#16 PRESENTATION: Rhonda Speaks at the Orthomolecular Congress in Bussum, Netherlands

And it's an honor to have her here speaking on the Congress at the moment. That is a mouthful. She's a scientist in the laboratory of Dr. Bruce Ames at Children's Hospital Oakland Research Institute. Een hele mond vol. And one of her goals is to encourage people for getting a healthy long life by using a proactive preventative approach. And today she will speak about micronutrients for the preventing of age-related diseases and brain dysfunction. Please welcome Dr. Rhonda Patrick. It's all yours.

You have to click. Goedemorgen. Good morning. That's about 95% of my Dutch vocabulary, so my talk will be in English today so that I do not butcher the Dutch language and you don't have to sit here for several hours. So, my name is Rhonda Patrick and I work with Dr. Bruce Ames in Oakland. California, United States. And I do a variety of different research with Bruce on how micronutrients can prevent age-related diseases and also neurodevelopmental, neuropsychiatric diseases as well. So I'm going to talk a little bit about that today. So metabolism is very complicated, as you can see here, very complicated. This is your metabolism. And your metabolism is running pretty much everything in your body.

Every cell in your body has this metabolism that is important for the organ to function properly. Whether we're talking about your immune cells to fight off an immune— you know, to fight off a disease or we're talking about your heart, your liver, it's your metabolism that is running your body. And many of the metabolic pathways require enzymes, and these enzymes are running your metabolism. Well, if we think about all the enzymes in a cell, about 22% of them require micronutrients as a cofactor. So that means they require micronutrients to function properly. And micronutrients are about 30 to 40 essential vitamins, minerals, amino acids, and fatty acids.

Okay.

that we must get from our diet because our body does not make them. And recommended daily allowances have been set to ensure that we get adequate levels of these vitamins and minerals and amino acids and fatty acids. The RDA you may be familiar with is usually set a couple of standard deviations above what would be required to prevent deficiency, which could cause acute death. But these micronutrients, these recommended daily allowances are not even being met. So in the United States alone, actually around 70% of the population does not meet the requirement for vitamin D, around 60% does not meet the requirement for vitamin E, 45% does not meet the requirement for magnesium, around 38% doesn't meet the requirement for calcium and vitamin K, 35% don't meet the requirement for— and it goes on and on.

You can see these These micronutrient deficiencies are widespread, at least in the United States. So what happens if you don't get enough of these essential vitamins and minerals? We don't see people walking around with scurvy, you know, if they're getting— don't get enough vitamin C. So around 24% of the population does not have adequate levels of vitamin C, but we don't see everyone walking around with scurvy. And that's where my mentor, Dr. Bruce Ames, comes in. He came up with something that he calls the triage theory, which I'll explain. So, throughout periods of evolution, humans were exposed to periods of food scarcity, food shortages.

We didn't always have food at our disposal. And so, Bruce thinks that there is a strategic rationing of vitamins and minerals in the body such that the proteins and enzymes that require vitamins and minerals as a cofactor to function, which are essential for short-term survival, which are required to make sure you live long enough to pass on your genes, those enzymes and proteins will get the micronutrients before other proteins and enzymes in the body which also require micronutrients but are not essential for short-term survival, they're more important to prevent diseases of aging, for example. Those proteins and enzymes will not get the micronutrients if you don't have enough of them.

And so this is what Bruce calls the triage theory. So what he posits is that when you have these micronutrient deficiencies or inadequacies, then this will lead to insidious damage that accumulates over time, over several decades, and can lead to age-related diseases. An example would be magnesium. I mentioned 45% of the U.S. population does not get enough magnesium. Well, magnesium is at the center of a chlorophyll molecule. Chlorophyll is what gives plants their green color. So spinach, kale, you know, things that are dark green leafy vegetables are very high in magnesium. Well, the RDA for magnesium is set around 400 milligrams a day. And people are not getting enough magnesium.

So what happens? What's the consequence? Well, magnesium is required for over 300 different enzymes in the body, and those enzymes include those that are important for the production of ATP, which is the energetic currency of the cell. So enzymes that produce ATP and enzymes that are needed to use ATP require magnesium to be bound to that ATP to use it. You can imagine that's a very essential function. If you can't make energy, you simply won't survive. So that would be something that is considered important for short-term survival. But there's also other proteins and enzymes in the body that require magnesium. Enzymes that repair damage to DNA also require magnesium as a cofactor.

So just normal living, breathing in oxygen, eating food, this is how we make energy. This process is coupled in a process called oxidative phosphorylation. Well, just that process of making energy causes reactive byproducts to accumulate called reactive oxygen species which can damage DNA. And this damage can cause double-stranded breaks in DNA. And this can lead, if it's not repaired, can lead to— usually, if it is repaired, that will make a happy cell. So your cell will not be damaged because you repair that.

