Unusual mitochondrial result.

justy

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Hi, i dont know if this is the right place to post this but i was wondering if anyone has any dea what this actually means or what could be causing it. I had my results back from the mitochondrial profile funtion testmany months ago, but am sill struggling to understand it. I think i have got my head round most of it but have 1 unusual result;
Krebs citric acid cycle:
"This is going borderline slow at 62.5% (normal range >60%). In order to assess the efficiency with which ADP is converted to ATP, an inhibitor is added and then removed to see how quickly ATP is reformed. Having added the inhibitor one expects levels of ATP and magnesium to drop below 0.3 if this does not happen this suggests there is blocking of the active sites. The acceptable percentage is up to 14% and Justines result is 37% (up to 14%). This suggests that there is significant blockage of the active sites (i.e. complexes I,II,III,IV and V on inner mitochondrial membranes). The likeliest reason for this is toxic stress and we could explore further by doing microrespirometry studies which look at oxidative phosphorylation in more detail.

"Justine has very unusual results with respect to ADP to ATP conversion efficiency. Although her conversion efficiency is just about normal at 62.5%, she has 37% blocking of active sites this is unusual normally one would expect such blocking of active sites to reduce ADP to ATP efficiency way below the normal figure of 60%. In this case Justine seems to be able to tolerate quite marked blocking of her active sites but still produce a reasonably normal result." Dr. Myhill.

any insights into this would be very greatly appreciated.
 

richvank

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Hi, Justy.

I think that the best way to shed light on this would be to run a Genova Diagnostics Metabolic Analysis Profile. This is a urine organic acids test, and I understand that this is available in the UK. This panel includes the Krebs cycle metabolites as well as pyruvate and lactate, fatty acids metabolites, hydroxymethylglutarate, and pyroglutamate. These will give information on what is going on inside the mitochondria.

In addition, I don't know if you ran other tests at AcumenLab, but Dr. McLaren Howard also offers tests that can identify the particular toxins that are present and could be blocking enzymes and complexes in the mitochondria. The translocator protein test is one of them.

For what it's worth, in my hypothesis (more information available at www.cfsresearch.org) the root causes of the mito dysfunction in ME/CFS are the glutathione depletion, coupled with the partial methylation cycle block. These are what allow the toxins and the oxidative stress to build up, and what deplete the production of L-carnitine, coenzyme Q10, phosphatidylcholine and creatine, all of which need methylation for their synthesis, and all of which are needed by the mitochondria.

If you should want to explore this, the methylation pathways panel is the best panel for this purpose, and the faster service is offered by the Health Diagnostics and Research Institute in New Jersey, USA. Here is the contact information. The panel costs $300 US.

Methylation Pathways Panel

This panel will indicate whether a person has a partial methylation cycle block and/or glutathione depletion. I recommend that this panel be run before deciding whether to consider treatment for lifting the methylation cycle block. I am not associated with the lab that offers this panel.

The panel requires an order from a physician or a chiropractor. The best way to order the panel is by fax, on a clinician’s letterhead.


Available from:

Health Diagnostics and Research Institute
540 Bordentown Avenue, Suite 4930
South Amboy, NJ 08879
USA
Phone: (732) 721-1234
Fax: (732) 525-3288

Lab Director: Elizabeth Valentine, M.D.

Dr. Tapan Audhya, Ph.D., is willing to help clinicians with interpretation of the panel by phone, or you can use the following interpretive comments:


Interpretation of the Health Diagnostics and Research Institute
Methylation Pathways Panel

by
Rich Van Konynenburg, Ph.D.


Several people have asked for help in interpreting the results of
their Health Diagnostics and Research Institute methylation pathway panels. Here are my suggestions for doing so. They are based on my study of the
biochemistry involved, on my own experience with interpreting more
than 120 of these panel results to date, and on discussion of some of
the issues with Tapan Audhya, Ph.D., at the Health Diagnostics and Research Institute.

