Understanding Methylation, Gene Regulation and the MethylDetox Profile

Narrator

Good evening. Thank you for joining us for tonight’s round-table lecture, “Understanding Methylation, Gene Regulation, and the MethylDetox Profile. As always, the information presented here is for your consideration and is in no way meant to replace specific consideration for your patient. Tonight’s round table is brought to you by Cell Science Systems. Cell Science Systems is a specialty clinical laboratory that develops and performs laboratory testing in immunology and cell biology, supporting the personalized treatment and prevention of chronic disease.

Presenting tonight is Amy Pieczarka. That’s “Ph.D” for us in the states. From the University of Heidelberg Germany and joining are Amy Pieczarka, RD/RDN, and Andrew Campbell, MD. If you are attending the A4M Annual Meeting in Decemeber, don’t miss Dr. Celeda’s lecture on methylation and its connection with metal intoxication in mitochondrial oxidative stress. To begin, we will turn to Amy Pieczarka.

Amy, RD, LDN, CCN, CDE

Thank you, Jason. Yes, I am a registered and licensed dietitian/nutritionist and also a clinical nutritionist. I have about thirty years of experience in this field, mostly in hospital nutrition, on an outpatient basis and as well as in private practice and now I am very happy to be the Director of Nutrition Services for PreviMedica. And Previmedica is a tele-health company, focused on nutrition and lifestyle. In addition to providing nutrition counseling via video we are providing exercise and culinary guidance this way as well. And I’m working with a wonderful group of health professionals who are dedicated and passionate about helping their patients promote optimal health. I’m thrilled to be working with them.

I’m also very excited about this webinar, as an introduction to the Methyl Detox Profile, because the nutrition professional actually may be the first healthcare practitioner that a patient is consulting with who would assess folate and the status of other key nutrients. And he or she can make the connections between the nutrient status, the lab values, whether the conventional lab values or some functional testing that they’re doing. That may be out of the optimal range and can connect that with the symptoms reported by the patient.

Now judging by the questions that we received pre-webinar, I think we have a wide range of knowledge with regard to this topic. Some, I think this information is quite new but for others, I think that others have had a lot of experience with methylation and working with clients in regard to this topic. I apologize to those of you who do have a lot of experience with this, if it sounds like I’m stating the obvious, but the consideration of genetic variance is now a very important part of the nutritionists’ assessment, thankfully. Nutrition and how it impacts gene expression and vice versa. Genes impacting nutrient needs knowing that information offers the nutritionist so much more information to tailor our nutrition plans to meet our patients’ individual requirements.

We can’t change genes, SNPs in those genes, but we can sort of work around them. And not knowing the SNPs can certainly be one reason we feel we are spinning our wheels with regards to symptoms that our patients report and the lack of progress they might be experiencing. So once again, I’m excited about this topic. I’m happy to be here with Dr. Celeda and Dr. Andrew Campbell who is the Medical Advisor to Cell Science Systems.

Dr. Campbell, M.D.

Thank you, Amy. My name is Dr. Andrew Campbell. I’m an immunologist and a toxicologist. The best way to find out what I do is to look me up on Pubmed.gov. Type in “Campbell, AW,” and the publications will light up. This Methylation Detox Profile is so vital now in these days. We used to die of infections. The Black Plague decimated one-third of Europe’s population in the Middle Ages. Pneumonia was the number one cause of death in the US in the early 1900s. Tuberculosis, cholera, leprosy were all familiar killers back then.

Then Sir Alexander Fleming of Scotland came along, gave us penicillin, the magic pill, starting in the 1940s. Medicine started using a pill for a cure. Fortunately we don’t have the old scourges that we suffered from for so long. Unfortunately, these have been replaced by chronic health problems, cardiovascular diseases, metabolic disorders, neurological disorders and cancers. But medicine has not shifted from focusing on using a pill to treat the symptoms of disease to preventing chronic disorders.

The importance of this Methylation Detox Profile is that it is predictive. You treat the abnormalities found in your patient with supplements and lifestyle changes. It gives you an insight into your patient’s future health, and you can monitor their progress a way from chronic disorders. I’ve had the privilege of working closely with Dr. Dino Celeda, who is now going to present his findings.

Dr. Celeda, Ph.D

Thank you very much, Dr. Campbell. Thank you very much, Amy. I am also very happy to be here in the United States and to work with both of you on our genetic – functional genetics. Little bit to my background.

I got my PhD or Dr. rer nat. how we call it at the University of Heidelberg in Germany, at the Institute of Human Genetics. I did a lot of research in genetics, regarding also, well, to some extent, telomere length so on and so on. After my scientific research, or let’s say I started to develop genetic testing for routine diagnostics. Right now, I am happy to be involved in a program of functional genetics, which we did a lot in Germany and which we continue to do right now here in the United States. Again, thank you very much for the invitation. Let us move on to our methylation detoxification cycle.

The objectives, actually what we want to present here right now, is not getting you in all this complicated reactions, metabolic processes, biochemical pathways. We just want to give you here, at the beginning, an introduction, a simple pathway of how methylation or how the homocysteine and methionine balance work. For this reason, we’re going to identify the most important genes involved in the methylation/homocysteine cycle, their mutations, SNPs controlling this homocysteine cycle. Let’s say influencing may be a better word for that.

We are looking at the most common SNPs very well described in literature, which actually affect the enzyme activity or the enzyme expression of the resulting gene. Then to look how the SNPs can impact homocysteine levels. After that, we can also give you an overview about a personalized intervention, according to the genetic test result. Here, I think, from my experience, what I had so far, via moving into functional genetics, we can give you targeted nutrition. We have all the . . . we can also give for targeted nutrition. I’m sorry. I’m German. I don’t find a word sometimes. Explanations. Then we also can give you some advice for targeted supplementation.