Okay.

But if the damage is not repaired, this can lead to potentially mutations. And these mutations in your DNA, depending on where they're at, can cause dysfunction of a cell. It'll lead to an unhappy cell, which can then, depending on where these mutations occur, can lead to cancer. And this happens over the course of several decades. There's also another protective response in the body. If you acquire these mutations, your cell goes, I don't want this mutation. I don't want it to lead to cancer. Therefore, I'm going to sacrifice myself. Myself. I'm going to kill myself through programmed cell death. And so the cell will kill itself. That can also have problems if this happens in stem cells.

It can lead to stem cell aging which leads to tissue degeneration, et cetera. But if cell death doesn't occur, that can also lead to cancer-causing mutations. So these DNA repair enzymes, they actually require magnesium as a cofactor to function. Without magnesium, these DNA repair enzymes do not work efficiently. But DNA repair is not required for short-term survival. You can get DNA damage and still survive. It doesn't— it's an insidious type of damage that accumulates over several decades and by the 5th, 6th, 7th decade, you may end up with cancer. So obese people are actually a perfect example of people that are not getting enough magnesium and also other micronutrients.

The obesogenic diet, as I like to call it, consist of a diet that is heavily processed foods, foods that are in packages, that are in boxes, very little micronutrients, very little vitamins and minerals, and a lot of refined sugars. So as a consequence, well, many things are happening, but these people— the obese people tend to be the most deficient in many different vitamins and minerals. Obesity increases the risk not only of type 2 diabetes and cardiovascular disease, but it also doubles the risk of many different types of cancer and neurodegenerative disease like Alzheimer's disease. Being obese is associated with taking 7 years off the lifespan and in extreme morbid cases, extreme morbid obesity, studies have shown that 14 years can be taken off lifespan which is very significant.

So I've been studying DNA damage as a marker, a biomarker of cancer in obese people and comparing that to people that are lean. And you can measure this in the lab because one of the first biological responses to damage in your DNA, as I mentioned, just normal living causes DNA damage. None of us can escape it, but if we're getting enough of these micronutrients to make sure that our enzymes that are able to repair that damage are functioning properly, we hope that we're going to repair this damage. One of the first molecular signaling events that occurs after a double-stranded break, which is the most deleterious type of DNA damage, it's the most difficult type of damage to repair.

One of the first events that occurs is a phosphorylation of histone H2AX. This is called gamma H2AX. And so this happens about an hour after the damage occurs. It peaks and the signal intensifies all around the site of the damage. And we can use an antibody with a fluorescent tag on it to actually biomark this double-stranded break by measuring gamma-H2AX, which is what I've been doing. So I've been taking peripheral blood mononuclear cells that are taken from people that are either lean or obese. So lean people are identified as people that have a body mass index of 25 or less. And people that are overweight or obese have a body mass index of around 28 or above.

And when we— when I measure the gamma-H2AX as a marker of DNA damage, I see that obese people have significantly more of this DNA damage than people that are lean, specifically of these double-stranded breaks because gamma-H2AX signifies a double-stranded break. In addition, I have some preliminary data that the obese people are not able to repair damage that I induce. So I induce a known amount of damage to their cells, to their PBMCs, and then I allow that damage to repair over time and measure it. And what I'm finding is that not only do they— the obese have higher DNA damage, but their capacity to repair that damage is also diminished. So the next step would be to measure the magnesium levels in the obese, which other people have done and have found that they are very deficient in magnesium, and then give them magnesium to see if you can boost their DNA repair capacity.

So that's in the future. Another example of triage theory which Bruce and my colleague Joyce McCann have published a few years ago is vitamin K. So vitamin K, I mentioned earlier about 35% of the U.S. population does not get adequate levels of vitamin K. Vitamin K is found also in green leafy plants because it's— vitamin K is required for the plant to photosynthesize, which is the way they make energy. So obviously, plants are high in vitamin K because they need it to undergo photosynthesis. There's 2 biologically active forms of vitamin K: vitamin K1, which is found in plants, and vitamin K2, which is produced by bacteria, including the bacteria in our gut and also other bacteria that ferment foods.

Natto, the Japanese fermented soybean, is probably the highest dietary source of vitamin K2. Vitamin K1 and vitamin K2 both serve as cofactors for a variety of proteins and enzymes in the body. They have very important role in activating proteins that are involved in coagulation and blood clotting. So you can imagine that this is probably an important function for short-term survival. Proteins that are important for coagulate need vitamin K to be activated. Well, coagulation is very important because if you damage You get damage to a tissue, you hurt yourself, you want to make sure that you clot to repair that damage. If you're unable to clot, then you have the potential of hemorrhaging, you know, bleeding out, and that could ultimately lead to death.

