Amino Acids The building blocks of life. Daniel Reheis

Amino Acids The building blocks of life Daniel Reheis February 1st, 2017

A`MI The biggest trend in the market is towards natural, raw, full spectrum, and high quality ingredients There is no amino acid product on the market that fulfills all these criteria like A MI Commerciality Although we just have hit the market, the response is huge The biggest response came from: People with chronic fatigue syndrome, who felt a significant change within 24 hours Professional athletes, who are very aware and sensitive of the effects and side effects of their supplements The response was so good that three premiere league football clubs, 2 world class skiers, 2 world class tennis players and one world class cycler decided to regularly use our product But they are not sponsored by us So 95% of the professional athletes who received test samples continued to use our product and are also paying for it Doctors, because of their feedback from their patients Marketing Besides professional athletes, the focus lies on middle aged persons with higher income and a focus on healthy lifestyle But we also offer protocols and lifestyle concepts for people who want to improve their health and energy levels, but have not much time or simply don t know how to start ur customer segment is not concerned with price, but demands uncompromising quality ur marketing tools are based around these demands Channels ur main channel is our own online shop ur customers start at doctors, therapists or coaches, who recommend the products to their athletes or other multipliers They then make the product known to their customers or social media followers Specific campaigns will be launched after further feedback from customers utra Pride There is an endless list of implications for amino acids Most of all, they seem to be a quick win for almost every chronic disease, mainly because of the amino acid consumption of the immune system Providing the first full spectrum, free form amino acid product in pharmaceutical grade quality, which has been formulated to fit a human s needs exactly, makes us proud This means that everyone at every age with all known allergies or intolerances can consume this product 1

ALIPHATIC ARMATIC ACIDIC BASIC HYDRXYLIC SULFUR CTAIIG AMIDIC H H 2 H H H H H ALAIE Ala GLYCIE Gly ISLEUCIE Ile LEUCIE Leu PRLIE Pro VALIE Val H H H AE H H H ME H H 2 H H H H PHEYLALAIE Phe TRYPTPHA Trp TYRSIE Try ASPARTIC ACID ASP GLUTAMIC ACID Glu ARGIIE Arg HISTIDIE His H 3 H H ME H H H AE HS H S H H 2 H H 2 H LYSIE Lys SERIE Ser THREIE THR CYSTEIE Cys METHIIE Met ASPARAGIE Asn GLUTAMIE Glu ALL THE REST AE ABSLUT ESSETIAL ME METABLIC ESSETIAL FE FUCTIAL ESSETIAL The totality of all the different proteins in a living organism is known as the proteome In humans, this numbers between 500,000 1,000,000 and all of them are made of long chains of amino acids The functions of proteins range from hormones and neurotransmitters for glucose handling (insulin) and influencing gene activity (mra) to the functions of sleep (melatonin), waking up (noradrenaline), dreaming (dopamine and serotonin), and above all, the structural components of our bodies muscles and connective tissue Figure 1: You may miss the classical categorization in essential and nonessential In this text we will explain our new classification Since 1947, amino acids were categorized as essential and non-essential 1 Essential was interpreted as the human organism couldn t make them, or couldn t make them in sufficient quantity on-essential was interpreted as that the body could manufacture those amino acids adequately Womack & Rose, however, clearly didn t mean it like that We have emphasized on several occasions that the classification of an amino acid like arginine or glutamic acid as dispensable or indispensable is purely a matter of definition 2

Essential and non-essential Manufactured amino acid products typically only contain the essential amino acids, given that it s widely accepted that the body can produce adequate amounts of the non-essential ones Essential would, by definition, imply that we must ingest these via our dietary intake However, we would argue that this actually applies only to threonine and lysine, because all of the other amino acids under the category of essential (valine, isoleucine, leucine (BCAA s), phenylananine (Pha), and methionine (Me), may be produced from alpha ketones Figure 2: The classification of essential and non-essential amino acids Taking a closer look at the non-essentials, glutamic acid and serine may be the only ones to fall under this category They can be built from a non-amino acid source (eg ammonium ions) and an appropriate carbohydrate source 2 If the production of these were independent of the availability of other amino acids, the internally manufactured levels in the body would be at a constant level irrespective of measuring before or after food Table 1 shows clearly that this is the case, and as it would be expected, glutaminic acid and serine also exhibit the highest levels of production If anything, then according to the definition, only these two should be considered non-essential 3 3