Then what is actually very good, we can also monitor the progress of the treatment or the progress of the supplementation. Call it a personalized approach by measuring, especially here, the homocysteine levels. Here, we are measuring homocysteine levels in other programs which we developed so far, which we are going to develop. We are measuring other markers, like also glutathione ratio, mitochondrial potential and so forth. Okay.

Basic stuff. It’s very basic. What is a SNP? A SNP is nothing else or also called genetic polymorphism. It’s a replacement of one nucleotide in the DNA. You know the DNA is made up of four nucleotides, adenine, thymine, guanine, and cytosine. The sequence of those four nucleotides makes a gene. What we have here right now on the left side, you’ll see, let’s say, the cytosine is replaced by adenine. Here, on the right side, we have, again, a chronological order of these nucleotides. The general population is, let’s say in this gene, guanine. Here, in the polymorphic population, where it’s polymorphic, the guanine is replaced by thyemidin.

So this is actually what we call SNPs. We imagine we have here two DNA strands. You have to be aware that we have two sets of genes, one from father and one from the mother. That’s the reason why we are all called diploid. What does a SNP do? A SNP . . . Sometimes it does nothing. We know there are companies around who measure a lot of SNPs. We have something estimated about 500,000 SNPs and something like about 25,000 genes. Yeah. So this is a big number.

So you can imagine that a lot of those SNPs are not doing any harm or not influencing any activity or expression. But yes, certain SNPs, which are described very well in literature, and we are using one of those SNPs, have some influence on enzyme activity in the metabolic processes. That’s what we are looking on.

The most simplest SNP and oldest SNP we know so far is actually sickle cell anemia. Sickle cell anemia have exactly here one replacement of one nucleotide. You see here on the left side, you see a red blood cell, a normal red blood cell. Here, everything is in order. You see here, on the left side, a sickle cell blood cell resulting from the SNP. Like we all know, the sickle cell shaped red blood cell doesn’t transport oxygen very well. So it is the simplest and first example of a so-called genetic polymorphism or, what we call it right now, SNP. Sorry.

So what is the function of genetic . . . What are the functional genetic test results doing? The individual functional genetic test results are shown on a level of functionality and/or expression of the corresponding enzyme. Again, I have to say we are not . . . The genes are not first responsible for the function. These are the enzymes, and all the enzymes in our body are metabolic processes, and our biochemical reactions are coded by genes. So genes code for an enzyme, and this influences our individuality so far. We all look a little bit different. No? It wouldn’t be good if I looked like Amy.

Amy

No

Dr. Celeda

If Amy looked like me. Sorry. The other way around. Okay. So we can divide that in the results of the SNPs. Remember what I said, kind of like we have one gene set from one parents, one gene set from the other parent. Homozygous positive is one of those kind of results, what we’re going to get. In genetic, we do everything the other way around. Homozygous positive here is called minus-minus, which means both alleles, both gene sets from both parents, show a mutation. Here, we can say, “Yes, we have a reduced functional expression of the corresponding enzyme.”

Heterozygous. This is plus-minus. This means one allele from one parent is normal. We call it wild-type. The other allele, 50% of the genes, showed a mutation. This 50% of the genes, the corresponding 50% of the enzymes, are going to have, in some cases, a reduced function or a reduced expression.

Then we have a situation of homozygous negative, which you see here, again plus-plus. Don’t get confused. I don’t know who did it. I think Mendel did that, yea this was about 150 years, yeah, I think about 150 years ago. So homozygous negative, both alleles from both parents show no mutation, and they are wild-type. This means 100% of these genes regarding investigated mutation sites are normal. This is the explanation to go into homozygous, heterozygous, and, so far, positive and negative. Sometimes I see a lot of results which say homozygous, and then I ask myself, “Well, fine. But is it positive or negative?” So we have to distinguish that a little bit.

Amy

Can I . . .

Dr. Celeda

Yes, please, Amy.

Amy

So just a comment. I know it’s pretty complex, and we’re trying to simplify it here, but the red, what we’re indicating in red here, the homozygous positive, with both alleles from both parents, the likelihood that there will be a decrease in enzyme activity would be higher than the heterozygous, plus/minus. However, both may or may not express a decrease in the enzyme activity, depending on environmental factors and lifestyle and nutritional intake. Then the one we’re showing in green, the homozygous negative, this person would be less likely to have any decrease in the enzyme activity, although lifestyle and nutrition and environmental factors are important to consider as well anyway.

Dr. Celeda

Yeah, yeah, yeah. You’re perfectly right. Kind of like you have also, when you have a normal set of genes, a lot of enzymes, especially the metalloenzymes. They need co-factors. When they don’t have this co-factor, they don’t work well. Compare this with a car, a beautiful car, a Mercedes. When a Mercedes doesn’t have gas, it doesn’t run. That’s the same thing like an enzyme, for example, in mitochondria.

We have a superoxide dismutase 2, which is also called manganese, dependent superoxide dismutase 2. This mutates. Yes, this enzyme requires manganese for proper function, which means when you even are a green, you need manganese for a proper function. Yeah. So this means also coming back not only to looking at the genetics, but looking at a broad spectrum of kind of lifestyle, of nutrition, also, yes, sometimes targeted supplementation.

