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+ | **Type:** Conference presentation\\ | ||
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+ | **See also:** [[https:// | ||
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+ | ===== Transcript ===== | ||
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+ | Thank you. I am going to talk about the VDR receptor. It’s one of the family of nuclear receptors, which includes the glucocorticoid receptor, the thyroid receptors and a number of other very important receptors. | ||
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+ | I just want to point out this statement from FDA commissioner Von Eschenbach to congress two years ago, pointing out that “New scientific discoveries are generating an emergent science of safety, where the new science combines an understanding of disease and its origins at the molecular level”. That is what I am going to talk about in this presentation: | ||
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+ | There are three types of biology in common use today, in-vivo in-vitro and the newer in-silico. The first time I came across in-silico biology was back in 1981. This is a photograph of myself and my colleagues at the Hospital for Sick Kids back in 1981, which was when IBM showed us the in-silico techniques that they had used for the synthesis of the first human insulin. The Humulin, the first human insulin. That was the first time I came across in-silico and realized the power of being able to emulate the operation of the human body at the level of individual atoms. | ||
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+ | Since then the in-silico work I think that everybody is most familiar with is the decoding of the human genome and probably more important now, but less well known, is the decoding of around 740 microbial genomes that have been fully decoded to this point in time. | ||
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+ | And in fact, NIH has just started a big Human Micobiome Project, with the idea that they wanted to characterize all the DNA that’s available from human sources, all the DNA in the human body. The normal infectious areas that we are aware of: the nasal, oral, skin, GI and urogenital cavities, but also within the cells of the body itself. Because NIH has estimated that around 10 percent of the total cells in the human body are human cells and as many as 90 percent of the cells could be bacterial cells. You will be hearing a lot about the Microbiome project over the next decade. | ||
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+ | Why is that important? Well it’s important because the bacterial cells, in many cases, perform functions that are very similar to those of the Homo sapiens itself, of the host. And what I have got here is a slide showing the E.coli Glucose Metabolism. And I know very few of you are going to follow every subtlety of it. Don’t worry; I just wanted to show you that this chart exists. I got this one from Vijay at the Bielefeld University in Germany. But it shows the way that the bacteria E.coli gets from Gluco-6-P substrate down to Pyruvate and produce the Serine Cystein Glycine amino acids, the Purine Nucleotides Adenine, the Tyrosine Phenylalanine, | ||
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+ | And that’s exactly what we have found. We have characterized that there is an intra-phagocytic metagenomic microbiota. Metagenomic: | ||
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+ | At the congress in 2004 in Budapest, I reported that sarcoidosis had succumbed to an antibacterial therapy that we had developed. And over the last six years our cohort of over 500 human subjects has demonstrated reversibility of many auto-immune diagnosis. Reversibility, | ||
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+ | But surprisingly, | ||
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+ | Well, what do these microbiota look like? Here is a monocyte, an infected monocyte and the cytoplasm has effectively exploded from pressure of the pathogens and it is throwing out these tiny biofilm tubules. I’m not quite sure what, it’s not showing up perfectly but you can see here is probably a dozen tiny biofilm tubules. They are very long and they’re also extremely small. You can compare them with the size of the cell, the standard cell diameter there of 4 - 5 microns or so. And extremely tiny little biofilm parlemor tubules are thrown out as the cell disintegrates. This is untreated blood, this is human blood put between cover slips and allowed to age for six to thirty-six hours. Look at the length of these; this is about 26 cell diameters long. This one is about ten diameters long. It’s amazing and you can see them very easily under light microscopy if you are looking for them. | ||
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+ | Under the electron microscope, they look a little bit different of course. Because you can actually look at them in the cells before they become heavily parasitized. This is an image from the Emile Wirostko TEM study in 1989 at Columbia University. This is a Juvenile Rheumatoid Arthritis lymphocyte. And what we have: we have the nucleus area of the lymphocyte. And then the cytoplasmic region outside, the nucleus and in the cytoplasmic region there is a staining artifact which is basically nucleic material, DNA material, inside some form of transparent biofilm type of protection and then a very thin exoskeleton to contain the whole lot. But what’s even more interesting are these tiny little elongated structures – which don’t come up all that well on the projection here, but when you look up the original paper -which was JRA Inflammatory Eye Disease, Parasitization of Ocular Leukocytes by Mollicute-like Organisms from 1989- when you look at the original photographs you can see the incredible detail of these transparent colonies that are living inside these lymphocytes. Of course, the lymphocyte should never allow this to happen. But it does allow it to happen because these microbiota have figured out how to overcome innate immunity. They have overcome the last line of defense. | ||
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+ | Well, how does it do that? In Homo sapiens the VDR nuclear receptor transcribes genes for the Cathelicidin, | ||
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+ | The microbiota changes the expressions of greater than 913 genes. And those are genes which do everything from create a parathyroid hormone precursor, right through to create the Missing in Metastasis protein. MTSS1 is the gene; Metastasis Suppressor number one. | ||
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+ | All of these genes, the 913 that have already been confirmed, are transcribed by the VDR. The VDR nuclear receptor is a very key nuclear receptor in Homo sapiens. But homeostasis of the other type 1 nuclear receptors isn’t directly upset by these pathogens. The VDR of course but the PXR, the Pregnane Xenobiotic Receptor, the Glucocorticoid Receptor, Thyroid-alpha-1 and Thyroid-beta-1 are all profoundly affected by the elevated levels of the seco-steroid that are caused by the VDR being knocked out. And obviously, note especially a loss of Glucocorticoid and Thyroid homeostasis leads to the diagnosis of hypothyroidism and adrenal insufficiency. We have demonstrated that both of these are reversible. | ||
