What were you doing during the start of the pandemic lock down? Did you build a puzzle for the first time in year? Did you get reacquainted with how stupid and long and frustrating monopoly is? How about sour dough bread? Did you make your first sour dough? Dr. Igor Kovalchuk and his wife, Dr. Olga Kovalchuk from the University of Lethbridge were locked out of their labs too. Their lock down side project was a little more ambitious. They had been researching hundreds of cannabis extracts to see if these extracts reduced inflammation and might help treat conditions like multiple sclerosis, psoriasis, inflammatory bowel disease and rheumatoid arthritis. During this forced pause in their research, they wondered in CBD rich cannabis extracts might be useful against SARS-CoV-2, the virus that causes COVID-19. You want to hear what they found out.
E53 - Does CBD have COVID-19 ACE'd?
- Dr. Kovalchuk's preprint article we discussed - In Search of Preventative Strategies: Novel Anti-Inflammatory High-CBD Cannabis Sativa Extracts Modulate ACE2 Expression in COVID-19 Gateway Tissues
- Dr. Matthew Elmes critique of Dr. Kovalchuk's paper - Cannabid & COVID-19: Breaking News or Bogus Science?
Music ByNelson Rudiak
(Yes we have a SOCAN membership to use these songs all legal and proper like)
E53_CBD and COVID.mp3
Trevor: Kirk. We're back. Well, I'm back. Where are you?
Kirk: Well, Trevor. Covid has made my life rather interesting. I'm right now currently sitting just north of Brandon and a little community called Forest on Highway 10, traveling to Brandon.
Trevor: All right. Well, Covid restrictions are lifting in Manitoba. So I can physically be in the studio. You've still got to be doing some socially distancing things. So you're not. And that's why it sounds a little different today.
Kirk: My world is basically for four weeks in community and four weeks home. And out of the four weeks home, I have to spend two weeks still socially isolating from friends. So, I basically have two weeks every six in which to live my life. Michelle and I are traveling to Brandon to get some commerce done, visit some grandchildren.
Trevor: That's good. They got to see the grandchildren. So, I'm Trevor Shewfelt I'm the pharmacist. Who are you?
Kirk: I'm Kirk Nyquist, I'm the nurse and we are Reefer Medness.
Trevor: So, Kirk, I brought Igor. I'm really excited about this one. I know it's a little dense and a little science on this one, but we'll do our best to sort of walk people through it. But so, we finally get to say we have a covid episode.
Kirk: Yeah, this has been a fun episode. I've been listening to your interviewed for the last couple of days, and I've actually been doing a lot of research, again, on my own here. And I guess the first thing to talk about this whole concept of pre-print. You and Igor to get into it.
Kirk: And, you know, as I was listening to them all, the pre-print is, to make it in layman's terms, it's basically a very public way to get the peers to review your paper.
Trevor: Yeah, it's a, pre-covid I'd never heard of the word pre-print. And like Igor said, he has published over 150 peer reviewed articles. But pre-prints, especially in this world of covid, seems to be a way to kind of dump your info out there for the world to read. But still, if you want to go into a journal later, it's still got to go through the peer review process. It sounds like you're still going to, they'll basically check your spelling and make sure it's in the right format. But they're not looking they're not doing any of the fact checking the peer review usually does. And like you're saying, and it's a speed thing. Right. Because he said, you know, when normally gets a peer reviewed paper, it is you know, you submit it, a paper and they like it. They don't like it. They send you. And, you know, the whole process can take, you know, literal years, one to three years to get your thing through peer review. So not surprising. Right now, we want as much covid info out there. And the quote I heard was there's like four thousand pre-print a week covid related going on. So that's great. We can you know, we the public can see all this info and the scientists and the doctors can see all this info. But the downside is basically, I think the easiest way is it hasn't been fact checked yet.
Kirk: Well, but, peers are reviewing these pre-print papers. So, you are getting your peer review done. You're just doing it in a very public way.
Trevor: Well, yeah, but by peer review, they specify and I've never published a peer review paper. So, all you publish, scientists can yell at me. My understanding is peer review is specifically going to be four or five people that publication has picked to look at your paper, to fact check it, to ask you a bunch of questions and bunch of correspondence goes back and forth and then the publication will choose to either accept or reject it. So and I don't think whether or not you've done a pre-print directly affects whether or not you're eventually going to be published. And, you know, let's say Nature, one of the other big journals.