Okay. What we are telling right now, targeted supplementation. How does functional genetics work? Like I said before, first a test, individual’s current status. We can combine that with SNP testing and also with corresponding metabolic markers in blood, like here. We measure the SNPs and the corresponding genes of the methylation or homocysteine/methionine balance. We’re going to get into that later. So we can also measure at the beginning, at the homocysteine level. So we have a whole picture of this person, patient, call it.

So with this picture we can do targeted intervention. This means personalized specific supplementation. Every one of us knows, or a few know, kind of what we are talking about, folic acid 5-Methyltetrahydrofolate in all the supplements, what are available. Also, of course, according also to the individual results of relevant SNPs and corresponding metabolic markers in blood.

Monitoring, again, after targeted supplementation, let’s call it, also targeted treatment. So far, we can also monitor the progress of this treatment by, again, metabolic markers in blood, what would be here, in our case, homocysteine.

Dr. Campbell

Let me bring in the practitioner’s aspect.

Dr. Celeda

Yes.

Dr. Campbell

That is that for an organism to live, it must create new cells as fast as cells die. This requires for the body to make millions of cells every minute, relying on DNA and RNA synthesis. Mutations in the methylation pathway can cripple the ability of the body to make the building blocks needed for new DNA and RNA synthesis. In particular, susceptible cells include bone marrow, lymphocytes, erythrocytes, and neural tissues. Remember that the nervous system is the highest concentration of RNA, and it has, therefore, the highest need for methylation.

Dr. Celeda

Yeah. Thank you very much, Dr. Campbell. I totally agree so far. Perfectly right. Dr. Campbell and me, we have been talking so much about developing this methylation cycle. I’m always happy. I was always very much happy. I’m still very happy for his comments and also his ideas of actually placing and targeted to certain conditions. So let’s call it also diseases.

Well, but let’s go a little bit further, the importance of personalized targeted nutrition. Again . . . Oh, sorry. Individual genetic SNPs, they say, may require personalized supplementation of specific vitamins, minerals, trace elements, and/or amino acids, even. This is important for the glutathione synthesis for the maintenance and optimization of metabolic processes. Fine.

But an overload of supplements . . . This is what we also see, in a lot of cases, can impact metabolic processes and lead to side effects. Side effects, like also the newest research, newest literature, shows it can influence a variety of functions in the organism. You have some literature describe certain overload of supplements with reduction and natural killer cell activity.

Amy

Yeah. It is. It’s so important to test. I think a lot of nutritionists and maybe some in the audience, myself included, we really try not to. But in some ways, we end up turning to OSFA. I stole that from somebody who was speaking last night. OSFA, one size fits all. Yeah, OSFA. Anyway, I liked it. I stole it. One size fits all. So one size fits all approach. Again, we try not to, but sometimes we turn to that. We’re leery about testing because, a lot of times, patients will come to us saying, “You’re my last resort. I’ve been through a whole bunch of tests as it is. I just . . . You really have to help me.” So dietitians and nutritionists might be leery about testing, and they might end up recommending a boatload of supplements and then just hoping for the best.

As we know, in the long run, that’s not going to work. So really nutrition experts should not be leery about running extra tests such as this. It’s so important to really know what we’re looking at and to monitor.

Dr. Celeda

Yeah, yeah. This is very good comment because in Europe, actually, about 200 years ago, each doctor who was studying medicine had to study botany too, just in order to learn about a side effect of certain, well, still, plants, certain plants of and side effects of kind of overload with drugs we all know. Okay. The genes. What we are looking in the homocysteine cycle and methionine/homocysteine cycle, let me say a couple . . . Let us say a couple of words to that. Naturally we are looking at MTHFR for this first, first, actually, enzyme kind of important in this whole game.

We are looking at a C677T mutation, which is very, very well-described for reduced activity, and also at the A1298C mutation also described to influence, in certain ways, the homocysteine levels. This means, in other words, the methylation potential.

Then methionine synthase, also coded by the gene MTR, methionine synthesis. Actually the second player in the whole homocysteine/methionine cycle, and we’re going to show it later on, on a chart, how this one works, but it’s actually a very important gene. Here, we are looking at a mutation at position C3518T. This is also described in literature with a reduced function of these enzymes.

Then as a third player in this ballgame, we have COMT. Everyone knows COMT, Catechol-O-Methyltransferase. Here, we are looking also at a very well-described mutation position, G472A, also called Val108/158Met, or Ala52/102 thorazine, Ala thorazine. This actually codes for the replacement of the amino acids and also the mutation position G304A. All those SNPs are actually investigated very well and described very well through enzyme activity and also highly distributed to the population.

Dr. Campbell

Let me bring in an important fact here.

Dr. Celeda

Yes, please, Dr. Campbell.

Dr. Campbell

What can make this, the methylation cycle, go wrong? Really there’s three areas. One is polymorphism that reduced the quantity or the effectiveness of any enzyme involved in the cycle. Second, a deficiency in any of the nutrients needed by these enzymes. Then third, a reduction in gene expression, which can be caused by prescription or recreational drugs, infections, or environmental factors.

Dr. Celeda

This was very, very good, Dr. Campbell, because right now you described, actually, the need of functional . . . medicine functional genetics. Look . . . Not only look at pieces. Look at it as a whole. Kind of imagine what you said and what we’re gonna also maybe discuss later on. You have no SNPs, and you have reduced . . . You have elevated homocysteine levels. Well, what does it say? Something is wrong with your nutrition.

Amy

Absolutely.