So vitamin K serves a very important function for short-term survival in that it activates proteins that are required for coagulation. But vitamin K is also required to activate proteins that pull calcium out of the vascular system, out of the blood vessels and arteries, and bring it to the bone where it's supposed supposed to go, bring it to other tissues where calcium is required for many different enzymes and metabolism. So this is also a very important function, but it's not important for short-term survival. Calcium can easily precipitate with the phosphate in the blood vessels and this can cause calcium plaques to build up. But these calcium plaques that build up take time, several decades, and ultimately can lead to things like atherosclerosis, vascular dementia, cutting off the circulation to the brain, So, this is also, you know, a very important function of vitamin K, but it's not important for short-term survival.

It doesn't matter if you have calcium building up until you enter your 6th, 7th decade in life when you have a risk to, you know, for a heart attack or vascular dementia. So, my colleagues Joyce and Bruce published a study where they posited and did a bunch of research in literature and found that Vitamin K appears to have a triage that is tissue-specific. So, all those proteins that are involved in coagulation, the short-term survival proteins occur in the liver. And so, vitamin K1 actually goes to the liver very readily. And that's the first place it goes to activate all those coagulation proteins. Once enough vitamin K1 goes to the liver to activate all those proteins involved in coagulation, it stays around in the periphery where then it activates other proteins involved in removing calcium from the blood vessels and bringing it to things like the bone or to other tissues.

And so what Joyce found is that when she looked at the literature, the proteins that are involved in coagulation, when you knock those proteins out in mice, they're embryonic lethal. Big surprise. It's required probably for short-term survival. The other proteins that are involved in removing calcium from the bloodstream, bringing it to the bone and tissues like osteocalcin, matrix globulin, protein, those phenotypes are not lethal when you knock them out in mice. However, they lead to buildup of calcium in the arteries, they lead to cardiovascular problems, and they lead to cancer, they lead to bone problems with osteoporosis, and also it leads to soft tissue problems.

Additionally, when you take a human and you deplete them of their vitamin K, not entirely, but you you cause them to undergo severe deficiency. What's very interesting is that there is no change in coagulation. Again, suggesting that whatever vitamin K you do get from your diet, it's going to the liver to make sure that coagulation is taken care of at the expense of the other proteins that are important for making sure calcium does not build up in the vascular system. Practical solution for this: eat your greens. Magnesium is high in spinach and kale and other green leafy vegetables. Vitamin K, calcium, vitamin C. This is me drinking my vegetable smoothie that I drink almost every day, which has kale and spinach and chard and more vegetables.

So I'm going to switch gears and talk a little bit about some of my current research on vitamin D. So as I mentioned earlier, Around 70% of the US population does not get adequate levels of vitamin D. Well, what is adequate levels of vitamin D? So the Endocrine Society has set a spectra of blood levels of 25-hydroxyvitamin D, which is the precursor to the active steroid hormone. Vitamin D gets converted into a steroid hormone. I'll show you that in the next slide. But Deficiency is considered at levels below 20 nanograms per milliliter. Sufficiency is considered at levels between 30 and 60, and being inadequate is considered at levels below 30 nanograms per milliliter.

So taking around 1,000 IUs of vitamin D per day can raise blood serum levels by about 5 nanograms per milliliter. So this is the standard based on all-cause mortality studies. There's about 31 different studies in the literature that show people that have vitamin D levels between 30 and 60, 40 to 60 nanograms per milliliter have the lowest all-cause mortality including cardiovascular disease, neurodegenerative disease, and cancer. So I like to be within that range. Too much vitamin D is also not good because vitamin D also allows you to absorb more dietary calcium. And as I mentioned, calcium can precipitate and form plaques in the vascular system.

So if you're taking megadoses of vitamin D and you don't have enough of your vitamin K, that could be a problem because now you're absorbing all this dietary calcium, but the calcium isn't going to the bones and other tissues where it's supposed to go because those proteins aren't being activated by vitamin K. The primary source of vitamin D is UVB radiation from the sun. UVB radiation hits our skin, it converts something called 7-dehydrocholesterol in our skin to vitamin D3. Vitamin D3 then gets released into the bloodstream where then it goes to the liver and is converted into 25-hydroxyvitamin D, which is the major circulating form of vitamin D and is the form that is measured when you get your vitamin D levels measured by your physician.

25-hydroxyvitamin D then goes to the kidneys where it's then activated into the steroid hormone. So vitamin D is a steroid hormone, much like estrogen or testosterone, that is able to— I'm stuck here on my slide here. It's not moving forward. Oh, there we go. The active steroid hormone. And it actually regulates the expression of over 1,000 different genes in the human body. And so if I can advance to the next slide. Whoops. Went a little too far. So vitamin D is a steroid hormone that regulates over 1,000 different genes in the body. So if you think about the entire protein-encoding human genome, that's around 5% of the human protein-encoding genome that vitamin D is controlling.

