppression, rears its head and still no-one "KNOWS" what it is caused by.
But SV40 is huge, (not just for GMO and recombinant work) but because of the genes it tweaks both on, and off. If there is even a single read let alone a double read of that, in any mRNA vaccine, that is a huge concern. Put LPS on top of SV40 turbo propelled MEP, AND ???? ... what then?
That leads to many more questions than answers. Like "What ELSE is in there that we haven't yet seen?"
All these topics are related. It is obvious when as a dinosaur you can see a few of the pieces.
The problem is how to find the ones you don't see, then put them all together to figure out the implications of it for those vaccinated now, and/or in the future..
The comment facility won't take the explanation to you in one comment so I will try to split it:
That is an extraordinary statement you have made. Let me explain why Anandamine's findings are potentially very important, and are possible the key to explaining what seems to be currently unexplainable, because if he is correct, the e.coli LPS is the first clinical step to much of what is being seen today..
But you won't see evidence of toxicity in clinical data if doctors haven't a clue the many and varied ways E.coli LPS can lead to death, or how it does that.
E.coli LPS death cannot be picked up on an autopsy unless you go into the body immediately, and take the right clinical samples. Who is going to even think of that, or allow it? And how many pathologists today, even know how to do that? Even back in the day, few pathologists understood what to look for and how. Besides which, any autopsy is normally a longish time after death, by which time the "evidence" is gone.
To understand the impact of e.coli endotoxaemia, you would have to go through all the old literature. It is a huge study. There are so many ways that e.coli endotoxin can affect cellular metabolism, that all possible scenarios are impossible to describe. The older books on "Intravascular Disseminated Coagulation" are more accurate than the newer ones, but in the more recent settings, Dr Paul Marik understands endotoxaemia better than anyone.
In terms of the body, the second defence the body has is vitamin C, because that detoxifies endotoxin in the blood. Marik knows that, and there is extensive literature on it, but big Pharma doesn't want to know, because that is too simple, and unpatentable..
Under endogenous circumstances the LPS from the gut is dealt with by the kupffer cells in the liver, but when LPS goes directly into the blood, that protection step is lost. And even under normal circumstances, with fever, E.coli multiplies much much faster so the curlin (envelope fragments that drop off with each bacterial mitosis) production ramps up hugely. Then there are other factors such as the insulin-like effects of bacterial lipopolysaccharides which provoke hypoglycemia, probably due to the inhibition of PEPCK. Another mechanism is where tiny amounts over a period of time, not detoxified by a temporarily dysfunctional reticulo-endothelial system results in removal of blood platelets and fibrinogen from circulating blood resulting in relatively large amounts of serotonin release, which can result in coronary chemoreflex (Bezold-Jarisch reflex) where there is inhibition of sympathetic outflow and increased activity of the cardiac (efferent) vagus leading to profound Bradycardia, hypotension and cardiovascular collapse.Endotoxin also has a more direct effect on cellular respiration, since it interferes with oxidative metabolism of mitochondria.
The pathogenesis of endotoxemia is complex depending on time, dosage and involves the release of norepinephrine, epinephrine, corticosteroids etc.
There is lots more, and if you are familiar with the extensive literature on the variables of endotoxins (which are not all equal) you will see the deficiencies of the above. But you will also know that E.coli endotoxin is one of the worst, clinically, - which is why Anandamine's findings here are crucial to clearly define, not just what is present, but based on the literature, what the physiological consequences could be, because there are many potential different scenarios, .....
There are a lot of "IFS" in all the possible scenarios, so lets look at just two which we can see in play..., otherwise I would write a book.:
First thing to remember is the E.coli toxin causes endothelial fragility so that blood can leak into places it shouldn't. Again, think meningitis blood spots, but there are other things that can happen apart from the most obvious.
Right now, one of the listed side effects is thrombocytopenia purpura. The proper name for that is endotoxemia, because e.coli endotoxin causes vascular capillary permeability, and varying amounts of oedema and hemorrhage by diapedesis. the vascular permeability is the start, which could lead to the end point of what you would see in meningitis where the blood spots don't disappear under the glass.
There very fact that the spike picks up e.coli LPS is a grave concern, because if it is delivered in the LNPs into the cells with the mRNA, it's right there for the spike to pick up as the ribosomes make it. And if spike attaches to endothelial ACE2 receptors along with an external trojan horse cargo of LPS, what will happen to those endothelial cells. And if you are looking a billions of spike with LPS, then you will see exhaustion, and all sorts of symptoms as the body tries to fight it off. That also applies with covid infection, which is why vitamin C needs to be part of the treatment protocol.
