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How to Plan and Carry Out a Simple Self-Experiment, a Single Person Trial of a Mitochondrially Targeted Antioxidant

This lengthy post walks through the process of setting up and running a self-experiment - a trial of one - with one of the various established mitochondrially targeted antioxidant compounds. Metrics are assessed beforehand and afterwards in order to shed some light on whether or not it worked, in the sense of improving one or more measures of cardiovascular health. The outline here is informed by a recently published small human trial of MitoQ, but cutting down the assessments to those that are cost-effective, easily carried out, and available without the aid of a physician.

The purpose in publishing this outline is not to encourage people to immediately set forth to follow it. This post, like others in this series, is intended to illustrate how to think about self-experimentation in the matter of interventions that might help to improve health or turn back aspects of aging: set your constraints; identify likely approaches; do the research to fill in the necessary details; establish a plan of action; perhaps try out some parts of it in advance, such as the measurement portions, as they never quite work as expected; and most importantly identify whether or not the whole plan is worth actually trying, given all that is known of the risks involved. Ultimately that must be a personal choice.

Contents

Why Self-Experiment with Mitochondrially Targeted Antioxidants?

Ordinary antioxidant supplements are thought to be, on balance, modestly harmful to long term health. They block signaling that is important to the beneficial response to exercise, for example. Mitochondrially targeted antioxidants, on the other hand, have been shown to slightly slow aging in short-lived species, and improve measures of health along the way. They also appear to be a viable treatment for some localized inflammatory conditions. The theory here is that mitochondria generate oxidative molecules in the normal course of operation that cause damage within the mitochondria themselves, and that in turn leads to dysfunctional cells in which the mitochondria produce a vastly greater amount of oxidative molecules. Delivering a constant supply of mitochondrially targeted antioxidants may either slow down the pace at which mitochondria damage themselves, or dampen the consequences of cells overtaken by damaged mitochondria, or both.

One of those consequences is the bulk export of oxidative molecules into surrounding tissues and the bloodstream, where they react with lipids. Oxidized lipids can cause further harm in all sorts of cellular processes, but of particular interest is the development of atherosclerosis. Oxidized lipids can cause inappropriate inflammatory reactions in blood vessel walls, and some forms can also cause the cells responding to that inflammation to become overwhelmed and die. This is how the fatty plaques of atherosclerosis form, then grow to weaken and narrow major blood vessels. Statin drugs, that reduce blood cholesterol, succeed in slowing atherosclerosis because they reduce the amount of oxidized lipids in the course of reducing the amount of all lipids.

Further, some degree of dysfunction in the vascular smooth muscle responsible for blood vessel contraction and dilation is thought to be caused by rising levels of oxidative stress in aging - too many dysfunctional mitochondria, too many oxidative molecules. This contributes to vascular stiffness and consequent hypertension, cardiovascular disease, and so forth. Suppressing the oxidative consequences of malfunctioning mitochondria may help here as well.

Mitochondrially targeted antioxidants don't solve the roots of these problems. At best, they somewhat compensate or attenuate ongoing mechanisms. They are cheap, however, and if they can produce effects on risk factors for cardiovascular disease that are, say, somewhere in the same order of magnitude as those achieved by statins or drugs that control blood pressure, with minimal side-effects, then they may well be worth using.

Caveats

While some mitochondrially targeted antioxidants are approved by regulators, widely used, and readily available, or otherwise come with an adequate set of human data to judge risk, one must still think about personal responsibility in any self-experiment. Firstly, read the relevant papers on the mitochondrially targeted antioxidant of choice - its effects, side-effects, and dosages - and make an individual decision on risk and comfort level based on that information. This is true of any supplement, whether or not approved for use. Do not trust other opinions you might read online: go to the primary sources, the scientific papers, and read those. Understand that where the primary data is sparse, it may well be wrong or incomplete in ways that will prove harmful. Also understand that older physiologies can be frail and vulnerable in ways that do not occur in younger people and that are sometimes not well covered by the studies.

Secondly, while work on mitochondrially targeted antioxidants hasn't moved all that rapidly over the past decade, it does move. This post will become outdated in its specifics at some point, as new knowledge and new mitochondrially targeted antioxidants arrive on the scene. Nonetheless, the general outline should still be a useful basis for designing new self-experiments involving later and hopefully better compounds, as well as tests involving more logistical effort.