Dr. Celeda

It’s easy like that. So this is actually our functional approach here. So last but not least, what I said, what we are looking in here and what we are also offering is the homocysteine for measurement of homocysteine levels to see the treatment progress. Okay. This is the whole cycle, what we are going to talk about. Again, what I said, please, this is a very simplified presentation of the cycle. We only want to highlight those one, two, three genes we are looking at, and also the homocysteine. We’re also going to show you also some ways for the glutathione synthesis, which is also connected to homocysteine.

Well, then let’s start. Okay. First, let’s start with the first step in this whole homocysteine/methionine cycle, methylenetetrahydrofolate reductase, MTHFR. We all know folic acid goes over certain genes, DHFR. We don’t want to go into that right now. It then becomes tetrahydrofolate. At the end, it becomes 5, 10-Methylene tetrahydrofolate. This is the first step which is important here in this whole cycle. 5, 10-Methylne tetrahydrofolate deems as a template for this enzyme, MTHFR, to produce the so-called bio-active folic acid. The bio-active folic acid or bio-active folic is nothing else then 5-Methyl tetrahydrofolate.

So in other words, when you have a SNP or when you have a reduced enzyme activity to a SNP here, what we recommend then bypass this SNP, just in order to take 5-Methyl tetrahydrofolate. This is a very, very normal thing to do, kind of. This is what we are always looking at, bio-chemical pathways, metabolic processes. When you can do something, when you like the enzyme activity to do something, give it or prescribe it as a supplement or as injection. You, Dr. Campbell, know much, much better what we can do about it in practice so far.

Okay. 5-Methyl tetrahydrofolate. Again, mutations at positions, these two positions we have been talking about. Presence of one or two SNPs affects enzyme activity, supplementation, 5-Methyl tetrahydrofolate.

Amy

So someone who does not have a SNP, if it’s MTHFR, would be more likely able to handle then folic acid from supplements and fortified foods, but obviously, as you said, the person with this SNP would not be able to convert that folic acid into the bio-active form. So the supplementation would be recommended, of course, because the build-up of the folic acid, without the conversion to its bio-active form, could contribute to a whole cascade of issues.

Of course, we would still recommend that people consume the folate from folate-rich foods, their leafy greens. Most the people in the audience, I’m sure, know the folate-rich foods, but leafy greens and the spinach and kale, legumes, asparagus, brussel sprouts, broccoli, avocado, and so forth. So that requiring fewer steps to be converted to the 5-Methyl tetrahydrofolate, but this person will likely need that supplementation, as you said.

Dr. Celeda

It’s easy when I can convert that. No? So I don’t need to supplement it. Why? First, let’s say it in a very, very normal way everyone understands. I can save the money to buy me that supplement. It’s very easy. Then I don’t have to burden my body with unnecessary supplementation. Sometimes in Europe, a lot of people are creating very expensive urine. Okay.

So this 5-Methyl tetrahydrofolate is not the end product of everything. It’s just the first step, the very first step to enter the homocysteine/methionine cycle. Here comes the player methionine synthase, which we were talking about before. I’ll call it MTR into play. The MTR so far is actually using the methyl group of 5-Methyl tetrahydrofolate and donates this methyl group to vitamin B12, creating methylcobalamin or kind of like or methylated vitamin B12. Yeah, so far.

This is the first step of this enzyme, MTR, using the methyl group. You see that 5-Methyl tetrahydrofolate is going to be recycled again to tetrahydrofolate, and we end up with methylcobalamin or methylated vitamin B12. In the next step, the same gene, all the MTR methionine synthase. That’s actually his name. Methionine synthase donates this methyl group to homocysteine, and the homocysteine is converted into methionine. So you see, this is also called the methylation, re-methylation. It’s described in literature all with a lot of names of methylation.

The only thing is the methyl group is actually donated further on, step by step, in order to create methionine. The methionine, like we know, is one of the universal donors in the body for methyl groups. Okay. Again, mutations at MTR or methionine synthase, so far, affect enzyme activity. We recommend methylcobalamin, but please be sure to take methylcobalamin. There’s also some other recommendations with other cobalamin or other vitamin B12 here. You really need methylcobalamin. Like the name says, you need the methyl group.

Amy

It’s important to remember those also who have an inadequate intake or an absorption of B12, even without the SNP. As people get older, the ability to absorb tends to decline. Digestion needs to be addressed, reduced stomach acid production. Maybe medications are contributing to that, of course, vegetarians as well, strict vegetarians, vegans who would not be consuming any sources of B12 from their pattern of eating because it’s only coming from animals. So it’s important to address that as well.

Dr. Celeda

This is a very good point, what you made, is people getting older. Right now, you see the whole process of aging is nothing else than oxidative stress. Through this oxidative stress, we are getting more so-called “not inherited” SNPs, SNPs which are made to our lifestyle, which we can detect, except that we sequence the DNA. This SNP, sure, they start also with the age affecting your enzymes’ activity. So when you’re getting older, and you have everything fine in your SNP situation, regarding mutation sites, but your homocysteine level is still too high, then you know, “Okay. I have to do something.”

So actually it’s also very, very good monitoring tests, especially in the field of, I hate to say, anti-aging. There is no anti-aging. We’re all going to die. But let’s say controlled aging.

Amy

Well, when we’re aging, we’re living longer. That’s a good thing.

Dr. Celeda

Yeah. Okay. So this is an important step right now, kind of. You see this is actually . . . Here, we have here now two supplements and two genes which are actually involved in the production of methionine from homocysteine. Okay.