Kirk: No, no, I agree with you Trevor, but I guess I see I see peer review as being a a cloistered event where, you know, hifalutin of like-minded read a paper and ask you a bunch of technical questions. From what I hear in this interview, Igor is getting some criticism through peers in this pre-print. So although it has to go through the process of peer review, peers are reading these papers now and giving him give me insights and giving him criticism now.
Trevor: They are, and I'm going to give Igor full credit that he agreed to our interviews. One of the things I did, and I'll make sure it's in the show notes is there was another article basically written about Igor's study in a more populous place, a place called Project CBD, a place I really quite like going for info. And basically it was another PhD biochemist saying how nicely saying how much he didn't like Igor's paper. So rather than me, try to come up with a bunch of questions for Igor, I just sort of sent him the paper and said, I will talk about this. And, you know, he graciously said, sure, we can talk about the problems in the paper. So I command Igor for, you know, happily talking about problems that have been pointed out.
Kirk: Yeah, no. And I guess that's the whole point of a pre-print, right is to get it out there because as you said right now with covid, scientists are having a tough time getting into the lab. And I guess I mean, could you imagine this this fellow's been studying cannabis for five years and all of a sudden in one of these experiments, he realizes that cannabis, you know, the exact CBD extract had an effect on an ACE2 inhibitor and it could be beneficial. Gee, I need to let people know about this? Although there are flaws, I need people look at this. Yeah. This is what he's done.
Trevor: Well, and I think this is a fascinating covid story. This is the part I really like, is, you know, we've got a bunch of people out there, you know, learning how to make sour dough bread because they're stuck at home. Igor and his wife, you know, they'd been studying cannabis and they'd been looking at things like MS and psoriasis and rheumatoid arthritis, you know, basically diseases, they're pro inflammatory. And he, they were thinking that, you know, something in these cannabis extracts, especially the high CBD ones, was anti-inflammatory and might help. And so, you know, they've already got this database. They've got, you know, four hundred different extracts and they're, you know, narrowed it down to two dozen that they think are really good anti inflammatories and that now suddenly they're stuck at home. And Igor's wife, Olga, who is another professor, finds some papers saying, you know, back when it was SARS one, you know, this ACE2 receptor on the cells was really important for the virus to get in. I wonder if it'll be important for this and then, you know, does anything in our database affect ACE2? And then then the scientists in China and Germany confirm that ACE2 is important for the sars-cov-2. And so they've now got an idea and they can't really get into the lab to test it. And then suddenly they can get into a lab like two people at a time and run a couple experiments. And they do and they publish it. So, you know, instead of learning to do sour dough, these two, you know, learned a little bit of a virology.
Kirk: What they're getting a lot of press on this. You know, if you Google, covid and ACE2 these guys come up. You know Trevor, the funny thing about, not the funny thing, but the observation when you're when you're in the medical field long enough, you're able to say you've seen it all or seen it before. I started I started nursing in 1980 just as the HIV AIDS epidemic was beginning. And in these 40 years from my time starting nursing, when, you know, the joke about doing CPR on an HIV patient was not a great joke. The thing was, nurses were refusing to see AIDS patients, HIV patients. In these 40 years now an HIV AIDS patients can actually live a long and fruitful life because we've done some incredible studies on HIV AIDS. Now we're covid and now we got these treatments. And, you know, vaccines that usually take five or six years to come to market are coming to market in six months. So I think we're living in an interesting time. And I find it fascinating that cannabis is there too. Helping cannabis help with covid. OK, Trevor I've been doing some studying. Let's go for let's help our listeners. Let's do a little review on an ACE inhibitors versus an ACE2 inhibitor.
Trevor: OK, well, so in pharmacy world, an ACE inhibitor is a blood pressure pill and ACE means Angiotensin Converting Enzyme. So that's just an enzyme that lives in your body. And a ACE inhibitor blocks that enzyme. But the enzyme it's blocking is called ACE one. So ACE one isn't, as far as we know, directly related to sars-cov-2. We think sars-cov-2, the gateway or the doorway that it gets into the cell with is a different enzyme or different receptor called ACE2. There's been some debate and study out there whether or not blood pressure pills called ACE inhibitors affect ACE2. There was some worry early on and in the pandemic that everybody on ACE inhibitors was going to get covid and die. That didn't happen. In fact, there's been some big studies published in JAMA and a few other places basically showing, no, it doesn't have any bad effects. And a paper to that you were pointing out this morning. Now there's some people out there looking to see if maybe ACE inhibitors might actually have some good effects covid wise, but that's far from proven. So right now, I think all we can say safely is ACE inhibitors seem to have no effect on whether or not you're going to get sick with covid-19 because they seem to have no effect on whether or not the sars-cov-2 virus is going to get into your cell. And like everything else, covid related subject to change.
