Intracellular magnesium defiency and vitamin B6 (and/or B2) deficiency in ME/CFS

Hi, all.

It has been known for quite a few years that in ME/CFS, there is often a deficiency of intracellular magnesium in the tissue cells. The best test for this is the EXAtest offered by Intracellular Diagnostics, Inc., in Oregon. People try various treatments to raise the intracellular magnesium level, including Epsom salt baths, injections with magnesium sulfate, magnesium oil applied to the skin, and oral magnesium supplements, and it has seemed to be difficult to raise the intracellular magnesium level and keep it up.

Another essential nutrient that I often find to be low when I analyze urine organic acids tests for PWMEs is vitamin B6 (and sometimes also B2, which is necessary to convert B6 into its active form, P5P). I think that the reason B6 (and maybe B2) tend to be low in ME/CFS is that P5P is very important in amino acids metabolism, enabling conversion of one amino acid into another, and this is necessary for feeding some of them into the Krebs cycle to be burned for energy. PWMEs burn amino acids at a higher rate than normal, because glutathione depletion puts a partial block in the Krebs cycle early on, and that inhibits the utilization of carbs and fats as fuel, according to my hypothesis.

I'm writing this post to point out that I think there is a connection between these two deficiencies, such that a B6 (or B2) deficiency will cause lowered intracellular magnesium.

Here's some history behind this issue: When the late Dr. Bernie Rimland started what became the Autism Research Foundation back in the 60s, he asked parents of autistic children what treatments they had found helpful for their children. Several replied that vitamin B6 was helpful, and Bernie started recommending it. He then received a phone call from the controversial pioneering nutritionist Adelle Davis, who told him that it is important to take magnesium with B6, because they work together in the metabolism. He started suggesting that to the autism parents, and significantly better results were found for this combination than when either of them was taken alone.

Since I heard Bernie tell this story at a DAN! conference several years ago, I've wondered what this connection is in the biochemistry. I've never seen it spelled out anywhere. And since we have found that autism and ME/CFS share much of the same pathophysiology, I've wondered how this might apply to ME/CFS.

Well, yesterday I think I came up with the connection. I was working on a case in which the person has high P5P in the blood, but appears to be unable to remove the phosphate group so that the pyridoxal can enter the cells to be rephosphorylated and utilized as P5P in its normal roles in the biochemistry. His urine organic acids panel showed lots of evidence of deficiency of P5P activity. But it also showed evidence of severe magnesium deficiency, even though he was getting magnesium injections. It occurred to me that perhaps intracellular P5P is needed to transport magnesium into the cells.

The mechanisms involved in the transport of magnesium have not been completely figured out by researchers yet, but I located a study published in 1981 (abstract below) in which vitamin B6 supplementation was found to raise the intracellular magnesium levels in the red blood cells. Since additional magnesium was not given, this indicates that vitamin B6 is important in the transport of magnesium into cells. Since vitamin B2 is necessary to form P5P (the active form of B6) from vitamin B6, I think that a deficiency in B2 would also impact intracellular magnesium.

I'm posting this to let PWMEs know of the importance of this connection.

Best regards,

Rich


Ann Clin Lab Sci. 1981 Jul-Aug;11(4):333-6.

Effect of vitamin B-6 on plasma and red blood cell magnesium levels in premenopausal women.

Abraham GE, Schwartz UD, Lubran MM.

Abstract

The effect of 100 mg of vitamin B6 twice a day on plasma and red blood cell (RBC) magnesium was evaluated in nine premenopausal subjects during the period of one month. According to reported normal ranges for plasma and RBC magnesium (1.7 to 2.3 and 4.7 to 7.0, mg per dl, respectively), three subjects had low plasma magnesium, and all subjects had low RBC magnesium during the control period. Following vitamin B6 administration, the mean plasma and RBC magnesium levels were significantly elevated, with a doubling of RBC levels after four weeks of therapy. These results support the postulate that vitamin B6 plays a fundamental role in the active transport of minerals across cell membranes.

PMID: 7271227

Very interesting. In your case, the person already has high levels of P5P in the blood, so I assume giving more B6 won't work. Do you have any ideas to how he can get the P5P dephosphorylated and into the cells?

Thanks.

Hi, adreno.

I'm hoping that injecting nonphosphorylated B6 by IV or IM will work. Without the phosphate, it should be able to diffuse into the cells. Time will tell.

Best regards

Rich

Rich, can you help me connect the dots here?