So it's turning genes on. It's turning genes off. I will get to that in just a second. There are many different factors that regulate the ability of our bodies to produce and use vitamin D. Because UVB radiation is the primary source of vitamin D, anything that blocks out UVB light like sunscreen or melanin, which is the this dark dark skin pigment that protects us from the burning rays of the sun, also impairs our ability to make vitamin D. In addition, body fat vitamin D is fat soluble. It's stored in the fat. And studies have shown that the more body fat, the less bioavailable vitamin D is to be released into the bloodstream to be converted into the active steroid hormone.

Age also, as we age, the enzymes in the skin that convert 7-dehydrocholesterol into vitamin D3 become less efficient, as everything does with age. And so a 70-year-old actually produces 4 times less vitamin D in their skin from the sun than their former 20-year-old self. Also latitude, depending on where you live. So living in a northern latitude also affects the ability to make vitamin D because at certain winter months of the year, UVB light isn't— or UVB rays aren't actually hitting the atmosphere. Many different factors that are regulating vitamin D. How does vitamin D regulate gene expression? Well, vitamin D, when you get it, it binds to the vitamin D receptor.

This causes the vitamin D receptor to heterodimerize with the retinoid receptor. And this complex then goes inside the nucleus of the cell where the DNA is, and it recognizes a little telltale sequence in the DNA called a vitamin D response element, which is essentially 2 repeats of 6 nucleotides separated by 3 nucleotides. And the sequence itself actually determines whether or not a gene is going to be turned on to be more active to do its function or going to be turned off. So when it's turned on, it recruits cofactors. This turns genes on. In the case of turning a gene off, the sequence in the DNA itself, even just one nucleotide change, can indicate to this complex off signal, and it recruits corepressors instead of coactivators.

And this will then silence the gene, turn it off, so that even though the gene is there, it's not functioning. So both activation and repression occur. So I recently found that the gene that encodes for the enzyme tryptophan hydroxylase, which is the rate-limiting enzyme in the conversion of tryptophan into serotonin, has a vitamin D response element in it. So most of you may be familiar with serotonin as being a neurotransmitter. I'm going to dive a little bit more into that in just a minute. But what's interesting is that humans and other mammals have 2 separate genes for tryptophan hydroxylase. We have tryptophan hydroxylase 1 and tryptophan hydroxylase 2, and they're localized to different tissues.

So tryptophan hydroxylase 1 is in the gut primarily, and it's also in the placenta and T cells. And this is where it takes tryptophan and converts it into serotonin in the gut. About 90% of the serotonin we make in our body is actually made in the gut. Serotonin does not cross over the blood-brain barrier, so the serotonin in the gut does not get into the brain. Okay. It's very important to make serotonin in the gut because that serotonin made in the gut is taken up by platelets and platelets then require serotonin to coagulate so that it's involved also in coagulation. But it's a double-edged sword because too much serotonin in the gut also activates T cells to proliferate and it's associated with hyperactive immune response in the gut associated with colitis and inflammatory bowel disease.

And other gut issues. So too much serotonin in the gut is involved in inflammation in the gut. In the brain, the gene that make— that converts serotonin— sorry, tryptophan into serotonin is called tryptophan hydroxylase 2, completely separate from the one in the gut. In the brain, tryptophan gets transported and is then converted into serotonin in the brain where it's a neurotransmitter and also much more than that. I'll get into that in just a minute. But what I found is that the 2 separate genes both have a vitamin D response element, but their response elements were different. They were functionally opposite to one another. The one in the gut had a vitamin D response element that was an off signal, meaning when vitamin D is present, it shuts down the production of serotonin in the gut.

Whereas the one in the brain had an on signal, meaning it activates or turns on produces the production of serotonin in the brain. So I published a paper with Bruce where I talk about how vitamin D hormone regulates serotonin and how this is relevant for autism. So putting together this autism puzzle, this explained 4 different characteristics associated with autism. The low vitamin D levels that are associated with autism explains the low serotonin levels in the brain but the high serotonin levels that are found in the gut and the blood of autistics. It also explains, we think, the high male prevalence and also the presence of maternal antibodies against fetal brain tissue in mothers of autistic children.

So the rise in autism has been exponential. I mean, it's risen about 600% over the last 40 years. And while, you know, while increased awareness definitely plays a role in this, There have— there's really been no underlying mechanism explaining why the prevalence has increased so rapidly. 1 in 68 children in the United States is diagnosed with autism. There have been some genetic factors identified to play a role in autism, but 70% of autistic cases cannot be linked to a genetic cause, meaning there must be something in the environment that is also playing a role in autism. So if you look at this at the same time period where autism is rising, vitamin D in the population is decreasing and that's largely as a consequence of awareness from skin cancer.

