The microbiome hypothesis: Lipkin's collaborative, part 1 (Simon McGrath blog)

[Learner1]

Did you take corticosteroids when the PEM already arrived? That apparently does not help at all to reduce PEM (explained in the PEM Busters thread).

Before, during, after.

I don't think those treatments would reduce mitochondrial ROS levels, as far as I am aware, anyway.

THE GLUTATHIONE DOES!!!!

 

[Hip]

THE GLUTATHIONE DOES!!!!

It may do if you can get the glutathione into the cell, but my understanding is that glutathione supplements don't really raise intracellular glutathione levels. I've read that intracellular levels are only increased by supplements like N-acetyl-cysteine, or cysteine-rich whey protein isolate.

 

[Learner1]

Look, I could write an entire book in glutathione, complete with a hefty bibliography of scientific references.

I have 10 years of experimenting with glutathione production and ingestion, with hundreds of labs, and symptom logs, working with doctors who are experts in usung nutrients and in personalizing medicine. And I've been researching why I've been sick and what to do about it. I will summarize here, in layman's terms:

1. Most, if not all, ME/CFS patients suffer from huge oxidative stress, with ROS created by mitochondrial production, infections, toxicity (mold, heavy metals, organophosphates, etc.), and other sources.
2. Glutathione is a powerful antioxidant and can help with all of these ROS. It must be made or recycled, but we can't keep up, so it becomes depleted, which had been noted by many researchers.
   
3. Taking glutathione isn't terribly efficient. It lasts for less than a day in the body. It is better to try to make it, by ensuring all of the necessary ingredients are available.
4. This is one reason why methylating nutrients and amino acids are critical - they lead to the production of glutathione. ME/CFS metabolomics research has found patients tend to be short of amino acids, B2, and B12, which are needed to make glutathione.
   
5. Glutathione works as part of an antioxidant network with vitamins A, C, and E and alpha lipoic acid. Shortages of any compromise the entire network, so ensuring all are there in adequate amounts is critical.
6. One can shortcut with whey protein or NAC, but this doesn't give you a robust, fully functioning system, resulting in other problems to solve down the line.

So, my labs have consistently shown shortages of glutathione for several years. Whey was not an option as it could cause anaphylaxis.

We increased all of my methylating nutrients and been able to improve my glutathione, but it was still a problem. I took lots of NAC, but it wasn't enough. We upped glycine to 4.5 g per day and made headway. We upped antioxidants as they all showed severely deficient - 9g vitamin C, 50,000 IUs vitamin A, 1g mixed vitamin E, Xymogen ALAmax plus 1 tsp PolyMVA for alpha lipoic acid.

Witb the oral supplementation, I also have done 2-3 injections of B complex + MB12 a week, plus an IV with Vs and glutathione once a week. I was still short of vitamin C and glutathione...

Adding in 500mg Seeking Health or Designs for Health liposomal glutathione pre-workout and taking 500mg Thorne R-glutathione, aling with 5g BCAAs has gotten rid of my PEM completely.

Now, you're probably wondering why I'm taking such crazy amounts of everything. I have some toxicity - mycotoxins from mold and a little arsenic, cadmium, lead, mercury, sbd platinum, even after chelating for years. I seem to be burning aminos for fuel as Fluge and Mella found. A recent test for peroxynitrites showed that they are high - this impairs complex I and damages mitochindrial membranes - my mitochondrial membranes seen to be leaky, and I'm inefficient at using the methylating nutrients - we are working to fix this with NT Factor and phosphatidyl choline. And Martin Pall recommends vitamin C, methylating nutrients and glutathione for peroxynitrites, which are partly caused by inadequate MnSOD.

But, I finally seem to have enough glutathione and am functioning the best I've been since 2015.

Having good labs and following up on the problems found is critically important. Following "what's helped other patients" can be a wasted effort and even dangerous. I realize your NHS isn't much help, but quite frankly, my US insurance doesn't pay for most of this either. Some of the tests are covered but the treatment has all been out of pocket. There are a few doctors in the UK who can do this, too.