Kirk: OK, so let's look at this a little bit. As a layman, we have skeletal muscles and we have smooth muscles. Skeletal muscles are obviously what move our arms and legs, right? We flex and we extend muscles. And those are the skeletal muscles. Now, ACE receptors are on the smooth muscle. Right. So when you talk about what's an example of an ACE inhibitor pill that you might that you might dispense to somebody and.
Kirk: The prills. The prills affect the blood vessels, right. To smooth blood vessels of our body. And that's how it affects blood pressure, is by dilating the blood, the arteries and therefore the heart doesn't have to pump as hard. So that's what ACE receptor is, right? It's found in the smooth muscle.
Trevor: Yeah. Does a few other things as well, including stuff in the kidneys and a few other things. But yes, it affects the blood pressure.
Kirk: But those are all smooth muscles, right?
Trevor: Well, yeah, we think some of the things it does in the kidneys are more hormone related and some of the things it does on the heart, it's a complicated pill that it seems like it's ACE the enzyme is literally all over the body. So it seems to have a lot of a lot of complicated effects.
Kirk: Kind of like CB2 receptors. all over the body.
Trevor: Something like that. Yes.
Kirk: Okay. So ACE2 is found in the tissues of your lungs. And the tissues of the kidney, tissues of a gastro intestinal tract so Igor is looking at how covid and CBDs affect these ACE2 receptors. Right. And that's where this is a little different.
Trevor: Right. And so what we think and Igor agreed I had this about right. The way I like to think about it is the ACE2 is a doorway into the cell and the sars-cov-2 virus uses that doorway to get into the cell. Now it needs a key to get in. And the key is this famous spike protein you keep hearing about on the news stuck all around the outside of sars-cov-2. But that key needs to be cut before it'll open the door. And that's this TMPRSS Protein. So we they were looking to see if the cannabis extract can make less doors, less of these ACE2 receptors and less key cutters. And so if you can make less of both expressed less of those on the cells, does that mean we can have less sars-cov-2 virus getting into the cells?
Kirk: OK, all right. That makes sense. So I guess we should like to hear what he has to say.
Trevor: All right. Let's go to Igor.
Igor: Nice to talk to you guys. Good morning I'm Igor Kovalchuk a Professor at University of Lethbridge in Alberta. And so are they in the university since 2001, research and medicinal herbs since 2008, working ? And for the past five years working on cannabis.
[00:13:33] Thank you, Igor. That's great. Now, you got you and your team, which has lots of Kovalchuk in it. I've got an Anna and Olga and Igor. You put out a paper talking about using extracts of cannabis that had lots of CBD in it and how that might affect ACE2 expression. So let's just to keep get our audience up to speed. First thing, I just want to ask because honestly, before covid-19, I don't think I'd ever heard of it before. Tell us what the difference is between a pre-print and what we normally consider a peer reviewed article.
Igor: So frankly, I also did not well, I heard about pre-print, but never used one before covid. So the preprint basically it's a research publication that is not peer reviewed, but that you can put online. So there are probably five or six journals that publish pre-print and they just basically check them for correct format and whatnot. But they don't check hypotheses or the data and whatnot. So technically that those journals that post the pre-print are not experts in the specific field, so that's why this research is not peer reviewed. Right. In contrast to other journals that peer reviewed. So typically for the paper, it goes to your peers. They criticize the paper, ask you to do something else, change this, change that. You either published in the same journal or elsewhere. And I have published over 150 peer reviewed articles. And on average, a submission goes through three to five journals right. You send to one, and we end up in journal number ten three years later. This is a very typical and very standard way how science is done. The only reason we've done the pre-print publications without the peer review because of the urgency we wanted to people know the flexibility. We wanted to expose ourselves to criticism and what importantly, we want it to attract the attention of researchers and, you know, media and whatnot so that we are able to get research money to conduct a proper clinical trial to actually confirm our findings.