I have many tests showing a need for B6. I have low intracellular magnesium per the Exatest test, in spite of lots of supplementation. I have low B2 (OAT) & lots of markers for phosphate are extremely high.
I also once ran a serum B6 test and the result was off the charts high.. maybe 10 or 20x the upper range of normal.

Thanks

Thanks Rich, this is interesting. Mg is very helpful for me. I suspect it works better when I sip the mg citrate in water with a meal. I often think about how we send massive doses of minerals into our guts that were never designed to process them that way. Mg Chloride spray on skin seems helpful also.

Hi, AQ.

I don't understand the part about lots of markers for phosphate being extremely high. Can you clarify this, or send me the test results? I think you have my email address, but it's richvank at aol dot com.

The serum B6 measurement is a measurement of P5P, because that's the dominant form of B6 in the serum. So it sounds as though your body is having difficulty taking off the phosphate from the PLP, just as in the case I mentioned, so that you aren't getting B6 into your cells, and hence, because it's needed to transport magnesium in as well, you have low intracellular magnesium.

Best regards,

Rich

This might tie in... here too. And if dog person is right & lack of B2 is interfering with iron metabolism .. then it would be a double whammy?
Lack of B6 & lack of iron?

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC137500/

Hi, AQ.

Thanks for this paper. This whole area is something I need to study--the heme metabolism and all the ramifications for ME/CFS, including the things Christine posted about, HPU, the porphyrins indicating heavy metal toxicity, porphyria, etc. I need to get my brain around all of this and understand how it all fits together.

And, oh, I just realized that you already sent me your GPL OAT results from May, 2011, and I see the high phosphoric acid. Is that what you meant? I'll look over these results.

Best regards,

Rich

Yes, it's high on the OAT. It's also high (but in range) on the Exatest results I sent you. I am scouring other tests too. I am emailing you the Vit B6 plasma results. The result was 230 (3.3-26). A later serum B6 tested showed B6 in range.. on the low end if anything. That was done thru ELN several years later. I have sent you that as well. I could not find any additional markers for high phosphorus beyond the OAT & Exatest.

So I just re-read your post & now I have another question.

I had been taking large quantities of injected B6 & magnesium I believe prior to the Exatest...but yet intracellular magnesium was still low. The test only reflects the tissue levels in the four days prior to the test from what I am told...& I don't have a record of what I took during those 4 days.. but I'm wondering whether the B6 failed to raise magnesium levels in my case..perhaps from a bottleneck in B6 metabolism.

By the way, my body seemed to like the B6. My blood pressure was up (a good thing!) and I had a general sense of well being. I increased very slowly and everything was going well.. until it went off the cliff, due to what I now think was an aggravation of already low B2.

Is low alk phos another variable here that could mess with B6?

FYI: susan costen owens posted this on the trying low oxolates yahoo group. She said it shows how high dosages of P5P can backfire.

Eur J Biochem. 1975 Aug 1;56(1):283-7.
Inactivation of rhodanese by pyridoxal 5'-phosphate.
Cannella C, Pecci L, Costa M, Pensa B, Cavallini D.
Abstract

Pyridoxal 5'-phosphate and other aromatic aldehydes inactivate rhodanese. The
inactivation reaches higher extents if the enzyme is in the sulfur-free form.
The identification of the reactive residue as an amino group has been made by
spectrophotometric determination of the 5'-phosphorylated pyridoxyl derivative
of the enzyme. The inactivation increases with pyridoxal 5'-phosphate
concentration and can be partially removed by adding thiosulfate or valine.
Prolonged dialysis against phosphate buffer also leads to the enzyme
reactivation. The absorption spectra of the pyridoxal phosphate - rhodanese
complex show a peak at 410 nm related to the Schiff base and a shoulder in the
330 nm region which is probably due to the reaction between pyridoxal
5'-phosphate and both the amino and thiol groups of the enzyme that appear
reasonably close to each other. The relationship betweenloss of activity and
pyridoxal 5'-phosphate binding to the enzyme shows that complete inactivation is
achieved when four lysyl residues are linked to pyridoxal 5'-phosphate.

PMID:
1236801
[PubMed - indexed for MEDLINE]

I'm a little confused about B6 metabolism and usage after reading this thread! ;(

Is PLP not able to easily raise intracelluar B6 levels? I always thought the cells wanted PLP!
So normal metabolism is B6 being converted to PLP on absorption into the serum, but the cells prefer plain old B6? Therefore if you bypass the enzymes that convert B6 -> PLP (injection) the cells get what they want?

Or have I got this completely wrong??

Hixxy82,

Good question. I started answering it and got confused myself... I leave it to Rich.