The panel consists of measurement of two forms of glutathione
(reduced and oxidized), adenosine, S-adenosylmethionine (SAM) , S-
adenosylhomocysteine (SAH), and seven folic acid derivatives or
vitamers.

According to Dr. Audhya, the reference ranges for each of these
metabolites was derived from measurements on at least 120 healthy
male and female volunteer medical students from ages 20 to 40, non-
smoking, and with no known chronic diseases. The reference ranges
extend to plus and minus two standard deviations from the mean of
these measurements.

Glutathione: This is a measurement of the concentration of the
reduced (active) form of glutathione (abbreviated GSH) in the blood
plasma. From what I've seen, most people with chronic fatigue
syndrome (PWCs) have values below the reference range. This means
that they are suffering from glutathione depletion. As they undergo
the simplified treatment approach to lift the methylation cycle
block, this value usually rises into the normal range over a period
of months. I believe that this is very important, because if
glutathione is low, vitamin B12 is likely unprotected and reacts with toxins
that build up in the absence of sufficient glutathione to take them
out. Vitamin B12 is thus “hijacked,” and not enough of it is able to
convert to methylcobalamin, which is what the methylation cycle needs
in order to function normally. Also, many of the abnormalities and
symptoms in CFS can be traced to glutathione depletion.

Glutathione (oxidized): This is a measurement of the concentration
of the oxidized form of glutathione (abbreviated GSSG) in the blood
plasma. In many (but not all) PWCs, it is elevated above the normal
range, and this represents oxidative stress.

Adenosine: This is a measure of the concentration of adenosine in the
blood plasma. Adenosine is a product of the reaction that converts
SAH to homocysteine. In some PWCs it is high, in some it is low, and
in some it is in the reference range. I don't yet understand what
controls the adenosine level, and I suspect there is more than one
factor involved. In most PWCs who started with abnormal values, the
adenosine level appears to be moving into the reference range with
methylation cycle treatment, but more data are needed.

S-adenosymethionine (RBC) (SAM): This is a measure of the
concentration of SAM in the red blood cells. Most PWCs have values
below the reference range, and treatment raises the value. S-
adenosylmethionine is the main supplier of methyl groups in the body,
and many biochemical reactions depend on it for their methyl
groups. A low value for SAM represents low methylation capacity, and
in CFS, it appears to result from a partial block at the enzyme methionine
synthase. Many of the abnormalities in CFS can be tied to lack of
sufficient methyation capacity.

S-adenosylhomocysteine (RBC) (SAH): This is a measure of the
concentration of SAH in the red blood cells. In CFS, its value
ranges from below the reference range, to within the reference range,
to above the reference range. Values appear to be converging toward
the reference range with treatment. SAH is the product of reactions
in which SAM donates methyl groups to other molecules.

Sum of SAM and SAH: When the sum of SAM and SAH is below 268
micromoles per deciliter, it appears to suggest the presence of
upregulating polymorphisms in the cystathione beta synthase (CBS)
enzyme, though this may not be true in every case.

Ratio of SAM to SAH: A ratio less than about 4.5 also represents low
methylation capacity. Both the concentration of SAM and the ratio of
concentrations of SAM to SAH are important in determining the
methylation capacity.

5-CH3-THF: This is a measure of the concentration of 5-methyl
tetrahydrofolate in the blood plasma. It is normally the most
abundant form of folate in the blood plasma. It is the form that
serves as a reactant for the enzyme methionine synthase, and is thus
the most important form for the methylation cycle. Many PWCs have a
low value, consistent with a partial block in the methylation cycle.
The simplified treatment approach includes FolaPro, which is
commercially produced 5-CH3-THF, so that when this treatment is used,
this value rises in nearly every PWC. If the concentration of 5-CH3-
THF is within the reference range, but either SAM or the ratio of SAM
to SAH is below the reference values, it suggests that there is a
partial methylation cycle block and that it is caused by
unavailability of sufficient bioactive B12, rather than
unavailability of sufficient folate. I have seen this frequently,
and I think it demonstrates that the “hijacking” of B12 is the root
cause of most cases of partial methylation cycle block. Usually
glutathione is low in these cases, which is consistent with lack of
protection for B12, as well as with toxin buildup.