Lastly, obtaining and using arbitrary compounds not yet approved in your country for human use - such as SkQ1 - in the manner described here is potentially illegal: not yet being a formally registered medical treatment in all jurisdictions, it falls into a nebulous area of regulatory and prosecutorial discretion as to which of the overly broad rules and laws might apply. In effect it is illegal if one of the representatives of the powers that be chooses to say it is illegal in any specific case, and there are few good guidelines as to how those decisions will be made. The clearest of the murky dividing lines is that it is legal to sell such compounds for research use, but illegal to market and sell them for personal use in most circumstances. This is very selectively enforced, however, and reputable sellers simply declare that their products are not for personal use, while knowing full well that this is exactly what their customers are doing in many cases.

Choosing to purchase and use SkQ1 would therefore likely be a matter of civil disobedience, as is the case for anyone obtaining medicines or potential medicines outside the established national system of prescription and regulation. People are rarely prosected for doing so for personal use in the US - consider the legions of those who obtain medicines overseas for reasons of cost, despite the fact that doing so is illegal - but "rarely" is not "never." If you believe that the law is unjust, then by all means stand up against it, but accept that doing so carries the obvious risks of arrest, conviction, loss of livelihood, and all the other ways in which the cogs of modern society crush those who disagree with the powers that be.

Choosing a Mitochondrially Targeted Antioxidant

There are two mitochondrially targeted antioxidants worth considering: MitoQ and SkQ1. These have the greatest human usage and data. A third, SS-31, also has useful data, but must be injected. It may well be that one or another of these compounds is significantly better than the others (and SkQ1 would probably be the one, if so), but there is no compelling reason to pick an injected compound over one that can be taken orally for the purposes of this self-experiment. Injections require a great deal more logistical organization than simply taking a pill.

Establishing Dosages

The only definitive way to establish a dosage for a supplement pharmaceutical in order to achieve a given effect is to run a lot of tests in humans. Testing in mice can only pin down a likely starting point for experiments to determine a human dose, but the way in which you calculate that starting point is fairly well established for most cases. That established algorithm is essentially the same for most ingested and intravenously (or intraperitoneally in small animals) injected medicines, but doesn't necessarily apply to other injection routes. The relationship between different forms of injection, dosage, and effects is actually a complicated and surprisingly poorly mapped topic, and we'll set that to one side here. Some compounds - as always - are exceptions to the rule, and the only way that scientists discover that any specific compound is an exception is through testing at various doses in various species.

Fortunately, some mitochondrially targeted antioxidants do have human data, so guesstimating an initial dose based on a mouse study can be skipped. Just use the amounts from the human studies. If lacking that data, the steps to figure out a suitable starting point for a human test based on data from a mouse study are as follows: firstly read the mouse studies for the compound in question, in order to find out how much was given to the mice and for how long. Doses for most ingested pharmaceuticals of interest will usually be expressed in mg/kg. Secondly apply a standard multiplier to scale this up to human doses, which you can find in the open access paper "A simple practice guide for dose conversion between animals and human". Do not just multiply by the weight of the human in kilograms - that is not how this works. The relative surface area of the two species is the more relevant scaling parameter. Read the paper and its references in order to understand why this is the case. Again, note that the result is only a ballpark guess at a starting point in size of dose. The duration of treatment translates fairly directly, however. For the period of treatment, start with the same number of doses, spacing of doses, and duration as takes place in the animal study.

MitoQ

All the work is already done in this case. The 2018 MitoQ human study used a daily 20mg dose for six weeks, and that seems a good place to start.

SkQ1

SkQ1 is a little more challenging, in that it was initially approved by regulators for the treatment of inflammatory eye disease under the name Visomitin; most of the human data focuses on delivery via eye drop. Dosage there isn't relevant at all to the ingested pill scenario, which is still earlier in the regulatory process. So retreating to one of the mouse life span studies, we find that the daily dose of SkQ1, delivered in drinking water, was in the 0.7-1.0 μmol/kg range. Scaling this up to a 60kg human comes to 3.5-5.0 μmol per day. The study ran for more than 150 days of treatment, as it was assessing life span, but mirroring the six weeks of the MitoQ study above seems reasonable when looking for short term effects.

Visomitin eye drops come in a 5ml container, with 0.155 mcg/ml dilution. To convert between micrograms (mcg or μg) and micromoles (μmol), one needs the molecular weight of SkQ1, which is where resources such as PubChem come in handy. That gives the figure of 617.608 g/mol. Thus a daily dose of 5μmol = 0.000005 x 617.608 = 0.0031g = 3100mcg = 3.1mg. Which suggests that people buying Visomitin to put in their drinking water are wasting their time - it is intended for point administration to a very small area of the body, the eye.

Over 6 weeks of daily administration, the above means a supply of about 130mg is needed.

Verify All of the Above

Assume that anything written anywhere other than the primary materials might be incorrect or misleading. Do not take my word for any of the above information; chase down the primary sources, run the numbers, and make the judgement calls yourself.