People wear sunscreen now a lot more. Also technology, everyone has a personal laptop. We're inside in offices on our laptop. Televisions are ubiquitous. People are playing video games. Children are playing video games. So vitamin D levels have been going down at the same time that autism levels have been rising. So there's sort of a correlation there.

Okay.

As I mentioned, vitamin D hormone has and activates the gene TPH2, which is in the brain, and it produces serotonin. Well, in addition to being a neurotransmitter, serotonin does much more. It's also what's called a brain morphogen. So during early brain development, serotonin is required to shape the wire and the structure of the developing brain. It tells neurons where to go in the brain and what to differentiate into. So, it plays a very important role for brain development. And, when you deplete serotonin in mice, it causes abnormal brain development and leads to autistic-like symptoms in mice, if you can imagine. Now, the developing fetus completely depends on the maternal levels of vitamin D.

So, the mother levels of vitamin D are very important because that's how the infant is getting vitamin D through the placenta. So vitamin— you can imagine if a mom is deficient in vitamin D, doesn't have enough vitamin D, there may not be enough vitamin D getting into the fetal brain to activate TPH2 to convert tryptophan into serotonin. As a consequence, this could lead to abnormal brain development. Since my paper was published, which sort of laid the groundwork for people that actually have been doing autism research. Another group independently confirmed biochemically that vitamin D hormone indeed does activate tryptophan hydroxylase 2 in various different neuronal cell populations.

So it does increase the expression of the gene. This has been validated biochemically. So I mentioned that it also explains the high male prevalence with autism. Well, in addition to vitamin D activating TPH2 to increase serotonin, estrogen also activates this gene. So there's sort of a backup mechanism in female fetuses. It's been shown from amniotic fluid that female fetuses have higher levels of estrogen. They also have higher levels of estrogen neonatally right after birth. So it's possible that if a mother is deficient in vitamin D and she's carrying a female child, there's a backup system because that estrogen is able to activate the same gene that vitamin D activates.

But if it's a male fetus, they don't have that backup system and they may be more susceptible to having the low serotonin, which would then affect the brain development and possibly lead to autistic-like behaviors. And lastly, I mentioned that we think that the vitamin D-regulated serotonin may explain the high maternal autoantibody to fetal brain tissue. So mothers of autistic children are 4 times as likely to have antibodies in their blood against proteins that are in the brain. So, you know, a developing fetus, then if you have an immune response, the immune cells then go and start to attack the fetal brain tissue and this can cause abnormal brain development.

And this has actually been shown in monkeys. So monkeys, this work is done at UC Davis, when they cause the monkeys to have this autoimmune response where these immune cells start to attack the fetal brain tissue, the monkeys then get autistic-like behaviors. So how does this occur? Well, I mentioned that in addition to vitamin D turning on the gene that makes serotonin in the brain, it turns off the gene that makes serotonin in the gut, also in the placenta. So tryptophan not only gets metabolized into serotonin by tryptophan hydroxylase, but there's another pathway called— an enzyme called IDO which also converts tryptophan into something called kynurenine which then gets converted into T regulatory cells.

T regulatory cells are very important immune cells that are essential to prevent your body from having an autoimmune response, to prevent your body's immune cells from attacking its own tissue. In mice, it's been shown that if you delete the enzyme IDO so that tryptophan cannot be metabolized into kynurenine or into the T regulatory cells, female mice have such a— when they're pregnant, they have such a strong autoimmune response against the fetus that their immune system actually kills the fetus. And it's obviously a very severe condition, but that's been shown to be dependent on the fact that these mice cannot make kynurenine and T regulatory cells.

So, tryptophan actually binds much more tightly to tryptophan hydroxylase 1 compared to IDO. So, we think under conditions of low vitamin D, the TPH1 is not being turned off, it's not being regulated normally. And, this leads to aberrant expression of this enzyme. You're making too much of it, which then acts as a tryptophan trap, a sink for tryptophan. So now, tryptophan is only getting metabolized into serotonin and it's not going to be producing enough of the T regulatory cells, potentially when this is happening during pregnancy, this could lead to maybe a little bit of an autoimmune response and possibly cause, you know, the mother's immune cells to then start making antibodies against fetal brain tissue.

So that needs to be tested. We're working with some people from UC Davis that are— that have been doing this type of research. Hopefully, they'll be interested in testing this out. But this all really leads to a very simple solution that is relevant for prevention. And that is, one, vitamin D levels should be measured prenatally. I mean, this should be part of a prenatal care package where much like folic acid— folic acid is emphasized to prevent neural tube defects— well, vitamin D levels should be measured And the levels of vitamin D should be within a certain range. And if these women do not have enough vitamin D, they need to be given a vitamin D supplement.