Cheers
SJB

As I understand it, pyridoxine gets phosphorylated in the gut to P5P before entering serum. Then P5P gets dephosphorylated before entering cells, and phosphorylated again inside the cells.

So taking P5P will bypass the first phosphorylation step, and will make it more easy to get P5P in serum. But it still needs to get dephosphorylated before it can enter cells, and then needs to get rephosphorylated inside.

So does any non-phosphorylated B6 get into the blood when taken orally?

Hope this is helpful:

"Vitamin B6 Forms: The vitamin B6 family includes pyridoxine (PN), pyridoxal (PL), pyridoxamine (PM), and their phosphorylated counterparts; pyridoxine phosphate (PNP), pyridoxal phosphate (PLP) and pyridoxamine phosphate (PMP). The vitamin is not synthesized in animal cells. However, mammalian cells can interconvert it. A series of phosphorylase, oxidase, and kinase enzymes effect the in vivo inter-conversion of pyridoxine and its derivatives.

In vivo, B6 compounds exist primarily as phosphorylated derivatives. In vitro, B6 compounds are provided in the media formulations as either pyridoxal or pyridoxine. The addition of serum or albumin to cell cultures may also provide pyridoxal and pyridoxal phosphate.

Pyridoxine (Pyridoxol) (PN):
Pyridoxine-HCl, C8H11NO3-HCl, molecular weight = 205.64, water-soluble;
It does not form a Schiff base and is relatively stable in culture media;
Cells do not use pyridoxine as an enzyme co-factor. They must convert pyridoxine into pyridoxal phosphate. This requires cells to have active pyridoxine kinase (EC 2.7.1.35) and pyridoxamine-phosphate oxidase (EC 1.4.3.5);
Pyridoxine may function as an antioxidant in cell culture.

Pyridoxal (PL):
Pyridoxal, C8H9NO 3-HCl, molecular weight = 203.64, water soluble;
This aldehyde form reacts with primary amines, such as amino acids, to form Schiff bases;
Pyridoxal Schiff bases non-enzymatically degrade amino acids (and other primary amines) in cell culture media:
Pyridoxal can form a relatively stable Schiff base with a lysyl residue of albumin;
Cells do not use pyridoxal directly as an enzyme co-factor. They must convert pyridoxal into pyridoxal phosphate. This requires cells to have an active pyridoxal kinase.

Pyridoxal-5-phosphate, (PLP):
This is the active coenzyme form in animal B6 enzymes, water-soluble;
This aldehyde form reacts with primary amines, such as amino acids, to form Schiff bases;
PLP Schiff bases can degrade amino acids non-enzymatically in cell culture media;
PLP is not added directly to cell culture media. However, it binds to albumin and may be fortuitously added to culture media that are supplemented with either serum or albumin. Pyridoxal phosphate can form a relatively stable Schiff base with a lysyl residue of albumin;
PLP does not cross cell membranes. It is dephosphorylated at the cell surface, transported into the cell, and then rephosphorylated."

http://www.sigmaaldrich.com/life-science/cell-culture/learning-center/media-expert/pyridoxal.html

Okay on second read through I do see rich mention that it's the dephosphorylation that is failing. Is this enzyme that dephosphorylates also B2 dependent? what is this enzyme??

That's exactly what I was looking for earlier. Seems my searching skills (or level of patience) needs improving.

***Hi, AQ.

Rich, can you help me connect the dots here?

***O.K. I've combined our posts together here.

I have many tests showing a need for B6. I have low intracellular magnesium per the Exatest test, in spite of lots of supplementation. I have low B2 (OAT) & lots of markers for phosphate are extremely high.

***O.K., I see these things in the test results you have sent me.

I also once ran a serum B6 test and the result was off the charts high.. maybe 10 or 20x the upper range of normal.

***Yes, I see that that was about 6 years ago (March 2006), and your more recent test (July 2010) showed P5P in the normal range. I'm not sure what was going on 6 years ago, and what you were supplementing then, so I will focus on the result of your more recent tests.

Thanks

Hi, AQ.

I don't understand the part about lots of markers for phosphate being extremely high. Can you clarify this, or send me the test results? I think you have my email address, but it's richvank at aol dot com.

The serum B6 measurement is a measurement of P5P, because that's the dominant form of B6 in the serum. So it sounds as though your body is having difficulty taking off the phosphate from the PLP, just as in the case I mentioned, so that you aren't getting B6 into your cells, and hence, because it's needed to transport magnesium in as well, you have low intracellular magnesium.