But many of the problems with endotoxemia can't be seen with the naked eye clinically in infection. Many people don't know that e.col LPS has an impact on rejection of organ transplants, but it is there in the literature. You are talking about a huge clinical field here, and there is NO WAY doctors have enough time in training to even understand the basics..
So here are two of the many possible scenarios after a covid vaccine IF there is LPS in, and the first determinant will be, if there is LPS there, what is the percentage in THAT particular vial? Some vials may have very little and some may have a lot based on what I've read here.
1) If there is E.coli LPS in the vaccine, first is injection technique. IF it turns out to be needle near or in a vein and the push creates a bolus dose which instantly floods through into the blood partially or fully, then if the vaccine vial has 10 - 30% e.coli LPS as Anadamine has hypothesised on rumble recently, you could have anaphylaxis very quickly - that is the simplest of endotoxemia mechanisms. But it would be labelled an "allergic response" and probably considered to be PEG related.
2) IF Anadamine is correct, and IF some of those plasmids integrate in any of his possible scenarios, and the person is able to overcome the bolus dose, but IF those plasmids start churning out either bacteria OR LPS, then over time, the kupffer cells will be stressed because the body's supply of vitamin C will have gradually decreased to the point of subclinical scurvy, and therefore the liver has more work to do. There comes a point where detoxification in the liver become dysfunctional.
People won't notice that slowly, the gums get redder and bleed more easily, or any of the other signs of subclinical scurvy. And in adults, unlike babies, autopsy X Rays won't show the tell tale Harris lines typical of subclinical scurvy in babies. And should any babies die of the vaccine and xrays show Harris lines, what will the death code be? Not the vaccine, that's for sure.
So in this second scenario, we have a slowly slowly slowly catchee monkey scenario where the first thing that has to be considered is blood flow, where the free endotoxin, is carried via the inferior vena cava into the right side of the heat and by the pulmonary artery into the lungs. Lesser amounts are carried into the left side of the heart and by way of the arterial circulation, into all parts of the body but in decreasing amounts. Extremely small amounts over a long time, will eventually exert adrenergic action in the lungs, and reduction of thrombocytes and fibrinogen. Alternating vaso constriction and dilatation, and increasing capillary permeability with decreased thrombocytes and fibrinogen levels, will result in varying amounts of edema and hemorrhage by diapedesis, primarily in the lungs where the greatest concentration of endotoxin occurs.
So lets take the sudden deaths on sports fields, which usually occur when the athletes core temperature is higher because they have warmed up and/or played a full game. Look at those athletes or referees who have died on the field. How long were they there for? they were all hot, which radically increases the speed of e.coli LPS production which is in effect another fast and sudden bolus dose. if the athlete is already down to zero virtamin C, then there is no safety net.
Imo, either face planking, or back planking clutching the chest, or just flat out unresponsive, is absolutely the end result of a constant build up on e.coli endotoxaemia by one mechanism or another.
And not all sudden deaths in sport will be vaccine induced, because we have always had a low level of sudden deaths in long distance runners. We all have e. coli in our guts because it does perform essential functions in the body. In the presence of e.coli under normal circumstances, sudden death can occur many ways. One of the most common but ignored by doctors is what they call an anaphylaxis response to antibiotics, when it's not. It's a result of sudden, massive die off of e.coli where all the envelope lipopolysaccharide becomes a huge bolus dose on it's own. "Oh they were allergic..."
Long distance runners who drop dead during an event, often near the end, will have a similar mechanism to the sudden deaths we see after covid vax, because they are addicted to running and having a regular high core body temperature they will have a higher level of LPS in their body than normal people. and if their body stores of vitamin C are low, they are a potential disaster waiting to happen.
However, what you don't see in the "old" sads were complaints of heart pain before hand, or the long list of "complaints" that many of those athletes were having beforehand, so the mechanisms back then, and now, have clinical differences which anyone who really understands e.coli endotoxemia, will understand properly, and be able to differentiate from the medical history, comments the patient has made, and the difference between the end point of vaccine induced LPS and exercise induced LPS. And it's interesting that even the medical literature recognised in the past that for athletes, vitamin C was a key to preventing those deaths.
I would guarantee that the vast majority of systematised doctors these days have very little, if any clinical understanding of e.coli LPS in the body.
Which might be why you see "no evidence of toxicity in clinical data" because what you don't understand and don't see, you won't report. And even if you did see it, would you report it?
It's not for you to tell any scientist, what and how they should proceed with what they are finding.
editted to add, that 2011 article was about student athletes only, and as I said before, there has always been a low level of SADS on athletes particularly runners. And I described one of the most common mechanisms.
That article has no relevance to what we are seeing today, either in principle or clinically.
Your tone above would indicate that you aren't here to learn anything, but simply to call Anandamine a whiner who won't follow your idea of appropriateness.