Obtaining Mitochondrially Targeted Antioxidants

MitoQ

MitoQ is cheap and readily available from MitoQ Limited via any number of reputable online storefronts. It really isn't worth the effort to find another, cheaper supplier, and then be tasked with verifying the quality of the product batch. Just buy it from a store.

SkQ1

Given a few years, pill forms will be readily available at a useful dosage for oral ingestion. For now, however, it is a matter of finding a manufacturer or supplier in the global marketplace, and then validating the product when it arrives. For individuals without suitable connections, the easiest way to obtain compounds that are not yet mass manufactured is to order them from manufacturers in China or other overseas locations.

As noted at the outset of this post, efforts to obtain, ship, and use a compound yet to be approved for human use in your country, such as SkQ1, may be to some degree illegal - it would be an act of civil disobedience carried out because the laws regarding these matters are unjust, albeit very unevenly enforced. Many people regularly order pharmaceuticals from overseas, with and without prescriptions, for a variety of economic and medical reasons, and all of this is illegal. The usual worst outcome for individual users is intermittent confiscation of goods by customs, though in the US, the FDA is actually responsible for this enforcement rather than the customs authorities. Worse things can and have happened to individuals, however, even though enforcement is usually targeted at bigger fish, those who want to resell sizable amounts of medication on the gray market, or who are trafficking in controlled substances. While the situation with an arbitrary compound such as SkQ1 isn't the same from a regulatory perspective, there is a fair amount written on the broader topic online, and I encourage reading around the subject.

Open a Business Mailbox

A mailbox capable of receiving signature-required packages from internal shipping concerns such as DHL and Fedex will be needed. Having a business name and address is a good idea. Do not use a residential address.

Use Alibaba to Find Manufacturers

Alibaba is the primary means for non-Chinese-language purchasers to connect to Chinese manufacturers. The company has done a lot of work to incorporate automatic translation, to reduce risk, to garden a competitive bazaar, and to make the reputation of companies visible, but it is by now quite a complicated site to use. It is a culture in and of itself, with its own terms and shorthand. There are a lot of guides to Alibaba out there that certainly help, even if primarily aimed at retailers in search of a manufacturer, but many of the specific details become obsolete quickly. The Alibaba international payment systems in particular are a moving target at all times: this year's names, user interfaces, and restrictions will not be the same as next year's names, user interfaces, and restrictions.

Start by searching Alibaba for suppliers of interest. There are scores of resellers and manufacturing biotech companies in China for any even somewhat characterized supplement, pharmaceutical, or candidate pharmaceutical. Filter the list for small companies, as larger companies will tend to (a) ignore individual purchasers in search of small amounts of a compound, for all the obvious economic reasons, and (b) in any case require proof of all of the necessary importation licenses and paperwork. Shop around for prices - they may vary by an order of magnitude, and it isn't necessarily the case that very low prices indicate a scam of some sort. Some items are genuinely very cheap to obtain via some Chinese sources.

Many manufacturers will state that they require a large (often ridiculously large) minimum order; that can be ignored. Only communicate with gold badge, trade assurance suppliers with several years or more of reputation and a decent response rate. Make sure the companies exist outside Alibaba, though for many entirely reliable Chinese businesses there are often sizable differences between storefronts on Alibaba, real world presence, and the names of owners and bank accounts. Use your best judgement; it will become easier with practice.

Arrange Purchase and Shipping via Alibaba

Given the names of a few suppliers, reach out via the Alibaba messaging system and ask for a quote for a given amount of the compound in question. Buy twice what you'll think you need, as some of it will be used to validate the identity and quality of the compound batch, and buy that much from at least two different suppliers present in widely separated regions. Payment will most likely have to be carried out via a wire transfer, which in Alibaba is called telegraphic transfer (TT). Alibaba offers a series of quite slick internal payment options that can be hooked up to a credit card or bank account, but it is hit and miss whether or not those methods will be permitted for any given transaction. Asking the seller for a pro-forma invoice (PI), then heading to the bank to send a wire, and trusting to their honesty is good enough for low cost transactions. It should work just fine when dealing with companies that have a long-standing gold badge.

To enable shipping with tracking via carriers such as DHL, the preferred method of delivery for Chinese suppliers shipping to the US or Europe, you will need to provide a shipping address, email address, and phone number. Those details will find their way into spam databases if you are dealing with more than a few companies, and will be, of course, sold on by Alibaba itself as well. Expect to see an uptick of spam after dealing with suppliers via Alibaba, so consider using throwaway credentials where possible.