Trevor: OK, no, that's great. And so and the other thing, I just I mean, again, I hadn't heard of pre-print before either. But I'm saying like just like you said, in this era of covid, there's now more than 4000 covid pre-print a week. So, you guys are certainly not the only people putting out pre-print in this in this covid time. So, let's talk about let's talk about ACE2 and TMPRSS. ACE2, jump in and correct me, I am just a pharmacist here, jump in and correct me when I'm going off the rails. But ACE2, we think is kind of the doorway into the cell that the sars-cov-2 virus uses and it sort of opens that door with the spiked protein, and we think that one of the things that kind of helps cut that key is the TMPRSS protease, the enzyme. So ACE2 lets the virus into the cell and this other protein, the TMPRSS helps make the spike protein open up that door might kind of on the right track.
Trevor: You nailed it. OK, good. Thank you. So, so, so now so the point of your paper and again tell me when I go off the rails, you wanted to see. What you found. See if we can with things like CBD, with cannabis extracts, if you can have less ACE2 doorways and possibly less TMPRSS key cutting protein. That's sort of what we're aiming for.
Igor: Absolutely. So, if you understand a lot of pathogens enter the human cells through specific interaction with receptors in the surface of the cell. The pathogens are typically tissue specific. Right? That's why we have viruses like cold causing viruses attack, your pharyngeal and lung tissues. We have hepatitis virus attacks, liver, HIV, immune cells and so on and so forth. So, tissue specificity is granted by a recognition of receptor on the surface of the cell. And that's what this virus does. So, it's a recognized ACE2. In addition to recognition for the virus to get in and starts replicating, it forces other cells to amplify the receptor. Right. So that means when viral particle aggravate the infected cell, they have better chances to, infect other cells around it. Right. So, when we have thought that cannabis extract can decrease the number of the receptors. We have two purposes. For one, we thought that there's a potential for some sort of reduction in infection disability. So, for people who take the extra. And second, the reduction off the possibility to develop a disease or develop a moderate or severe form of disease, because even if the person that is infected that partake in the extract, there is a possibility that it is not checked yet. Not proven yet. But there is a possibility that taking the extract would reduce the actual disease development because simply it reduces the receptor expression on other cells, the virus has a difficult time to infect you systemically.
Trevor: OK, no, that's good. Now, how about let's take a step back. Why would you or someone on your team have thought that cannabis extracts might affect ACE2 receptors at all?
Igor: Right. So basically, that comes from our previous work where we have tested various extracts for their ability to reduce inflammation, for variety of conditions. Whether the chronic or acute, different type of chronic inflammation. And we were just preparing the clinical trials on diseases like rheumatoid arthritis, multiple sclerosis, skin conditions like psoriasis. So we accumulated a lot of data. Profiles over 400 cultivars and basically decrease the number of affected cultivars to about two dozen. So we knew that cannabis extracts, again on our models, simple cell tissues have a very strong potential to regulate the variety of inflammatory process. So our initial idea was just to use the cannabis extract to reduce inflammation and therefore severity to the disease. But when Olga Kovalchuk, my wife and business partner and colleagues started reading more on how the sars-virus itself from back 2004 or five interacts with, so she found a couple of articles showing the importance of ACE2 and she just suspected, back in early February that this virus, even just because of similarity, may do the same. And simply what we have done without any experiment, we started profiling our data for the ACE2 expression an expression of other receptors and identified that in many targeted tissues that we have analyzed before, these receptors can be reduced. And then I believe in two or three weeks after that came confirmation from Germany and Chinese groups that ACE2 would be a critical too. To target, and then they recorded on the second protein as well. And that's when we tested a second protein and also found that it is reduced not to the same degree, but also substantially. So that's basically part serendipity, part basically we have something to do because of the covid, we could not go to the to the lab. We only got a granted permission to get back to the lab, and only two people, in April. And that's when we started to do some additional experiments. And even now, we are still on a very reduced load. So only three people are allowed to work because university is still in shut down.
Trevor: Well, I'm not sure if I'm going to say I'm glad the scientists have the same restrictions as the rest of us. Or maybe we'll just commiserate that like everybody else, you've been more or less stuck at home with small outings. OK? Other things that I again made because I don't read these things often, I had never heard of 3D tissues. So, you were using something called Epi-airway, Epi-oral and epi-interstitial tissues. So, this isn't just a few like mucosal cells in a petri dish. This is something different. Can you sort of describe to our audience what these things are that you were doing your tests on?
