Glutathione in the brain in CFS/ME

Discussion in 'Fibromyalgia Main Forum' started by richvank, Nov 20, 2009.

  1. richvank

    richvank New Member

    Hi, all.

    Professor Basant Puri et al. recently published a paper in which they reported measurement of glutathione in the cerebral cortex of the brain of patients with chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) using proton magnetic resonance spectroscopy.

    The abstract of this paper is below. Their conclusion was that they found no evidence that cerebral glutathione levels are decreased in CFS/ME relative to normal levels.

    I was surprised at this result, given the large number of test results I have reviewed showing glutathione depletion in blood plasma, as well as some results from red blood cells or whole blood. There are also publications reporting glutathione depletion in CFS/ME patients, some of which were cited by these authors.

    On further examination of this paper, I noted that the mean values of the parameter used to quantify glutathione in this study were 2.703 for 13 CFS/ME patients, and 5.191 for 13 matched normal controls. (There are no units on this derived parameter.) However, the standard deviations of the data for these two mean values were 2.311 and 8.984, respectively. Because of these large standard deviations, the difference between the mean values was not statistically significant (p=0.361).

    I wrote to Professor Puri and asked if it would be fair to say that their study lacked sufficient statistical power to determine whether glutathione is indeed depleted in the brains of CFS/ME patients. He responded that this potential explanation of their results “does make sense.” He agreed that a further, larger study with greater statistical power is needed to examine this question, and expressed his willingness to carry out such a study if funding could be found.
    I very much appreciated his response.

    In view of this, I think it is correct to say that this study does not provide any evidence contradicting the hypothesis that glutathione is depleted in CFS/ME patients, and in particular in the brains of these patients. I’m hopeful that a larger study can be done in the future to test this hypothesis.

    Rich Van Konynenburg

    Prostaglandins Leukot Essent Fatty Acids. 2009 Nov 9

    An in vivo proton neurospectroscopy study of cerebral oxidative stress in myalgic encephalomyelitis (chronic fatigue syndrome).

    Puri BK, Agour M, Gunatilake KD, Fernando KA, Gurusinghe AI, Treasaden IH.

    MRI Unit, Imaging Sciences Department, MRC Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Hospital, Du Cane Road, London W12 0HS, England, UK.

    A particularly important family of antioxidant defence enzymes in the body are the glutathione peroxidases, which remove H(2)O(2) by coupling its reduction to H(2)O with oxidation of reduced glutathione (GSH) to oxidised glutathione (GSSG). There are suggestions that GSH in the peripheral blood may be reduced in myalgic encephalomyelitis, which is a highly disabling neurological disease of unknown aetiology. Since many of the symptoms relate to cerebral functioning, it would seem probable that peripheral blood GSH findings would be reflected in lower cerebral GSH levels. The aim of this study was to carry out the first direct assessment of cerebral GSH levels in myalgic encephalomyelitis; the hypothesis being tested was that cerebral GSH levels would be reduced in myalgic encephalomyelitis. Cerebral proton neurospectroscopy was carried out at a magnetic field strength of 3T in 26 subjects; spectra were obtained from 20x20x20mm(3) voxels using a point-resolved spectroscopy pulse sequence. The mean cerebral GSH level in the myalgic encephalomyelitis patients was 2.703 (SD 2.311) which did not differ significantly from that in age- and gender-matched normal controls who did not have any history of neurological or other major medical disorder (5.191, SD 8.984; NS). Therefore our study does not suggest that GSH is reduced in the brain in myalgic encephalomyelitis. At the present time, based on the results of this study, there is no evidence to support the suggestion that, by taking glutathione supplements, an improvement in the brain-related symptomatology of myalgic encephalomyelitis may occur.

    PMID: 19906518 [PubMed - as supplied by publisher]
  2. mbofov

    mbofov Active Member

    Thanks for posting this, but I'm having trouble understanding. Could you translate this paragraph into layperson's language:

    "On further examination of this paper, I noted that the mean values of the parameter used to quantify glutathione in this study were 2.703 for 13 CFS/ME patients, and 5.191 for 13 matched normal controls. (There are no units on this derived parameter.) However, the standard deviations of the data for these two mean values were 2.311 and 8.984, respectively. Because of these large standard deviations, the difference between the mean values was not statistically significant (p=0.361)."