Flux synthesis Amino acid Fed state % Fasted state % Glutamate 1 98 98 Serine 1 94 95 Aspartate 1 76 78 Alanine 1 45 46 Table 1 (based on Stoll, 1999 3 ) Amounts of synthesis of a selection of non-essential amino acids In all instances glutamate and serine have the highest production Proline, in contrast, is not produced in the fed state, and must therefore be consumed via the diet Glycine 1 35 35 Arginine 2 14 10 Proline 0 7 n this point, we differ from other experts Indeed, essential amino acids are not easily formed from their ketone precursors In this sense they remain essential But there is also a wide range of reasons that may result in inadequate endogenous production of the non-essentials In this text we would like to discuss the principle that all amino acids are essential for health, and an optimal supply must be consumed via the diet We will now demonstrate why we should still consume adequate amounts of amino acids that we can manufacture internally ourselves 1 Lack of precursors The amino acids arginine, cysteine, proline, and perhaps glycine can be made from carbon donors such as glutamate and serine, or other amino acid precursors, or ammonia (table 2) Product Carbon precursor ther precursor Arginine Glutamate/glutamine/proline Ammonia and aspartate Proline Glutamate/glutamine Glutamate/glutamine Cysteine Serine Methionine (S) Glycine Serine Table 2 (based on Stoll, 1999 3 ) The precursors of arginine, cysteine, proline, and glycine A deficiency of the precursors may lead to a lack of these amino acids The nutritional intake can therefore augment the availability Under these circumstances, they would no longer be considered non-essential 2 Insufficient production A certain number of amino acids may be produced in certain organs arginine and proline can be made exclusively in the liver and kidneys 4 (figure 3) 4

H Kidney Liver PRLIE Pro Arg Arg Arg H 2 H H 5 rn 6 ARGIIE Arg Cit Pro 7 Pro H 2 H Cit GLUTAMIE Glu Pro CITRULIE Cit GSA 4 rn 3 2 1 Glu RITHIE Small intestine Figure 3 shows how arginine is produced from the byproducts of the glutamine cycle (citrulline) in the kidney Following that, arginine can also act as a precursor of proline 4 It shows that arginine and proline can only be produced from glutamine which must be consumed in the diet 3 Digestive problems and glutamine deficiency may therefore lead to deficiencies of arginine and proline Shortcomings in these processes can lead to a number of disturbances in the synthesis and function of connective tissue (proline), at the energy level (creatine which is formed from arginine), or neurological disturbances (due to a lack of nitric oxide () also formed from arginine) A nitric oxide deficiency can also be a reason for high blood pressure, immune and cardiovascular disturbances 5 Glutamate, which is built from glutamine is only metabolized in the gut The same applies for one third of all the nutritionally sourced amino acids threonine, leucine, lysine and phenylalanine 6 Figure 3 (based on Brosnan, 2003 4 ) The formation of arginine (Arg) and proline (Pro) by means of the de-novo synthesis of glutamine, (Glu) and arginine (Arg) Primary pathways of arginine and proline synthesis The numbered enzymes are: 1 pyrroline-5- carboxylate, 2, spontaneous reaction; and pyrroline-5 carboxylate reductase 3, ornithine aminotransferase; 4, ornithine transcarbamoylase; 5, argininosuccinate synthetase; and argininosuccinate lyase; 6, pyrroline- 5-carboxylate reductase; and arginase 3 ot fast enough In many cases, amino acid production does not adjust quickly enough to an acute increase in demand 7; 8 This applies in particular to people with injuries 9; 7, and also to children during periods of increased growth At these times, children may not be able to produce sufficient enzymes to convert citrulline into 5