Igor: Right. So, the 3D tissues are basically engineered human tissue. So, they come from real patients. Right. So they are made from layers of cells to determine whether it's lungs or intestines. We literally mimic tissues of our organs, right. And so, there are many companies that produce that. And ?? is one of them. And that's where we use the tissue of this company. And so basically they can make it sex specific, right, male or female. They can make it age specific or even a condition specific so they can mimic colitis, psoriasis and many other diseases. Right. So basically, 3D tissue, it's engineered human tissue. Right.
Trevor: Thank you very much. And then before you started applying your extracts to it, you basically caused them to become artificially inflamed with things like tumor necrosis factor. So that was sort of to mimic the tissues being sick. Is that kind of the idea?
Igor: Absolutely. So that's exactly what's happening with manufacturing process. There are a number of questions, whether it’s a bacterial, viral fungal. There is inflammation. So it's the natural response of how our immune system to fight off pathogens. And so we want to mimic invitro, the inflammation process. So please understand this inflammation process is not specific to the virus. It occurs in many different diseases, but specific molecules that are targeted. So, for example, with covid causing virus, it's TNF alfa and interleukin-6. So they're probably most important ones. So when we focus on what molecules are targeted, we focus on these two. But in addition, we profile close to hundreds of different inflammatory molecules and even develop our own computer algorithm, that the computer basically searches for a pattern that is specific for the covid infection. And that was in part was reported in the second paper.
[00:26:08] OK, and might be a detail both just because of the chemical I recognize. So DMSO, was your carrier now as a compounding pharmacy? I actually use DMSO in the lab too and I just thought that was an interest and tell me if I'm wrong. Interesting choice because as a drug, DMSO, we actually treat things like cystitis with it and it is sort of an anti-inflammatory all on its own. So does using DMSO as a carrier sort of interfere with the anti inflammatory effects that you're hoping to have from the cannabis extracts?
Igor: It's an excellent question and that's why a proper control has to be always done and we will see this right. The DMOS cells is basically, as said, it's a carrier that three molecules to cross, let's say, extracellular matrix, cell membrane and whatnot. So, because we're dealing with artificial tissue, we don't have a blood supply, you know, that allows the molecules come in proximity of receptors and whatnot on petri dish. In order to apply the molecules, you need to use something that would allow the active ingredient to efficiently. And so when we get all the comparisons, obviously, we compared the effect of extra DMSO. Right. So there DMSO constituency and there is DMSO extra. And when we explain that there is a reduction by 50, 70 or 80 percent, it means reduction against the control of the DMSO
Trevor: OK, OK, now, so you want to find out if there is going to be less ACE2 expressed. Now, how does that work? I read in the paper you both looked at the amount of RNA and the amount of protein to try to figure out how much less ACE2 was expressed after you after you put on the cannabis extract. So what are you looking at? Like you how do you figure out how much ACE2 is in there and how does the RNA and protein help you figure that out?
Igor: Well, it's an excellent question and it's always a dilemma for many scientists how to correctly presented data on reduction of expression, and that's where a lot of criticism come from as well for many papers, including potentially ours. So if you talk about the expression, the expression in the process of making a transcript from a gene. So, a gene, is a portion of DNA where a specific sequence is located. It encodes a specify protein lets say, right? Not always. So there's genes made from RNA in the process of transcription. And RNA often is an indication that lets say how much of that molecule is in the cell? Right. And then in the process of translation, the RNA is converted to proteins. Right. And protein is basically what the receptor is. So the protein acquires a certain character of what is the restructure and gets embedded into the plasma membrane that becomes trans membrane protein. And it's, of course, additional modifications that make the protein active inactive let's skip that part. But very often the linearity between the level of RNA and the level of protein. But there are many cases where it's not right. And so we just monitor the RNA, which is easy because there are methods that you can do it in bulk for thousands and thousands of molecules.
Igor: It gives you an indication of the entire pathway. But the protein, you have to be one by one, the so-called left-hand block and the quantification is just the action of intensity of the image of the protein that comes from, you know, stayed in the jell and the technicality itself.
