Rejuvenation for experimenters

This long article describes the process of planning and conducting an experiment on yourself using antioxidants with a proven effect that have a targeted effect on mitochondria. In order to evaluate the effect of the antioxidant, the indicators of the state of the cardiovascular system are evaluated before and after the experiment. The experimental scheme described in the article is borrowed from a recently published small clinical study, however, the range of evaluation methods has been narrowed to cost-effective, not requiring special skills and suitable for independent approaches.

This scheme is not published in order to encourage people to implement it immediately. The purpose of the publication is to illustrate an approach to conducting experiments on oneself dedicated to testing interventions that can potentially improve health or eliminate the manifestations of aging. To do this, the experimenter must: establish his limitations, determine the most appropriate approaches, conduct a study to clarify the necessary details, draw up an action plan, perhaps analyze some aspects beforehand (for example, dosages, since they often do not have the expected effect) and, most importantly, taking into account all possible risks, establish whether whether "the game is worth the candle". Ultimately, the decision to conduct an experiment is a personal choice.

Why experiment on yourself with antioxidants that target the mitochondria?

It is generally believed that conventional antioxidant dietary supplements have a moderately pronounced harmful effect on health in the long term. For example, they block signaling pathways that are important for the manifestation of beneficial reactions to physical activity. On the other hand, the ability of antioxidants acting on mitochondria to slightly slow down the aging process in short-lived species, while improving health indicators, has been demonstrated. Their use has also proved to be a viable method of treating a number of localized inflammatory conditions. This is explained by the theory that mitochondria in the course of their vital activity generate oxidative molecules that cause damage in the mitochondria themselves, which, in turn, leads to the appearance of non-functional cells, whose mitochondria produce a huge number of oxidative molecules. Ensuring a constant supply of antioxidants targeting mitochondria can slow down the rate at which mitochondria damage themselves, reduce the consequences of the appearance of cells with damaged mitochondria, or have both effects.

One of these consequences is the entry into the surrounding tissues and bloodstream of a large number of oxidative molecules reacting there with lipids. Oxidized lipids can cause even greater harm to all cellular processes, but the development of atherosclerosis is of the greatest interest. Oxidized lipids can trigger the development of inadequate oxidative reactions in the walls of blood vessels, and some of their forms can also lead to the fact that cells responding to this inflammation are depleted and die. Thus, the formation of lipid plaques occurs in atherosclerosis, leading to weakening of the main blood vessels and narrowing of their lumen. Cholesterol-lowering drugs of the statin class slow down the progression of atherosclerosis by reducing the amount of oxidized lipids, which is a consequence of a decrease in the total amount of lipids.

It is believed that in the future, violations of the ability of the smooth muscles of the vascular wall, responsible for the contraction and expansion of blood vessels, are to a certain extent due to the increasing level of oxidative stress during aging (the more damaged mitochondria, the more oxidative molecules). This contributes to the sealing of vascular walls and the development of hypertension, cardiovascular diseases, etc. Suppression of the oxidative effects of mitochondrial dysfunction can also have a positive effect on the body.

Antioxidants targeting mitochondria do not eliminate the root cause of the problem. At best, they somewhat compensate or weaken the ongoing processes. However, they are cheap and if they are able to influence the risk factors for developing cardiovascular diseases to about the same extent as statins, then their use may be a good idea.

Warnings

While some antioxidants affecting mitochondria have already been approved for use, are widely used and are freely available or are accompanied by clinical data that are sufficient to assess possible risks, when conducting an experiment on oneself, a person should remember about personal responsibility. First, it is necessary to read the literature describing the selected antioxidant, namely its effects, possible side effects and dosages. After that, based on the information received, you should make your own decision regarding the possible risk and the level of comfort of the experiment. This applies to any dietary supplement, regardless of whether it has received official approval for use or not. Do not trust the various opinions that can be read on the Internet. It is better to turn to primary sources and scientific articles. It should be understood that in the case of a limited number of primary data, they may be erroneous or incomplete, which is fraught with undesirable consequences. It should also be understood that, due to physiological characteristics, older people may be more sensitive and susceptible to various influences, which is not always described in studies.

Secondly, although research in the field of antioxidants affecting mitochondria has not made significant progress over the past decade, they are slowly, but still moving forward. After some time, this article will lose its relevance, as new data and new anti-dants will appear on the scene. However, the general scheme described in it will remain a useful tool for planning new experiments on yourself using newer and possibly more effective compounds, as well as methods for evaluating the results.