Well, let’s get us to the last step. The methionine here, like I said, is a universal methyl donor. The methionine is converted by gene methyltransferase, to SAMe. Methyltransferases actually has no such SNPs, and we can neglect here something, but S-Adenosyl-methionine is the major substance, also called SAMe, is one of the major substances, not the major substance, one of the major substances acting as a methyl donor for degradation or, let’s say, for the synthesis of specific substances by COMT, Catechol-O-Methyltransferase.

This means SAMe, in connection with COMT, are involved, let’s say, in methylation of nucleic acids, this is nothing else than DNA methylation, which controls our gene regulation. In another transmitter regulation, here we have dopamine, serotonin, epinephrine, norepinephrine in xenobiotics. What are xenobiotics? Nothing else than toxic substances from the environment. As also, drugs, yes. Methylation is involved in phase two detoxification of certain drugs.

Also, in the degradation of certain or activation of certain phytochemicals . . . So you see, methylation processes are very, very important in the body. This cycle here, byhomocysteine/methionine is the major or one of the major cycles to donate methyl groups, which can be used further on by COMT or other methylating enzymes, in order to process certain methylation process in the body, involved in certain fields, detoxification and so on and so on. Yes, Dr. Campbell, please.

Dr. Campbell

So let me ring in about this Methylation Detox Profile, the certain clinical aspects. One is cardiovascular diseases that we mentioned earlier. You’ve described this profile beautifully. So clinically, coenzyme q10 has been used as part of the treatment for angina, heart failure, re-profusion injury after coronary bypass, cardiomyopathy and various other disorders.

The synthesis of CoQ10 in the body requires components of the methylation pathway. In particular, the last area that you mentioned, the SAMe, is what is . . . It needs adequate levels of that same SAMe that is generated in this methylation cycle. The statin drugs decrease the levels of CoQ10 in the body. So practitioners need to be aware of the methylation status of their patients taking a statin drug.

Also, the relationship between elevated homocysteine levels and an increased risk for heart disease and the genetic risk associated with MTHFR, C677T that was mentioned, this mutation in the methylation pathway has been recognized for some time. In this cardiovascular area, there are two types of white blood cells that must be methylated: lymphocytes and monocytes. Low methylation in these cells lead to excessive blood clotting, resulting in thrombosis and stroke.

When you step over into the neurological area, folic acid is converted to monoglutamate entities by the enzyme, alpha L-glutamyl transferase in the intestinal wall. Once absorbed, monoglutamate entities are converted to MTHF, the fat soluble form of folate that crosses the blood/brain barrier and is utilized to facilitate the synthesis of neurotransmitters, in particular, dopamine, epinephrine and norepinephrine. In patients with methylation deficiencies, the level of MTHF is . . . Production is reduced, thereby limiting the production of these neurotransmitters.

When you step into what was mentioned before, the xenobiotics at the very top there, that’s our environmental toxins. We’re constantly and persistently exposed to environmental toxins. There’s artificial colorings, flavorings, sweeteners and preservatives in our foods and beverages, which we test for in our laboratory in patients. The US manufactures about 5 trillion pounds of chemicals annually. That’s a five with 12 zeros. We use 2.3 trillion pounds of pesticides annually. So methylation helps clear these environmental toxins from the body.

This process involves the conjugating of methyl groups to the toxin prior to removal, as well as supporting the production of glutathione. Now as many of you already know, in the audience, that glutathione is a protein involved in inflammatory control and reduction reactions, and it is important in producing and controlling the leukotrienes. Glutathione is the major endogenous antioxidants produced by cells, very important, and involved in the neutralization of free radicals and reactive oxygen species. It also maintains exogenous antioxidants, such as vitamin C and vitamin E, in their reduced form, which is their active form.

Through direct conjugation, glutathione detoxifies many of the xenobiotics or environmental chemicals and carcinogens, both organic and inorganic. This includes the heavy metals, such as mercury and lead and arsenic. Every system in the body can be affected by the levels of glutathione available, especially the immune system and the nervous system, the gastrointestinal system, and the lungs.

So here, we see how important this cycle that we test for in the Methylation Detox Profile can become. Lastly, I want to mention . . . I don’t want to leave cancer out of this. Abnormal DNA methylation, both hyper-methylation and hypo-methylation, is associated with cancer. In particular, low levels of white blood cell DNA methylation is associated with many types of cancers.

So we can see how this . . . The importance and health of this entire cycle, in this particular test . . .

Dr. Celeda

Thank you very much, Dr. Campbell, especially mentioning the hypo-methylation. Yes, the hypo-methylation is very well-known, actually described also as global hypo-methylation so far, due to not right nutrition, what you have today. But I also thank you very much, Dr. Campbell, that you went into the glutathione cycle. You see what you said. Right? Methylation is a conjugation reaction. We have a lot of conjugation reactions for the detoxification that this is acetylation. But also, yeah, kind of glutathione. Glutathione is nothing else than a conjugation reaction. Let me finish that up. Then I’m going to show you something, how this whole cycle is interconnected with glutathione.

Again, when we have, here, mutations in the COMT so far, or when we have genetic SNPs in the COMT, we actually recommend an S-adenosyl-methionine, or SAMe, so far, in order well to, again, let’s say to bypass this whole situation. So now comes this big thing, Dr. Campbell. What I’m actually also going to present in about, I think, three weeks, how is this whole thing connected, homocysteine to glutathione. Well, homocysteine is nothing else than the template, first template also for glutathione synthesis.

Homocysteine, in connection with vitamin B6, produces the very important amino acid, L-cysteine, and cysteine is the major component with its sulfur group, the major component of the glutathione, used by the so-called glutathione S-transferase. This means transferring the sulfur group so far, by conjugation, in order to detoxify or to neutralize substances, toxic substances, not only toxins, but also, yes, like in mitochondria, also hydrogen peroxide.