10-Formyl-THF: This is a measure of the concentration of 10-formyl
tetrahydrofolate in the blood plasma. It is usually on the low side in PWCs.
This form of folate is involved in reactions to form purines, which
form part of RNA and DNA as well as ATP.

5-Formyl-THF: This is a measure of the concentration of 5-formyl
tetrahydrofolate (also called folinic acid) in the blood plasma.
Most but not all PWCs have a value on the low side. This form is not used
directly as a substrate in one-carbon transfer reactions, but it can
be converted into other forms of folate. It is one of the
supplements in the simplified treatment approach, which helps to
build up various other forms of folate.

THF: This is a measure of the concentration of tetrahydrofolate in
the blood plasma. In PWCs it is lower than the mean normal value of 3.7
nanomoles per liter in most but not all PWCs. This is the
fundamental chemically reduced form of folate from which several
other reduced folate forms are made. The supplement folic acid is
converted into THF by two sequential reactions catalyzed by
dihydrofolate reductase (DHFR). THF is also a product of the
reaction of the methionine synthase enzyme, and it is a reactant in
the reaction that converts formiminoglutamate (figlu) into
glutamate. If figlu is high in the Genova Diagnostics Metabolic
Analysis Profile, it indicates that THF is low.

Folic acid: This is a measure of the concentration of folic acid in
the blood plasma. Low values suggest folic acid deficiency in the
current diet. High values are sometimes associated with inability to
convert folic acid into other forms of folate, such as because of
polymorphisms in the DHFR enzyme. They may also be due to high
supplementation of folic acid.

Folinic acid (WB): This is a measure of the concentration of folinic
acid in the whole blood. See comments on 5-formyl-THF above. It
usually tracks with the plasma 5-formyl-THF concentration.

Folic acid (RBC): This is a measure of the concentration of folic
acid in the red blood cells. The red blood cells import folic acid
when they are initially being formed, but during most of their
approximately four-month life, they do not normally import, export, or use
it. They simply serve as reservoirs for it, giving it up when they
are broken down. Many PWCs have low values. This can be
caused by a low folic acid status in the diet over the previous few
months, since the population of RBCs at any time has ages ranging
from zero to about four months. However, in CFS it can also be
caused by damage to the cell membranes, which allows folic acid to
leak out of the cells. Dr. Audhya reports that treatment with omega-
3 fatty acids can raise this value over time.




Best regards,

Rich
 

curry

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In addition, I don't know if you ran other tests at AcumenLab, but Dr. McLaren Howard also offers tests that can identify the particular toxins that are present and could be blocking enzymes and complexes in the mitochondria. The translocator protein test is one of them.
As far as I understand this test is part of the 'mitochondria function profile test' by Dr Myhill.
So justy may have already the results?


************************************************************************************

Rich, I am getting completely confused which is the best test for mitochondria function.

The following seems to be available:

1. The 'mitochondrial function profile', offered by a UK CFS doctor. The test includes the following:

- "ATP profiles" and has been developed by Dr John McLaren-Howard at Biolab in London. It measures the rate at which ATP is recycled in cells.
- The second part of the test measures the efficiency with which ATP is made from ADP.
- The third possibility is that the protein which transports ATP and ADP across mitochondrial membrane is impaired and this is also measured.
- To get the full picture of mitochondrial function the profile also measures levels of Co-enzyme Q10, SODase, NAD, L-carnitine and cell free DNA


2. Genova Diagnostics, Metabolic Analysis Profile
- This is a urine organic acids test. This panel includes the Krebs cycle metabolites as well as pyruvate and lactate, fatty acids metabolites, hydroxymethylglutarate, and pyroglutamate. These will give information on what is going on inside the mitochondria.


3. The methylation pathways panel
- This panel will indicate whether a person has a partial methylation cycle block and/or glutathione depletion. I recommend that this panel be run before deciding whether to consider treatment for lifting the methylation cycle block.