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+ | So why haven’t we seen this microbiota before? There has been so much study of pathogens in mankind. Well there are a few reasons. The first one is that the VDR homology, the shape, the amino acids that go together to make up the VDR, is a little bit different in Homo sapiens to what it is in all of the other mammals and all of the other fish etc. as well which have VDR. And it transcribes different genes from the VDR of the mammals. And you know how much of our work we have been doing in animal models. Well, a very key-function of the bacteria that have evaded the human immune system do not appear in mammals because the VDR homology is so unique to Homo sapiens. The VDR from the murine and canine genomes for example doesn’t transcribe Cathelicidin or the Defensins, at all. So a human metagenomic microbiota won’t survive if it was transfected into, for example, a mouse. Because different species and different mutations would be necessary if the microbiota was to knock out the different gene pathways needed for survival in a mouse. | ||
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+ | Further, the microbiota is only stable in-vivo. It defies extraction using standard techniques. You saw how that cell had disintegrated in about six hours of aging. You can imagine what it does under centrifuge. Further, most of these species in the biofilm microbiota defy attempts in-vitro (culture). This is a study from Dempsey et al, which was a study of biofilm from prosthetic hip-joints, which were removed during revision arthroplasties. And they did gene sequencing and tried to match up the 740 known genomes that we have for bacteria against what they found, the DNA that they found in the biofilm. And this is what they found. Lysobacter. Lysobacter was about 44 percent of the clones that were sequenced. Proteobacterium, | ||
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+ | Until the genome was cracked, we only had the postulates of Koch as a guide. They caused us to search for Koch’s singular pathogenic species. We kept looking for one pathogenic species. As a result you can find a paper that will blame poor old EBV for just about every disease known to man. And CMV and HHV as well. Because Koch’s singular pathogenic species in this age of the genome really means very little. It sidetracks science from understanding horizontal transfer of DNA within the microbiota. Sharing of genes between the organisms, which we now know occurs very, very much faster than we ever could have dreamed to be the case. Science became fixated on the co-infections, | ||
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+ | Another reason is that Vitamin D is the primary ligand that activates the VDR receptor. And at some stage during the twentieth century mankind decided that Vitamin D was nutrient. Well, Vitamin D is not a nutrient. It is a seco-steroid transcriptional activator. And its concentrations are very closely controlled by a very complex control system which involves not only the VDR but also the Pregnane X Receptor, the Pregnane Antibiotics Receptor, the P300/CBP PKA pathways and feedback via a number of enzymes. CYP24, 27, A1, 27B1. There is transrepression from VDR activation, feedback path, there is transrepression or actually antagonism, receptor antagonism from the metabolites. And there is also feed-forward pathways. Quite a complex mechanism… If Vitamin D was a nutrient we would see a simple first order mass action metabolism. We do not see that. We see the complex control system of a hormone. Of a seco-steroid transcriptional activator which is what it is. When we concentrate on the concentration of this intermediate substrate, 25-hydroxy-vitamin-D, | ||
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+ | But, there is another problem too. Only 1, | ||
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+ | Well, luckily there is an agonist that works in-vivo. It’s a drug called Olmesartan. Here we have a molecular dynamics emulation of the human VDR with Olmesartan sitting in the binding pocket in an activated position. As you know, all proteins are in motion, at all times. | ||
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+ | And, here we have the same thing in rat (Rattus Novegicus). It’s a very, very similar VDR and a very, very similar ligand, positioned, but they are not quite the same. And when we put them side by side you can really see the difference. In particular: look at this Tetrasol. The tetrasol-ring is a totally different orientation because it’s binding to totally different amino acids in the VDR of the rat and the VDR of the human being. | ||
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+ | It’s only by getting down to the level of the molecules that we can really understand the difference between our animal models and Homo sapiens. | ||
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+ | Then, once the innate immune system has been activated again, we can use very low-dose bacteriostatic antibiotics to block protein synthesis. Here I’ve got Azithromycin blocking the 70S-ribosome which translates RNA into proteins in the bacterial organisms. They are being blocked by Azithromycin. | ||
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+ | The rate of bacterial death when you are using bacteriostatic antibiotics is controlled by inhibiting the protein synthesis and we can use sub-inhibitory, | ||
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+ | But just remember that one bacterium is weakened if just one antibiotic molecule is bound into one ribosome (because these antibiotics actually block the functioning of protein generation). So intermit low doses can proportionally control the rate of bacterial death. And that’s very fortunate. Because recovery isn’t easy. There’s a huge bacterial cellular load, whether it’s ninety percent of the body or not I would not know but it is a huge load. As the intra-cellular bacteria are killed, some of the infected cells undergo apoptosis, some even disintegrate. | ||
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+ | The loss of cells -both white and red cells- and the cytokine storm which is concomitant with that, has to be controlled so it doesn’t become life-threatening. The damage is called immunopathology. And people who are seriously ill carrying a heavy bacterial load (which isn’t about every individual with an autoimmune diagnosis), they need to spread the therapy over many years if the immunopathology is to be kept at a tolerable level. | ||
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+ | You can’t just give the patient the anti-biotics, | ||
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+ | Finally, I want to point out that what we have done is evolutionary. It’s not revolutionary. We have built on the work of people like Wirostko, Yahooda from back over the last couple of decades… Many of you, I’m sure, too in the audience… Our model, the molecular in-silico model of the disease fits your data! Please seek out my colleagues over the next couple of days. Talk with us about your data, your studies, especially if you think that our model does not stand up to scrutiny. We would love to discuss it with you. It does fit your data! | ||
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+ | And, finally, we’ll end this presentation contemplating Newton. Thank you. | ||
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