Glutathione depletion blocks conversion of OH-B12 to methyl-B12: new evidence

[richvank]

Hi, all.

Will Marsden recently called my attention to a recent paper from Prof. Richard Deth's group (abstract below) that provides evidence from a rat experiment that major glutathione depletion blocks the conversion of hydroxocobalamin to methylcobalamin.

This has been one of the main propositions of the pathogenesis model that I have proposed for ME/CFS, i.e. the Glutathione Depletion--Methylation Cycle Block hypothesis. So far, this model has continued to be supported as more research is being completed.

One of the things this has brought home to me is that in cases in which glutathione or SAMe are extremely low, it will be difficult to get the methylation cycle going using hydroxocobalamin as the form of B12. This is the form included in the simplified protocol I have suggested, and it was found to be helpful for more than two-thirds of the people in our clinical study, but this may explain why some of the people did not receive benefit from this protocol.

Note that the protocol recommended by Freddd uses methylcobalamin as one of the forms of B12. Methylcobalamin is also used by Dr. Amy Yasko in some cases, depending on genomic polymorphisms. It is also used by Dr. Neubrander and other physicians participating in the DAN! project for treating autism.

Recently I have been suggesting that if the simplified protocol does not produce benefits within three months, either testing should be performed to determine why, or a change should be made in the protocol used. One possibility would be to add methylcobalamin, starting at low dosage and working up, as tolerated.

Best regards,

Rich



Alcohol Clin Exp Res. 2011 Feb;35(2):277-83. doi: 10.1111/j.1530-0277.2010.01343.x. Epub 2010 Dec 1.

Ethanol lowers glutathione in rat liver and brain and inhibits methionine synthase in a cobalamin-dependent manner.

Waly MI, Kharbanda KK, Deth RC.
Source

Department of Food Science and Nutrition, Sultan Qaboos University, Muscat, Sultanate of Oman.
Abstract
BACKGROUND:

Methionine synthase (MS) is a ubiquitous enzyme that requires vitamin B12 (cobalamin) and 5-methyl-tetrahydrofolate for the methylation of homocysteine to methionine. Previous studies have shown that acute or chronic ethanol (ETOH) administration results in the inhibition of MS and depletion of glutathione (GSH), and it has been proposed that GSH is required for the synthesis of methylcobalamin (MeCbl).
METHODS:

We measured GSH levels and investigated the ability of different cobalamin cofactors [cyano- (CNCbl), glutathionyl- (GSCbl), hydroxo- (OHCbl), and MeCbl] to support MS activity in liver and brain cortex from control and ETOH-treated rats.
RESULTS:

In control animals, MS activity was higher in liver than in cortex for all cobalamins and MeCbl-based activity was higher than for other cofactors. S-adenosylmethionine (SAM) was required for OHCbl, CNCbl, and GSCbl-based activity, but not for MeCbl. Feeding an ETOH-containing diet for four weeks caused a significant decrease in liver MS activity, in a cobalamin-dependent manner (OHCbl ? CNCbl > GSCbl > MeCbl). In brain cortex, OHCbl, CNCbl, and GSCbl-based activity was reduced by ETOH treatment, but MeCbl-based activity was unaffected. GSH levels were reduced by ETOH treatment in both liver and cortex homogenates, and addition of GSH restored OHCbl-based MS activity to control levels. Betaine administration had no significant effect on GSH levels or MS activity in either control or ETOH-fed groups.
CONCLUSIONS:

The ETOH-induced decrease in OHCbl-based MS activity is secondary to decreased GSH levels and a decreased ability to synthesize MeCbl. The ability of MeCbl to completely offset ETOH inhibition in brain cortex, but not liver, suggests tissue-specific differences in the GSH-dependent regulation of MS activity.

Copyright 2010 by the Research Society on Alcoholism.

PMID:
21121936
[

 

[Tristen]

Thanks Rich,

I have made significant progress on your Simplified Protocol over the last 1.5 years. The HydroxB12 has been very helpful. I don't tolerate MethylB12. Currently, I seem to have plateaued in my progress, but rather than switch over to Methylcobalamin, I'm wondering what you think of adding a good source of Glutathione to my regimine. I know this isn't a very good idea until one has done some work lifting the partial block, but I've been at it for some time now. So, if raising the GSH levels increases ones response to B12, it seems that keeping my HydroxyB12, and adding GSH supplementation, may get things moving again?