***I still think this is probably the explanation.

Best regards,

Rich

Yes, it's high on the OAT. It's also high (but in range) on the Exatest results I sent you. I am scouring other tests too. I am emailing you the Vit B6 plasma results. The result was 230 (3.3-26). A later serum B6 tested showed B6 in range.. on the low end if anything. That was done thru ELN several years later. I have sent you that as well. I could not find any additional markers for high phosphorus beyond the OAT & Exatest.

***I don't know why the phosphorus was high, both intracellularly and in the urine. The high urine phosphate can result simply from high phosphate in the diet, or it can be due to very high vitamin D, hyperparathyroidism, and a few other things. If you've had a standard blood comprehensive metabolic panel lately, was the calcium at a normal level?

So I just re-read your post & now I have another question.

I had been taking large quantities of injected B6 & magnesium I believe prior to the Exatest...but yet intracellular magnesium was still low.

***Were you also doing this prior to the OAT in May 2011?

The test only reflects the tissue levels in the four days prior to the test from what I am told...& I don't have a record of what I took during those 4 days.. but I'm wondering whether the B6 failed to raise magnesium levels in my case..perhaps from a bottleneck in B6 metabolism.

***Yes, I think that is probably what happened.

By the way, my body seemed to like the B6. My blood pressure was up (a good thing!) and I had a general sense of well being. I increased very slowly and everything was going well.. until it went off the cliff, due to what I now think was an aggravation of already low B2.

***I think that could be what happened.

Is low alk phos another variable here that could mess with B6?

***Yes. There are several isotypes of alkaline phosphate. The one that is supposed to removed the phosphate from P5P so that the resulting pyridoxal can diffuse into the cells is called "tissue nonspecific alkaline phosphatase." It is actually bound to the cell membrane, on the outside. If this enzyme is not functioning properly, it will prevent the B6 from getting into the cells, and the metabolic testing will show evidence of low P5P function. From what I've read, if the tissue nonspecific alk phosphatase is low, it should show up as low serum alk phosphatase in the comprehensive blood metabolic panel. The serum alk phosphatase is composed of several isotypes of alk phosphatase that come from various specific organs, also. It is a zinc enzyme, so low serum alk phosphatase activity can be due to low zinc.

***In looking through the OAT from May 2011 that you sent me, I noted some other things:

Maybe a yeast infection (based on elevated arabinose), though it can be caused by the diet.

Glyceric and glycolic were elevated, while oxalic was normal, and I'm not sure what to make of this.

High-normal pyruvic with low-normal citric. This suggests a bottleneck in the pyruvate dehydrogenase complex, which can be caused by low B1, B2, B3, magnesium, or lipoic.

High malic, which suggests low B3.

High aconitic, which suggests low glutathione (this also agrees with the high pyroglutamic).

A drop from HVA to VMA, which could be due to low copper or low vitamin C (Later, Vitamin C was found to be low, so this probably explains it. Low Vitamin C would be consistent with low glutathione, because the latter normally recycles the former.)

Low 5-HIAA with normal quinolinic. This could be due to the tryptophan metabolism shifting toward making B3 instead of serotonin, because B3 was deficient.

Looks like you were not oxidizing fatty acids much by either beta- or omega-oxidation. This could have been caused by low B3.

The high pyridoxic suggests that you were taking B6 in high dosage at the time of the OAT. Is that true?

B5 was a little low.

B2 was definitely low, and this could have affected your ability to convert B6 to P5P.

As noted earlier, Vitamin C was very low.

The low level of HMG suggests that coenzyme Q10 was low, also.

The low values of the first four amino acid metabolites suggest very low B6 (and/or B2).

***I hope this is helpful.

***Best regards,

***Rich

Hi Rich

I don't think I was supplementing much of anything at the time of that first B6 test back in '06, certainly not mega dosages. People on pyroluria groups tell me serum B6 readings are unreliable.

I didn't take any supplemental B6 between 2007 & 2011, bearing in mind Amy Yasko's cautions about B6 & the CBS SNPs, so no I was not taking any B6 at the time of that OAT in 2011, so that result is a puzzle.

Which raises the question, was I wasting B6 in the urine, and if so why? KPU maybe?

Calcium is in range on my recent CBCs.

Alk phos is low in range. I've had my eye on it since hearing Klinghardt talk about it.. I think it has got lower over the years. A plasma amino acids test in 2010 was strongly suggestive of hypochloridia .. so that could be a clue to low zinc, I guess.

Thanks for taking the time to look at these.

Hard to know where to go next. I have been going around in circles for a while now.

Cheers
AQ

ETA: So how useful is it to supplement magnesium if it can't get into the cells because of some B6 bottleneck?