Here is my summary of his reasons, and if I've made a mistake then Anandamine can correct them.
Right now, the most important thing is to get other labs to reproduce the results and as quickly as possible. There is already another laboratory on board, and the fact that the primers are being made available, would indicate than others have contacted the company.
for the sake of sick people, Anandamine and others, are doing what they can to try to figure out why so many vaccinated are having problems and then once that is verified and understood, to try to help those people get better.
The system as it currently stands does not have the ability or will to do that.
Peer Review is now politically captured, and takes around 12 months and sometimes longer. It also costs 3 - 5 thousand dollars, and even if you get through the peer review process, it is way too common for someone to come along, complain and the article to be retracted, because it's not the correct narrative..
Waiting around for 18 months for the system to return to scientific principles (or maybe never) does not help those who are suffering right now. Real science is about reproducibility, not sidelining the messenger.
So it comes down to this. Is your priority patients' well-being, or satisfying the blinkered system?
Mar 12, 2023·edited Mar 12, 2023Liked by Anandamide
Well done, again! Not surprising you have confirmed the presence of circular plasmid by sequencing given your earlier results, but it did need to be done. Also nice to have an improved assembly of their vector sequence and solve the polyA "mystery".
A stand out for me is still the quantity of DNA you detected in your previous Substack. Circular or linear, I still think this will happily transfect human cells, in some cases permanently altering the genome.
I was left wondering how the heck it could be that there is that much DNA left there, and also what kind of QC parameters they have for residual DNA in the first place. After perusing some Australian TGA FOI documents yesterday, and quickly reading through the EMA document you've linked here, this is basic summary as I see it:
1) The manufacturing process uses a linearised DNA plasmid together with T7 polymerase and other reagents to generate the RNA.
2) The removal of the DNA is reliant on the DNase I step performed after RNA product is complete. The EMA document suggests sizable (10×) batch-to-batch variation in the amount of residual DNA present (10-220 ng/mg) in production batches, even if all of these were below their specification of 330 ng DNA/mg RNA. One development batch did have much higher template concentration (815 ng/mg) due to an "incorrect DNase I stock solution", as you noted.
3) The assay to detect residual DNA is a qPCR-based assay with primers on the T7 promoter and Kozak sequence. Presumably the motivation for this assay is high sensitivity to detect what is expected to be a rather trace impurity. This method will not quantify any DNA not containing the target sequence, but should detect the input plasmids sequenced here (I checked and the primers do match).
4) The residual template check is done after the Drug Substance production stage, prior to the LNP Fabrication stage.
6) The is an oversight in this process that I can see, which is there is no guarantee that the subsequent steps used in product will not effectively concentrate the DNA relative to the RNA. For example, the DNA will be obviously a lot more stable than RNA, so will be more resistant to degradation. In addition, what if the LNP is more efficient at packaging DNA than RNA?
The last point could alone potentially bring the residual amount of DNA to above the specification of 330 ng/mg RNA, if it was already close to the spec. I was trying to dig up some figures on what the RNA losses are going from Drug Substance production to final product to estimate worst case scenario but haven't found any yet. Nevertheless, I assume the losses are <25% at worse case, so in order for a large amount of DNA to be present the DNAse would have to fail, and the residual DNA assay would have to fail or be ignored, implying extremely shoddy standards. In any case, residual DNA testing should really by part of the QC for the final product, but it's not. That's lax in itself.
As for the specification itself, the EMA document has this justification:
"The specification for residual DNA template was based on the WHO recommendation of not more than 10 ng DNA/dose. Based on these considerations, and assuming a maximum dose of 30μg, the commercial acceptance criterion at release is ≤330 ng DNA/mg RNA. The approach is endorsed. "
I haven't looked at the WHO justification for this and I'm not sure I really agree 10 ng is acceptable. This is more than enough DNA to transfect some cells, with potential genomic integration, when multiplied over enough cells exposed and enough recipients.
Indeed. But their vector map matches our sequence derived map and the length is only 14 bases different. Probably polyA differences which are hard to sequence.
But not providing the exact sequence enables them to change the vector on the fly which we now have evidence they are infact doing within a single lot. That 72bp insertion shouldn’t occur within the same lot.
I am unclear who is supplying the vectors/plasmids. Are they doing it in-house (Andover) or are they getting it elsewhere. If the latter could they have more than one supplier?