Trevor: OK, no, I think we're at least grasping why you did both. And even though the RNA is an easier test to do, it doesn't always correlate exactly to how much ACE2 receptor there is. And the Western blot for the protein is harder to do, but possibly more accurate for the further amount of protein. That makes sense to me. So, a few critiques out of the paper that I sent you there and again, we'll try not to get too bogged down to details for our listeners because no one's looking at graphs in front of us. But just a couple, I guess they were pointing out some and maybe these are just hard things to do in the lab. Like one of your runs, the control solution, DMSO, it's figured 2A and 2B. The amount of DMSO in run A seem to have more RNA and then in run B and you would think that you know, if it was the control it should be the same or very similar. And another one was talking about the protein end of things that your control protein, the GAPDH which looked like it was overloaded, like there was basically too much in too many cells had been run through. So it then hard to tell if you had more ACE2 to protein or less EACE2 protein because the control there was so much protein in it. Can you talk to either of those criticisms and just try to walk through for non-lab people if these are valid or just, you know, problems that pop up in labs?
Igor: Right. It's a great question. And this has details that are typically addressed in the battle continues years, the battle between reviewers and authors. Right. So when you talk about individual DSMO and they don't have the paper in front of me, but I'm just thinking that you probably refer to the fact that seen between different tissues in between different experiments. So when you do an experiment, the only valid comparison is within the same experiment. So even if you talk about the same tissues, given the same proliferation of cells, so in order to make it even so, you distribute them well. So hopefully that your pipett is ideal, look at any instrument and make mistakes. It's unavoidable. So that's how loading becomes an unequal, for example. Right. And when we talk about staining of the gel, using specific, you know, chemical reactions or whatnot, it depends on what side of the gel that is because edges are not adequately covered by the staining or detection solution yada yada. So all these technicalities, there were three or four steps that easily can introduce an error or bias. That's why we do three, four or five whatever experiments. And that time, as I mentioned before, we only had to do two experiments. And that's why there are only two. And because we were not allowed in the lab for a long time and really wanted to have the story out, we addressed the criticism and on the expose ourselves to experiment. So when we give the average, we give the average of two experiments and when you give an image, it was just a representation. So it's kind of saying, well, my calculations are done on these images, so you should trust me. And of course, you automatically expose yourself to criticism, because it's not perfectly like in the case of being overloaded. It would be very easy, like overload means, like so when we basically had this, we could use instead of 10 minutes, it's only 10 seconds. And then you will see a perfect picture exposure without any overload for every scientist now that. Right. So, we didn't want to use this trick like that and just show whatever it was. Right. So, both in both cases, I got the agent, basically a very abundant protein. So, it's very easy to, quote, overload it. And in most of the publications, when you see that people use the trick of light staining to show to send it back. Right. So, I don't want to uncover all the mechanisms. So, you beautify the image because image is irrelevant for confiscation purposes. Right. So, I can debate every point of criticism. But at the same time, I agree with an individual who wrote it, right. Right now, in fact, we are addressing the criticism from both papers from both journals. So, we are in the process of answering the questions for some very constructive criticism. So, there are many questions where they agree with us. And I am certain that this paper will be published, but I don't know whether it will take half a year or year or two.
Trevor: I appreciate how honest you've been with us and I think you've addressed all the questions I had. Now, just kind of an interesting thing. So, you did find that some of your cannabis strains seemed to reduce the expression, but it also looked like you had a couple strains that seemed to increase the expression of ACE2 possibly like, you know, we are currently thinking that cigarette smoking increases the expression of ACE2. Is it possible that some cannabis extracts actually might make it more likely then, for you to get sars-cov-2, or is that something for another or another experiment or another day?
Igor: I think it is possible. I would not I would not use the argument of smoking per say, because the argument of ACE2 increase in smoking was given by tobacco smoke. Right. So, yes, cannabis smoking in part is the same, but combustion products are very different. And we have modulating the fact of active ingredients in terms of cannabinoids and Terpenoids until the experiment of smoking cigarettes that are different on the active ingredients and showing the ACE2 is differently regulated, I would not buy the argument. However, at the same time, absolutely that the point of what we're showing them with different cultivars is playing a very different in their capacity. And mind you, we only show a profile the about 20, 24 cultivars out of 400 that have been preselected already. Right. So when we arrived and the idea of using the cultivars for what we did, we used those that were extremely active against other similar processes with inflammation. So that means that the percentage of cultivars is actually very good, is very small. Right. And in the second paper, we very clearly show anti-inflammatory properties of several lines and present two lines, one where there was no difference and one where the completely opposite effect from not one, not two genes, but on the entire pathway. Some very strong pro inflammatory cannabis line. And mind you that best line with all of the high CBD line. So, again, cannabis is not generic.