    I don't know what it means that the mean values for CFS/ME patients was 2.703 and 5.191 for normal controls. The two numbers are so different - could you explain what they mean? And also the bit about standard deviaions being 2.311 and 8.984 ...

    Thank you!


  3. mbofov

    mbofov Active Member

    What I find puzzling is this, and this is what I need explained: What does the number 2.703 for CFS/ME patients mean? Why is it so much less than 5.191 for non-CFS persons? Why do CFS patients have a different parameters? I thought the study concluded there was no difference between the 2 groups.

    I understand the two groups are too small to be statistically significant, but I don't understand what those 2 numbers above mean.

    I hope this explains my question more clearly -

    Thanks -

  4. richvank

    richvank New Member

    Hi, Mary.

    I happen to believe that the average glutathione level in the brain in CFS really is lower than in the normal population, and that this difference really is meaningful. However, from a statistical point of view, because there is so much scatter in the data, and the two data sets overlap so much, it's not possible to be sure that this difference in mean values is not simply due to the randomness of the data.

    Whenever you measure something in a group of people, there will be a range of values, partly because everyone will have a somewhat different value, and partly because the measurement technique will have some uncertainty associated with it. For example, suppose you measured people's heights, and suppose you wanted to compare the average heights of men to those of women. If you just measured a few people, some taller, some shorter, you could take an average for the men and an average for the women, but there would be some scatter in each data set, which is quantified by the standard deviation. It's possible that if you measured only a few people, you could have had some very tall women and some very short men in your groups, and then you might even have come out with the average height of your women being higher than the average height of your men. But you would have big standard deviations in your data sets, so you couldn't say for sure that your groups really represented the larger populations of men and women. But if you measured 1,000 each of men and women, and you selected them at random, you would end up with the average for the men being higher than the average for the women, and your standard deviations would be small enough that you could say with more certainty that the average man is taller than the average woman.

    I hope this explains the situation.

  5. mbofov

    mbofov Active Member

    I don't think I'm making myself clear. What does the number 2.703 mean? Is it a measure of the average amount of glutathione in CFS patients? If not, what does it measure? I have the same qusetion for 5.191 - is it a measure of the amount of glutathione in "normal" controls? If not, what is it a measure of?

    Maybe I would need a course in statistics to make sense of this?

    I probably would need a course in statistics to undertand the second part of my question, which is, waht the heck is a standard deviation?

  6. richvank

    richvank New Member

    Hi, Mary.

    O.K., I think I'm finally getting what your asking! :)-)

    Ideally, we would like to know the concentration of glutathione in the brain in units such as millimoles per liter of volume. Something like that is possible if you are doing chemical analysis of brains from autopsies, and in fact, I understand that some of that has been done in autism, but is not yet published. However, when you are dealing with living people, cutting chunks out of their brains is usually frowned upon. :)-) So more indirect methods of study have to be used. This study used magnetic resonance spectroscopy, which is somewhat similar to running an MRI scan of the brain, but is more sophisticated, in that it can look for certain chemical species, in this case glutathione.

    However, it is not easy to calibrate an MRS measurement on an absolute basis, so what is often done is to look at ratios of the various peaks that are observed, and determine relative values instead of absolute values. By doing this, you can end up with a value of a parameter that is proportional to the concentration of the species, but is not actually equal to the concentration. By using the same approach on CFS patients and normal controls, you can obtain a value for this parameter for both, and then take the ratio. The parameter itself does not have an absolute meaning in terms of concentration, but the ratio of the parameter value between the CFS patients and the normal controls should represent the ratio between their concentrations. If the ratio came out to be 1.0, that would mean that they had the same concentration of glutathione. As you know, the ratio was quite different from 1.0, suggesting lower glutathione concentration in the CFS patients. This may very well be meaningful, but because the scatter in the data from one person to another was so great, this difference could also be due to the randomness of the data.

    I might add that not many details are given in the paper I cited about the calculations that were done to arrive at the parameter values that were given, so it's difficult to identify the uncertainties involved in their calculational method.

    The standard deviation is a measure of how widely the set of data varies from its mean value.
    It is determined by first calculating the mean (simple average of the numbers), and then calculating the difference between each number and the mean value. These differences are then squared (i.e., multiplied by themselves) and then the squares are added together.
    The sum of the squares is then divided by one less than the number of data points, and then the square root is taken of this quotient. This is the standard deviation of the set of data points.

