Detoxicification

&

Oxidative stress

scavenging

 

Presented at;

The 22nd & 23rd Scientific Meeting of the Japanese Society of Anti-Aging Medicine (17-19 June 2022, Osaka, Japan, 9-11 June 2023, Tokyo),

The 2nd Edition of Food Science and Technology Virtual (V-Food 2022) (15-16 April 2022, Online),

The 30th Annual Meeting of International Congress on Nutrition and Integrative Medicine (ICNIM 2022)  (9-10 July 2022 Sapporo, Japan),

The 3rd International Electric Conference on Food: Food, Microbiome, and Health (FOODS 2022) (1-15 October 2022, Online),

The 8th International Electronic Conference on Medicinal Chemistry (ECMC 2022) (1-31 November 2022, Online)

Joint Research Partner : O’Atari Inc.

   Recently, more and more people around me have been saying that they are feeling “sluggish” and “don’t have the motivation to do anything.” I often meet people who tell me “Maybe it’s my age, but I’m suddenly not feeling well these days”, meaning their physical condition is deteriorating. Even younger people often say, “My body temperature has been low lately…” or “I have an itchy rash on my head and hairline. Do you know any good medicines?” I have also been asked many times recently as this. I don’t think it is a disease, but something strange.

   When I thought about the causes of these problems, I felt that, in addition to aging, we apparently lack the ability to eliminate oxidative stress generated by particle matters (PM), plastics, and other nanoparticles entering into our bodies, or by a lifestyle that exposes us to cell phones and other electromagnetic waves. Therefore, I wanted to “protect my body” with food, and “efficiently reduce oxidative stress and toxins in my body” with food, which was the impetus for the development of this product.

The environment around us these days is full of dangers!?

   Humans cannot live without air, water and food. However, the air is filled with chemicals and electromagnetic waves including nanoparticles such as PM [1-8]. Water is filled with nanoparticles including micro and nano plastics, environmental hormones including pesticides and pharmaceuticals [3, 9-12]. Food is filled with nanoplastics, additives including nanoparticles and residual pesticides [10, 13-16], and even pharmaceuticals are filled with foreign substances, including nanoparticles [17-20].

   Various problems have recently been pointed out, and it is not easy to avoid all of them. In particular, nanoparticles can easily enter into the body not only by breathing but also by simply touching the skin [6, 21]. The common problem here is that all of them cause oxidative stress and even inflammation, but the symptoms depend on the type (compound) of the nanoparticle (Fig. 1) [2, 4, 5, 6, 16, 21, 22-27]. The scary part of this situation is that it is usually perfectly normal, but at some points it can develop into a sudden hypersensitivity [28-30]. It is like hay fever and is called environmental hypersensitivity. It does not lead to illness, but it also causes what is called a poor condition, such as feeling tired, unmotivated, or depressed.

Fig. 1    Problems in the living environment

What is oxidative stress?

   The atmosphere contains about 20% oxygen, and even when we are not doing anything, we use about 430L of oxygen per day to generate energy in the mitochondria [31]. Reactive oxygen species (ROS) are metabolized and converted into something more reactive from oxygen.

   It is because of their high reactivity (oxidative capacity) that ROS can play important roles in several physiological processes, such as cell signaling, gene regulation, and modulation of hormone action [32]. In addition, ROS can modulate the immune system [33, 34], and activated neutrophils and macrophages use ROS to cause an oxidative burst, destroying viruses and adjacent cells [35]. However, excess ROS can overwhelm the antioxidant defenses of cells and cause cell death [36], creating conditions such as protein denaturation and lipid peroxidation (oxidative stress) [37], which can lead to various diseases including cancer and cardiovascular disease [38-43] (Figure 2).

   For this reason, ROS are normally maintained at optimal levels in the body, eliminated by many ROS scavenging enzymes, including superoxide dismutase (SOD), which protects the organism from ROS injury, and antioxidants such as glutathione [44]. However, with age, the production of biological defenses decreases, and mitochondria are no longer able to scavenge the excess ROS generated [45-47]. This makes us more susceptible to various diseases.

Fig. ２  Diseases resulting from oxidative stress

What is environmental hypersensitivity?

   Environmental hypersensitivity is a medical condition in which the body is unable to tolerate various external environmental factors, such as chemicals and electromagnetic radiation, which induce oxidative stress at levels that are normally not problematic [28].

   The idea has been proposed that the mechanism of environmental hypersensitivity can be explained in three stages of genetic, exposure and trigger (onset) [28, 48, 49]. First, there are genetic factors that are congenitally determined. Genetic factors related to the stress response to exposure to the external and internal environment (hypothalamus, hypothalamic-pituitary-adrenal and limbic system), innate immunity and the response of the thrombic system are thought to control the susceptibility to develop environmental hypersensitivity. In other words, they are constitutionally, genetically strong or weak with respect to the environment.