If I understand correctly, either test 1. or 2. should be done to test mitochondrial performance, and in addition, if necessary test 3. should be done?
Sorry, for all those questions. :ashamed:
 

richvank

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As far as I understand this test is part of the 'mitochondria function profile test' by Dr Myhill.
So justy may have already the results?


************************************************************************************

Rich, I am getting completely confused which is the best test for mitochondria function.

The following seems to be available:

1. The 'mitochondrial function profile', offered by a UK CFS doctor. The test includes the following:

- "ATP profiles" and has been developed by Dr John McLaren-Howard at Biolab in London. It measures the rate at which ATP is recycled in cells.
- The second part of the test measures the efficiency with which ATP is made from ADP.
- The third possibility is that the protein which transports ATP and ADP across mitochondrial membrane is impaired and this is also measured.
- To get the full picture of mitochondrial function the profile also measures levels of Co-enzyme Q10, SODase, NAD, L-carnitine and cell free DNA


2. Genova Diagnostics, Metabolic Analysis Profile
- This is a urine organic acids test. This panel includes the Krebs cycle metabolites as well as pyruvate and lactate, fatty acids metabolites, hydroxymethylglutarate, and pyroglutamate. These will give information on what is going on inside the mitochondria.


3. The methylation pathways panel
- This panel will indicate whether a person has a partial methylation cycle block and/or glutathione depletion. I recommend that this panel be run before deciding whether to consider treatment for lifting the methylation cycle block.


If I understand correctly, either test 1. or 2. should be done to test mitochondrial performance, and in addition, if necessary test 3. should be done?
Sorry, for all those questions. :ashamed:
Hi, Curry.

I think there are different schools of thought on this.

Dr. Myhill would opt for number 1, I think. These tests, when run on PWMEs/PCWs almost always come back showing that the mitochondria are dysfunctional because of oxidative stress and buildup of toxins. They usually also show deficiencies in some of the substances needed by the mitochondria. In response, Dr. Myhill prescribes her mitochondrial package, which is intended to support the mitochondria and add substances that are deficient. She may also recommend FIR sauna to help remove toxins. These measures usually help to some degree. Dr. Myhill also recommends methylation cycle treatment to some of her patients, I think.

I like to see results of the other two tests, because I think they address the more fundamental issues that are causing the mito dysfunction (as well as addressing a number of other aspects of the overall metabolism in the case of the MAP profile).

Namely, I believe that glutathione depletion and the partial methylation cycle block are the more fundamental issues. The methylation pathways panel gives direct measurements of reduced and oxidized plasma glutathione, red blood cell SAMe and SAH, and several of the folate forms. These will tell you directly if you have these more fundamental issues. They also will give you baseline data so that when you are doing treatment to lift the partial block, you can run the panel again to see how the treatment is going. I believe that if these more fundamental issues are addressed, the mitochondrial dysfunction will be corrected, because the oxidative stress will be removed and the detox system will begin taking out toxins. Methylation will be available to produce several of the substances needed by the mitochondria.

I have no objection to using Dr. Myhill's mito package, and in fact, I think it can help significantly. However, I don't think it will correct these more fundamental issues by itself.

Incidentally, I have corresponded with both Dr. Myhill and Dr. McLaren Howard repeatedly over the past few years, and I hold them both in high regard. They have done a great deal for the ME/CFS community, and particularly in emphasizing the importance of mito dysfunction in this disorder.

Best regards,

Rich
 

curry

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Thank you Rich.

May I ask if you have been in contact with:

- Dr med. Wilfried Bieger, Munich, Germany
- Dr sc. med. Bodo Kuklinski, Rostock, Germany

Bieger actually also has the opinion that glutathione depletion is one of the main factors responsible for low mitochondrial performace.

They're the most renowned capacities in orthomolecular medicine in Germany, and I think it would be interesting what their take on is about the methylation cycle block theory.
(Maybe they could also help with funding for studies on patients?)
 

