Das Spike-Protein hat diverse Homologien zu menschlichen Proteinen, somit auch zu den entsprechenden Genen. Nun müsste man schauen, was und warum gewisse Codons “optimiert” wurden, um möglicherweise einen solchen miRNA Effekt zu erzeugen.
Fragmente reichen, schätze ich. Auch DNA Fragmente reichen, siehe Antisense Oligonucleotide bei Nusinersen. dsRNA, ssRNA, ssDNA, alles kann problematisch sein, würde ich sagen. Nicht, dass wir das alles wirklich verstanden hätten. Sollbruchstellen müssen nicht sein, das Zeugs wird aktiv von Dicer zerschnippelt.
Es gibt ein Patent, wo BioTNech versucht dsRNA im Produkt zu reduzieren, wodurch mehr DNA Probleme entstehen. It's a mess.
Regarding the "vaccine" spike protein, may I ask a question?
EMA assessment report states that: "The technical procedure for the quantification of the poly(A) tail is considered, in general, sufficiently described but the suitability of this method for the intended purpose remains to be confirmed." What does this mean? Are we sure that, among all these fragmented mRNAs (degraded mRNA? It really doesn't take much to inactivate it) the spike protein can actually be expressed?
I read that the vaccine Spike would be detected in the form of an immunecomplex produced by an artificial antibody (which refers to software that integrates sequence gaps on a statistical basis, correct?) in culture of cancer cells, not healthy cells. Correct? Does it make a difference?
Ogata's study used the SiMoA method to detect the vaccine spike, however I read that the capture antibody is ideally specific only for our protein of interest and does not bind to other things (like what?) Then the beads are "washed" to eliminate anything that binds only weakly to the capture antibodies. After the washes, the hope is that only things that bind strongly to the capture antibodies will remain. Then another "detection" antibody is used and numerous washes follow. In addition, Ogata et al. explicitly state this: "Our Simoa antigen tests cannot detect antigen-antibody immune complexes". My doubt is, based on these methods and what the EMA wrote, what certainties can we have about the presence of a "vaccine" spike protein?
Yes, the spike protein can be detected by immunolgical staining. In German there is a pathological atlas available showing some 60 cases: https://www.histo-atlas.com/
In many cases the spike protein was detected but not die nucleocapsid protein.
Immunostaining works by detecting the target with a specific antibody and then detecting this antibody either with a precipitation of a chemical or a secondary antibody which can be detected with horse raddish peroxidase or a fluorescent marker. This is old school and which secondary antibody is used depends on the detection method used in the lab. The Mörz paper uses the method Arne Burkhard used. The paper descibres the sample preparation.
BUT
you will not be able to detect misfolded spike protein missing the antibody recognition sides. There will be species which will be not detectable due to the +1 frame shift or alternative splicing (as described by JJ). We can only detect more or less correctly folded spike protein.
Another doubt I had concerns the cold chain to maintain the construct correctly. I've read that mRNA-LNP can be studied using the cryoTEM image technique which involves temperatures at - 153° Celsius. "Even though in the majority of approaches in electron microscopy one tries to get the best resolution image of the material, it is not always the case in cryo-TEM... That is why the probabilistic approaches become more powerful in this type of investigation" (Wikipedia).
The stability of the structure is very important. If the structure degrades, the delivery of the active ingredient does not occur.
How can we be sure about the whole structure? And that the whole structure is maintained throughout the manufacturing, transport, pre- and post-injection process of this drug, up until the moment the mRNA is released into the cell? The contaminations detected, the aldehydes that are generated in the ALC, the hydrolysis, can inactivate the mRNA? Thanks in advance!
Also for Moderna. There is something going on with that 3' UTR and they have patents on using miRNAs in LNPs I saw somewhere.
I was at an RNA conference last week and presented a poster regarding the regulatory assessment, or lack thereof including transcription, translation, dsRNA the SV40 sequences and lipid issues. Right next to mine was a researcher working in Ambros's lab. And he had bad Bell's Palsy from the jab though he had been gaslight that it was from Covid. We discussed the jab over the 3 days he was there. Hope he brings it back to his the lab. The world is very small, isn't it?
Yea, I read you substack on that. Brave to go into the lions den. How can it be that those "Experts TM" are so ignorant?
non-coding RNA ist a very dangerous area to mess with. I worked as administration for the Hess Lab in Freiburg. They worked on non-coding RNA in cyano bacterie. Stefanie Klein was in that group. She is the master of the RNA-Library of BioNTech.
Ich kann es mir nicht vorstellen, dass ich, als alleinstehende Forscherin, so viele Probleme finde, und dass die "Zustaendigen" das nicht auch wissen!!!!