I have right now a question, asking me kind of why manganese . . . Yeah. What’s this . . . Yeah, we can go into that right now because it makes interconnections very good. Why using manganese so far for the superoxide dismutase 2. Very, very easy. Kind of like we have in our bodies so-called superoxide dismutases, which have only the task to neutralize or further process oxygen super oxide radicals. The superoxide dismutase 2 or the manganese depend on super oxide dismutase has to test in mitochondria to convert a superoxide ion to hydrogen peroxide. Then this hydrogen peroxide is, again, converted to water, yes, but a glutathione peroxidase or also catalase.

Here, you see glutathione peroxidase in order to defeat or fight oxidative stress. The other superoxide dismutases are the so-called SOD-1, superoxide dismutase and superoxide dismutase requires copper and zinc as co-factor. It’s found in cells so far.

Last but not least, you have the superoxide dismutase 3. This superoxide is found in arterial walls so far, and it also needs, as a co-factor, copper and zinc. So I think I answered that. Please, for further questions, you can also email questions to us, or we’re going to answer it by email, also by phone call and so on.

But anyhow, I wanted to show you here the way homocysteine not only for methionine synthesis, but also for glutathione synthesis so far. Then you can also imagine when you have high homocysteine levels. It’s bad, actually, for your blood or for your arteries or, let’s say, for your plugs. But you have also a high level of homocysteine for glutathione synthesis so far.

Amy

But it does serve good purposes. So a little bit is good. Too much, no.

Dr. Celeda

It depends. See, again, here we have, again, the connection. Don’t look at one cycle. Look at the interconnection of these different metabolic cycles so far. Again, what we said, homocysteine monitoring and maintaining progress here, this cycle here, for personalized treatment. We also see in our lab reports kind of, when you do everything right here, support, give the right nutrition, give the right supplementation, your homocysteine levels are still elevated or reduced so far. Then try it with vitamin B6, co-factor of glutathione for glutathione synthesis, L-cysteine, L-glycine and glutamic acid.

With other words, let’s make them this way. You did everything right here. Supplementation is right, and you still have elevated homocysteine levels. Maybe something is wrong in this pathway, which have the genes CBS and CSE so far involved in bio-synthesis of L-cysteine or maybe further on in the whole bio-synthesis of glutathione. So this would be our recommendation. Yeah.

Amy

Dr. Celeda, with regards to the monitoring of the homocysteine, what is your recommendation for how often to monitor that homocysteine?

Dr. Celeda

Well, let us come also to the results of what lab report looks like, but Dr. Campbell can answer this much better. I am the scientist. He is the doctor. But we would recommend two months or three months. What do you think, Dr. Campbell?

Dr. Campbell

I would say, depending on the patient’s compliance and the severity, I would check it every three to four months.

Dr. Celeda

Yeah. You see, this is also what we are . . . We are a lab. We know your patient only from the paper and his birthday. That’s about it. So you know the person. You know his habits. You know how he eats and his diet. You know whether he smokes, drinks, all these little things which influence, actually, our controlled aging.

Dr. Campbell

So I would mention and recommend that you do the same homocysteine test in the same laboratory every time. Don’t go jumping around because each lab is its own reference range.

Dr. Celeda

Yeah. Exactly. This is very, very important. Sometimes we get . . . We measure the homocysteine. We get then from the doctors “but the other lab, we have different results.” You see, Dr. Campbell, you’re totally right. Everyone has its own reference ranges and kind of like we would be happy that you would measure the homocysteine with us. Very nice. When you started measuring homocysteine in another lab, keep it at that to get consistent results. Okay. Good. Let’s go a little bit into our . . . at the end, into our lab report.

So far, you see this is the first page of our lab report, and we also tried to make it very simple. We use the whole cycle, actually, as a lab report. Here, we have yellow COMT. Like you know, yellow is . . . 50% is normal. 50% has mutation in the mutation site. Green is 100% negative, what we call, so far, what we call homozygous negative plus-plus. Again, here, MTHFR. All the genes from father and mother are okay.

On the second page, we’re going to get a little bit of description, background description, what to avoid when a SNP is there, what to do, kind of what you have to be aware of, for example, taking anti-folates. That’s also here for COMT, a little bit more to diet, avoiding or being aware that quercetin can also inhibit COMT activity. Those little things are what you have to know, whether your patient also has Parkinson’s treatment and so forth.

On the third page of the lab report, we’re going to give you like here. You remember we had in COMT yellow. This means 50% of the enzymes are normal. We’re going to give you additional information, what you can do, recommendation for supplementation, as others here, in this case, COMT, SAMe, s-Adenosyl Methionine. Also, if homocysteine levels are still elevated, and everything is . . . diet is normal, also try vitamin B6, L-cysteine, L-glycine, and L-glutamic acid. Like we said before, homocysteine is also a template for glutathione synthesis.

Last but not least, you have here homocysteine levels in the serum. So far here, the person is age 48, 11.63 micro milliliters. So according to this range, what we are using here, and this is the last range, actually, which is recommended here in the US. It’s still in the normal range.

Amy

So I wanted to mention the target range that CSS has here. I know some practitioners are looking at a tighter range and look for a range of between four and 10 as an optimal range. So this particular guideline came from . . .

Dr. Celeda

American Heart . . . Yeah, American Heart . . . This is 2015 recommended. You see? Sure, we want to be on the sure side and something you can’t get attacked for, but you see, everyone knows . . . Every practitioner knows his ranges so far, and this is the most important thing, what we are giving here as a guideline of, yeah, the latest literature.