Joopiter76

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I dont believe so, Ive been to Dr. Bieger with no helping results at all !!! So I injected GSH several months and check GSH and GSSG levels by laboratory. They were in the normal range under treatment. So as I was very very weak during this time GSH may be just an additional factor. One thing is, that I believe is an issue is that phosphorylation is impaired in CFS. In this case for example people who take enough vitamines still have a metabolic deficiency of these vitamines. The key factor may be, that B2, B1, B6 and so on have to become phosphorylated to become active, this phosphorylation is mainly done in the gut mucosa. This may explain why treating the gut helps so many people. A loss of phosphorylated vitamines leads to a genaral dysfunction of important metabolic processes and so on to a slowing down of them. I myself have high oxidated GSSG low GSH although I take 600mg of B2 a day !! So im I take FMN which is phosphorylated B2 together with Niacin the GSH level raises into the normal range and GSSG improves much. So the limiting factor of the mitochondria may be FAD generation. So it would be very interesting doing the organic urine profile because then you can see where the metabolic problem is located. Additionally you will see if you have deficiencies of other vitamines and dysbiosis as well as detoxification problems. I think these tests are very helpful if you understand the results.
 

justy

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Thank you Rich for your input. This is all the science stuff that makes my brain go wobbly! but i am slowly undersanding.
Dr Myhill suggested i take L Glutathione 250mgs a day due to this result;

Red cell glutathione peroxidase (GSH-PX) - 53U/gHb (67 – 90) – very poor result
Red cell glutathione (GSH) – 1.78mmol/l (1.7 – 2.6) – low normal result

to be honest i dont really understan what this test/result is all about, but i have read somewhere on this forum that there is no point in supplementing with Glutathione. Anymore thoughs on this very greatly appreciated.

Once again thanks to those who have helped so far with replies.
 

justy

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yes i have Selenium to take 500mcg a day recommended for 4 months then dropping to 200mcg maintenance dose - i havent started them yet as i am introducing supplements gradually and keeping a diary.
 

richvank

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yes i have Selenium to take 500mcg a day recommended for 4 months then dropping to 200mcg maintenance dose - i havent started them yet as i am introducing supplements gradually and keeping a diary.
Hi, justy.

I agree on taking the selenium to boost the activity of glutathione peroxidase.

With regard to suppementing glutathione, if it is taken orally, most of it is broken down in the gut, but it will supply amino acids, some of which will go to the liver and help it to make more glutathione.
Probably the best way to support glutathione directly is by use of the liposomal glutathione products. They are able to deliver glutathione directly to the cells of the body by passing through lipid membranes.

The methylation protocol will automatically build glutathione in most cases, without directly boosting it. You can see the evidence for this at www.cfsresearch.org by clicking on CFS/M.E. and then on my name. Read the last entry, on the Yasko Protocol conference talk that I gave recently. It has plotted data out to nine months of treatment.

Best regards,

Rich
 

richvank

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Thank you Rich.

May I ask if you have been in contact with:

- Dr med. Wilfried Bieger, Munich, Germany
- Dr sc. med. Bodo Kuklinski, Rostock, Germany

Bieger actually also has the opinion that glutathione depletion is one of the main factors responsible for low mitochondrial performace.

They're the most renowned capacities in orthomolecular medicine in Germany, and I think it would be interesting what their take on is about the methylation cycle block theory.
(Maybe they could also help with funding for studies on patients?)
Hi, curry.

I haven't been in contact with these doctors, but if you have email addresses for them, I would be happy to correspond with them.

I think there are 3 or 4 published papers that report that glutathione depletion causes mitochondrial dysfunction, and I'm glad that Dr. Bieger shares this view.