Habe das Buch gelesen. Die miRNA Frage spukt mir schon lange im Kopf, zumal ich da von der australischen FOIA vorgetriggert bin. Der Nobelpreis war einfach DIE Gelegenheit das PEI damit zu trollen.
Was passiert eigentlich mit der modRNA nach dem Ablesen? Speziell das pseudo Uridin... wird das ausgeschieden oder - Gott bewahre - im Zellkern zur RNA Synthese verwendet? Gibts da was dazu? Danke für die Hilfe!
BioNTech patent (google WO2021213945A1) includes (quote)
In one embodiment, the removal of dsRNA from non-immunogenic RNA comprises a removal of dsRNA such that less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.3%, or less than 0.1% of the RNA in the non-immunogenic RNA composition is dsRNA.
Frage:
Ist es überhaupt nötig, dsRNA für Existenz und Bedeutung von RNAi zu postulieren?
Ich denke, es wäre relativ leicht, Sollbruchstellen in die modRNA einzubauen. Dann hätte man rasch RNAi.
Das Spike-Protein hat diverse Homologien zu menschlichen Proteinen, somit auch zu den entsprechenden Genen. Nun müsste man schauen, was und warum gewisse Codons “optimiert” wurden, um möglicherweise einen solchen miRNA Effekt zu erzeugen.
Das beantwortet noch nicht die Frage:
"Ist es überhaupt nötig, dsRNA ..."
Ginge das nicht auch via Sollbruchstellen?
Fragmente reichen, schätze ich. Auch DNA Fragmente reichen, siehe Antisense Oligonucleotide bei Nusinersen. dsRNA, ssRNA, ssDNA, alles kann problematisch sein, würde ich sagen. Nicht, dass wir das alles wirklich verstanden hätten. Sollbruchstellen müssen nicht sein, das Zeugs wird aktiv von Dicer zerschnippelt.
Es gibt ein Patent, wo BioTNech versucht dsRNA im Produkt zu reduzieren, wodurch mehr DNA Probleme entstehen. It's a mess.
Generell sind wir hier im Bereich der non-conding RNA, die man zusammenfassen kann mit "Anzahl und Funktion unbekannt".
Regarding the "vaccine" spike protein, may I ask a question?
EMA assessment report states that: "The technical procedure for the quantification of the poly(A) tail is considered, in general, sufficiently described but the suitability of this method for the intended purpose remains to be confirmed." What does this mean? Are we sure that, among all these fragmented mRNAs (degraded mRNA? It really doesn't take much to inactivate it) the spike protein can actually be expressed?
I read that the vaccine Spike would be detected in the form of an immunecomplex produced by an artificial antibody (which refers to software that integrates sequence gaps on a statistical basis, correct?) in culture of cancer cells, not healthy cells. Correct? Does it make a difference?
Ogata's study used the SiMoA method to detect the vaccine spike, however I read that the capture antibody is ideally specific only for our protein of interest and does not bind to other things (like what?) Then the beads are "washed" to eliminate anything that binds only weakly to the capture antibodies. After the washes, the hope is that only things that bind strongly to the capture antibodies will remain. Then another "detection" antibody is used and numerous washes follow. In addition, Ogata et al. explicitly state this: "Our Simoa antigen tests cannot detect antigen-antibody immune complexes". My doubt is, based on these methods and what the EMA wrote, what certainties can we have about the presence of a "vaccine" spike protein?
Concering the problems on the protein level, there are a few more problems than "just" the modRNA https://drbine.substack.com/p/ugurs-grenzdebile-schwachsinnsideen-b2b
Yes, the spike protein can be detected by immunolgical staining. In German there is a pathological atlas available showing some 60 cases: https://www.histo-atlas.com/
In many cases the spike protein was detected but not die nucleocapsid protein.
Dailyclout also has some articles on that. Michael Mörz published a paper on an interesting case with stainings. https://pubmed.ncbi.nlm.nih.gov/36298516/
Immunostaining works by detecting the target with a specific antibody and then detecting this antibody either with a precipitation of a chemical or a secondary antibody which can be detected with horse raddish peroxidase or a fluorescent marker. This is old school and which secondary antibody is used depends on the detection method used in the lab. The Mörz paper uses the method Arne Burkhard used. The paper descibres the sample preparation.
BUT
you will not be able to detect misfolded spike protein missing the antibody recognition sides. There will be species which will be not detectable due to the +1 frame shift or alternative splicing (as described by JJ). We can only detect more or less correctly folded spike protein.
Thank you for your kind reply.