Finally, obtaining and using random compounds that have not yet been approved for clinical use in your country, such as SkQ1, according to the plan described in this article, is potentially illegal. However, in many cases, the sale of such products is allowed for research purposes. Therefore, sellers with a good reputation often declare that the products they sell are not intended for personal use, realizing that some buyers purchase these products for this purpose.

Selection of antioxidants that target mitochondria

There are two noteworthy antioxidants that target mitochondria: MitoQ and SkQ1. Their effects are confirmed by the largest amount of scientific data, including the results of clinical use. The effectiveness of the third compound, known as SS-31, is also confirmed by scientific data, but it is injected. It may well be that one of these compounds acts much better than the others (with a high probability it is SkQ1), however, when conducting an experiment on yourself, there is no convincing reason to give preference to an injectable drug over oral drugs.

Determination of dosages

The only current method of selecting the optimal dosage of a pharmaceutical additive is to conduct large–scale clinical trials. The results of studies on mice can only indicate the most promising reference point for conducting experiments on the selection of dosages for humans, however, adequate approaches to calculating this reference point in most cases do not exist. The established algorithm is generally similar for most drugs for oral administration and intravenous (or intraperitoneal in the case of small animals) administration, but they are not always applicable to other methods of administration. The relationship between the various forms of administration, dosage and the effects exerted is in fact a complex and unexpectedly poorly understood issue. Some compounds are exceptions to the rule, and the only way for specialists to identify this exclusivity is to test different dosages on different animal models.

Fortunately, the effects of antioxidants acting on mitochondria have been confirmed by the results of clinical studies, so the stage of selecting the initial dose based on experiments on mice can be skipped. Just use the dosages used in clinical trials. In the absence of these data, it will be necessary to calculate the initial dosage based on experiments on mice. To do this, first you need to read the articles describing the studies conducted on mice of the selected drug. Doses of drugs for oral administration are usually indicated in mg / kg of body weight. After that, you need to use a special tool to convert the indicated dosage into acceptable doses for humans, described in the open access article A simple practice guide for dose conversion between animals and human. A simple recalculation of the dosage per person's body weight in kilograms is not applicable in this case. The relative surface area of the body of representatives of different species is a more adequate conversion factor. In any case, it should be understood that it is very difficult to predict the result at the initial stage of dose selection. However, the duration of therapy is very clearly reflected in the result. When conducting an experiment, you should start with the number of doses of the drug, the duration of the intervals between doses and the duration of therapy used in animal studies.

MitoQ

In this case, all the preparatory work has already been done. In a clinical trial conducted in 2018, participants received 20 mg of the drug daily for 6 weeks, and this dosage can be taken as the initial one.

SkQ1

The case of SkQ1 is more complicated, since this compound was initially approved as a drug for the treatment of inflammatory eye disease called visomitin, and most of the clinical data was obtained for its use as eye drops. In this case, the dosages used cannot be extrapolated to a drug for oral administration, which is currently at a very early stage of the regulatory process. In one of the lifelong studies on mice, the daily dose of the drug added to drinking water varied in the range of 0.7-1.0 micromole/kg. When calculated for a person with a body weight of up to 60 kilograms, the dosage is 3.5-5.0 micromoles per day. The duration of therapy in the study was 150 days, but the study itself did not end there, since its purpose was to estimate the life expectancy of animals. However, to assess the short-term effects of the drug, it is more appropriate to use a 6-week protocol from a study using MitoQ.

Visomitin eye drops are sold in 5 ml vials in the form of a solution with a concentration of 0.155 micrograms/ml. To convert from micrograms to micromoles, it is necessary to know the molecular weight of SkQ1, as indicated on the PubChem website, which is 617.608 g/mol. Thus, the daily dose of 5 micromoles = 0.000005 x 617.608 = 0.0031 g = 3100 mcg = 3.1 mg (4 vials, in Russia the cheapest cost almost 500 rubles). Therefore, the purchase of visomitin for oral administration is impractical.

Confirmation of everything mentioned above

Assume that all publications, with the exception of primary sources, may be incorrect or misleading. Don't take anyone's words for granted, find the primary sources, recalculate the numbers and make your own conclusions.