Chinese manufacturers active on Alibaba are familiar with international shipping practices, and smaller companies will, on their own initiative, apply whatever description to packages will most likely get it past customs. Since declared pharmaceuticals may well be taken aside and confiscated, the description will therefore not involve pharmaceuticals. This is as much motivated by dealing with customs at the Chinese end as pushing things past the US authorities; it is again a form of widespread civil disobedience that reflects a popular disdain for petty laws and regulators where they act as impediments to useful activity.

Storage of Mitochondrially Targeted Antioxidants

Both MitoQ and SkQ1 have a long shelf life of years if kept in a freezer. For more convenient use over shorter periods of time, say a few weeks to a few months, should be kept in the dark in a refrigerator for shorter periods of time. SkQ1 is less resilient than MitoQ and will degrade after few weeks at room temperature.

Validating the Purchased Mitochondrially Targeted Antioxidants

While it is reasonable to trust MitoQ Limited to deliver what was ordered, that may not be the case for other sources. A compound may have been ordered, but that doesn't mean that what turns up at the door is either the right nondescript powder or free from impurities or otherwise of good quality. Even when not ordering from distant, infrequent suppliers, regular testing of batches is good practice in any industry. How to determine whether a compound is what it says it is? Run the compound through a process of liquid chromatography and mass spectrometry, and compare the results against the standard data for a high purity sample of that compound. Or rather pay a small lab company to do that.

Obtain the Necessary Equipment

Since this process will involve weighing, dividing up, and shipping powders in milligram amounts, a few items will be necessary: spatulas or scoops for small amounts of a substance; a reliable jeweler's scale such as the Gemini-20; sealable vials; small ziplock bags; labels; and shipping and packing materials. All of these are easily purchased online. The recommended shipping protocol is to triple wrap: a labelled vial, secured within a ziplock bag and tape, and then enclosed within a padded envelope.

Use Science Exchange to Find Lab Companies

Science Exchange is a fairly robust way to identify providers of specific lab services, request quotes, and make payments. Here again, pick a small lab company to work with after searching for LC-MS (liquid chromatography and mass spectrometry) services. Large companies will want all of the boilerplate registrations and legalities dotted and crossed, and are generally a pain to deal with in most other ways as well. Companies registered with Science Exchange largely don't provide their rates without some discussion, but a little over $100 per sample is a fair price for LC-MS to check the identity and purity of the compound.

Work with the Company to Arrange the Service

The process of request, bid, acceptance, and payment is managed through the Science Exchange website, with questions and answers posted to a discussion board for the task. Certainly ask if you have questions; most providers are happy to answer questions for someone less familiar with the technologies used. Service providers will typically want a description of the compounds to be tested and their standard data sheets, as a matter of best practice and safety. It is good enough to provide the name for established pharmaceuticals, as the data sheets, mass spectrometry profiles, and other detailed information are freely available online from databases such as DrugBank.

Ship the Samples

Measure out a small amount (1mg is more than enough) from each separate order as a distinct sample, label it carefully, make sure you have a record linking the sample label to the specific supplier, and package it up. More in the sample is better than less, as several attempts might be needed to get a good result out of the machines used, but each attempt really only needs a very tiny amount of the compound. Ship the sample via a carrier service such as DHL, UPS, or FedEx. Some LC-MS service companies may provide shipping instructions or recommendations. These are usually some variety of common sense: add a description and invoice to the package; reference the order ID, sender, and receiver; clearly label sample containers; and package defensively with three layers of packing; and so forth.

Examine the Results

Once the LC-MS process runs, the lab company should provide a short summary regarding whether or not the compound is in fact the correct one and numbers for the estimated purity. Also provided are the mass spectra, which can be compared with the standard spectra for the compound, which can be found at DrugBank or other sources online.

Ingestion Logistics

MitoQ comes prepackaged in a convenient pill form, so there is no need to do any additional work here. Just take the pills in the appropriate amount. If ordered from a source other than MitoQ Limited, it may arrive dissolved in an ethanol solution, or as a solid. Similarly, an order of SkQ1 will arrive as a tiny amount of powder that needs to be dissolved in solution in order to be ingested. Neither MitoQ nor SkQ1 is soluable in water. In dry form they should be dissolved in a minimal amount of ethanol or dimethyl sulfoxide (DMSO) - very little is needed, and can be applied gradually with a dropper. Then pour the resulting solution into a glass of water and drink it. This will roughly replicate the delivery mode used in many of the animal studies.

Establishing Tests and Measures

The objective here is a set of tests that (a) match up to the expected outcome based on human trials of mitochondrially targeted antioxidants, and (b) that anyone can run without the need to involve a physician, as that always adds significant time and expense. These tests are focused on the cardiovascular system, particularly measures influenced by vascular stiffness, and some consideration given to parameters relevant to oxidative stress and the development of atherosclerosis.