adequate amounts of arginine Enzymes involved would be arginine succinate synthetase, and arginine succinate lyase Breast milk contains an optimal composition of amino acids to support the organs and optimal human function That s why our formula is based on the composition of late-stage mother s breast milk (hindmilk) with some adaptions to our modern lifestyle 4 o storage The body has no mechanism to store amino acids This means there is a continuous need for amino acids, in order to be able to produce any of those 500,000+ proteins 5 They are used as an energy source Almost all amino acids act like glycogen that is, they can be transformed into glucose, which is the most important substrate for energy delivery, and in the case of the activated immune system is the only source 12 During higher demand like during an infection, or starvation - first the glycogen stores of the liver are used, lasting about 6 hours In the next 3 days, the stores of amino acids in muscle and connective tissue are used Primarily, glutamine and alanine are used as energy sources for the immune system providing a faster source of fuel for the immune system than fat nly after 3 days the body begins to consume its fat reservoirs 12 If the immune system is chronically activated (eg low grade inflammation, chronic stress, intestinal permeability, poor food quality, etc) there is a corresponding increase in demand for amino acids 15 Figure 4 (adapted from Lochmiller, 2000 15 ) Increases in protein consumption during different types of immune system activity Low grade inflammation is a state that more or less affects almost every human in industrialized countries 15 Hence: Excess amino acids are catabolized (converted into energy) Deficiency of amino acids in terms of dietary intake potentially leads to impaired production capability (see under 5) From a functional perspective all are essential 10; 11 6

So where does that bring us? In summary, it can be said, all amino acids are functionally essential A deficiency of one or more amino acids in dietary intake produces in the first instance a shift in the production of classically defined non-essential amino acids, whereby the function of the classic essential amino acids is also put at risk Long-term deficiency of amino acids due to high turnover, reduced intake, or reduced absorption leads to an absolute deficiency in amino acids and disturbance of all functions This affects in particular the two most important life critical materials which may not be produced to adequate levels: creatine for energy and glutathione for detoxification This may also explain why people with chronic diseases often show more and more symptoms Any disturbances in the relationship between the branched chain amino acids (BCAAs - leucine, isoleucine, valine) and the aromatic amino acids (tryptophan, phenylalanine, tyrosine) may lead to problems in the neuromuscular, immune and central nervous system 19; 20 The classical distinction between essential and non-essential is no longer valid Because all may be considered essential, we d like to suggest some different classifications or descriptions: Absolutely essential Threonine, Lysine, Tryptophan We really can t produce these Metabolically essential Glutamine, Serine We cannot build them fast enough when demand increases Functionally essential all the others We cannot build enough, if the turnover is chronically high for an extended period of time Why were carnitine, taurine and choline included in the formula? At any time, a certain number of amino acid compounds are not produced in sufficient amounts in the body 2 This applies to carnitine and its related compound acetyl-l-carnitine L-carnitine is formed from l-lysine and methionine in the liver and kidneys Cofactors for the production of carnitine include vitamin C, vitamin B6, niacin and iron Carnitine and acetyl-l-carnitine (figure 5) are responsible for a variety of functions in the human body The current understanding regarding the functions of carnitine and acetyl-l-carnitine include energy homeostasis of the heart (in particular fat metabolism, beta oxidation, acetyl CoA), mitochondrial detoxification (2-acetyl-carnitine transferase and 1-carnitine-acetyl-carnitine translocase) and the regulation of the im- 7

mune system (phagocyte activity, natural killer cell activity) Less well known is the fact that carnitine, and above all, acetyl-l-carnitine are important precursors for the neurotransmitters glycine, asparatate, GABA and glutamate 13 Figure 5 (from Kuratsune, 2002 13 ) It has been shown that people with Chronic Fatigue Syndrome have reduced uptake of acetyl-l-carnitine across the blood brain barrier This means that these people suffer from an insufficiency of the Glutamate/ GABA system underlying a large number of the symptoms associated with this syndrome 8