 

[Hip]

I am not saying glutathione supplementation does not have benefits; glutathione also works extracellularly, which may offer benefits. However, glutathione supplementation does not have much impact on intracellular glutathione.

Earlier you were theorizing that ROS from mitochondria are the cause of PEM. Well, that's an interesting theory, but as far as I can see, glutathione supplements are not going to help target those mitochondrial ROS, because glutathione supplementation does not efficiently raise intracellular level of glutathione, from what I have read at least.

 

[Learner1]

Well, you are wrong.

 

[Hip]

Well, you are wrong.

Well if you want to make a strong definitive statement like that, you really need to back it up with some studies that support your view that glutathione supplementation is effective in raising intracellular level of glutathione.

 

[Learner1]

This is from 2011, but Rich's talk, halfway down the page, with downloadable slides, are helpful as general info on glutathione and ME/CFS.

https://phoenixrising.me/treating-c...e-mecfs-glutathione-and-the-methylation-cycle

I am not a fan of his simplified methylation protocol - it has led too many people into trouble. An individualized approach, based on labs, is preferable.

I have to go to work now, but when I have time, I will pull up more specific reference to Hip's concern.

 

[Learner1]

@Hip There is a plethora of info on glutathione and how it works inside and outside of cells. This is just a minute sampling...there are over 13,000 articles on PubMed...

First, an article describing where ROS come from:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3454471/

Reactive oxygen species (ROS) are generated during mitochondrial oxidative metabolism as well as in cellular response to xenobiotics, cytokines, and bacterial invasion. Oxidative stress refers to the imbalance due to excess ROS or oxidants over the capability of the cell to mount an effective antioxidant response. Oxidative stress results in macromolecular damage and is implicated in various disease states.

Paradoxically, accumulating evidence indicates that ROS also serve as critical signaling molecules in cell proliferation and survival. Cellular ROS sensing and metabolism are tightly regulated by a variety of proteins involved in the redox (reduction/oxidation) mechanism.

Next, oxidative and nitrosative stress and glutathione depletion in ME/CFS causing mitochondrial dysfunction:

https://www.ncbi.nlm.nih.gov/m/pubmed/24669210/

Oxidative stress results from an imbalance of O&NS species and antioxidants and can result from impaired cellular anti-oxidant defenses. Thus diminished activity of superoxide dismutase [117], zinc [118] and glutathione homeostasis [119,120] may contribute to increased O&NS. Prolonged O&NS is one of the major causes of mitochondrial dysfunction in neurological diseases [121]. Impaired antioxidant defenses and impaired glutathione homeostasis are found in Parkinson's [122], Alzheimer's [123] and Huntington's disease [124] and Amyotrophic Lateral Sclerosis [125]. We have reviewed elsewhere that ME/CFS is accompanied by lowered levels of key antioxidants, such as coenzyme Q10, zinc and glutathione [2].

This has a lengthy discussion of ROS production in mitochondria and the role of glutathione in dealing with them, including how glutathione gets into the cell to do this.

https://www.hindawi.com/journals/jaa/2012/736837l/

Due to the pivotal role of mitochondria in programmed cell death (apoptosis) as well as extensive ROS involvement in this process, and adding the fact that mitochondria produce over 90% of cellular ROS, the role of GSH in cell protection cannot be overestimated. GSH may either directly bind some ROS species or serve as a source of reductive power for certain antioxidant systems.

The inner mitochondrial membrane is particularly rich in cardiolipin, whereas it is virtually absent from other membranes and only the outer mitochondrial membrane contains minor amounts of this phospholipid. When mGSH levels are compromised, cardiolipin is one of the important targets of oxidative damage. Due to its unique chemical structure among phospholipids, cardiolipin confers stability and fluidity to the mitochondrial membrane. In addition, cytochrome is normally bound to the inner mitochondrial membrane via its association with cardiolipin.