The cardiovascular health measures in that list are those that are impacted by changes in the elasticity or functional capacity of blood vessels, such as would be expected to occur to some degree in a treatment that compensated in some way for the effects of aging on the smooth muscule cells in blood vessel walls - as is thought to be the case for mitochondrially targeted antioxidants. Positive change of the average values in most of these metrics are achievable with significant time and effort spent in physical training, so movement in the numbers in a short period of time as the result of a treatment should be an interesting data point.

Bloodwork

There exist online services such as WellnessFX where one can order up a blood test and then head off the next day to have it carried out by one of the widely available clinical service companies. Of the set of test packages offered by WellnessFX, the Baseline is probably all that is needed for present purposes. But shop around; this isn't the only provider.

Oxidized LDL Cholesterol

The more mainstream blood test services such as WellnessFX don't offer as wide a range of testing as some of the specialists. For example, the Life Extension Foundation maintains a blood test service that includes a test for oxidized LDL cholesterol. Again, shop around. There are others.

Resting Heart Rate and Blood Pressure

A simple but reliable tool such as the Omron 10 is all you need to measure heart rate and blood pressure. It is worth noting here a couple of general principles for cardiovascular measures. Firstly, the further away from the center of the body that the measurement is taken, the less reliable it is - the more influenced by any number of circumstances, such as position, mood, stress, time of day, and so forth. Fingertip devices are convenient, but nowhere near as useful as something like the Omron 10 that uses pressure on the upper arm. Secondly, all of the above-mentioned line items also influence every cardiovascular measure, so when you are creating a baseline or measuring changes against that baseline, carry out each measure in the same position, at the same time of day, and make multiple measurements over a week to gain a more accurate view of the state of your physiology. The Omron 10 is solid: it just works, and seems quite reliable.

Heart Rate Variability

Surprisingly few of the numerous consumer tools for measuring heart rate variability actually deliver the underlying values used in research papers rather than some form of aggregate rating derived by the vendor; the former is required for any serious testing, and the latter is useless. Caveat emptor, and read the reviews carefully. As an alternative to consumer products, some of the regulated medical devices are quite easy to manage, but good luck in navigating the system to obtain one. The easiest way is to buy second hand medical devices via one of the major marketplaces open to resellers, but that requires a fair-sized investment in time and effort - which comes back to the rule about keeping things simple at the outset.

After some reading around the subject, I settled on the combination of the Polar H10 device coupled with the SelfLoops HRV Android application. I also gave the EliteHRV application a try. Despite the many recommendations for Polar equipment, I could not convince either setup to produce sensible numbers for heart rate variability data: all I obtained during increasingly careful and controlled testing was a very noisy set of clearly unrealistic results, nowhere near the values reported in papers on the subject. However, plenty of people in the quantified self community claim that these systems work reasonably well, so perhaps others will have better luck than I. Take my experience as a caution, and compare data against that reported in the literature before investing a lot of time in measurement.

Pulse Wave Velocity

For pulse wave velocity, choice in consumer tools is considerably more limited. Again, carefully note whether or not a device and matching application will deliver the actual underlying data used in research papers rather than a made-up vendor aggregate rating. I was reduced to trying a fingertip device, the iHeart, picked as being more reliable and easier to use than the line of scales that measure pulse wave velocity. Numerous sources suggest that decently reliable pulse wave velocity data from non-invasive devices is only going to be obtained by measures at the aorta and other core locations, or when using more complicated regulated medical devices that use cuffs and sensors at several places on the body.

Still, less reliable data can be smoothed out to some degree by taking the average of measures over time, and being consistent about position, finger used for a fingertip device, time of day, and so forth when the measurement is taken. It is fairly easy to demonstrate the degree to which these items can vary the output - just use the fingertip device on different fingers in succession and observe the result. All of this is a trade-off. A good approach is to take two measures at one time, using the same finger of left and right hand, as a way to demonstrate consistency. While testing an iHeart device in this way, I did indeed manage to obtain consistent and sensible data, though there is a large variation from day to day even when striving to keep as many of the variables as consistent as possible. That large variation means that only sizable effects could be detected.

DNA Methylation

DNA methylation tests can be ordered from either Osiris Green or Epimorphy / Zymo Research - note that it takes a fair few weeks for delivery in the latter case. From talking to people at the two companis, the normal level of variability for repeat tests from the same sample is something like 1.7 years for the Zymo Research test and 4.8 years for the Osiris Green tests. The level of day to day or intraday variation between different samples from the same individual remains more of a question mark at this point in time, though I am told they are very consistent over measures separated by months. Nonetheless, as for the cardiovascular measures, it is wise to try to make everything as similar as possible when taking the test before and after a treatment: time of day, recency of eating or exercise, recent diet, and so forth.