 

[aquariusgirl]

so is that 2 strikes v. hydroxy? It gets oxidisd by toxins and it can't be converted to methyl B12? and also where do you stand now on the issue of methyl B12 overdriving the methylation cycle..\
\\
a couple of softball questions for you!!
Thanks

 

[anne_likes_red]

For some people, if glutathione depletion is the cause of their failure to convert, then perhaps MethylB12 is only required as a short term measure anyway?
(And perhaps there is a role for supplementary glutathione in the short term too?)
Anne.

 

[Sallysblooms]

I have noticed a difference with the addition of the Lipo. GSH and ATP with all of my other supplements. All have to work together for sure.

 

[aquariusgirl]

sally... i think u said u are using the stabilsied ATP.. what brand is that and where do u get it?
thanks

 

[Sallysblooms]

It is called ATP-20 by Douglas Labratories. I get it from my doc or you can get it online. I get all my supplements online or from my doctors. They tell me the brands and amounts to use, I am on a schedule of supplements three times a day.

The ATP is a little pill for under the tongue. I take that once or twice a day. B12 under the tongue also.

 

[aquariusgirl]

thanks! how many do u take a day?

 

[Sallysblooms]

Started with one, sometimes two now.

 

[Dreambirdie]

Hey Rich--

I have run out of the Perque (the hydroxo B12). I know you recommend another brand now (in drops?).
What's it called and where do I get that one?

thanks.

 

[anne_likes_red]

Hi Dreambirdie )
It's Hydroxy B12 Mega Drops from Holistic Health: 2 drops under the tongue daily.
HTH.

 

[richvank]

Thanks Rich,

I have made significant progress on your Simplified Protocol over the last 1.5 years. The HydroxB12 has been very helpful. I don't tolerate MethylB12. Currently, I seem to have plateaued in my progress, but rather than switch over to Methylcobalamin, I'm wondering what you think of adding a good source of Glutathione to my regimine. I know this isn't a very good idea until one has done some work lifting the partial block, but I've been at it for some time now. So, if raising the GSH levels increases ones response to B12, it seems that keeping my HydroxyB12, and adding GSH supplementation, may get things moving again?

Hi, Tristen.

I think that's probably worth a try. Liposomal glutathione seems to be able to deliver glutathione to the cells pretty well. Lately I've been hearing more about acetyl glutathione as another effective way to do this, but I don't know much about it yet.

If this doesn't help, it may be necessary to do some lab testing to try to figure out what is holding up your recovery. There could be a deficiency of something, or there could be a problem with the recycling of glutathione. There could also be a "stressor" of some sort that is placing a demand on glutathione and keeping it from coming up to normal (this could be any of a variety of things, such as heavy metals, infections, mold illness, or something else). A methylation pathways panel would tell you what your glutathione status actually is, since it measures both the reduced and oxidized forms.

Best regards,

Rich

 

[richvank]

so is that 2 strikes v. hydroxy? It gets oxidisd by toxins and it can't be converted to methyl B12? and also where do you stand now on the issue of methyl B12 overdriving the methylation cycle..\
\\
a couple of softball questions for you!!
Thanks

Hi, aquariusgirl.

Yeah, thanks for the slow pitches!! -)

I continue to struggle with the issue of what should be the form or forms of B12 that a person should try first as part of a methylation-type treatment, and I still don't have a very satifying answer. Different people do seem to respond differently. Please bring me up to date on your experience. I know that the initial simplified protocol with hydroxo B12 did some good things for you at the beginning, and then I recall that you hit a plateau and that switching to methyl B12 was beneficial. Is that correct? What is your situation now with respect to these two forms? Are there things that aren't being resolved by taking either of them as a component of methylation treatment?

It's true that hydroxo B12 does have these downsides, but it did seem to help over two thirds of the women in our clinical study. I realize that for many people it is not feasible to run much in the way of lab tests, either for reasons of finances or inability to get to a physician or phlebotomist, or to find one who will order the tests that require doctor's orders. For those who are able to run lab tests, the picture can be made somewhat clearer, but as you know, there are usually still some unknowns.