By protecting cardiolipin from oxidative damage, GSH prevents changes in the physicochemical properties of the mitochondrial inner membrane that lead to membrane destabilization and the dissociation of cytochrome .

ROS also induce an increase in permeability of the internal mitochondrial membrane for calcium. Enhanced ROS and calcium levels, acting in concert, may trigger the cell death machinery via apoptosis or necrosis. Hence, mitochondrial GSH clearly has an important role in preventing apoptosis triggered by cytochrome release from the inner membrane.

Not surprisingly, therefore, a decrease in mGSH levels is closely associated with certain pathologies in both humans and animals.

And this describes how glutathione moves around throughout cells, including carriers that transport it across cell membranes.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4079069/

While synthesized exclusively in the cytosol from its constituent amino acids, GSH is distributed in different compartments, including mitochondria where its concentration in the matrix equals that of the cytosol. This feature and its negative charge at physiological pH imply the existence of specific carriers to import GSH from the cytosol to the mitochondrial matrix, where it plays a key role in defense against respiration-induced reactive oxygen species and in the detoxification of lipid hydroperoxides and electrophiles.

This describes onion extract and quercetin gerting through cell membranes to increase intracellular glutathione:

https://www.ncbi.nlm.nih.gov/m/pubmed/11864778/

Our results demonstrate that extract from onion and various flavonoids induce the cellular antioxidant system. Onion extract and quercetin were able to increase the intracellular concentration of glutathione by approximately 50%. Our data strongly suggest that flavonoids are important in the regulation of the intracellular glutathione levels.

And this is a US University hospital clinic providing IV glutathione to patients:

https://www.gwcim.com/services/intravenous-therapies/

This deficiency causes increased free radicals that further the disease process. Mitochondrial cells, which produce energy, are the most vulnerable to low glutathione and can die when glutathione levels are below 70%.

 

[percyval577]

Thank you @Learner1 for the nice articles all of which I didn´t know I think.
(Maybe everybody may put the year or an author to it, the reader then may know "Ah this article", or when rereading the message may coordinate his knowledge (though me/cfs may hinder such memories ...). I need to remember myself I guess.)

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A generel assessment to the use of "ROS" (still somehow abused by scientists)
Since around 2000 the idea has come up that ROS are not only a detrimental byproduct but are a (critical) byproduct that has become utilizised, and therefore would serve for different roles under different circumstances. Its more a product, may a "inevitable product".

Before I come to the critic: Scince around 2000 then there has been often spoken of ROS as "signaling molecules" which is inaccurate because a signaling molecule is defined as a specie that doesn´t change itself when acting. But it might be also look a bit strange to talk of them as "metabolites", because their generation has - as increasing amounts of articles show - become regulated for special purposes. So, it my be best to say "metabolite xy" or to come up with a new, third term.

The saying of "ROS" is still connected to te first and only detrimental sight, which had been justified because little was known, and you ´d to deal with the unknown by some terms of course. But with further knowledge:


Especially the first step of changes that occurs - the dismutation of the ROS superoxide to the ROS H2O2 - shows nowadays that the term "ROS" itself cannot indicate a generel properity. You have the choice which ROS you want to have. I "admitt" that also the production itself seems to be regulated, but that´s not in question here.

Superoxide will react fast with iron-sulfur centers, which are (almost ridiculous) high in complex I and then become less often in complex II and III. It is not difficult to guess that less dismutation of superoxide will "damage" these iron-sulfur centers and therefore will "damage" the complexes I-III to different extent (and can lead to a slowed down oxphos).

H2O2 then, that will be promoted by dismutation, will do many other things, for example it can react to HOCl, damaging pathogenes. Or to OH(-), the most dangerous one (so far), which will damage human DNA.

It would be nice if all (researchers and others) would say then "H2O2 and downstream molecules" instead of "ROS", or that superoxide might get excluded from the term "ROS".