An Example Set of Daily Measures

An example of one approach to the daily cardiovascular measures is as follows, adding extra measures as a way to demonstrate the level of consistency in the tools:

  • Put on the Polar H10; this is involved enough to increase heart rate a little for a short period of time, so get it out of the way first.
  • Sit down in a comfortable position and relax for a few minutes.
  • Measure blood pressure and pulse on the left arm using the Omron 10.
  • Measure blood pressure and pulse on the right arm using the Omron 10.
  • Measure pulse wave velocity on the left index fingertip over a 30 second period using the iHeart system.
  • Measure pulse wave velocity on the right index fingertip over a 30 second period using the iHeart system.
  • Measure heart rate variability for a ten minute period using the Polar 10 and Selfloops.

Consistency is Very Important

Over the course of an experiment, from first measurement to last measurement, it is important to maintain a consistent weight, diet, and level of exercise. Sizable changes in lifestyle can produce results that may well prevent the detection of any outcome using the simple tests outlined here. Further, when taking any measurement, be consistent in time of day, distance in time from last exercise or meal, and position of the body. Experimentation with measurement devices will quickly demonstrate just how great an impact these line items can have.

Guesstimated Costs

The costs given here are rounded up for the sake of convenience, and in some cases are blurred median values standing in for the range of observed prices in the wild.

  • Business mailbox, such as from UPS: $250 / year
  • Baseline tests from WellnessFX: $220 / test
  • Oxidized LDL test from LEF: $170 / test
  • MyDNAage kits: $310 / kit
  • Osiris Green sample kits: $70 / kit
  • Omron 10 blood pressure monitor: $80
  • Polar H10 heart monitor: $100
  • iHeart monitor: $210
  • American Weigh Gemini-20 microscale: $90
  • Miscellenous equipment: spatulas, labels, vials, pill capsules, etc: $60
  • SkQ1 via Alibaba: $200
  • A bottle of dimethyl sulfoxide (DMSO): $20
  • MitoQ capsules from MitoQ Limited: $190
  • Shipping and LC-MS analysis of samples: $120 / sample

Schedule for the Self-Experiment

One might expect the process of discovery, reading around the topic, ordering materials, and validating the pharmaceuticals to take a couple of months. Once all of the decisions are made and the materials are in hand, pick a start date. The schedule for the self-experiment is as follows:

  • Day 1-10: Once or twice a day, take measures for blood pressure, pulse wave velocity, and heart rate variability.
  • Day 10: Bloodwork and DNA methylation test.
  • Day 11: Start on the six week program of daily doses.
  • Day 53-62: Repeat the blood pressure, pulse wave velocity, and heart rate variability measures.
  • Day 62: Repeat the bloodwork and DNA methylation test.

Where to Publish?

If you run a self-experiment and keep the results to yourself, then you helped only yourself. The true benefit of rational, considered self-experimentation only begins to emerge when many members of community share their data, to an extent that can help to inform formal trials and direction of research and development. There are numerous communities of people whose members self-experiment with various compounds and interventions, with varying degrees of rigor. One can be found at the LongeCity forums, for example, and that is a fair place to post the details and results of a personal trial. Equally if you run your own website or blog, why not there?

When publishing, include all of the measured data, the compounds and doses taken, duration of treatment, and age, weight, and gender. Fuzzing age to a less distinct five year range (e.g. late 40s, early 50s) is fine. If you wish to publish anonymously, it should be fairly safe to do so, as none of that data can be traced back to you without access to the bloodwork provider. None of the usual suspects will be interested in going that far. Negative results are just as important as positive results. For example, given the measures proposed in this post it is entirely plausible that positive changes in a basically healthy late 40s or early 50s individual will be too small to identify - they will be within the same range as random noise and measurement error. Data that confirms this expectation is still important and useful for the community, as it will help to steer future, better efforts.

Comments

I highly recommend to publish such experiments in https://www.biorxiv.org -- this will be much easier to find and analyze all info in one place. And more useful for peer review not just by a few LE enthusiasts but by many realy bench researchers.

Posted by: Ariel at May 1st, 2018 1:17 PM

Hello Reason,
Since the whole setup is quite involving one could expect a considerable placebo effect. The more expensive placebo or e one requiring a higher degree of effort works much better.

Also those supplements should be compared against basic alternatives like exercise, calorie restriction, and regular periodic fasting.

Ideally there should be a group/covenant that will have a pact to share the same protocol to generate comparable data points.

From personalized medication point of view, you are the most relevant dataset for your health
But making sense out of hard to compare data which has a high level of noise will be a gargantuan effort.