Amy

Can you comment on two low homocysteine?

Dr. Celeda

Well, too low homocysteine, what we would say . . . Again, what I said in connection with the glutathione synthesis, maybe there’s a detoxification going on. Maybe there’s really a need of glutathione for, let’s say, oxidative stress. But also, again, what I’m going to maybe go a little bit into in December at A4M, also kind of in . . . They’re metal detoxification. The glutathione transferase M1, the specific enzyme is also T1, glutathione transferase T1. It’s involved in metal intoxication. This has been known for a long time. Yeah, it can be that the person kind of has a metal sensitivity. So there are different approaches to go around that, kind of also specific monitoring testing, but also in combination with genetic testing. Here, you really have also possibility of targeted supplementation.

Okay. Let us move to another . . . I think we have five minutes. Yeah. Let us move to the last lab report as an example. Here, we choose . . .

Dr. Campbell

This is a different patient.

Dr. Celeda

This is a different patient. Yeah, yeah, yeah. Exactly. Very, very good. You see here, we have the whole story, yellow, green, red, kind of in COMT, yellow, kind of 50% of the enzymes. Again, fine. 50% show mutation. Again, MTR SNP is okay. 100% of all the enzymes contain MTHFR. Here, we have here, in both positions, from father and mother, as a mutation, a SNP which is affecting really the enzyme activity so far.

Again, second page, we give you all of this. I say always what we say, Europe blah blah. What you have to know, in short, comprehensive tables and short comprehensive form so far. We give you here, again, the recommendation for the targeted supplementation. You see here, we have here COMT, we have COMT, s-Adenosyl methionine and for the MTHFR, what we said before, 5-Methyl tetrahydrofolate bypassing, what we say, bypassing the SNP.

Again, additional information if homocysteine levels are still elevated after targeted supplementation, take, again, the vitamin B6, as also the components of glutathione.

Amy

Pre-webinar, we did get a lot of questions regarding dosing, and I think it’s, of course, impossible to come up with a one-size-fits-all approach with regard to that. There are practitioners that are recommending quite a bit more than what we have here. But for our purposes, we are sticking with this. I think a good rule of thumb is to start low and go slowly and follow up to monitor for side effects.

Dr. Celeda

Yes, exactly. That’s what . . .

Amy

Follow up within the first week.

Dr. Celeda

That’s what we are recommending. Dr. Campbell and me discussed this a lot of times, what we’re recommending so far. I would, yeah, say start with . . . We advise start with a small dosage, and you see also the recommendation on dosage, what we did here. Some might think, “Oh, that’s too low.” But again, we have to follow certain rules so far. We have to follow certain rules. This is only what we say is . . . The lab report is nothing else than to give the practitioner an orientation so far. Again, the practitioner knows his patient, knows the person, and we are actually only giving him hints or supporting him in the treatment because, again, like I said, we don’t know his background, nothing so far. So we are actually only giving orientations for targeted, let’s say, treatment or supplementation, whatever you want to call it.

Okay. Let’s go to . . . Oh, yeah, yeah. Here. Let’s go to the homocysteine levels and see this person, age 23. Remember, we have COMT yellow, MTFHR red. Here, we have really elevated homocysteine levels, 34.2626 micro milliliters. What would you recommend, Dr. Campbell, other kind of all the monitoring?

Dr. Campbell

Again, this one I would monitor probably every three months.

Dr. Celeda

Yeah, yeah.

Amy

She needs to do what she’s told. She needs to follow the recommendations. We know she isn’t, and she’s here with us.

Dr. Celeda

Yeah, yeah. This is . . .

Dr. Campbell

Practitioners know the importance of patient compliance. How that play a huge role, especially in someone 23 years old.

Dr. Celeda

Yeah, yeah. Naturally, sure, for this presentation, we picked up some kind of very, very nice cases for what we say, not so nice for the person who has the test, kind of extreme cases.

Amy

Right.

Dr. Celeda

But here, you can see really kind of like impacts. Then like Amy said, she’s a coworker of us, a colleague of us. Let’s see how this is going to look like in about two or three months, when she follows our advice.

Dr. Campbell

So let us kind of get into what happens when your body is not methylating efficiently. To cap it up, you can accumulate abnormal levels of toxic, could be metals, which are found in foods and the environment. You can develop signs of cardiovascular disease, including hypertension, cardiomyopathy, chest pain, tachycardia, et cetera. However, you may not have any symptoms at all until you suffer from an acute myocardial infarction.

You may develop thrombosis or a stroke. You’re more likely to develop neurological disorders. You’ll be at an increased risk for cancer. You may be more susceptible to a wide range of inflammatory conditions, including autoimmune disorders, and you are likely to have a weakened immune system and will be unable to mount a robust immune defense against infections, which goes to show the importance of this Methylation Detox Profile. The application of it is vast, many, many patients. I know in my practice, most of the patients need this test.

Dr. Celeda

Thank you, Dr. Campbell. I think we are right now . . . We are over the time for a couple of seconds. No?

Amy

Well, I know we want to be really respectful of everyone’s time, but there are some pre-webinar questions that came in, and if we . . . I think we have them on the next slide. No? Yeah.

Dr. Celeda

No? On the next slide?

Amy

Oh, okay. Actually, yes. Can we go back?

Dr. Celeda

This was the pre-webinar question.