Acquisition of antioxidants that affect mitochondria

MitoQ

MitoQ is inexpensive and can be purchased from MitoQ Limited through several reputable online stores. The search for cheaper sources is not worth the trouble, since then you will have to confirm the quality of the product. Just buy the drug in the store.

SkQ1

The appearance of tablet forms of the drug for oral administration is a matter of several years. However, today the task is to find a manufacturer or supplier on the world market with subsequent product quality control. For people who do not have the right connections, the best way to purchase products that have not yet entered mass production is to order them from manufacturers in China or other regions. The main tool for finding Chinese manufacturers is the Alibaba marketplace. [The original article describes in detail the process of searching and ordering drugs on Chinese trading platforms.]

As mentioned above, such a purchase of drugs that are not approved for human use in your country is to a certain extent illegal. The worst case scenario in this case is the confiscation of the product at customs. However, the situation may be different in different countries, so before ordering a drug from China, you should carefully study this issue.

Storage of antioxidants affecting mitochondria

Both MitoQ and SkQ1 have a long shelf life when stored in the freezer. For more convenient use for short periods of time – from several weeks to several months – they can be stored in the dark in the refrigerator. SkQ1 is less stable than MitoQ and decomposes at room temperature within a few weeks.

Quality control of purchased antioxidants

There is no need to worry about the quality of the MitoQ Limited product, but other sources are not always trustworthy. Having received a drug ordered from abroad, you cannot be sure that its composition corresponds to the declared one, that it does not contain impurities and does not have proper quality in any other parameters. This can only be verified by sending a sample of the drug to the laboratory for analysis using liquid chromatography and mass spectrometry. After that, the results obtained must be compared with the standard data for a sample of a high degree of purification. [The article describes in detail the process of packaging and sending samples of the drug to the laboratory for analysis.]

Methods of taking medications

MitoQ Limited produces the MitoQ drug in a convenient tablet form that does not require additional manipulations. It remains only to take the required number of tablets. When ordering from other manufacturers, you can get both an alcohol solution and a powder. SkQ1 is usually a powder that will need to be converted into a solution for use. Neither MitoQ nor SkQ1 are soluble in water. In dry form, they can be dissolved in a minimum amount of ethyl alcohol. The resulting solution will need to be pipetted into a glass of water and drink it. This approach allows us to approximately reproduce the method of drug administration into the body used in most studies on animal models.

Determination of necessary tests and estimated parameters

In this case, it is necessary to determine a set of tests that (a) correspond to the expected result based on the data of clinical studies targeting antioxidants affecting mitochondria and (b) can be carried out without the participation of a doctor. These tests should assess the state of the cardiovascular system, especially the parameters that depend on the elasticity of blood vessels. In addition, some attention should be paid to the parameters for assessing the level of oxidative stress and the degree of development of atherosclerosis.

• standard blood test with markers of inflammation;
• assessment of the level of oxidized cholesterol of low-density lipoproteins;
• resting heart rate and blood pressure;
• heart rate variability;
• pulse wave propagation velocity;
• determination of biological age by DNA methylation profile.

The parameters of the state of the cardiovascular system included in this list depend on changes in the elasticity or functionality of blood vessels, which are the expected result of therapy, presumably compensating for the effect of aging on the smooth muscle cells of the walls of blood vessels, in this case, antioxidants that target the mitochondria. Positive changes in the average values of most of these parameters can be achieved with the help of long-term active training, and reducing this period with the help of pharmacological intervention is a very interesting prospect.

Laboratory blood test

[The original article provides online services that allow you to quickly conduct laboratory tests in the United States.]

Resting heart rate and blood pressure

When assessing the parameters of the state of the cardiovascular system, a number of general principles should be followed. Firstly, the further the change is carried out from the center of the body, the less adequate the readings are taken and the more they are influenced by external factors, such as body position, mood, stress, time of day, etc. Devices for taking readings from the fingertips are convenient to use, but they are far from accurate devices for registering pressure on shoulder level. Secondly, all of the above factors have an impact on each parameter of the state of the cardiovascular system, therefore, when determining the basic values of the parameters and evaluating their changes relative to the basic values, each change must be made in the same position at the same time of day. It is also important to take readings repeatedly throughout the week, which will allow you to get a more accurate picture of the state of your body.