Taurine ne of the most comprehensive review studies on taurine is entitled Taurine: A very essential amino acid, by Harris Ripps 22 Ripps says: one of the few amino acids not incorporated into proteins, taurine is one of the most abundant amino acids in the brain, retina, muscle tissue, and organs throughout the body He highlights how important it is to consume a sufficient amount of taurine through food or supplementation Indeed the more this not yet fully understood substance is studied the more important it becomes nly very few of its functions can be presented here Taurine deficiency is associated with cardiomyopathy, renal dysfunction, developmental abnormalities, and severe damage to retinal neurons 8, 25 28 Taurine plays an essential role in systemic, metabolic inflammatory diseases, via its anti-oxidant activity and indirect regulation of inflammation Taurine also plays an essential role in systemic, metabolic, and inflammatory diseases 31; 32 Taurine acts as a form of neurotransmitter, and also helps to regulate the neurotransmitters glutamate and GABA 22 Amino acids and anti-aging Calorie restriction is the only scientifically supported anti-aging mechanism The main reason is the reduction of growth factors Dietary amino acids can activate growth factors, such as methionine and leucine Joel Brind and his team showed that a reduction in methionine increased life expectancy of Fischer Rats (similar to humans in terms of metabolism) by 30-40 % 14 This is highly significant Glycine is responsible for the clearance of methionine via the enzyme glycine -methyltransferase (GMT) The study showed that life expectancy increased with increased levels of glycine intake The other side of the coin would have been a decrease in growth factors o significant detrimental effect on growth factors was seen Another study showed a clear association between methionine concentration in mammals and life expectancy 35 9

Figure 6 (from Pamplona 2006 35 ) Inverse correlation between protein methionine content and maximum life span (MLSP) in eight mammalian species (R2 = 093, P< 0001, y = 288 x026) Values are means SEM In industrial countries, far too many growth factors are consumed along with growth factor stimulants such as starch, sugar, milk, alcohol, etc A reduction of methionine and a concurrent increase in glycine can be considered a modern anti-aging intervention Many publications support this hypothesis 16 18 Choline instead of methionine ne disadvantage of reducing methionine consumption is a resultant reduction in methyl groups However, this may be compensated for with choline which also has synergistic effects with amino acids From a chemistry point of view, choline (2-Hydroxyethyl)-trimethylammonium chloride) is an ammonium salt that has three different methyl groups These can replace the methionine methyl groups Choline is a component of the lipoproteins, which plays a role in the transport of fat to and from the liver Choline deficiency in diet increases the risk of fatty liver disease More than 50% of the populations of industrial countries are impacted by this 21 10

Day 30 Day 60 Day 90 Figure 7 (from Al-Humadi, 2012 36 ) verview of the hematoxylineosin (H E) staining of liver sections obtained from rats exposed to long-term (30, 60, and 90 days) dietary choline deprivation (CD), where evidence of the gradually developing steatosis is provided With regard to cholesterol transport, if choline is deficient, it can be replaced by apolipoprotein B-100, which makes the lipoproteins (LDL and HDL) more susceptible to oxidation This is the scenario where oxidized cholesterol may have a damaging effect in the body TMA trimethylamine oxide A handful of studies have shown that humans with high risk of cardiac disease also had high levels of TMA 23; 24 Two possible sources are carnitine and choline - in humans with certain gut flora But can TMA really cause heart disease? And if so, are carnitine and choline the significant sources? The amount of choline in our products per 10g daily dose is equivalent to two eggs and 800 mg carnitine This is a significant amount However, to put this in context, a study in 1999 showed that 19 out of 21 seafoods and fish contributed to a significant increase in TMA following consumption Halibut increased TMA by 53 times more than eggs 29 In so far as anything in nutritional science can be known, it is accepted that increased regular fish consumption lowers risk of cardiovascular disease For 40 years carnitine has been one of the most studied amino acids, with long-term studies and large sample sizes o serious negative side effects have been reported In contrast, many studies of regular supplementation showed lower cardiovascular risk 30 Even if carnitine and choline did result in an increase in TMA, the strongest proponents of this hypotheses commented on : the high correlation between urine and plasma levels of TMA argues for effective urinary clearance of TMA It is quickly and efficiently excreted 23 11