   Second, exposure to environmental pathogenic factors can lead to the acquisition of environmental stress hypersensitivity, resulting in a hypersensitive response.

   Finally, it is considered that even environmental changes at a level not normally felt by normal healthy people can cause generalized ill health and hypersensitivity.

How to protect our body?

   Foreign substances such as nanoparticles attack the mitochondria in cells, causing oxidative stress due to the increased electrons leaking [23-25]. For this reason, at a certain point, the condition suddenly becomes as bad as hay fever.

   In fact, we eat out, and it is impossible to do anything to clean the air outside… Therefore, it is important to detoxify the body to remove toxins and foreign substances, and to eliminate oxidative stress caused by ROS. Glutathione is the most important factor in the defense of the body against oxidative stress. [44-46].

What is glutathione?

   Glutathione is a tripeptide consisting of glutamic acid, cysteine, and glycine bonded in this order. It is not degraded by most proteolytic enzymes [50-52].

   Glutathione is present in relatively high intracellular concentrations of 0.5-10mM. Extracellular concentrations are about 1/100 to 1/1000 of that [50-52]. Glutathione is available in both reduced and oxidized forms. Normally, the oxidized glutathione that has already reacted is converted back to reduced glutathione by glutathione reductase, so that more than 98% of glutathione is present in the reduced form [50-52].

   It is known that the production of glutathione in the body decreases with age, and that the activity of glutathione peroxidase also decreases due to genetic mutations [53, 54]. This has been reported to increase oxidative stress and cause various diseases, as shown in Fig. 2.

There are many functions of glutathione, but detoxification and scavenging of oxidative stress (ROS) are the most important ones [50-52].

Detoxification effect of glutathione

   Detoxification of foreign substances in the body is divided into three phases: denaturation, conjugation, and elimination [55]. These allow foreign substances to be expelled from the body and detoxified. Among these, glutathione plays an important role in detoxification because it can deal with nanoparticles and other foreign substances that cannot be denatured (metabolized) [56].

   Glutathione detoxifies foreign substances such as antibiotics and drugs, poisons, heavy metals, and transmitters such as prostaglandins by removing them from the cells, mainly in the liver [50-52]. In particular, nanoparticles and foreign substances increase the production of ROS, so detoxification of these is crucial at the source [16-27].

   Specifically, glutathione is detoxified by glutathione S-transferase, which binds a thiol group to a cysteine residue and expels it along with glutathione out of the cell [50-52, 56] (Fig. 3). In particular, it also prevents water-insoluble nanoparticles, etc., from entering the cell as large water-soluble molecules [56].

   The bindings then travel through the bloodstream to the kidneys, where cysteine and foreign substances are converted to mercapturic acid, which mixes with bile and urine and is excreted out of the body [22, 56].

Fig. ３  Detoxification effect of glutathione

Oxidative stress (reactive oxygen species: ROS) scavenging effect of glutathione

   Glutathione gives electrons to molecules such as ROS that have lost electrons in the cell and become unstable, returning them to stable molecules, eliminating oxidative stress and preventing cell damage (Fig. 4) [50-52].

   Specifically, superoxide (a type of weak ROS) is converted to hydrogen peroxide H₂O₂ (a type of moderate ROS) by superoxide dismutase in mitochondria, and glutathione, together with glutathione peroxidase, converts this hydrogen peroxide into water and oxygen, making it nontoxic [50-52].

   In addition, superoxide loses more electrons with copper and iron ions, as well as nanoparticles and foreign substances, and becomes the hydroxyl radical OH• (a potent form of ROS) [57, 58]. This can be highly toxic, but glutathione traps and scavenges it [50-52].

   Furthermore, although not a major pathway, superoxide can also be directly scavenged by glutathione.

   Catalase, which scavenges reactive oxygen species as does glutathione, is rarely found in mitochondria, whereas 5-15% of glutathione is found in mitochondria [59]. This is believed to be because glutathione makes it easier to cope with oxidative stress.

   As described above, glutathione can eliminate ROS, especially those increased by nanoparticles and foreign substances, as well as those that have become more potent.

Fig. ４ Oxidative stress scavenging effect of glutathione

How to take glutathione?

   Glutathione is used medicinally as an antidote for drug and metal poisoning [60]. Unfortunately, it has side effects, and rashes, vomiting, and liver dysfunction have been reported [61, 62]. Furthermore, if we create a situation where the body always receives large amounts of glutathione, the human body will become lazy to make it, resulting in side effects that make the drug unstoppable. Therefore, we thought it would be better if we could take it from foods without side effects, and we found that it is contained in tomatoes, cabbage, and liver of livestock [63]. However, even liver, which is the most abundant, must be consumed at least 100 g every day, which is quite difficult.