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Angajala, Lim et al. 2018: "Diverse Roles of Mitochondria in Immune Response ..." a nice review, S.14:

:https://www.frontiersin.org/articles/10.3389/fimmu.2018.01605/full​

"Natural antioxidants (vitamine E, curcumin, ginko biloba, melatonin) in addition to target TPP-based antioxidants (MitoQ, Mito-VitE, Mito-alpha-lipoic acid, Mito-PBN), some small peptide-based molecules (SS31,SS02,SS19,SS20), choline esters of glutathione, and N-acetyl-L-cysteine neutralize mtROS which further maintains normal MMP ( ). These molecules are preferentially taken up by mitochondria due to different charge (negative charge in mitochondria and positive charge on the molecules)."

 

[Hip]

There is a plethora of info on glutathione and how it works inside and outside of cells.

Sure, but nothing you posted there that indicates glutathione supplementation will increase intracellular glutathione to any significant degree.

This describes onion extract and quercetin gerting through cell membranes to increase intracellular glutathione:

https://www.ncbi.nlm.nih.gov/m/pubmed/11864778/

That's an interesting study, but note it is in vitro, so its results may not apply in vivo.

Quercetin I don't think will work to raise intracellular glutathione in vivo, by a quick calculation that I just did now. My calculation showed you cannot get sufficient levels of free quercetin in the blood to match the concentrations of 5 μM and 25 μM of free quercetin used in the in vitro study.

My calc showed an oral dose of 500 mg quercetin daily will only result in a 0.06 μM peak free concentration in the blood, which is far too low, compared to 5 μM.


The onion extract looks like a potentially interesting way to increase intracellular glutathione. I don't have the data on the onion compound to perform the calculation, so cannot say whether it will work or no in vivo, but one could certainly try it.


One individual "hidir" on my website posted a comment about an onion treatment devised by Prof Ibrahim Adnan Saraçoğlu in Turkey. Hidir said he and his wife used this onion treatment with some beneficial effects. In the recipe he used, the onions were boiled for 10 minutes:

hi hip,
I did this onion cure 10 days with my wife. I felt like my contagious level become very low. i am more comfortable around people. if something working on this virus I understand from my throat. red spots in throat. disappear. in this onion cure red spots become blurry . in my opinion i dont see them .
i got some positive results. my gastrointestinal tract problems become smaller. I got more energy.
i sleep better. my memory more clear. onion cure is helpful.
this approach turn regular food into medicine is powerful. if you use same food every day large amount turn them into medicine. very low side effects.
i like to give onion cure again here.

onion, brown onion, golden brown , regular onion ,common onion .(not red ,not blue ,not white)
cut whole onion 1/4 parts and put in 1 and 1/2 cup water , bring water to boil and boil them around 5 minutes (my wife boiled more than 10 minutes)
drink whole water warm
do it 2 times a day for 15 days (1 before noon , 1 after noon )
if you working person you can do it when come from job and before bed.
for women start this cure 2-3 days after menstural cycle over.
you can do it twice a year.
ulcer people may be dont want do it.
ref.. prof Ibrahim Adnan Saracoglu

http://en.wikipedia.org/wiki/İbrahim_Adnan_Saraçoğlu

I tried the Saracoglu onion remedy myself, but after two days had worsened brain fog and depression so stopped. But this might have just been a coincidence, as I naturally get days with worsened symptoms anyway.

But it might be worth trying the Saracoglu onion remedy, especially combined with N-acetyl-cysteine, which is known to raise intracellular glutathione.

 

[Learner1]

Sure, but nothing you posted there that indicates glutathione supplementation will increase intracellular glutathione to any significant degree

I've been working with doctors to actually put this into practice and have been steadily improving, using multiple strategies to increase my intracellular glutathione.

What I've learned in practice is that neither onions nor NAC ingestion is sufficient to raise my intracellular glutathione levels adequately, as I've been ingesting both for over 5 years. It has taken adding a large amount of methylating nutrients, amino acids, and direct glutathione supplementation to do this. And, because its been done in incremental steps, its been possible to track progress with labs.