With regard to overdriving the methylation cycle, I continue to have concern about that when high dosages of sublingual or injected methyl B12 and high dosages of methylfolate are used together. It makes sense biochemically that this would occur, and the data I've seen from I think three people now does confirm that this does happen. In the cases I've had data from so far, the result of it seemed to be low flow of homocysteine into the transsulfuration pathway and thus low rate of synthesis of glutathione and other sulfur-containing substances. Overdriving the methylation cycle causes the methylation cycle and the folate metabolism to go into a "futile cycle," in which excess methyl groups are passed back and forth between the two by sarcosine and 5-methyl THF. So long as the person has functioning glycine N-methyltransferase, sarcosine dehydrogenase, and MTHFR enzymes, which nearly everyone does, though some have SNPs in MTHFR, this is all that happens. I still have concerns that some may have genomic abnormalities that could cause other effects, though.

I don't know how low the dosages of methylfolate and methyl B12 would need to be to prevent this futile cycle and allow flow into the transsulfuration pathway, but my guess is that a few hundred micrograms per day of the folates and somewhere in the low milligrams per day of methyl B12 would probably be the ranges.

I do think that it is important to get some flow going into the transsulfuration pathway, while also restoring the methylation cycle to more normal flow.

My current suggestion is that if it is not feasible for a person to do lab testing, but if they clearly do have ME/CFS, then they should try the current version of the simplified treatment approach, which uses hydroxo B12 as its B12 form. If they do not experience improvements within 3 months, and they are still not able to do testing, then I suggest that they switch to methyl B12 and see if that helps.

If it is feasible to do lab testing, then I suggest as a minimum that they run the Health Diagnostics and Research Institute (or European Laboratory of Nutrients) methylation pathways panel and the Metametrix plasma 50-amino acids panel, and from these decide whether hydroxo- or methyl-B12 would be the best form to use, based on the levels of SAMe, SAH and glutathione, and decide on whether additional supplements (such as methionine, serine, magnesium, B6 and others) are needed based on the amino acids panel.

In all cases, I continue to recommend that people be monitored by a physician while on this type of treatment, to make sure that if any serious adverse effects arise, they will be recognized early and dealt with promptly and properly.

Best regards,

Rich

 

[Tristen]

Hi, Tristen.

I think that's probably worth a try. Liposomal glutathione seems to be able to deliver glutathione to the cells pretty well. Lately I've been hearing more about acetyl glutathione as another effective way to do this, but I don't know much about it yet.

If this doesn't help, it may be necessary to do some lab testing to try to figure out what is holding up your recovery. There could be a deficiency of something, or there could be a problem with the recycling of glutathione. There could also be a "stressor" of some sort that is placing a demand on glutathione and keeping it from coming up to normal (this could be any of a variety of things, such as heavy metals, infections, mold illness, or something else). A methylation pathways panel would tell you what your glutathione status actually is, since it measures both the reduced and oxidized forms.

Best regards,

Rich

Thanks Rich, I wanted to add that I hadn't been taking the Serine due to bad reactions to it in the past. I did see an obvious improvement after adding the lecithin a few months back, but have since then plateaued. Because of this nudge in progress after adding the Lecithin, I think you may be right about other deficiencies.

I'll continue with the current regimen, add some GSH (liposomal or nasal), and see how it goes. My last holdout would be the testing (financial reasons). Maybe I can tolerate SAMe now.

I saw a dramatic change when I added 250mg/day Acetyl-L-Carnatine. At first I thought it quite diagnostic of a deficiency, which may be true, but I made the mistake of thinking that I could push my stress boundary farther (based on the idea that mito malfunction is the cause of PEM). I'm planning to restart it, but next time force myself to take it very slow, in spite of feeling it's not necessary.

The Hydroxy B12 mega drops from Holistic Health is what I use too.

 

[DANEL]

hi rich,

i am new here and trying to take all this in-a bit overwhelming.

i tried methy b12 sub 1 mg and folate but the fillers/sugar/futose, mannitol, sobitol, were making me feel sick, nausea immed-don't think it was detox b/c this has also happened with homeopathic pellets with fuctose and lactose. i have heavy metal, mold illness, infections, etc

looked at holsitic health for the hydo and methyl drops also contain sugar in the form of tae...(sp)

i see you first recommend the hydo and folinic acid, then it not better in 3 months switch to the methy b12

i can try that first,BUT

was wondering if the methyl b12, COMPOUNDED without fillers (except cellulose in a gelitin capsule), no sugar, no lactose, 1 mg, would be okay. it is capsule form, not drops or sublin,
so didn't know how taking a capsule would work

or if taking the methyb12 first in my case was preferable due to the sugar in the hydo drops.