So glad I've subscribed to this substack, amazing detailed work (much beyond my limited uni biology studies two decades ago!) ... A few substacks have mentioned the DNA plasmids, but not in the detail you have explored... Do you think these plasmids are replication competent due to the sheer amount of linearized DNA? Given they are less replication competent than circular plasmids, is there still enough potential replication possible, at the high volumes measures? There are indications of circular plasmids detected as mentioned, in which case is this a potential long term expression of spike for the vaccinated? As they appeared to be replication competent in E.Coli in the lab... and therefore also could go onto contribute high endotoxin loads in the jab if there's any contamination at the production stage of the vaccine (anaphalxis potential/toxic shock) due to E.Coli based plasmids detected in the Pfizer jab ?
Excuse my poor dropout level of understanding, but this article compliments and refined the other substack from Jessica Rose I'd read on the subject.
Given there are serial conditionals (If (gets to GI) AND if (Transforms) AND if (Antibiotic Selection)), I have tried to not speculate much on the linear DNA's transformation efficiency. Lucigen has linear vectors for E.coli transformation. With Trillions of DNA molecules and some portion of them competent with proof of circularity... and the lot numbers dont seem to mean anything... Im having a hard time believing any 'data not shown' portion of the EMA document.
Thank you very much for that, and yes "data not shown" ... For their benefit... Scary that this lack of scrutiny passed by the gatekeeper (although I suspect almost "anything goes" these days with the level of funding regulators receive from the very industry they are meant to regulate)
The Endotoxin in the Pfizer jabs comes from the E coli cell wall and is not related to the Plasmid synthesis of circular DNA which is then chopped up to linear segments for mRNA synthesis. Lipid A is about 50,000 times more toxic than the larger LPS fragments.
Thanks for that link Geoff, avid fan and follower of your substack (although I am playing catch up with many of your articles since discovering your work)... I had noted your endotoxin notes regarding use of E.Coli in the production process of the Pfizer jabs too. I'll check out your above link... Keep up the great work, I had shared one of your articles in a comment or too regarding anaphylactic shock. Can't believe you still have a Twitter ban, you must be too close to the truth!
Anandamide, you says that the plasmid contamination is a proxy for LPS contamination.
As a dinosaur in age, I remember back to the 60's when the NCI scientists were making L-asparaginase to be used against cancer, primarily leukemia, lymphomas, melanomas, breast cancer etc...
Except it was causing huge problems for patients, including the liver, kidneys, blood clotting system and the brain, and central nervous system issues.... There were a lot of sudden deaths....
A bright spark suggested that it was E.coli LPS toxicity to which they laughed and said that "the amounts wouldn't hurt a monkey" Dr Gordon Zubrod was in charge...
Anyway, Zubrod wouldn't listen, until three years later Dr Tibor Borsos proved that 13 of 15 commercial lots of L-Asparaginase were contaminated with E.Coli Endotoxin.
all that time, they were still using it, and killing patients with it. All during that time, none of the oncologists could see the LPS toxic syndrome for it punching them on the nose.
At some point the light-bulb lit, and they changed the manufacturing process and used Erwinia Carotovora instead, and suddenly all the "mysterious" toxic deaths stopped.
Another story buried in the "no fishing" cupboard to remind us just what can happen when people don't appear to understand or take note of - or perhaps just ignore.... the basics of E.coli LPS toxicity.
Wow, thanks for this bit of science history.
Back in the day there was a huge amount of information on e.coli endotoxin. Lots of medical articles... everyone seemed to understand how dangerous LPS is and was, even in terms of organ rejection. Marathon runners who drop dead on the run = lps endotoxaemia... work was done looking at SIDS and massive amounts of endocab was found, then funding pulled. Why?
I worked with someone who worked in the NCI in the 60's and 70's and they knew that LPS was a core factor in cancer.
In front of me is a letter he sent to the NCI Associate Director on December 29, 1976. On page 3 it says:
5) DPT (diphtheria-pertussis-tetanus) vaccine, alum absorbed (the pertussis factor of which is known to contain endotoxin) when inoculated into mice, resulted in a tumor rate of 10.2%, while mice inoculated with the alum adjuvant alone, developed no tumors."
He told me he was told to do no further research using any vaccines. He also worked with the salk polio vaccine and was friends with Bernice Eddy, the lady who first found the SV40 virus in the vaccine in 1956. Think how many years it took to actually pin SV40 down, and be able to identify it, and then make a test for it? What else was missed along the way? How does this apply today?
Even worse, by the time SV40 was pinned down, the virus had learned to get into adeno and other viruses and form stealth viruses which are almost impossible to figure out, but when they did, they found that the stealth viruses all had different cancer fingerprints. So much research was stopped at the NCI and DBS in the 60's and 70's. Some of that was part of a really big senate committee hearing in April/May 1972 (S.3419) which dealt with why Bernice Eddy was hung out to dry alongside other DBS (later FDA) scientists who spoke out against corruption in the DBS. Most of the scientists just walked. Only two stayed to fight, winning courtcases that lasted years.