Trevor: No, I agree. So where does your research go next? Like you are involved with a couple startup companies? We have Swished.inc and Pathway's research is the hope that after some of your findings have been sort of verified. And is it to go on to some sort of animal or clinical trials? Where do you think this is going to go?
Igor: Right. So, yes. So, we have started animal research last fall with several varieties that were strong anti-inflammatory. We have some preliminary behavioral studies because obviously we want to make sure that there's no changes impairment psychology. We in process right now of another two animal studies primarily again on the safety rather than the efficacy. And so right now, we engaged with two groups from the U.S. preparing clinical trials on prophylactic. Right. So is one specific type of asterisk and for treatment that gave you the specific concentrations and specific varieties to hopefully to prevent side effects and whatnot. So we would not make any claims that this cannabis can be used for Covid until we've done phase two trial and have the results.
Trevor: Thank you Igor. This has been fascinating. Anything else you wish to ask? Thought I was going to ask. Anything else you think are our listeners need to need to hear about the need to hear about your work on cannabis extracts and the sars-cov-2 virus?
Igor: Well, yes and no. So what will probably be the most difficult part and I don't want to call myself a pioneer, but the to expose as a preliminary research is the criticism. And, of course, the vigilantes, like the ones who have made extensive comments of our work and all the nasty comments. So I would probably accept specific comments if they were not peppered by personal comments. But she thinks she is doing the right job so they want to make people aware that person is also a businessman and CEO of the company. That made that comment very wishy washy, in my opinion.
Trevor: OK, Kirk, I really like that I really enjoyed my talk with Igor, I know it was a little dense. What else what do we need to talk about now that we're back out of Igor and giving our heads a shake a bit?
Kirk: Well, it was very dense. But you know what? I appreciated going back and doing my pathophysiology review on the ACE receptors. But here's the thing that I marvel at. You know, fast food restaurants are now giving you beyond meat burgers. So, is this making beyond petri dish burgers like he's basically cloning, growing human tissues in the lab?
Trevor: Yeah, well, no, I thought that part was fascinating too. He actually bought these 3D printed bits of, you know, basically lung and throat and mucosal tissue. And I didn't even know those were the things. So, yeah, it's not just a couple little mucosal cells in a petri dish that it's like a pre-grown section of trachea pre-grown section of lung. And to the point, you know, it can be male, it can be female. And like you said in some of their other trials, because before this, they were looking at all sorts of inflammatory diseases. You know, they can be, you know, one that looks like it has inflammatory bowel disease. So that I didn't know these things existed before talking to Iger.
Kirk: Well, and that's I really enjoyed that part of the conversation because I kept thinking to myself, well, this is this is really curious. So, they can actually now experiment on actual human tissue that has been grown in the lab. But yet he also made a comment that was interesting. When you're dealing with the inflammatory process in the human body, circulation of fresh blood and the immune system, the T cells and, you know, the immune system response, it's very critical. So, it's almost a wonder, I wonder if having human cells cuts both ways. You had the advantage of having the experiment on human cells, but you have the disadvantage of the human cells not behaving the way they should in the real environment. I found that fascinating.
Trevor: Well, yeah, and that's where and I know I might be getting into minutia, but that's where I was talking to him about DMSO. So that was the carrier that was taking his cannabis extract through the tissue. And we use DMSO in in the pharmacy lab and the compounding lab because, well, one of the things I do with it is I'll put an antifungal in DMSO and the reason I'll do that is the DMSO will let the antifungal pass through your toenail. So if you have toenail fungus, DMSO will let you sort of paint on this toenail fungus treatment. And it's usually very difficult to get stuff through toenails, so DMSO will let it pass through. So he's sort of using DMSO in a similar way as it'll sort of drive stuff through tissue. But yeah, so that was neat. And the other thing I really liked that I think he nailed down, sort of for the for the criticisms, you know, most of the criticisms were, you know, he didn't do enough trials and he didn't do you know, this the pictures of the stains weren't right. And I think he really nailed down, look they weren't allowed in the lab. You know, this was you know, they were allowed to run in the lab, two people run, you know, a handful of experiments and got an interesting result. And he says, of course, we could have, you know, redone them later when we could get back in and do dozens or hundreds of trials. And, yes, we could have made the pictures look prettier by doing a bunch of techniques to it. But we really thought we had an interesting result. So we want to just sort of show it to the world, warts and all, just to say, hey, there's something really interesting going on here. So I thought that was very honest of a man. You know, like you said, he knew if we put this up early, we're going to be opened ourselves to criticism and they were. But it's not like they were being sloppy. They just, it was covid. They weren't allowed in the lab. This was this was as much as they could do before the pre-print.