Posted by: Cuberat at May 1st, 2018 9:13 PM

While MitoQ is a synthetic supplement, there is a potent mitochondria-directed antioxidant found in food, especially mushrooms: ergothioneine.

Cheah and Halliwell, 2012. Ergothioneine; antioxidant potential, physiological function and role in disease. Biochim Biophys Acta (BBA)-Mol Basis Dis, 1822(5), pp.784-793.
Gruber et al, 2013. Mitochondria-targeted antioxidants and metabolic modulators as pharmacological interventions to slow ageing. Biotech adv, 31(5), pp.563-592. https://www.sciencedirect.com/science/article/pii/S0925443911002201

> Few naturally occurring antioxidant compounds have been reported to penetrate into the mitochondria and fewer still are actively transported there. Ergothioneine (EGT) is a unique naturally occurring, sulfur-containing, amino acid acquired by higher order plants and animals only through dietary means. In humans, EGT has been shown to specifically accumulate in certain cells and tissues at up to millimolar concentrations. Interestingly, EGT has been found to accumulate in the mitochondrial fraction of hepatic cells following injection of radiolabelled EGT into rats. Despite the ubiquitous presence of EGT in many cells, the intrinsic impermeability of plasma membranes to EGT dictates the need for a mechanism of uptake and accumulation in cells. Indeed, an organic cation transporter (OCTN1) has been found to facilitate the specific and Na+ -dependent transport of EGT across the membrane. http://www.ribobio.com/uploadfiles/20140709114002464.pdf

> Consistent with the observation of EGT in the mitochondrial fraction, was the co-localization of green fluorescent protein tagged OCTN1 expression with a red fluorescent protein conjugated mitochondrial marker (cytochrome c oxidase subunit) in several mammalian cell lines, likely indicating the facilitated transport of EGT into the mitochondria. The result was further confirmed by OCTN1 immunoblot of isolated mitochondria from HepG2 cells and also transmission electron microscopy of murine heart incubated with gold-labeled OCTN1 targeted antibodies indicating mitochondrial localization of this transporter.

> While the exact function of EGT has yet to be elucidated, antioxidant and cytoprotective attributes have been often cited with EGT established in vitro, as a powerful scavenger of reactive species including hydroxyl radicals (•OH), hypochlorous acid (HOCl) and peroxynitrite (ONOO-) . EGT was also shown to deactivate singlet oxygen at a higher rate (kΔ= 2.3 × 107 M-1 s-1 than other simple thiols including glutathione and reactions of EGT with •OH and peroxyl radicals were determined by pulse radiolysis to have almost diffusion-controlled rate constants of 1.2 × 1010 M-1 s-1 and 1.2 × 109 M-1 s-1 , respectively. In vitro assays have demonstrated that EGT offers complete protection of DNA and α1-antiproteinase upon exposure to •OH (Cu2+ in the presence of H2O2/ascorbate) and HOCl, respectively. It has been previously established that ONOO- can lead to mitochondrial dysfunction and cell death. EGT demonstrated a dose-dependent inhibition of tyrosine nitration and inactivation of α1-anti-proteinase by ONOO-. Furthermore, EGT prevented H2O2-induced cytotoxicity and DNA damage (observed by the comet assay) in PC12 cells.

> Studies revealed that addition of 20 μM and 100 μM EGT inhibited alloxan induced lipid peroxidation of phosphatidylcholine liposomes by 67% and 100%, respectively, double the protective effect of CoQ10. These antioxidant functions suggest that EGT may act by protecting vulnerable mitochondrial constituents from damage by ROS generated by the electron transport chain. Supporting this theory are the findings by Paul and Snyder, demonstrating increased mitochondrial DNA damage (measured by quantitative real-time PCR of D-Loop mutations) in response to OCTN1 silencing when HeLa cells were exposed to hydrogen peroxide. Furthermore, the authors found that depleting the cells of EGT also led to an increase in protein carbonyls, "lipid peroxidation" and susceptibility to H2O2-induced cell death. Although the protective ability of EGT in mitochondria has yet to be established in vivo, in vitro assays have demonstrated that EGT is able to protect DNA from damage against various reactive species.

> Furthermore EGT has been demonstrated to form redox-inactive complexes with divalent metal cations such as Cu2+ and Fe2+ . Harnessing this ability, EGT was found to protect both DNA and protein against copper-induced oxidative damage (caused by Cu2+ , H2O2 and ascorbate). In contrast to the generation of ROS by GSH in the presence of Cu2+ (via formation of a redox-active Cu(I)-[GSH]2 complex), the complex of EGT with Cu2+ is relatively stable and hence does not decompose to generate radicals, suggesting another means of protection in the mitochondria.