Best regards,

Rich
 

richvank

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I dont believe so, Ive been to Dr. Bieger with no helping results at all !!! So I injected GSH several months and check GSH and GSSG levels by laboratory. They were in the normal range under treatment. So as I was very very weak during this time GSH may be just an additional factor. One thing is, that I believe is an issue is that phosphorylation is impaired in CFS. In this case for example people who take enough vitamines still have a metabolic deficiency of these vitamines. The key factor may be, that B2, B1, B6 and so on have to become phosphorylated to become active, this phosphorylation is mainly done in the gut mucosa. This may explain why treating the gut helps so many people. A loss of phosphorylated vitamines leads to a genaral dysfunction of important metabolic processes and so on to a slowing down of them. I myself have high oxidated GSSG low GSH although I take 600mg of B2 a day !! So im I take FMN which is phosphorylated B2 together with Niacin the GSH level raises into the normal range and GSSG improves much. So the limiting factor of the mitochondria may be FAD generation. So it would be very interesting doing the organic urine profile because then you can see where the metabolic problem is located. Additionally you will see if you have deficiencies of other vitamines and dysbiosis as well as detoxification problems. I think these tests are very helpful if you understand the results.
Hi, Joopiter.

I very much appreciate reading your insights and experiences with this treatment. I had not considered the possible problems with phosphorylation of the vitamins. It's certainly true that the gut often has a lot of problems in people with M.E./CFS. I note that Dr. Yasko puts emphasis on treating the gut before boosting the methylation cycle, and I think it is important to do this in cases in which the gut is not functioning well.

I might note that FAD is available in Japan as an approved drug, called Flavitan. I think it is also made in one of the Scandanavian countries and in Russia. It is not available in the U.S., but I was able to get it for one person whose body was not able to make it from riboflavin (B2) by having her doctor send a request to the Tokyo Medical and Surgical Clinic, and having her pay by wire transfer to a bank in Japan. The Health Diagnostics and Research Institute in New Jersey, USA, offers lab tests for riboflavin, FMN and FAD, so it is possible with these tests to determine whether the phosphorylation of riboflavin is working well or not.

I am away from home for a few weeks and am not able to be on the computer as much for a while, so I appreciate others helping to answer questions that are addressed to me.

Best regards,

Rich
 

rlc

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hi for anyone interested in trying to get their mitochodrial working, i have read Dr mayhills book, but a bit of information that appears to be missing is that vitamin D is responsible for the absorbtion of calcium, and phosperous, lack of phospherous leads to lack of ATP therefor constant fatigue. i know she mentions taking vitmin d in her book, but if you are seriously deficent some people can need very large suplement which should be given by a doctor. Dont try large doses without knowing your deficiency status it can be dangerous and you need your calcium tested to make sure your not overdosing. vit D deficiency also causes muscle, joint and bone pain and muscle twitching (tetany) if your short on magnesium you wont be able to absorb vit d and it also causes musle twitching as well hope this helps useful vit d information here http://www.ncbi.nlm.nih.gov/pubmed/19835345 and here http://www.vitamindcouncil.org/ Please be carefull with vit d hope this helps best of luck
 

aquariusgirl

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I would like to point out that in some, or maybe all cases, oxidative stress seems to get worse with methylation supplementation before it gets better.

Rich has made the point that if people can support undenatured whey, (glutathione precursors) they may want to supplement this as they start to support their methylation cycle.
I think whey contains casein so it's not an option for many of us.
I guess that leaves the other forms of glutathione ..liposomal, nebulized etc.
But those of us who have CBS upregs, cannot tolerate a lot of sulphur amino acids either...so it's a bit of a juggling act.
Molybdenum can help. and stuff that binds ammonia.
 
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I agree with richs comment that Dr myhills protocol helps a little.
I agree that taking niacin is very important_at high doses I would argue and maybe that is the road to go down?
 

Joopiter76

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@AQ: search for riboflavine-5-phosphate which is FMN the phosphorylated form of B2 (I have it from Thorne). From FMN its only one step to FAD. I take niacinamid. the branch doesnt matte I think, but NADH is one active form of Niacin. But ad the active forms only if the first detox signs of the methalytion cycle suppot supps has gone, because the will invrease by supplementation of active forms in my experience.