Another doubt I had concerns the cold chain to maintain the construct correctly. I've read that mRNA-LNP can be studied using the cryoTEM image technique which involves temperatures at - 153° Celsius. "Even though in the majority of approaches in electron microscopy one tries to get the best resolution image of the material, it is not always the case in cryo-TEM... That is why the probabilistic approaches become more powerful in this type of investigation" (Wikipedia).
The stability of the structure is very important. If the structure degrades, the delivery of the active ingredient does not occur.
How can we be sure about the whole structure? And that the whole structure is maintained throughout the manufacturing, transport, pre- and post-injection process of this drug, up until the moment the mRNA is released into the cell? The contaminations detected, the aldehydes that are generated in the ALC, the hydrolysis, can inactivate the mRNA? Thanks in advance!
There are two articles on this on my substack
https://drbine.substack.com/p/die-plorre-ist-stabiler-als-gehofft
https://drbine.substack.com/p/die-plorre-ist-wohl-innen-flussig
Also for Moderna. There is something going on with that 3' UTR and they have patents on using miRNAs in LNPs I saw somewhere.
I was at an RNA conference last week and presented a poster regarding the regulatory assessment, or lack thereof including transcription, translation, dsRNA the SV40 sequences and lipid issues. Right next to mine was a researcher working in Ambros's lab. And he had bad Bell's Palsy from the jab though he had been gaslight that it was from Covid. We discussed the jab over the 3 days he was there. Hope he brings it back to his the lab. The world is very small, isn't it?
Yea, I read you substack on that. Brave to go into the lions den. How can it be that those "Experts TM" are so ignorant?
non-coding RNA ist a very dangerous area to mess with. I worked as administration for the Hess Lab in Freiburg. They worked on non-coding RNA in cyano bacterie. Stefanie Klein was in that group. She is the master of the RNA-Library of BioNTech.
Makes you think.
Publikations of the Hess Lab: https://pubmed.ncbi.nlm.nih.gov/?term=Hess%20WR%5BAuthor%5D
Freiburg RNA tools: a central online resource for RNA-focused research and teaching
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6030932/
The Link between Hess Lab and BioNTech Stephanie Hein
https://pubmed.ncbi.nlm.nih.gov/?term=Hess+WR%5BAuthor%5D+HEIN+S&sort=pubdate
https://pubmed.ncbi.nlm.nih.gov/?sort=pubdate&term=Hein+S&cauthor_id=37164012
Wow, thanks.
That Thomas Cech was creepy. Wanted non-coding RNA to work on our telomeres and so become immortal. Gah.
It's easier with BCAA. But why easy if you can have it dangerous and complicated.
At Annelise's suggestion I discuss interactions with a few lncRNAs, including:
LncRNA H19 upregulation
LncRNA FENDRR
LncRNA HOTAIR
LncRNA MIAT
LncRNA APOA1-AS
LncRNA LINC01505
https://doorlesscarp953.substack.com/p/covid-19-long-non-coding-rnas-and
The lack of understanding or interest is mind blowing. This is not good!
J&J found an alternative splicing side. Did anyone check for alternative splicing?
Wie immer, danke!
Ausgezeichneter Artikel ueber RNAi:
https://www.sciencedirect.com/science/article/pii/S0160412019306038?via%3Dihub
Hab ich als Basis fuer mein Buch verwendet, und im Zusammenhang der mRNA "Impfungen" - dsRNA, Bruchstuecke, ... analysiert.
Fuer mich, wirft extreme Fragen auf!!!! Auch bzgl. Umweltbelastung... (https://www.calameo.com/read/006649539dfd6e1625dc6).
Ich kann es mir nicht vorstellen, dass ich, als alleinstehende Forscherin, so viele Probleme finde, und dass die "Zustaendigen" das nicht auch wissen!!!!
Habe das Buch gelesen. Die miRNA Frage spukt mir schon lange im Kopf, zumal ich da von der australischen FOIA vorgetriggert bin. Der Nobelpreis war einfach DIE Gelegenheit das PEI damit zu trollen.
Endotoxine > microRNA > über 10.000 verschiedene Schäden
Was passiert eigentlich mit der modRNA nach dem Ablesen? Speziell das pseudo Uridin... wird das ausgeschieden oder - Gott bewahre - im Zellkern zur RNA Synthese verwendet? Gibts da was dazu? Danke für die Hilfe!
BioNTech patent (google WO2021213945A1) includes (quote)
In one embodiment, the removal of dsRNA from non-immunogenic RNA comprises a removal of dsRNA such that less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.3%, or less than 0.1% of the RNA in the non-immunogenic RNA composition is dsRNA.
So that’s as clear as muck.