Kirk: But once again, once again, cannabis, it confounds us because at the very end of the interview, you raised the issue that did they not find that some of the strains actually work the opposite. So you have the high CBD. You know, I think we've had this conversation. When I think of CBD, I think of Ibuprofen. When I think of THC, I think a Tylenol, you know, Tylenol, the analgesics, ibuprofen, NSAIDs tend to be an anti inflammatory. So I know I know that it's a very, very flat you know, there's flaws in this. But that's the way I sort of look at CBD and Tylenol and THC. But what at the very end of his episode and this discussion, he says that one strain actually may have actually progressed covid. And one strain, while the other strain inhibited covid. So, again, cannabis confounds us.
Trevor: Yeah, no, I found that part fascinating. Yeah. So they just like you is I, I put the word CBD together with anti-inflammatory like that's what it does. And this was a high CBD strain that both showed more inflammation and actually increased the amount ACE2 receptors. What does that mean? What we don't know, other than like he said right at the end, is, you know, cannabis isn't generic like, you know. And we hear that from the medical patients all the time. You know, this strain helped me. This one did not. So, you know, it is a as usual, a fascinating plant.
Kirk: So there we go, Trevor. We have our covid, we have our covid episode. Cannabis could actually help us fight covid if you get the right strain.
Trevor: Right. And I want us to be careful and I know our listeners are all smart. And will this, as fascinating as Igor and his team's research is? This is bench science. So this is a long, long way away from us saying that this is going to help people in a clinical trial. Like if you look at like hydroxy chloroquine, you know, we thought it was good. There's lots of theoretical reasons to think it was good and then it turned out not to be. And it's not quite the same. But one of the, so in cancer research, we think that despite, you know, a bunch of successful preclinical work, about 85 percent of the early clinical trials never make it to phase three. And phase three is the one where they put it into humans. And only about 50 percent of the ones that in cancer that get to phase three actually make it to market. So as fascinating and well done, et cetera, as the research is on the bench, in the lab, it's a long way from saying that, you know, cannabis fights covid. This is fascinating and I hope it goes somewhere. But there's a lot of trials left to go before we put it for sure, into humans.
Kirk: Well, on this can't possibly be our last covid episode either, because, you know, with all due respect to this first wave and second wave, my observations and experience and studying suggests that we're not in a wave of covid, we are in a tsunami of covid. It's just one giant waves. We're going to be talking about covid for another year anyway. So I'm sure there's going to be more opportunity to bring cannabis into the covid story.
Trevor: Oh, absolutely. Kirk, that was another really good one. What should people do if, you know, they say, I want to have my message in front of all these smart, canna-curious people?
Kirk: Well, you know, Trevor, our project Reefer Medness has been built on word-of-mouth. I've always asked people, do you like or if you like our podcast, tell a friend. We would like to have all sorts of people listening to us. We try to we try to create stories that follow the pillar of Reefer Medicine, Reefer Madness and Reefer Mellow. I think there's something for everybody. This episode we just finished might be a little bit more of a clinician's episode, whereas, you know, we've also had episodes with musicians in the Key of Weed, which is open to anybody. So I think people should listen to our episodes and tell their friends. And, you know, if you want to have an international audience listening, listening to your about a product you want to sell, let us know and we will. We're looking for sponsors.
Rene: That was great, guys. I guess it's my turn now to add a piece of music to the end of the episode and trying to keep it local. We, of course, produced this this podcast out of Dauphin, Manitoba, in the studios of CKDM Radio. And just north of here is a small fishing village called Winnipegosis. And there is an artist named Nelson Rudiak. He's actually quite the accomplished jazz musician out of New York, and his father is from Winnipegosis, and he came out with an album called Winnipegosis and seeing that's its summer. I figured I'd play a song from that album, and it's called Barbecue Boogie. So here is Nelson Rudiak, whose father was born in Winnipegosis to finish off the Reefer Medness podcast.
Kirk: Oh, you go ahead.
Trevor: No, I was clearing the throat.
Kirk: Yeah, that was good Trevor. I enjoyed that story. I have nothing more to say, I guess.