Then there is Dr Robert Hull's monograph on the monkey viruses SV1 - 39 (40) and SV41 to 146, which also came out of vaccine supernatant...
I put the simian viruses like that, to emphasise the fact that there were far more monkey viruses which jumped species via vaccines, than just SV40. SV40 was the gold mine, because without it, so much that you could loosely call GMO, would never have been possible. Hull's work didn't include the Mason-Pfizer Monkey virus, or the other viruses which have jumped species as well, which were far more important than people realised. There were people at the NCI at the time, who believed that Kawasaki's Disease was (and still is) an unidentified monkey virus which was in both the SALK and Sabin polio vaccines. They too were told to zip it. They said the culprit was never identified but clearly followed the oral polio vaccine in its emergence. Whatever Kawasaki's is, it was thoroughly embedded vertically and horizontally, and still to this day, under immune suppression, rears its head and still no-one "KNOWS" what it is caused by.
But SV40 is huge, (not just for GMO and recombinant work) but because of the genes it tweaks both on, and off. If there is even a single read let alone a double read of that, in any mRNA vaccine, that is a huge concern. Put LPS on top of SV40 turbo propelled MEP, AND ???? ... what then?
That leads to many more questions than answers. Like "What ELSE is in there that we haven't yet seen?"
All these topics are related. It is obvious when as a dinosaur you can see a few of the pieces.
The problem is how to find the ones you don't see, then put them all together to figure out the implications of it for those vaccinated now, and/or in the future..
Wow, you are blowing my mind!!
Just spend a few years studying the old medical literature, because it's all there hidden in plain sight.
The comment facility won't take the explanation to you in one comment so I will try to split it:
That is an extraordinary statement you have made. Let me explain why Anandamine's findings are potentially very important, and are possible the key to explaining what seems to be currently unexplainable, because if he is correct, the e.coli LPS is the first clinical step to much of what is being seen today..
But you won't see evidence of toxicity in clinical data if doctors haven't a clue the many and varied ways E.coli LPS can lead to death, or how it does that.
E.coli LPS death cannot be picked up on an autopsy unless you go into the body immediately, and take the right clinical samples. Who is going to even think of that, or allow it? And how many pathologists today, even know how to do that? Even back in the day, few pathologists understood what to look for and how. Besides which, any autopsy is normally a longish time after death, by which time the "evidence" is gone.
To understand the impact of e.coli endotoxaemia, you would have to go through all the old literature. It is a huge study. There are so many ways that e.coli endotoxin can affect cellular metabolism, that all possible scenarios are impossible to describe. The older books on "Intravascular Disseminated Coagulation" are more accurate than the newer ones, but in the more recent settings, Dr Paul Marik understands endotoxaemia better than anyone.
In terms of the body, the second defence the body has is vitamin C, because that detoxifies endotoxin in the blood. Marik knows that, and there is extensive literature on it, but big Pharma doesn't want to know, because that is too simple, and unpatentable..
Under endogenous circumstances the LPS from the gut is dealt with by the kupffer cells in the liver, but when LPS goes directly into the blood, that protection step is lost. And even under normal circumstances, with fever, E.coli multiplies much much faster so the curlin (envelope fragments that drop off with each bacterial mitosis) production ramps up hugely. Then there are other factors such as the insulin-like effects of bacterial lipopolysaccharides which provoke hypoglycemia, probably due to the inhibition of PEPCK. Another mechanism is where tiny amounts over a period of time, not detoxified by a temporarily dysfunctional reticulo-endothelial system results in removal of blood platelets and fibrinogen from circulating blood resulting in relatively large amounts of serotonin release, which can result in coronary chemoreflex (Bezold-Jarisch reflex) where there is inhibition of sympathetic outflow and increased activity of the cardiac (efferent) vagus leading to profound Bradycardia, hypotension and cardiovascular collapse.Endotoxin also has a more direct effect on cellular respiration, since it interferes with oxidative metabolism of mitochondria.
The pathogenesis of endotoxemia is complex depending on time, dosage and involves the release of norepinephrine, epinephrine, corticosteroids etc.
There is lots more, and if you are familiar with the extensive literature on the variables of endotoxins (which are not all equal) you will see the deficiencies of the above. But you will also know that E.coli endotoxin is one of the worst, clinically, - which is why Anandamine's findings here are crucial to clearly define, not just what is present, but based on the literature, what the physiological consequences could be, because there are many potential different scenarios, .....
There are a lot of "IFS" in all the possible scenarios, so lets look at just two which we can see in play..., otherwise I would write a book.:
First thing to remember is the E.coli toxin causes endothelial fragility so that blood can leak into places it shouldn't. Again, think meningitis blood spots, but there are other things that can happen apart from the most obvious.