Heart rate variability

Ideally, special medical devices are required to evaluate this parameter, which can be very difficult to get. Alternatively, you can use a sports heart rate monitor in combination with a special smartphone application. However, not all users manage to get reliable data using this system. [For more details, see the original article.]

Pulse wave propagation velocity

As for the pulse wave propagation speed, the choice of applicable devices is even more limited. According to the author, the most objective data is provided by the iHeart device. However, the values obtained with its help differ significantly for different days of changes, even if the values of the maximum possible number of variables are kept constant. [For more details, see the original article.]

DNA methylation

A test for determining biological age by DNA methylation can be ordered from Osiris Green or Epimorphy/Zymo Research. According to company representatives, the normal level of variability for repeated tests of the same sample is about 1.7 years for the Zymo Research test and 4.8 years for the Osiris Green test. The level of variability for different days or within one day is still questionable, but experts say that they are comparable for measurements separated by several months. As for the indicators of the state of the cardiovascular system, for these parameters it is advisable to observe the reproducibility of the maximum number of external conditions (time of day, the period that has passed after eating or physical activity, diet, etc.) when undergoing testing before and after therapy.

Example of a daily measurement plan

Below is one of the possible options for daily taking readings of the state of the cardiovascular system, which includes several additional parameters to demonstrate the level of accuracy of measurement instruments:

• put on a heart rate monitor, since this manipulation causes a short-term increase in heart rate, do it first;
• sit in a comfortable position and relax for a few minutes;
• measure the blood pressure and pulse on your left hand;
• measure the blood pressure and pulse on your right hand;
• measure the pulse wave propagation velocity at the tip of your right index finger for 30 seconds using the iHeart system;
• measure the pulse wave propagation velocity at the tip of your left index finger for 30 seconds using the iHeart system;
• Measure the heart rate variability for 10 minutes using a heart rate monitor and a smartphone app.

Reproducibility of parameters is very important

Throughout the experiment, from the first measurement of parameters to the last, it is very important to maintain a stable body weight, diet and exercise level. Significant lifestyle changes can lead to consequences that make it difficult to register any results using the simple methods proposed above. Moreover, when carrying out measurements, it is necessary, as already mentioned above, to reproduce environmental factors as much as possible. Experiments with different measuring devices will quickly demonstrate how great the influence of these factors can be.

[The approximate costs of conducting a study in the USA are given in the article.]

The scheme of the experiment on yourself

The process of studying the issue, ordering and purchasing the necessary materials and devices, as well as checking the quality of the drug can take several months. After all the decisions have been made and the materials have been purchased, select the start date of the experiment. After that, follow the scheme below:

Days 1-10: Measure blood pressure, pulse wave propagation rate and heart rate variability 1 or 2 times a day.

Day 10: Take a blood test and a DNA methylation test.

Day 11. Start a 6-week course of daily medication.

Days 53-62. Repeatedly measure blood pressure, pulse wave propagation velocity and heart rate variability.

Day 62. Re-take a blood test and a DNA methylation test.

Where can I publish the results?

If you conduct an experiment on yourself and do not share the results with anyone, you will only help yourself. The results of a rational thought-out experiment can bring real benefit only if many members of the community post the data obtained in open access, which will facilitate the planning process of official clinical trials and indicate current research and development directions. There are many communities of people conducting experiments on themselves using different compounds and interventions. One of the suitable resources for posting descriptions and results of personal experiments are the LongeCity forums. You can also create your own website or blog.

The publication should include all measurement results, names and doses of medications taken, duration of therapy, as well as the age, weight and gender of the experimenter. The age can be specified approximately, in the range of 5 years. If you want to publish the results anonymously, this is quite realistic, since it is impossible to track your data without access to the database of the laboratory service that conducted the analyses. Negative results are no less important than positive ones. For example, when using the parameters listed in this article to evaluate the results, it is quite possible that the positive changes for people aged 45-55 years will be so insignificant that they will not go beyond the range of random noise and measurement errors. The data confirming this assumption is also important and useful for the community, as it will help in planning future, more effective experiments.

Evgenia Ryabtseva, portal "Eternal Youth" http://vechnayamolodost.ru based on Fight Aging!: How to Plan and Carry Out a Simple Self-Experiment, a Single Person Trial of a Mitochondrially Targeted Antioxidant.