ur Formula ur main differences: Free amino acids from fermentation Pharmaceutical grade Composition similar to late mother s hindmilk Less methionine and leucine, more glycine With carnitine, taurine and choline o sweeteners or lecithin Why amino acids from fermentation? Fermentation is a process that takes place everywhere, all the time Fermented food has always been a part of human food Besides that, it takes place within the body itself By way of example, some amino acids are metabolized that way by bacteria The end product is familiar to our bodies and can be known as artgerecht (translated as: right for, or known to, our species) Fluid bed granulation together with fibre gum shows significant advantage over amino acids in powder form The combination melts on your tongue, as compared with a dusty powder feel in the mouth, and it does not clump Because of this, the product can be consumed straight off the spoon And because there is already absorption of amino acids via the mouth s mucous membranes, this offers the advantage that it goes to work on the spot, and the bioavailability and absorption is improved Besides that, the fibre gum naturally masks the naturally bitter taste of the amino acids, and also acts as a prebiotic in the gut 33; 34 It was very important for us not to use any sweeteners to avoid any disturbance to the human metabolism signaling pathways The formulation was based on the amino acid requirements for optimal growth and function of all systems and organs in the human body A similar composition is found in the breast milk of tribes that still live as hunter/gatherers 37 The demands associated with the growth and development of a child would indicate in principle that all amino acids are essential Therefore, this formula contains all of those The formula has been slightly adjusted to account for the typical diets of industrial nations, which contain too many growth factor drivers like milk products, cereal starch and sugars For that reason, the leucine portion is below a certain threshold, and the proportion of methionine has been reduced These two are the strongest activators of the growth factor mtr (mechanistic target of rapamycin) 38; 39 12

At the same time, we increased the glycine content, which acts like an antagonist 16 18 Many studies have identified reduction of growth factors as one of the few effective anti-aging methodologies Allergies and intolerances are massively increasing The immune system can react to proteins, which are long chains of amino acids, and often have other substances attached Because our product only contains free amino acids, and not proteins, an allergic reaction is highly unlikely Test our product for yourself and enjoy the benefits References 1 Womack M, Rose MC: The role of proline, hydroxyproline and glutamic acid in growth J Biol Chem 1947; 171 2 Reeds P J: Dispensable and Indispensable Amino Acids for Humans J utr 2000; 130 3 Stoll B, Burrin D G, Henry J, et al: Substrate oxidation by the portal drained viscera of fed piglets The American journal of physiology 1999; 277 4 Brosnan J T: Interorgan Amino Acid Transport and its Regulation J utr 2003; 133 5 Boger R H: When the endothelium cannot say anymore ADMA, an endogenous inhibitor of synthase, promotes cardiovascular disease European heart journal 2003; 24 6 Fuller M F, Reeds P J: itrogen cycling in the gut Annual review of nutrition 1998; 18 7 Yu Y M, Ryan C M, Castillo L, et al: Arginine and ornithine kinetics in severely burned patients: increased rate of arginine disposal American journal of physiology Endocrinology and metabolism 2001; 280 8 Beaumier L, Castillo L, Am Ajami, et al: Urea cycle intermediate kinetics and nitrate excretion at normal and ther- apeutic intakes of arginine in humans The American journal of physiology 1995; 269 9 Scibior D, Czeczot H: Arginine-metabolism and functions in the human organism Postepy higieny i medycyny doswiadczalnej (nline) 2004; 58 10 Reeds D, Cade W T, Patterson B W, et al: Whole-body proteolysis rate is elevated in HIV-associated insulin resistance Diabetes 2006; 55 11 Reeds P J, Burrin D G: Glutamine and the bowel The Journal of nutrition 2001; 131 12 Straub R H, Cutolo M, Buttgereit F, et al: Energy regulation and neuroendocrine-immune control in chronic inflammatory diseases Journal of internal medicine 2010; 267 13 Kuratsune H, Yamaguti K, Lindh G, et al: Brain regions involved in fatigue sensation: reduced acetylcarnitine uptake into the brain euroimage 2002; 17 14 Brind J, Malloy V, Augie I, et al: Dietary glycine supplementation mimics lifespan extension by dietary methionine restriction in Fisher 344 rats FASEB J 2011; 25 15 Lochmiller R L, Deerenberg C: Trade-offs in evolution- ary immunology: just what is the 13 cost of immunity? ikos 2000; 88 16 Zimmerman J A, Malloy V, Krajcik R, et al: utritional control of aging Experimental gerontology 2003; 38 17 Sun L, Sadighi A A, Miller R A, et al: Life-span extension in mice by preweaning food restriction and by methionine restriction in middle age The journals of gerontology Series A, Biological sciences and medical sciences 2009; 64 18 rentreich, Matias, JR, DeFelice A, et al: Low methionine ingestion by rats extends life span The Journal of nutrition 1993; 123 19 Fernstrom J D: Branchedchain amino acids and brain function The Journal of nutrition 2005; 135 20 Manjarrez G, Herrera R, Leon M, et al: A low brain serotonergic neurotransmission in children with type 1 diabetes detected through the intensity dependence of auditory-evoked potentials Diabetes care 2006; 29