   In contrast, it has been reported that eating foods high in cysteine (an organosulfur compound) increases glutathione production in the liver, along with glutathione S-transferase activity [64-67]. The reason for this is that the synthesis of cysteine in glutathione is the slowest, and this determines the speed of glutathione synthesis in our body. Garlic and leeks were found to have this effect [68-69]. In particular, jumbo leek has a higher active oxygen scavenging capacity (oxygen radical absorption capacity) than others [70], and its components have been reported to detoxify toxins by denaturing them and increasing the gene expression of antioxidant and detoxification metabolic enzymes [71-74], making it the most likely candidate.

What is jumbo leek (French garlic)?

   Jumbo leek, also called French garlic, Russian garlic, elephant garlic, jumbo garlic, jumbo garlic, wild leek, and odorless garlic, is a member of the leek genus [75-82]. It is a separate species from garlic and is a different variant of the same species as leek, which is called western leek. It is native to the Mediterranean region and has been used as a traditional remedy since 3,000 BC. It was especially used by pyramid builders. Its medicinal effects are much higher than those of ordinary garlic.

   It has been reported to prevent obesity, diabetes, and arteriosclerosis (suppresses the breakdown and absorption of carbohydrates after meals and lowers cholesterol), improve liver function (suppresses alcohol absorption and strengthens the liver), improve antibacterial, anti-inflammatory, gene mutation suppression, improve immunity, neutralize toxins, enhance tonic effect, and anti-cancer effect. Consequently, it has been called the “Magic herb”.

   It is rich in vitamins A, C, and E, minerals, folic acid, and dietary fiber, and has more amino acids and polyphenols than ordinary garlic. Allicin, the source of the smell, is only 1/60th that of regular garlic, so it does not leave a lingering odor and does not cause stomach or abdominal pain.

How can we take full advantage of jumbo leek (French garlic)?

   The jumbo leek (French garlic) bulb is a baby. In early summer, the bulb goes to sleep, wakes up in the fall, germinates, and goes to sleep again [83]. As winter passes and spring arrives, jumbo leek wakes up again from sleep, sprouts, and the bulbs enlarge and grow. For this reason, the components of jumbo leek (French garlic) are insoluble in water when they are asleep, so that they do not run off when it rains. When it awakens from sleep, it transforms itself into a water-soluble form and uses it for its own growth [84]. In this process, carbohydrates, proteins, and fats are broken down to make sugars, amino acids, fatty acids, minerals, growth hormones, and repair hormones, as well as components that boost immunity to prevent disease in preparation for sprouting.

   The same is true in humans; if some components are hard to dissolve in water, they must be converted into water-soluble ones in the body, which can be less efficient and more toxic [85].

   Extracts are not bad, but based on human long eating habits, we wanted to consume them as foods in their natural state, without extraction or concentration.

How do we wake jumbo leek (French garlic) up?

   If the awakening is not simultaneous, the ingredients will be uneven… Therefore, we used our proprietary Grandir recipe™ for rapid and synchronized dormancy-breaking jumbo leek (French garlic) grown in Shimabara, Nagasaki Prefecture, Japan within two days to create the ingredients for the baby’s life force. The whole balls were then heated and freeze-dried (Fig. 5).

Fig. ５  Experimental scheem for rapid and sychronized dormancy-breaking junbo leek (French garlic) 

Glutathione production enhancing effect of rapid and synchronized dormancy-breaking jumbo leek (french garlic) bulbs in the liver

   The glutathione production enhancing effect of the rapid and synchronized dormancy-breaking jumbo leek (French garlic) bulbs in the liver was investigated by measuring glutathione production in human liver cells, assuming oral administration (eating). The results showed that the amount of glutathione increased 9-fold in the heat- and freeze-dried samples that had not been subjected to rapid and synchronized dormancy breaking (untreated), compared to the control.

   In contrast, the rapid and synchronized dormancy-breaking sample showed an even greater increase, approximately 35-fold higher than the control and 4-fold higher than the untreated sample (Fig. 6).

   Therefore, it is considered that the increase in the amount of glutathione in the liver of the rapid and synchronized dormancy-breaking jumbo leek bulbs promotes detoxification of foreign substances such as nanoparticles in the body and elimination of oxidative stress caused by ROS (Fig. 7).

Fig. ６  Change of glutathione production in liver by rapid and synchronized dormancy-breaking treatment

Fig. ７  Detoxicifacation and oxidative stress scavenging effects of rapid and synchronized dormancy-breaking jumbo leek (french garlic) bulbs

Rapid and sychronized dormancy-breaking jumbo leek (French garlic) bulb

   We have succeeded in creating a safe and highly functional product without extraction or concentration, and without changing the long-established eating habit of the food. Along with the prevention and improvement (anti-aging) of cerebral infarction, cancer, wrinkles, and blemishes caused by foreign substances (toxins) and oxidative stress in the body, it is expected to maintain important stem cells [86], promote intestinal activity by increasing butyric acid bacteria [87], and also prevent obesity and diabetes.

   Youthfulness and health decline with age. The life force of the French garlic baby works on the body’s natural power and helps to keep it in good condition.

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