As I mentioned, about 1,000 IUs of vitamin D raises serum levels by about 5 nanograms per milliliter. If you have a woman that's severely deficient in vitamin D, less than 20 nanograms per mL, and you only give her 400 IUs, you're not even going to raise her blood levels by 2.5 nanograms. So it's just not enough. So I think measuring and making sure you have the right dose, not to mention there are many common gene polymorphisms in the enzymes that convert vitamin D3 into 25-hydroxyvitamin D, very common. In fact, I know several people that have these polymorphisms that make the enzyme less active. And so people with these polymorphisms have lower vitamin D levels and they'll never know that.

They may be taking a vitamin D supplement, but if they don't get their levels measured, they won't know. So, in those cases, people actually need a higher dose to get normal levels of vitamin D, like I said, which are above 30 nanograms per milliliter. So, a little bit of a follow-up on my first study, I published another study recently that elaborates on my first study and talks about the role of vitamin D in producing serotonin in the brain and activating the enzyme in the brain. And also how omega-3 fatty acids, specifically the marine omega-3 fatty acids, eicosapentaenoic acid, EPA, and docosahexaenoic acid, DHA, also regulate the serotonin system and how this is relevant for brain function and brain dysfunction, particularly for bipolar disorder, for schizophrenia, depression, anxiety, ADHD, these types of neurodevelopmental and neuropsychiatric disorders.

So, vitamin D hormone I mentioned, you know, activates this gene TPH2— I'm really driving this home— to make serotonin in the brain. Well, serotonin in the brain, in addition to being a neurotransmitter that most people associate with mood, does much more. It also regulates impulse control, executive function, aggression, anxiety, memory even. So, these studies have been done by psychologists and other neurobiologists where They've taken normal people and depleted their serotonin by giving them a shake of branched-chain amino acids like leucine and isoleucine, which outcompete tryptophan to be transported into the brain. So, what happens is after you give someone a big shake of these branched-chain amino acids without tryptophan, their tryptophan levels drop in their brain and subsequently their serotonin levels drop by about 90%.

And what happens is people become very impulsive. Their short-term thing— their short-term gratification kicks in. Their long-term planning and memory, those long-term functions, they shut down. They go for the instant gratification. People become anxious, depressed. Their mood changes. They have a hard time interpreting facial expressions of anger or happiness, which is related to empathy. They also become a little bit more impulse-aggressive as well. So, you know, and in addition, their sensory gating, so the ability to filter out, you know, different auditory and visual stimuli and just focus in on one thing, that also changes and that they have problems with that, which is very relevant for schizophrenia and ADHD.

So, serotonin is doing a lot in the way we behave and the way we feel. In the way our brain is functioning. And if vitamin D is important for the production of serotonin and people are not getting enough vitamin D, what does that say about our brain functioning? It may not be optimal. So in this study, I talk about the interaction between gene polymorphisms, people that have polymorphisms in serotonin-related genes that already predispose them to low serotonin, and how those people in addition to having that polymorphism, if they are also deficient in vitamin D, that may precipitate brain dysfunction. It may precipitate schizophrenia or in combination with other stressful factors in life.

So I think it's really important to optimize what we have control over. And what we have control over is our vitamin D levels. In addition to vitamin D, omega-3, the marine omega-3 fatty acids also regulate serotonin function. So, tryptophan gets converted into serotonin by the enzyme TPH2, which is what vitamin D regulates. But, in the neuron, in the presynaptic neuron, serotonin has to be released into the synapse. And, the release of serotonin into the synapse depends on inflammation. So, when inflammation is high, whether that's because you're not eating a good diet, you have gut issues, whatever the cause, many different things cause inflammation, overactive immune system, not getting the right micronutrients.

nutrients, this causes the production of something called the E2-series prostaglandins. E2-series prostaglandins have been shown if you're making them in the gut, they cross over the blood-brain barrier, they get into the brain, and they prevent the release of serotonin. So, serotonin cannot be released from the presynaptic synapse or neuron into the synapse when inflammation is high. Well, eicosapentaenoic acid, the marine fatty acid EPA, stops the production of E2-series prostaglandins. And because it dampens that production of E2-series prostaglandins, it allows serotonin to be released from the presynaptic neuron. That's been shown in animals.

In addition, once the serotonin is released from the presynaptic neuron, it has to bind to the postsynaptic neuron. And there's a receptor on the postsynaptic neuron, the serotonin receptor, that binds to serotonin in order for the action of serotonin to happen so that So, you're releasing serotonin and then the function occurs because it binds to the receptor. Well, docahexaenoic acid, DHA, is very important for the cell membrane and particularly in neurons. The cell membrane has a certain fluidity to it and this fluidity is very important because receptors are embedded in that cell membrane. And, if you disrupt that cell membrane fluidity, it changes the structure and ultimately the function of those receptors.