Amy

All right. Yeah, let me address this. Okay. So some of the nutrition experts in the audience may have noticed on the second page of the sample test results that there’s a whole chart, list of foods that are considered high in quercetin. Of course, at first glance, you might think, “Are you kidding me?” That’s how I reacted, actually, because when you look at this list, you just would feel uncomfortable suggesting people avoid all these things because we’re normally telling people to hit the body hard with plants, plants, plants, and as many vegetables as your colon can handle. Of course, a lot of these things on here are good sources of folate as well. The important thing here to recognize is that quercetin is a COMT-inhibitor.

Dr. Celeda

One thing I can tell you . . . If you remember, I showed you a pharmacological paper from 1996 . . .

Amy

Yes.

Dr. Celeda

. . . saying the quercetin not very well-known as the COMT-inhibitor.

Amy

Yes.

Dr. Celeda

This was in the introduction.

Amy

Right. But nutritionists . . . We’re suggesting quercetin all over the place. We have been. It is in supplements as well. So I don’t . . . I think the important thing here to know is that we’re not saying . . . It’s not recommended to avoid all these things. First of all, if the person has a COMT SNP, just be aware of the person’s consumption here and monitor progress. If something is not . . . If the patient is not progressing, and you’re thinking what . . . You want to maybe investigate further. Possibly, the person might be consuming quercetin.

But now, my suggestion would be, though, if you look at this whole chart, not to restrict the person with regard to all these, but maybe look towards the left column, the first maybe 20 foods, to determine that those high-quercetin foods, if the person is consuming a lot of those, but it’s not at all to give the message that they should be restricting these or avoiding these completely. It might be a good idea, though, for someone with a COMT SNP to be avoiding quercetin in supplements, however. So I assume it’s on the next? Yeah. Yes. Okay.

So I think a red flag for a practitioner to prompt the order of the Methyl Detox Profile, I think that that’s pretty much been covered. Dr. Campbell went over quite a few, pretty much touches on every organ, pretty much on every organ system in the body. How do you know when someone is under or over methylating? The under-methylating, I think we already discussed, but over-methylating . . .

Dr. Campbell

Just order the test.

Dr. Celeda

Order the test. That’s what I wanted to say. Under-methylating . . . You see, we have, in Europe, a lot of pediatricians who are using actually COMT. COMT is also the methyl detox order to . . . Using COMT and also the other genes, what we were talking about, yes, in autistic patients. You see, the fact is it’s also very important or very convenient, let’s say, just for pediatricians, because the genetic testing is done with saliva samples. So yeah, this is also one thing.

Does someone with a SNP require lifelong supplementation, from a standpoint of a supplement industry? Yes. From our standpoint too, kind of, and that’s what we are telling about bypassing the SNP. Like we showed here, you can produce something, a specific bio-chemical substance in your body. Okay. Try it with supplementation. Yes, the genes are never . . . The SNPs are never going to be changed, never, never.

Dr. Campbell

Yep.

Amy

How does the standard American diet, which unfortunately is very sad, versus the Paleo plan affect this? Dietary factors overall . . . As you know, the methylation can be affected by dietary levels of methyldone, or obviously, as we’ve been talking about this whole presentation, folate, B12, and other vitamins and minerals as the co-factors, and, of course, amino acids.

So a plant-based pattern of eating, including plenty of vegetables and leafy greens and legumes, nuts and seeds, grass-fed beef and chicken, shellfish, and so forth, that would, of course, be the way to go. That would give you the nutrients necessary for the methyl donors. But with someone, the standard American diet is typically not plant-based. It’s an inflammatory pattern of eating. There’s not enough intake of plant-based foods and leafy greens, for the most part, with a consumption of way too many packaged and processed foods overall, not providing the key nutrients needed for methylation.

But also, on the other hand, with the consumption of those packets and processed foods, the foods that . . . A person could be consuming foods fortified with folic acid. Then of course, the SNP, the MTHFR, without that bio-active form of folate, could be contributing to a whole cascade of issues. So of course, the other thing to keep in mind is the SAD, standard American way of eating, is not only methyl-deficient, but contributes to oxidative stress.

With regard to the Paleo plan, I know that there are a lot of versions of the Paleo plan out there. But overall, with the emphasis on vegetables and protein sources from beef and chicken and fish, it can provide those much-needed nutrients. But for vegetarians, the plan would be quite a challenge, of course. So it’s a nutritionist’s job to make sure that all the key nutrients are represented in the plan, the food and supplementation plan for each one of their patients.

Dr. Celeda

Okay. Let’s go to the . . . Thank you very much, Amy. Let’s go to the next question. How does one decide which methylating supplement to use?

Dr. Campbell

Well, we already described that, in the lab test results.

Dr. Celeda

Yeah.

Dr. Campbell

When you order this test, we send the recommendations right alongside, depending on the individual’s testing.

Dr. Celeda

What all else did we point out? We had a lot of things, but again, like I said, email us for further questions. 3-methyl-glycine . . . Yeah. It was also a question. Can you use that? Yes, you can use it because 3-methyl-glycine is actually betaine actually and in the liver the methylation of homocysteine to methionine. It’s used with betaine. So much for that.

How do I specifically use the MTHFR results on pharmacogenomics testing to help mental health patients? Here, you need, actually, the COMT. This is the most important gene we said before, what we just stated before, pediatricians so far, autism, but also to a certain circumstance. We are also working with labs together, which are specialist neurotransmitter testing. Here, you can also marry those two tests. You can use sometimes the neurotransmitter testing as a follow-up, as a monitoring test.

Amy

Excellent.

Dr. Celeda

Okay. I think we are it. Thank you for joining us.