    This rather complicated procedure produces a measure of how widely scattered the data points are. You can have the same mean value for a set of data that is tightly grouped, or one that is widely scattered, so long as they average to the same value. But the reliability of the mean value for comparison to another one depends on how scattered the data points were. That's what the standard deviation tells you.

    Using the means and the standard deviations of two data sets, one can go on to do more statistical calculations that will eventually give what is called the p-value for the comparison of the two. The p-value is a measure of how likely it is that the two sets of data are really different from each other. The lower the p-value, the more likely they are to actually be different, and not just appearing different because of the randomness of the data. Usually a p-value below 0.05 is interpreted as showing statistical significance, but some people use stricter standards than that. In the study I cited, the p value was 0.361, so it was concluded that no significant difference was found.

    By using more people, the standard deviations associated with the mean values will decrease, because the calculation involves dividing by the the total number of data points minus one, and this quantity will rise more rapidly than the sum of the squares of the deviations from the mean.

    I hope this helps.

  7. mbofov

    mbofov Active Member

    I appreciate your very thorough response.

    I just looked at the verbiage in the original study which says:

    The mean cerebral GSH level in the myalgic encephalomyelitis patients was 2.703 (SD 2.311) which did not differ significantly from that in age- and gender-matched normal controls who did not have any history of neurological or other major medical disorder (5.191, SD 8.984; NS).

    So the study did show lower GSH in CFS/ME patients, although concluded that it was not significant. I wonder what they would consider significant? (your don't need to answer me again) And I understand the group was too small.

    I don't think I'll ever take statistics! :)

  8. Ales

    Ales New Member

    do you happen to have the actual data? Thank you

  9. richvank

    richvank New Member

    Hi, Ales.

    No, I don't. I have a copy of the full paper, but they didn't give the data. It's a pretty brief paper, a little over two pages, and the journal in which it was published is not one that specializes in MRS or glutathione.

  10. munch1958

    munch1958 Member

    Hi Rich:

    Have you applied the GD-MCB model to MCS patients? I'm interested in your findings. MCS seems to be another one of those alphabet soup illnesses like CFS, FM, GWI, PTSD, LD or chronic Lyme Borreliosis, that is supposed to be "All in Our Heads." But I'd sure like to know how every single one of us managed to make up the same exact symptoms.

    Here are a few protocols by Dr Grace Ziem:

    I've been nebulizing GSH for over 3 years now and have NO more symptoms of MCS. I've never tried the GSH drops in my nose for "sudden exposures" because of the light and temp instability of GSH. I just don't see how a spray bottle of saline and GSH could retain it's potency if carried around in my purse.

    Low adrenal function is part of the MCS problem. But I do believe that GSH was the missing part or the reason why I could never quite get rid of my MCS no matter how much my Lyme, CFS, FM symptoms improved.

    I'm lactose intolerant and allergic to Goat's milk so I have never tried undenatured whey. I've also got a severe allergy to sulfites, sarcoidosis and an asthma diagnoses. I've also had GSH IV treatments at one of the FFC clinics and found them to be very helpful. But I did have to load up on anti-histimines before infusions because of my sulfite problem. Getting GSH IVs at home can be a bit of a problem.

    I've had NO problems with inhaling the GSH but did start out with a low dose and titrated upwards. I was never quite positive that any supplements that I tried to boost GSH ever really did what they were supposed to because of my leaky gut issues.

    Some of us have to take very large quantities of things just to see an improvement. I fall into the more than average category so supplementation gets to be very costly. Just how much turmeric is enough anyway?

    In Suzanne Somer's last two books, "Breakthrough" and "Knockout" ,she reported on some nano technology glutathione patches from a company called Life Wave. I was considering buying them but 30 patches cost $79.90. Have you heard of these? Tried them? Any thoughts?

    Because of cost issues, we all need to pick and choose what supplements we can buy. Then also figure out what our insurance companies will cover. For a $25 copay, my insurance does cover twelve 3 ml vials of compounded 200 mg/ml GSH which is a month supply. It does need to be shipped in a cooler overnight from the pharmacy to me. My insurance also covers 30 ml of methylcobalamin for IM injection at home too.