Right now, one of the listed side effects is thrombocytopenia purpura. The proper name for that is endotoxemia, because e.coli endotoxin causes vascular capillary permeability, and varying amounts of oedema and hemorrhage by diapedesis. the vascular permeability is the start, which could lead to the end point of what you would see in meningitis where the blood spots don't disappear under the glass.
There very fact that the spike picks up e.coli LPS is a grave concern, because if it is delivered in the LNPs into the cells with the mRNA, it's right there for the spike to pick up as the ribosomes make it. And if spike attaches to endothelial ACE2 receptors along with an external trojan horse cargo of LPS, what will happen to those endothelial cells. And if you are looking a billions of spike with LPS, then you will see exhaustion, and all sorts of symptoms as the body tries to fight it off. That also applies with covid infection, which is why vitamin C needs to be part of the treatment protocol.
But many of the problems with endotoxemia can't be seen with the naked eye clinically in infection. Many people don't know that e.col LPS has an impact on rejection of organ transplants, but it is there in the literature. You are talking about a huge clinical field here, and there is NO WAY doctors have enough time in training to even understand the basics..
So here are two of the many possible scenarios after a covid vaccine IF there is LPS in, and the first determinant will be, if there is LPS there, what is the percentage in THAT particular vial? Some vials may have very little and some may have a lot based on what I've read here.
1) If there is E.coli LPS in the vaccine, first is injection technique. IF it turns out to be needle near or in a vein and the push creates a bolus dose which instantly floods through into the blood partially or fully, then if the vaccine vial has 10 - 30% e.coli LPS as Anadamine has hypothesised on rumble recently, you could have anaphylaxis very quickly - that is the simplest of endotoxemia mechanisms. But it would be labelled an "allergic response" and probably considered to be PEG related.
2) IF Anadamine is correct, and IF some of those plasmids integrate in any of his possible scenarios, and the person is able to overcome the bolus dose, but IF those plasmids start churning out either bacteria OR LPS, then over time, the kupffer cells will be stressed because the body's supply of vitamin C will have gradually decreased to the point of subclinical scurvy, and therefore the liver has more work to do. There comes a point where detoxification in the liver become dysfunctional.
People won't notice that slowly, the gums get redder and bleed more easily, or any of the other signs of subclinical scurvy. And in adults, unlike babies, autopsy X Rays won't show the tell tale Harris lines typical of subclinical scurvy in babies. And should any babies die of the vaccine and xrays show Harris lines, what will the death code be? Not the vaccine, that's for sure.
So in this second scenario, we have a slowly slowly slowly catchee monkey scenario where the first thing that has to be considered is blood flow, where the free endotoxin, is carried via the inferior vena cava into the right side of the heat and by the pulmonary artery into the lungs. Lesser amounts are carried into the left side of the heart and by way of the arterial circulation, into all parts of the body but in decreasing amounts. Extremely small amounts over a long time, will eventually exert adrenergic action in the lungs, and reduction of thrombocytes and fibrinogen. Alternating vaso constriction and dilatation, and increasing capillary permeability with decreased thrombocytes and fibrinogen levels, will result in varying amounts of edema and hemorrhage by diapedesis, primarily in the lungs where the greatest concentration of endotoxin occurs.
So lets take the sudden deaths on sports fields, which usually occur when the athletes core temperature is higher because they have warmed up and/or played a full game. Look at those athletes or referees who have died on the field. How long were they there for? they were all hot, which radically increases the speed of e.coli LPS production which is in effect another fast and sudden bolus dose. if the athlete is already down to zero virtamin C, then there is no safety net.
Imo, either face planking, or back planking clutching the chest, or just flat out unresponsive, is absolutely the end result of a constant build up on e.coli endotoxaemia by one mechanism or another.
And not all sudden deaths in sport will be vaccine induced, because we have always had a low level of sudden deaths in long distance runners. We all have e. coli in our guts because it does perform essential functions in the body. In the presence of e.coli under normal circumstances, sudden death can occur many ways. One of the most common but ignored by doctors is what they call an anaphylaxis response to antibiotics, when it's not. It's a result of sudden, massive die off of e.coli where all the envelope lipopolysaccharide becomes a huge bolus dose on it's own. "Oh they were allergic..."
Long distance runners who drop dead during an event, often near the end, will have a similar mechanism to the sudden deaths we see after covid vax, because they are addicted to running and having a regular high core body temperature they will have a higher level of LPS in their body than normal people. and if their body stores of vitamin C are low, they are a potential disaster waiting to happen.