the folinic looks pretty pure while the folate by solgar has mannitol.

do most here start with the hydo, methyl or both?


also would taking the urine amino acid test be as good/tell me as much, as the plasma 50 aa panel?

is sam-e prefered over tmg or dmg? to start or does your protocol not suggest any tmg or dmg ONLY sam-e?

any help is appreciated

denise

 

[aquariusgirl]

Hi Rich

Well, as you know I started the simplified protocol in Feb of 07 & it helped globally with almost all my symptoms, but the progress was painstaking slow, even when I eventually upgraded to large injectable dosages of hydroxycobalamin.

I was injecting 1cc or ml of 25mg/ml a day,and sometimes I would use some methyl B12 sublinguals as well.

The injectable B12 was running a $150 a month. of which insurance paid all but $30.

On my first MAP, in March 2008, my MMA was 17.3. Figlu was 3.3

I just checked and my MMA on a MAP I ran in August 2010 was 10.4. Figlu was 4.3. So I still had a significant B12 deficiency at that point.

(I know these are not straight comparisons.because the baseline numbers do not reflect the true deficiency because the cycle is not functioning properly. But anyway just for thoroughness...)

(I tell you what though. My Krebs cycle looks a lot better though! If I knew then what I know now ...I think I would have chelated...but having said that we have better chelation options now. Thank you Boyd Haley!!)

I suspected that a lot of this B12 was being oxidised. I did try the glut + hydroxy B12 combo once to try and prevent this...but I got awful burning sensations in my brain that went on for several days. I have never been brave enough to repeat that experiment with lower dosages.

I think part of the problem was I wasn't getting flow down my transsulfuration pathway due to lack of B6 & therefore I wasn't making glutathione. I had avoided all B6 because of my CBS upregulations.

So even though I was supporting my methylation cycle, I wasn't doing enough to support the transulfuration pathway, so I was deprving myself of glutathione while increasing my need for it.

I noticed that I did better when I made sure to include all the B vits, esp B2+3 +6.

Having said that, I was getting some good metal pulls on UTMs and FTMs on a random basis.. but it never translated into real functional improvement.

Eventually though,I, like a lot of the old timers hit a point where excitotoxicity became a real problem. Even small amounts of folate & methyl B12 created really unpleasant brain sensations.

I also got supplement fatigue & so I have switched tacks.

I have wondered if it would be good to combine therapies ...something like LDN that tamps down inflammation, that spares glutathione, + methylation.

I recall that a woman called Sue B or Sue T on the yasko list did much better when she switched to methyl B12 from hydroxy and went on Valtrex. I assume the Valtrex helped reduce oxidative stress.

Of course, I fall into the very sick category. Sick 16 years. Positive for chronic EBV even on regular labs. Presumably huge burden of toxins, metals, pathogens.

I have been detoxing for a couple of years now, chugging down the charcoal, & it still keeps coming.

This is a very long way of saying that I am kinda sceptical that the hydroxy B12 is getting to where it needs to go to. Even mega dosages don't seem to cut it. Some of it is getting through,,, but I don't know it's practical, or cost-effective.

I'm not sure if I would have tolerated methyl B12 ..in the early days. Maybe I would have done. You have mentioned that Dr Amy Yasko choses the type of B12 based on genetics.

She has also said that she initially favours hydroxy for the adults on her program.. because they have higher body burdens of toxins, and a lower threshold for detox .. and hydroxy is gentler in this regard.

As you have mentioned the capability to detox is kind of the rate limiting factor. Methyl is going to mobilise a lot of nasty stuff into the bloodstream.

In one videotaped presentation I saw, Dr Dietrich Klinghardt recommended that people kind of tough it out.. by throwing every chelator they could at it.

To be clear, he was talking about the KPU protocol, which can also cause massive detox, & his advice was to keep going when things got rough but have masses of chelators on hand to mop it up. DMSA, DMPS, Microsilica yatta yatta.

Well those are my thoughts for just now. A bit disjointed but I got distracted!

Thanks for this thread.. I had seen that study.. but I don't thnk I had connected the dots like Will had.

ETA: I think I have KPU....that could explain why I had a hard time making headway because as you explained I was lacking cofactors..specifically B6 & Zinc.