    I wish your GD-MCB protocol would address how some items can be purchased via a doctor's prescription for items which may or may not be covered by insurance. Most holistic doctors are willing to work with patients as long as something does no harm. It would be helpful to know if Intrinsic factor is included in methylcobalamin for instance.

    Do you have any data on the return of HPA function in people who have tried your GD-MCB protocol. At some point, is the hypothalamus just too damaged to turn back on?

  11. lukro

    lukro Member

    In March you said that you no longer had mcs due to nebulizing glutathione. Now, several months later, do you still think that nebulizing the glut cured your mcs?

    This is really important information and I wish you would share what cured your mcs. Thanks.
  12. richvank

    richvank New Member

    Hi, munch.

    I'm happy to hear that the glutathione has stopped your MCS symptoms. I did put together a hypothesis that may fit some cases of MCS, and perhaps yours is one of them. It is pasted below. I need to update it with some new information, but I think it is still essentially correct.
    It fits cases of MCS that came on as part of CFS. It may not fit some other cases, such as the ones that started with a large acute exposure to some chemical. In those cases, I suspect that there has been damage to the barrier in the olfactory epithelium, that can't be repaired (except possibly by stem cell therapy). I think there are other cases that result from inherited polymorphisms in the detox enzymes. It may be that in some cases the chemicals enter the brain from the blood, by crossing the blood-brain barrier, rather than coming in through the olfactory epithelium in the nasal cavity. I think it's a heterogeneous disorder, so that one model probably doesn't fit all the cases.

    I'll try to address the other questions you raised later.


    February 15, 2008

    Hypothesis for the Pathogenesis of Multiple Chemical Sensitivity

    Rich Van Konynenburg, Ph.D.

    The olfactory epithelium in the “ceiling” of the nasal cavities is a small patch of tissue that is made up of two major types of cells, the olfactory neurons and the supporting or sustentacular cells, which are interspersed with the neurons. The olfactory neurons have axons that are connected into the olfactory nerve, which is the shortest pathway to the brain from the outside.

    The olfactory neurons have cilia on their surfaces facing the nasal cavities, and these incorporate receptors that are specific for the various substances that can be smelled.

    In addition to the olfactory neurons and the supporting cells, the olfactory epithelium also contains cells that secrete mucus, which covers the cilia.

    The supporting cells contain cytochrome P450 enzymes at concentrations higher than are found in the liver. It seems clear that their role is to break down (perform Phase I detoxication on) substances that are inhaled.

    In normal operation, inhaled substances are dissolved in the mucus, and those for which there are sensitive neurons elicit nerve impulses that travel to the brain and give the sensation of particular smells. The supporting cells apparently then break down and dispose of these substances, maintaining the sensitivity of the olfactory neurons to substances that are newly arrived, and protecting them from the entry of toxic substances.

    It is known that glutathione plays a major role in detoxication. Not only does it conjugate certain toxic substances in one of the major Phase II pathways, but it also serves to quench hydrogen peroxide that results from the action of superoxide dismutase on the superoxide free radicals that are generated by the action of the cytochrome P450 enzymes in Phase I detoxication.

    If the supporting cells are deficient in the reduced form of glutathione, I suggest that two adverse effects would occur. First, a state of oxidative stress would arise, because of the buildup of reactive oxygen species. These species would be likely to damage the membranes of both the supporting cells and the olfactory neurons. Second, toxic substances would not be broken down, but instead, in the presence of the damaged cell membranes, would likely enter the neurons, and from there would have a short path to travel into the brain, thus producing symptoms such as headache.

    It would seem that this model for multiple chemical sensitivity would explain several of the observed features of this disorder. One is that people with MCS are sensitive to a range of volatile substances. Another is that symptoms appear very rapidly upon exposure to inhalation of these substances. Another is that temporary relief can often be obtained by using a glutathione nasal spray.

    If this model is correct, then it would seem that a cure for MCS might be brought about by restoring the levels of reduced glutathione in the sustentacular cells on a longer term basis. In autism and in chronic fatigue syndrome, it appears that glutathione levels are held down by a vicious circle mechanism that involves a partial block in the methylation cycle, at methionine synthase. Since MCS often occurs together with chronic fatigue syndrome, perhaps this same biochemical mechanism is responsible for MCS. If this is true, the same treatments that are being found helpful in autism and CFS may be helpful for MCS.