ELI15: Ergothioneine accumulates in fairly high concentrations (in the mM range), especially in mitochondria. This is orders of magnitude greater than most dietary antioxidants. Against 3 of the 4 major reactive oxygen species, its a very potent antioxidant (carotenoids are much better against the 4th, singlet oxygen which is generated by UV). Unlike other antioxidants like endogenous glutathione or vitamin C, it forms stable complexes with copper and iron ions, so doesn't become a pro-oxidant in their presence.

Ergothioneine is fairly unique, in that we possess a transporter to get it across cell membranes (and into mitochondria), which among dietary components is usually restricted to essential nutrients like vitamins. Indeed, the only thing preventing its designation as a vitamin is the lack of a known deficiency disease. But it may be "conditionally essential", essential in disorders that cause oxidative stress.

great sources: king trumpet mushrooms, king bolete mushrooms, oyster mushrooms
good sources: other mushrooms incl. portabello/champignon/white button, shitake; black/kidney beans, liver, oat bran, garlic
has some, but relatively little: meat, egg yolk, pumpkin seeds, whole wheat

Posted by: John at May 1st, 2018 10:53 PM

sad part is both are not available in India

Posted by: Salman at May 1st, 2018 11:37 PM

Reason, where did you come up with the recommendation to refrigerate MitoQ? Instruction on the bottle say, "After opening, store in a dry place below 25℃ /77℉."

Posted by: NY2La at May 2nd, 2018 2:32 AM

It would be interesting to add a section on what to expect from the supplement, is it only a long-term benefit or some short-term effects are noticeable beside changes in the inflammation blood markers?. Also, taking epigenetic age measurements on a six-week window seems to be too short to see any significant change in the aging rate, a six-month period I think is the shortest reasonable window for that.

Posted by: Guillermo Fernandez at May 2nd, 2018 3:23 AM

interesiting thanks reason. i have taken mitoq on a daily basis for over 2 years. it would be nice to try out skq1. is it legal to import into the uk if its intended for research use only?

Posted by: scott emptage at May 2nd, 2018 5:49 AM

@scott emptage: That is definitely a question to run past a lawyer.

Posted by: Reason at May 2nd, 2018 6:33 AM

@NY2La: From one of the chemical supplier product pages.

Posted by: Reason at May 2nd, 2018 6:37 AM

@Guillermo Fernandez: Expectations can be managed by looking at the referenced MitoQ study. I would expect little to no result for younger people, for some definition of "younger", and reduced pulse wave velocity and reduced oxidized blood lipids in some older people after six weeks.

Posted by: Reason at May 2nd, 2018 6:42 AM

@John
An economical and convenient source of ergothioneine is porcini mushroom powder. I consume it in dietary amounts (or perhaps a bit higher) in the form of a broth. The most noticeable effect has been longer eyelashes (so apparently it affects cell cycle control). Let's hope there are no downsides.

Posted by: CD at May 2nd, 2018 12:14 PM

@Reason, what do you think -- does that make sense to use MitoQ for 25 - 30 yo people as a preventative approach? The same for senolitics. Because the goal of rej bio not only reverse high level of accumulated damage but also keep it as low as possible to prevent forming ageing pathology. Maybe better to begin rej bio therapy at 30 and never let damage accumulate to some mesuarable level than await when you are 50 - 60 and remove it in big 'chunks' when you already have some pathology.

Another reason may be that now all alleged rej bio therapies are not very effective in removing damage when organism is already damaged and old. Hovewer, they may be more effective as preventative approach in more young people, they may slow accumulation of the damage. For example, MitoQ is not SENS approach per se -- it doesn't fix mt dna mutations -- only can slightly slow their accumulation.

Posted by: Ariel at May 2nd, 2018 2:54 PM

@Ariel: it is hard to say. To make the argument for prevention while levels are low, you need the real numbers of cells present or problems present and cells removed or problems removed. I think we're a long way away from being able to produce reliable numbers at lower levels of damage.

Posted by: Reason at May 2nd, 2018 3:21 PM

Hi Ariel,
You are a healthy 25 then your body is its prime, so for sure you will witness no improvement. And since ask the housekeeping system work very well you can somehow abuse your body and get away without much damage. At 30 there might be some marginal waste accumulation.
So after 30 you might actually witness some effects, especially if you have some risk factors like obesity or bad lifestyle

Posted by: Cuberat at May 3rd, 2018 6:15 AM

Would reaction time testing be a useful assessment method?

There are free tests on the web, though I doubt their validity. I tried a few and was really shocked at how well I did considering I can't concentrate on anything anymore, I forget to buy things at the grocery store, I forget to put food back in the refrigerator, etc. These tests surely cannot be valid.

Posted by: CD at June 22nd, 2018 12:22 PM

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