21 Pissios P, Hong S, Kennedy A R, et al: Methionine and choline regulate the metabolic phenotype of a ketogenic diet Molecular Metabolism 2013; 2 22 Ripps H, Shen W: Review: taurine: a very essential amino acid Molecular vision 2012; 18 23 Tang W H, Wang Z, Levison B S, et al: Intestinal microbial metabolism of phosphatidylcholine and cardiovascular risk The ew England journal of medicine 2013; 368 24 Koeth R A, Wang Z, Levison B S, et al: Intestinal microbiota metabolism of L-carnitine, a nutrient in red meat, promotes atherosclerosis ature medicine 2013; 19 25 Zulli A: Taurine in cardiovascular disease Current opinion in clinical nutrition and metabolic care 2011; 14 26 Yamori Y, Taguchi T, Hamada A, et al: Taurine in health and diseases: consistent evidence from experimental and epidemiological studies Journal of biomedical science 2010; 17 Suppl 1 27 L Amoreaux W J, Cuttitta C, Santora A, et al: Taurine regulates insulin release from pancreatic beta cell lines Journal of biomedical science 2010; 17 Suppl 1 28 Schmidt S Y, Berson E L, Hayes K C: Retinal degeneration in cats fed casein I Taurine deficiency Investigative ophthalmology 1976; 15 29 Zhang A, Mitchell S, Smith R: Dietary Precursors of Trimethylamine in Man: A Pilot Study Food and Chemical Toxicology 1999; 37 30 Diicolantonio J J, Lavie C J, Fares H, et al: L-carnitine in the secondary prevention of cardiovascular disease: systematic review and meta-analysis Mayo Clinic proceedings 2013; 88 31 Marcinkiewicz J, Kontny E: Taurine and inflammatory diseases Amino acids 2014; 46 32 Aruoma I, Halliwell B, Hoey B M, et al: The antioxidant action of taurine, hypotaurine and their metabolic precursors The Biochemical journal 1988; 256 33 Su P, Henriksson A, Mitchell H: Selected prebiotics support the growth of probiotic mono-cultures in vitro Anaerobe 2007; 13 34 Michel C, Kravtchenko T P, David A, et al: In vitro prebiotic effects of Acacia gums onto the human intestinal microbiota depends on both botanical origin and environmental ph Anaerobe 1998; 4 35 Pamplona R, Barja G: Mitochondrial oxidative stress, aging and caloric restriction: the protein and methionine connection Biochimica et biophysica acta 2006; 1757 36 Al-Humadi H, Zarros A, Kyriakaki A, et al: Choline deprivation: an overview of the major hepatic metabolic response pathways Scandinavian journal of gastroenterology 2012; 47 37 Zhang Z, Adelman A S, Rai D, et al: Amino Acid Profiles in Term and Preterm Human Milk through Lactation: A Systematic Review utrients 2013; 5 38 Zhou Y, Ren J, Song T, et al: Methionine Regulates mtrc1 via the T1R1/T1R3- PLCβ-Ca2+-ERK1/2 Signal Transduction Process in C2C12 Cells International Journal of Molecular Sciences 2016; 17 14 39 Deldicque L, Sanchez C C, Horman S, et al: Antagonistic effects of leucine and glutamine on the mtr pathway in myogenic C2C12 cells Amino acids 2008; 35