One key marker is 8 OHdG - which is a marker of DNA damage caused by oxidative stress from the hydroxyl radical coming from peroxynitrite production resulting from too much superoxide reacting with NO. As a stage 3 cancer survivor, I am very interested in reducing DNA damage, and have been pleased to see that this marker, which was high a year ago has now normalized with increased glutathione in my system.

And, my PEM is gone. I believe the results of excessive oxidative stress have a lot to do with the development of PEM.

 

[Hip]

It is clear you either didn't read or didn't understand the papers that described the many ways glutathione is transported into cells.

Perhaps you could do your own further reading to satisfy yourself with some of the other 12,994 articles in PubMed.

Do you really expect someone to read the 13,000 articles on PubMed you mentioned about glutathione!? That's ludicrous.

It's incumbent on you to provide the references to support your statements, that's normal scientific etiquette. It's no good going around making all sorts of assertions which you cannot support with factual evidence, and then saying to people that it is up to them to find the evidence to support your statements.

They are your statements, so you provide the references. Etiquette please.

 

[Learner1]

Since around 2000 the idea has come up that ROS are not only a detrimental byproduct but are a (critical) byproduct that has become utilizised, and therefore would serve for different roles under different circumstances. Its more a product, may a "inevitable product".

Yes, superoxide is an inevitable product as mitochondria make energy.

It's the balance of these ROS vs our natural antioxidant mechanisms that is critical to health. Healthy people will have a more even balance between the two.

But in people with inflammation, infections, toxicity, etc., this balance is usually tipped to too many ROS vs the amount of antioxidants available - this is shy we see depletion of glutathione.

Especially the first step of changes that occurs - the dismutation of the ROS superoxide to the ROS H2O2

This requires having enough superoxide dismutase in the mitochondria, or MnSOD. There are different reasons, like SOD2 SNPs, manganese deficiency, etc. that less MnSOD may be available to tackle superoxide radicals.

The superoxide radical doesn't only go to hydrogen peroxide. Theres an instantaneous reaction with NO creating peroxynitrites, which damage mitochondrial membranes, impair complex I function and lead to crrating hydroxyl radicals which can damage DNA.

Superoxide will react fast with iron-sulfur centers, which are (almost ridiculous) high in complex I and then become less often in complex II and III.

These, too. The iron containing molecules can do a lot of damage too.

H2O2 then, that will be promoted by dismutation, will do many other things, for example it can react to HOCl, damaging pathogenes.

This is how high dose vitamin C works to kill pathogens (even Ebola) and cancer.

Or to OH(-), the most dangerous one (so far), which will damage human DNA.

Yes, it will. As I mentioned previously, labs csn mrssure 8 OHdG, which a marker of this damage.

Patients with ME/CFS are known to have huge oxidative stress. Trackng antioxidant status, including glutathione (and its precursors), vitamins A, C, and E, alpha lipoic acid, selenium, zinc, and CoQ10 is very useful, and supplementing to balance oxidants vs antioxidants can be very helpful in reducing the damage being done daily by too many oxidants.

 

[Learner1]

It's incumbent on you to provide the references to support your statements, that's normal scientific etiquette. It's no good going around making all sorts of assertions which you cannot support with factual evidence, and then saying to people that it is up to them to find the evidence to support your statements.

I provided ample references with factual evidence to support my position.

 

[Hip]

I provided ample references with factual evidence to support my position.

you need to understand that it is not attacking someone just because you ask them for a link to a paper which supports the assertions they make.

If you are not able to find the reference yourself, perhaps ask one of your doctors to provide you with one.

 

[Hip]

I provided ample references with factual evidence to support my position.

Asking for references to support a statement being made is not attacking another member. It's simply trying to verify whether the statement is true or not. We should not need to get into this stressful type of dialogue over questions like this.