  13. lukro

    lukro Member

    Then if this is correct, nebulizing glutathione should rid one of mcs? Am I reading this correctly? Although, you did say that it would not work for everyone. But should work for those w/ cfs?

    Thanks, again. I need to share this info.
  14. richvank

    richvank New Member

    Hi, lukro.

    Yes, it does seem to work for at least some of the cases. Even a simple nasal spray can work, because it just has to get to the nasal cavity to correct the problem I described, at least temporarily. As you can see from munch's post above, it has worked in her case.

    This treatment approach was pioneered by Dr. Grace Ziem, M.D., with the help of Jim Seymour, a pharmacist at Key Pharmacy in Kent, Washington. I first heard about it from a talk Jim gave a few years ago at an MCS workshop held in a hotel near the San Francisco airport. The Key Pharmacy supplies a glutathione nasal spray. They also supply a glutathione solution for nebulizing.

    In my view, these approaches are helpful, but temporary in nature. To correct the glutathione depletion on a more permanent basis, I recommend testing to see if there is a partial methyation cycle block, and if so, to treat this block using the Simplified Treatment Approach. When the block is lifted, glutathione will come up by itself on a more permanent basis.

    Best regards,

  15. Ales

    Ales New Member

    it is strange that the journal that published it didn't make the authors to reveal the actual data. As I understand it there should be 2x13 values and with it one can make serious statistical analysis. It seems to me that to just state the two mean values and the two standard deviations in this kind of research is quite nonstandard.
  16. richvank

    richvank New Member

    Hi, all.

    I just received an email from Dr. Dikoma Shungu of Cornell, Columbia and Mt. Sinai medical schools. He does research involving magnetic resonance spectroscopy and is currently funded by the CFIDS Assn. to work on ME/CFS. He wrote that he agrees with me about the Puri et al. study, and that he is currently evaluating the levels of glutathione in the brains of PWCs himself by a more precise method. He is not ready to comment publicly on his results yet, but he wrote, "it is safe to conclude that to date, there is no valid study that has rejected your hypothesis about decreased glutathione in ME/CFS." Needless to say, I was very happy to receive his comments.

    Best regards,

  17. lukro

    lukro Member

    and it did not work for me. I still nebulize glutathione and do feel as though I'm better (cfs/fibro,mcs); however, perhaps I have resigned myself to my ailments and have simply "normalized" my symptoms. Who knows....

    I need to try the methylation cycle block protocol but am not sure when that would be--I'm trying to remain gainfully employed, which does not allow for much experimentation.

    Thank you for your answer.
  18. richvank

    richvank New Member

    Hi, ales.

    I agree with you. Perhaps if the study was reporting that the glutathione level came out significantly different in the brains of ME/CFS patients from the level in normal controls, the paper would have gone into more detail. Reporting negative results is not a very dramatic thing to do, and perhaps the editor of the journal didn't want to devote too much space to it. There are no page charges to the authors for this journal, so the editor probably limits the length of the papers. Some journals charge hefty fees per page, so that if the author has the funding, he or she can write a longer paper.

    I look forward to seeing what Dr. Shungu will report from the study he is doing. He specializes in the technique called "spectral editing" that is needed to detect signals from substances such as glutathione that are very weak and are buried by the main peaks that appear in MRS spectra from the brain. I think we will see a much more credible report from his work.

    I sent him copies of my papers last May, and he is aware of the significance of glutathione levels in the brain. He has already reported observing elevated lactate there, and that suggests that the mitochondria are dysfunctional. He has reported that he believes that is due to oxidative stress, with which I agree, and if that is indeed true, I think it is almost a forgone conclusion that glutathione is depleted, because glutathione is at the basis of the antioxidant enzyme system. If there is oxidative stress, it is going to show up as a deficit in the antioxidant system.

    So I am really looking forward to his results. He has quite a bit of credibility in the CFS community, so if he comes up with low glutathione, perhaps the folks in the leadership of the IACFS/ME as well as other researchers and clinicians associated with ME/CFS will start to take an interest in glutathione. I have been trying to get their attention to this since the 2005 conference, with little response. No doubt XMRV will be in the limelight now, but I think we will be able to connect XMRV to glutathione depletion. So hopefully these various pieces will come together.

    Thanks for your interest.