However, what you don't see in the "old" sads were complaints of heart pain before hand, or the long list of "complaints" that many of those athletes were having beforehand, so the mechanisms back then, and now, have clinical differences which anyone who really understands e.coli endotoxemia, will understand properly, and be able to differentiate from the medical history, comments the patient has made, and the difference between the end point of vaccine induced LPS and exercise induced LPS. And it's interesting that even the medical literature recognised in the past that for athletes, vitamin C was a key to preventing those deaths.
I would guarantee that the vast majority of systematised doctors these days have very little, if any clinical understanding of e.coli LPS in the body.
Which might be why you see "no evidence of toxicity in clinical data" because what you don't understand and don't see, you won't report. And even if you did see it, would you report it?
See the reasoning above.
It's not for you to tell any scientist, what and how they should proceed with what they are finding.
editted to add, that 2011 article was about student athletes only, and as I said before, there has always been a low level of SADS on athletes particularly runners. And I described one of the most common mechanisms.
That article has no relevance to what we are seeing today, either in principle or clinically.
Your tone above would indicate that you aren't here to learn anything, but simply to call Anandamine a whiner who won't follow your idea of appropriateness.
Frankly, that is trolling.
Anandamine has explained several times why not.
Here is my summary of his reasons, and if I've made a mistake then Anandamine can correct them.
Right now, the most important thing is to get other labs to reproduce the results and as quickly as possible. There is already another laboratory on board, and the fact that the primers are being made available, would indicate than others have contacted the company.
for the sake of sick people, Anandamine and others, are doing what they can to try to figure out why so many vaccinated are having problems and then once that is verified and understood, to try to help those people get better.
The system as it currently stands does not have the ability or will to do that.
Peer Review is now politically captured, and takes around 12 months and sometimes longer. It also costs 3 - 5 thousand dollars, and even if you get through the peer review process, it is way too common for someone to come along, complain and the article to be retracted, because it's not the correct narrative..
Waiting around for 18 months for the system to return to scientific principles (or maybe never) does not help those who are suffering right now. Real science is about reproducibility, not sidelining the messenger.
So it comes down to this. Is your priority patients' well-being, or satisfying the blinkered system?
Well done, again! Not surprising you have confirmed the presence of circular plasmid by sequencing given your earlier results, but it did need to be done. Also nice to have an improved assembly of their vector sequence and solve the polyA "mystery".
A stand out for me is still the quantity of DNA you detected in your previous Substack. Circular or linear, I still think this will happily transfect human cells, in some cases permanently altering the genome.
I was left wondering how the heck it could be that there is that much DNA left there, and also what kind of QC parameters they have for residual DNA in the first place. After perusing some Australian TGA FOI documents yesterday, and quickly reading through the EMA document you've linked here, this is basic summary as I see it:
1) The manufacturing process uses a linearised DNA plasmid together with T7 polymerase and other reagents to generate the RNA.
2) The removal of the DNA is reliant on the DNase I step performed after RNA product is complete. The EMA document suggests sizable (10×) batch-to-batch variation in the amount of residual DNA present (10-220 ng/mg) in production batches, even if all of these were below their specification of 330 ng DNA/mg RNA. One development batch did have much higher template concentration (815 ng/mg) due to an "incorrect DNase I stock solution", as you noted.
3) The assay to detect residual DNA is a qPCR-based assay with primers on the T7 promoter and Kozak sequence. Presumably the motivation for this assay is high sensitivity to detect what is expected to be a rather trace impurity. This method will not quantify any DNA not containing the target sequence, but should detect the input plasmids sequenced here (I checked and the primers do match).
4) The residual template check is done after the Drug Substance production stage, prior to the LNP Fabrication stage.
5) Key point: After this stage, no further testing for residual DNA is performed. The amount of residual DNA is assumed to be within spec after that in the initial test. Many of the methods used to check RNA will happily detect DNA as well, such as the RT-PCR you highlighted. I have found no information on the amount of DNA present in the batch QCs reported in the TGA FOI requests (e.g. FOI 3471 from https://www.tga.gov.au/resources/publication/publications/documents-released-under-section-11c-freedom-information-act-1982-jul-2021-jun-2022).
6) The is an oversight in this process that I can see, which is there is no guarantee that the subsequent steps used in product will not effectively concentrate the DNA relative to the RNA. For example, the DNA will be obviously a lot more stable than RNA, so will be more resistant to degradation. In addition, what if the LNP is more efficient at packaging DNA than RNA?
The last point could alone potentially bring the residual amount of DNA to above the specification of 330 ng/mg RNA, if it was already close to the spec. I was trying to dig up some figures on what the RNA losses are going from Drug Substance production to final product to estimate worst case scenario but haven't found any yet. Nevertheless, I assume the losses are <25% at worse case, so in order for a large amount of DNA to be present the DNAse would have to fail, and the residual DNA assay would have to fail or be ignored, implying extremely shoddy standards. In any case, residual DNA testing should really by part of the QC for the final product, but it's not. That's lax in itself.