Including the serotonin receptors, which cross the membrane 7 times. And, it's been shown that when you deplete mice of DHA that their serotonin receptor changes in structure and subsequently, they can't bind— it does not bind to the serotonin as well. So, DHA deficiency also changes the serotonin system by by altering the function of serotonin. So what this says to me and what I talk about in the paper is that under conditions of low vitamin D, which we know is ubiquitous, at least in the United States, and also it's globally, people are, you know, they're not getting enough vitamin D for the reasons I mentioned. But also, fish consumption is down and people don't eat enough fish and are not getting enough omega-3 fatty acids.

So if you're not getting enough vitamin D, you're not getting enough omega-3, then what's happening in the brain in terms of the serotonin system, it may not be working optimally. And, you take a person then that also has gene polymorphisms that already predispose them to low serotonin production or metabolizing the serotonin too quickly or the serotonin receptor not working, on top of that they're deficient, then this is a bad combination. And then, you add a stressful life event and things like this, you're talking about the perfect storm for neuropsychiatric disease, for depression, anxiety, schizophrenia. So I think that this may be an underlying mechanism.

There's probably many things going on, but it's something that explains why people can be treated by high doses of fish oil and vitamin D. It's been shown to help with symptoms of schizophrenia. It's been shown to help with anxiety, depression. It's been shown to help with ADHD. Both vitamin D and omega-3 have been shown. And I think this may be partly— part of the underlying mechanism by which both of these micronutrients are working. And I think that also it offers a very simple and easy lifestyle change and that is make sure you're getting enough of these micronutrients. You can take a vitamin D supplement. It costs a penny a pill. And you can take a fish oil supplement.

They're extremely important and it's an easy change to make. It's often hard to get people to make lifestyle changes as It's hard to get people to change their diet most of the time because they're not— their brain isn't working optimally already, so it's hard to make the change. But something like taking a fish oil pill or a vitamin D pill is easy enough to do. And once they start to feel better and, you know, executive function is working and they start to notice the change, I think other lifestyle changes are then easier to implement. So I think that, you know, vitamin D and omega-3 are simple solutions. to complex problems. And with that, I would like to end and thank you all for inviting me.

Dank u wel. And thank my mentor Bruce Ames for all the guidance.

Thank you, Dr. Rhonda Patrick. Questions? Are there any questions? I come with the mic. Is it on? Did you turn it on?

Yeah. Yeah. When you give a patient vitamin D supplements, can you think of causes that the body doesn't take it in or accept it, or that there are problems with that?

So the question is, if you're going to give a patient a vitamin D supplement, are there problems with not absorbing it, bioavailability, and things like that? And to answer the question, yes, there are— there can be. So, you know, vitamin D is absorbed better with fat. It's, you know, a fat-soluble vitamin. And vitamin D3, is better than vitamin D2, which is— vitamin D2 is found in plants and fungi. They also make vitamin D upon UVB light. So vitamin D3 is actually better. It's more easily converted into the vitamin D hormone compared to vitamin D2. Also, the dose you give also regulates how much vitamin D is converted into the hormone. So it's been shown that the higher the vitamin D dose, the more bioavailable it is.

Now, you don't want to give someone too much. Like, too much vitamin D can be toxic. The National Institute for Medicine has set the upper tolerable intake at 4,000 IUs a day. So, I personally take 4,000 IUs a day and I get my measure— my vitamin D levels measured ideally quarterly. And, my levels hover around 50 nanograms per mL.

Okay.

So, I think that giving a patient a supplement should be done along with measuring their levels because you won't know also if they have a polymorphism that changes the conversion of D3 into 25-hydroxy. I have 3 friends that have this and they did 23andMe genetic test and found indeed they do have a polymorphism in the CYP2R1 gene, which is the gene that converts that makes the enzyme that converts D3 into 25-hydroxy. But I think that measuring the blood levels also very, very important along with taking the supplement.

Okay, any more questions? When we describe vitamin D, should we also describe vitamin K and how much?

Great question. So the question is, if you prescribe vitamin D, should you also co-prescribe vitamin K2? And the question is, how much? And I say K2 because vitamin K2 does not— I mentioned there's 2 biologically active forms of vitamin K, K1 and K2. Vitamin K1 is found in plants and readily goes to the liver. Once it goes to the liver and has activated all the blood clotting proteins, then it stays around in the periphery in the blood system and activates the calcium-dependent proteins that pull calcium out of— I'm sorry, the vitamin K-dependent proteins that pull calcium out of the bloodstream, bring it to the bone, etc. Vitamin K2, which is made in bacteria in our gut if we have a healthy gut and also in other bacteria that ferment foods, does not go to the liver as readily.