Saying that "there are over 13,000 articles on PubMed" about glutathione does not mean anything. There could be a million articles, but that still does not prove your statement (or your doctor's statement?) that oral glutathione gets into cells.


Here is an example of how to provide a supporting reference:

Glutathione does not cross cell membranes directly. It is synthesized in every cell of the body and largely remains sequestered within cells. Glutathione administered exogenously thus has little effect on intracellular levels, and this limits the therapeutic value of this agent.

Source: The Veterinary ICU Book, by Wayne Wingfield, Marc Raffe, Page 33.

So you can see from the above that the glutathione you take orally or intravenously is not going to get into cells. However, because glutathione is broken down into L-cysteine, and because L-cysteine supports the cells' own synthesis of glutathione, supplementing with glutathione may have some effects via that route.

But if you are just taking glutathione to provide L-cysteine, you may as well take L-cysteine directly, in the form of N-acetyl-cysteine for example, which is actually cheaper that glutathione.

 

[percyval577]

Yes, superoxide is an inevitable product as mitochondria make energy.

Rather no. This would be an inevitable byproduct, but insofar it is used for purpurses and is under some circumstances be wanted to be higher (or even high?), its a real product. Still a dangerous one of course.

That includes that a reaction is not in itself a damage, but should be part of an equilibrium. So, superoxide will be used to destroy iron-sulfur clusters ... and it´s so far neither bad nor good. It depends on what is going on. Or to say it from the other side, the account of iron-sulfur clusters is acting for an equilibrium, eg to slow down complex I more easily than complex III. (In complex I there are 8-9 in the electron transduction-channel, wheras in complex II there are three, and finally in complex III only one).
Correct me if this is nonsense or my memory works wrong.


Finally I have to really critisize you. The review "Glutathione and mitochondria" Ribas et al. 2014 does not indicate that glutathione supplement should work (in respect of the illnesses looked at).
Glutathione might get into the cells, it [apparently will] pass through the outer mitochondria membrane, but it is not able to cross the inner mitochondria membrane. Active transporters are needed and here different tissues behave different.

Maybe you suffer from a glutathione-synthase defect, and then it would help you.

 

[mariovitali]

@Learner1
I had a look about Glutathione synthesis and -as expected - Liver can be the problem :

Outside of polymorphism, decreased GSH synthesis occurs during aging, diabetes mellitus, fibrotic diseases (including cystic fibrosis and pulmonary fibrosis), endotoxemia, and several hepatic disorders such as cholestatic and alcoholic liver injury

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3549305/


Please see the several mentions on Cholestasis. Have you ever performed a Total Bile Acids test?

Also due to low levels of GSH, Liver Fibrosis is possible so you may consider to have a Fibroscan test

 

[msf]

Now where have I heard this before? Oh, that's right, in Belgium.

 

[Learner1]

@Learner1
I had a look about Glutathione synthesis and -as expected - Liver can be the problem :

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3549305/

Please see the several mentions on Cholestasis. Have you ever performed a Total Bile Acids test?

Also due to low levels of GSH, Liver Fibrosis is possible so you may consider to have a Fibroscan test

According to the Mayo Clinic, the liver removes and breaks down most drugs and chemicals from your bloodstream. Breaking down toxins creates byproducts that can damage the liver. Although the liver has a great capacity for regeneration, constant exposure to toxic substances can cause serious, sometimes irreversible harm.

So, yes, this is a concern, not only for me, but for most ME/CFS patients, for whom oxidative stress and glutathione depletion are ongoing problems.

In my case, my doctors check my liver numbers every 3 weeks, along with lab testing for homocysteine, glutathione and other antioxidants every few months. I also had a CT scan - I was able to stand next to the radiologist, with whom I shared my concerns, and he found it to look robustly healthy, likely due to the liver support nutrient protocol I've been on.

A Fibroscan would be a good next step if we were to be concerned...far less invasive than a liver biopsy.