 

[Vegas]

She has also said that she initially favours hydroxy for the adults on her program.. because they have higher body burdens of toxins, and a lower threshold for detox .. and hydroxy is gentler in this regard.

That was precisely my experience. Start with hydroxy progress to methyl. As I previously stated, methyl is more "spiky;" its effects are more marked. Isn't it still likely most will benefit from hydroxo because even if there are conversion limitations secondary to glutathione scarcity, a conversion to methyl would continue to take place, albeit at a slower rate?

I have "graduated" to approx 3 x's week dosage of 1mg methyl + 800 mg metafolin. Interestingly, I don't have a symptomatic response to either anymore. This seemed to change pretty quickly after 10 or so rounds of chelation, although I have to mention it (the lessening of any "detox" symptoms) had progressively diminished beginning six months or so after starting the protocol. I think I still have a "need" for these nutrients; however, if what recently happened to me is any indication. I stopped taking methylfolate for about 10 days and I had a recurrence of angular chelitis. I seem to recall that Freddd hypothesized that methylfolate deficiencies produce this symptom.

 

[aquariusgirl]

Vegas
so you take 3mg of methyl a week? I am surprised you even notice a benefit from such a low dose..esp. since you need methyl b12 for teh cells to utilize all that metafolin.

Thanks for posting.

 

[drex13]

I have a difficult time tolerating both the Jarrow mb12 and Solgar Metafolin, because of what I have decided, are the sweeteners used. The #1 ingredient in the Solgar Metafolin is mannitol and xylitol in the Mb12. These cause severe digestive upset for me ( to put it politely). I have known for years that I can't eat sorbitol, and I also suspect I now have a fructose malabsorption problem. The hydroxy b-12 drops are sweetened with trehalose, which doesn't cause a problem for me, so I'm thinking of sticking with the drops for the hydroxy and methylfolate and also buying some mb12 drops as well. So far, anytime I have tried to add any AD b12, I get a rapid increase in anxiety that lasts for 1 1/2 to 2 days. I've actually made good progress using hydroxy b-12 drops and methylfolate drops : noticeable increases in stamina, no muscle fatigue, and no brain fog, but seem to have stalled. However, I have no co factors in place, and am in the process of adding those individually, instead of using the Yasko Multi. (I don't know if folic acid is a problem for me, but thought I would just avoid it). The jury is still out on the folinic acid.

Rich - Are the dosages for Hydroxy b12, Methylfolate, and the Folinic Acid in the SMP starting dosages only or can / should they be increased if tolerable ? If so, how high would you recommend ?

Thanks, Drex

 

[DANEL]

I have a difficult time tolerating both the Jarrow mb12 and Solgar Metafolin, because of what I have decided, are the sweeteners used. The #1 ingredient in the Solgar Metafolin is mannitol and xylitol in the Mb12. These cause severe digestive upset for me ( to put it politely). I have known for years that I can't eat sorbitol, and I also suspect I now have a fructose malabsorption problem. The hydroxy b-12 drops are sweetened with trehalose, which doesn't cause a problem for me, so I'm thinking of sticking with the drops for the hydroxy and methylfolate and also buying some mb12 drops as well. So far, anytime I have tried to add any AD b12, I get a rapid increase in anxiety that lasts for 1 1/2 to 2 days. I've actually made good progress using hydroxy b-12 drops and methylfolate drops : noticeable increases in stamina, no muscle fatigue, and no brain fog, but seem to have stalled. However, I have no co factors in place, and am in the process of adding those individually, instead of using the Yasko Multi. (I don't know if folic acid is a problem for me, but thought I would just avoid it). The jury is still out on the folinic acid.

Rich - Are the dosages for Hydroxy b12, Methylfolate, and the Folinic Acid in the SMP starting dosages only or can / should they be increased if tolerable ? If so, how high would you recommend ?

Thanks, Drex

i thought i was crazy with the sugar, dyes, filler thing b/c of the small amts in the small pills,
so good to know about the hydro drops, even with the trehalose and even the mb12 drops. have you done the mb12 drops at all yet?

so you are not on solgar metafolin (folate), and you are on the methylfolate drops but is this folinic acid? are they not the same. did you get the drops from holistic health?
what doses are you on?
how long have you been on the protocol?

i am confused by the methylfolate and folinic acid?

i am going to go back to the suggestions of the protocol and the website for holistic and try to figure out

thanks for your help
denise