So, vitamin K2 is kind of like a backup insurance for vitamin K1. I don't know how much vitamin K2 should be taken. So if you get the natural form of vitamin K2 from natto, studies have shown that there is— even though it is a fat-soluble vitamin, there is no upper level of toxicity. So there's been no limit that has been set for vitamin K2. You can take a very high dose of it and have no toxicity issues. I personally do take vitamin K2, and I take around 50 micrograms. a day. And, I even think that's probably more than enough. So, there have been some studies showing that taking between 25 and 50 micrograms a day of vitamin K2 does decrease calcium buildup in the arteries.

So, I wish I had a more direct answer. And, the answer is I don't know. Those studies haven't been done. But, at the very least, we know that the natural form I say natural because there's menaquinone and then they have this synthetic form they make called menadione, and that does have a toxicity level. So, I think that getting vitamin K2 from natto, which is where a lot of the supplements, it's MK-7 or MK-4. MK-7 is menaquinone-7, which has a longer half-life than MK-4. MK-4, most of the clinical studies that have been done looking at the effects of vitamin K2 supplementation on improving cardiovascular problems have been done with MK-4, but they're essentially very similar except for that MK-7 has a longer half-life.

I personally take MK-7. I take 50 micrograms a day.

Okay. Just one more question.

Thank you for your presentation. I have a question about the marine fatty acids. In the Netherlands, we have a recommended allowance of 450 milligrams of EPA and DHA together per day. Do you suggest in your paper or otherwise any specific dose?

So the question is, you know, what dose of omega-3 fatty acids are recommended for some of the effects that I talk about in the paper regulating the serotonin system and also helping improve symptoms of ADHD, bipolar, schizophrenia. And the answer is it is— it varies between studies. So most of the studies that I cite in the paper are using high doses of omega-3, of fish oil. And they're using between 3 to 6 grams a day. It's a clinical dose. You know, sometimes you have to take higher doses to get a therapeutic benefit. And also, there are some very important studies showing the ratio of EPA to DHA is very important, particularly for the anxiety and depression, for eliminating anxiety and depression.

It's been shown that a 2-to-1 ratio of EPA to DHA DHA is very important. If you have too much DHA, then the effects are not found. So the depression was not— the symptoms were not treated. And the scientists that are studying this don't really know why. They have some speculations. For one, they think possibly that the DHA and EPA are competing for a similar enzyme. And so if you have too much of the DHA, the EPA, you know, isn't you know, there's a competition going on. Another speculation is that DHA is getting metabolized into something that's interfering with EPA effects. Personally, what I think is important is I think that because EPA is more of the anti-inflammatory, it's involved in preventing the prostaglandin production and inflammation plays a major role in depression.

And we know this because of several studies that have shown even injecting people with a pro-inflammatory cytokine like interferon, they inject people with that and they immediately become depressed. But if they co-administer EPA with that, they don't get depressed. We know that this, you know, inflammatory cytokines, inflammatory molecules produced anywhere in the body, main source of it is gut. Endotoxin gets released when we have— our gut barrier becomes compromised, you know, and then inflammation is happening. all these molecules, they get into the brain and that messes up the serotonin system and messes up other neurotransmitters, not just serotonin, dopamine as well.

So, I think that the EPA itself has a very important role just because it's negating the inflammation. So, it really depends on the person. And, I think that for a therapeutic effect, you probably, at least in the studies that I've read, doses have to go much higher than 450 milligrams. They were taking—

Yeah.

pretty high doses that were, you know, 3 grams a day.

So, okay, thank you, uh, very much. Oh, there's still another question. It's the last question. We have to go to the break.

Your name, please?

Marion.

Doctor, like Miss Marion has said before, are we part of the whole environment And is it not true that we supposed to know a little of the physical body consciously? Can it be controlled by us, our sensory and mechanical force? Is it possible?

So, I think the question is, can we control our physiology?

Yes.

Biologic. And to answer that question, I don't have the answer to that question, so, to start off. But I'm very interested in the answer to that question. I think that if you look in the literature and see how people taking a placebo pill, how they can achieve therapeutic benefits from taking a placebo pill, which they don't know is a placebo. I think that's direct evidence that indeed, yes, we can control our physiology. We can control our immune system. You know, we can control dopamine production in our brain. And evidence of that really is the placebo effect. I mean, the fact that that works tells us that, yes, we can control our physiology to some degree.

Now, how that's happening, I don't know.

I can't give the correct answer, but some of the answers will be after lunch.

Yeah, there you go.

I know, you told me. Okay, uh, thank you very much, Dr. Rhonda Patrick.

Thank you.

It was very interesting.