As for the specification itself, the EMA document has this justification:
"The specification for residual DNA template was based on the WHO recommendation of not more than 10 ng DNA/dose. Based on these considerations, and assuming a maximum dose of 30μg, the commercial acceptance criterion at release is ≤330 ng DNA/mg RNA. The approach is endorsed. "
I haven't looked at the WHO justification for this and I'm not sure I really agree 10 ng is acceptable. This is more than enough DNA to transfect some cells, with potential genomic integration, when multiplied over enough cells exposed and enough recipients.
We are seeing 10ng/ul DNA and the dose is 300ul.
There is a place in the EMA doc where they convince themselves there is no DNA by RNAse A treatment and running a gel.
There are ways this can eliminate DNA if you are not careful listed in the limitations.
I bet they see a different result with RNase R.
wow, quite good work.
Excellent work, thanks very much.
Wow. Amazing work.
Something tells me the methylpseudouridine replacement is going to have many many far reaching, yet to be revealed ripples. Yikes.
From the EMA doc
“Neither the source and generation of the pST4-1525 plasmid used are documented ”
Indeed. But their vector map matches our sequence derived map and the length is only 14 bases different. Probably polyA differences which are hard to sequence.
But not providing the exact sequence enables them to change the vector on the fly which we now have evidence they are infact doing within a single lot. That 72bp insertion shouldn’t occur within the same lot.
I am unclear who is supplying the vectors/plasmids. Are they doing it in-house (Andover) or are they getting it elsewhere. If the latter could they have more than one supplier?
I think Charles River and one other company was mentioned in the doc.
So glad I've subscribed to this substack, amazing detailed work (much beyond my limited uni biology studies two decades ago!) ... A few substacks have mentioned the DNA plasmids, but not in the detail you have explored... Do you think these plasmids are replication competent due to the sheer amount of linearized DNA? Given they are less replication competent than circular plasmids, is there still enough potential replication possible, at the high volumes measures? There are indications of circular plasmids detected as mentioned, in which case is this a potential long term expression of spike for the vaccinated? As they appeared to be replication competent in E.Coli in the lab... and therefore also could go onto contribute high endotoxin loads in the jab if there's any contamination at the production stage of the vaccine (anaphalxis potential/toxic shock) due to E.Coli based plasmids detected in the Pfizer jab ?
Excuse my poor dropout level of understanding, but this article compliments and refined the other substack from Jessica Rose I'd read on the subject.
https://open.substack.com/pub/jessicar/p/follow-up-on-dna-contamination-of?utm_source=share&utm_medium=android
Apologies again for being almost kindergarten level compared to your knowledge base, hopefully my questions make sense...
Given there are serial conditionals (If (gets to GI) AND if (Transforms) AND if (Antibiotic Selection)), I have tried to not speculate much on the linear DNA's transformation efficiency. Lucigen has linear vectors for E.coli transformation. With Trillions of DNA molecules and some portion of them competent with proof of circularity... and the lot numbers dont seem to mean anything... Im having a hard time believing any 'data not shown' portion of the EMA document.
Thank you very much for that, and yes "data not shown" ... For their benefit... Scary that this lack of scrutiny passed by the gatekeeper (although I suspect almost "anything goes" these days with the level of funding regulators receive from the very industry they are meant to regulate)
The Endotoxin in the Pfizer jabs comes from the E coli cell wall and is not related to the Plasmid synthesis of circular DNA which is then chopped up to linear segments for mRNA synthesis. Lipid A is about 50,000 times more toxic than the larger LPS fragments.
https://geoffpain.substack.com/p/epigenetics-of-endotoxin-poisoning
The process of plasmid isolation from Ecoli is notorious for LPS contamination.
We have evidence their linearization reaction with Eam1104i isn’t complete and is leaving behind circular plasmid.
I have no reason to believe their LAL assay results given all the other discrepancies in the document.
Let's hope someone is successful getting all the actual numbers from the LAL assays.
Thanks for that link Geoff, avid fan and follower of your substack (although I am playing catch up with many of your articles since discovering your work)... I had noted your endotoxin notes regarding use of E.Coli in the production process of the Pfizer jabs too. I'll check out your above link... Keep up the great work, I had shared one of your articles in a comment or too regarding anaphylactic shock. Can't believe you still have a Twitter ban, you must be too close to the truth!
Aaaauuugh! (Charlie Brown gif here)
Thanks for all your persistent work Kevin. In my head I now call you Hercule Poirot, for solving one of the greatest mysteries of our time.
The work you are doing is of historic significance and important to humanity. #TheLittleGreyCells