384 replies to this topic

[Delta Gamma]

Sorry for the lack of a pubmed study, but its more the fact that increased oxygen consumption is only beneficial if there is proper oxygenation of all tissues. I'm more saying this would be a issue in individuals with poor circulation or lung conditions, MB is quite safe in healthy individuals.

Hope that makes sense.

Na, I don't get the link how mitochondria will change based on uneven distribution of oxygen consumption. It doesn't really make sense. It just seems like a fear, but fear is sometimes a "God-given" thing - an intuition. Intuition is not scientific (yet), but everyone has experienced it. I still have both my lungs and circulation in every organ, so maybe I can't sense your fear as intuitive.

I just don't want people thinking that because they have one lung or poor circulation in one arm, that MB is going to mutate their mitochondria into little swamp monsters who want to take over the world using methylene red, the evil methylene blue that only exists in my imagination. I just don't want anybody to dream about that, or even think about the possibility of methylene red outside my imagination. Nobody should be thinking about mitochondrial swamp monsters protruding from arms and lungs.

Well upregulation of complex IV may cause your cells to operate under hypoxic conditions if you have a disorder like Raynaud's. I said it was safe in healthy individuals, but in some people with pre-existing conditions may lead to problems however slight. Its not a good idea to completely brush a potential contradiction under the table, because in all reality if someone dies or experiences a severe reaction with this stuff its a shame to think that there was nothing we could do to prevent it.

What I'm saying is that if you have extremely poor circulation or oxygenation issues, MB may not be for you.

[Isochroma]

From ICAD: Tau-Targeted Therapy Slows Alzheimer's Progression for 19 Months:

"The drug, Dr. Wischik said, was effective when it dissolved in the stomach, but was not effective when the drug was absorbed through the intestines.

This was an issue for the 100-mg dose, which had "absolutely no activity because it didn't dissolve in the stomach."

Moreover, because the 100-mg dose dissolved in the intestines, it was more likely to cause diarrhea, which was reported by about 30% of patients and was the most common adverse event."

Edited by Isochroma, 17 July 2011 - 08:09 PM.

[aaron43]

To get this back on a progressive subject:

Cerebral vasodilators and metabolic activators (enhancers) for the treatment of cerebrovascular disorders

Methylene Blue an effective metabolic activator, with the ability of cognitive enhancement

___


Creatine— protects ATP during transport Study Shows Intelligence, Memory Improvements With the Dietary Supplement (but still hoping to clear up if this is partially a cerebral vasodilator, I personally have felt that I feel more sharp when I take creatine before working out, but did not make the connection that creatine could be synergizing)

I found this interesting as well:
Oral creatine monohydrate supplementation improves brain performance: a double-blind, placebo-controlled, cross-over trial.

Creatine supplementation is in widespread use to enhance sports-fitness performance, and has been trialled successfully in the treatment of neurological, neuromuscular and atherosclerotic disease. Creatine plays a pivotal role in brain energy homeostasis, being a temporal and spatial buffer for cytosolic and mitochondrial pools of the cellular energy currency, adenosine triphosphate and its regulator, adenosine diphosphate. In this work, we tested the hypothesis that oral creatine supplementation (5 g d(-1) for six weeks) would enhance intelligence test scores and working memory performance in 45 young adult, vegetarian subjects in a double-blind, placebo-controlled, cross-over design. Creatine supplementation had a significant positive effect (p < 0.0001) on both working memory (backward digit span) and intelligence (Raven's Advanced Progressive Matrices), both tasks that require speed of processing. These findings underline a dynamic and significant role of brain energy capacity in influencing brain performance.

L-Arginine- Vasodilator
Pyritinol HCL- Vasodilator
Vinpocetine- Vasodilator
Hydergine- Vasodilator, but a fungus, MB is an anit-fungal, so it would have to be seperately timed doses.


This sounds logically correct, could it work is the question

Edited by aaron43, 20 July 2011 - 08:22 AM.

[thedevinroy]

To get this back on a progressive subject:

Cerebral vasodilators and metabolic activators (enhancers) for the treatment of cerebrovascular disorders

Methylene Blue an effective metabolic activator, with the ability of cognitive enhancement

___


Creatine— protects ATP during transport Study Shows Intelligence, Memory Improvements With the Dietary Supplement (but still hoping to clear up if this is partially a cerebral vasodilator, I personally have felt that I feel more sharp when I take creatine before working out, but did not make the connection that creatine could be synergizing)

I found this interesting as well:
Oral creatine monohydrate supplementation improves brain performance: a double-blind, placebo-controlled, cross-over trial.

Creatine supplementation is in widespread use to enhance sports-fitness performance, and has been trialled successfully in the treatment of neurological, neuromuscular and atherosclerotic disease. Creatine plays a pivotal role in brain energy homeostasis, being a temporal and spatial buffer for cytosolic and mitochondrial pools of the cellular energy currency, adenosine triphosphate and its regulator, adenosine diphosphate. In this work, we tested the hypothesis that oral creatine supplementation (5 g d(-1) for six weeks) would enhance intelligence test scores and working memory performance in 45 young adult, vegetarian subjects in a double-blind, placebo-controlled, cross-over design. Creatine supplementation had a significant positive effect (p < 0.0001) on both working memory (backward digit span) and intelligence (Raven's Advanced Progressive Matrices), both tasks that require speed of processing. These findings underline a dynamic and significant role of brain energy capacity in influencing brain performance.

L-Arginine- Vasodilator
Pyritinol HCL- Vasodilator
Vinpocetine- Vasodilator
Hydergine- Vasodilator, but a fungus, MB is an anit-fungal, so it would have to be seperately timed doses.


This sounds logically correct, could it work is the question

It is an effective treatment under the trade name "Rember" for DAT. http://pipeline.cora...es_comeback.php And I would recommend it for Dementia sufferers of all types since it increases brain energy associated with aging. Since most people with Dementia have aged quite a bit, this would be helpful in every day-to-day activities. Even though Rember is primarily for DAT, it is probably prescribed for the other types.

[aaron43]

To get this back on a progressive subject:

Cerebral vasodilators and metabolic activators (enhancers) for the treatment of cerebrovascular disorders

Methylene Blue an effective metabolic activator, with the ability of cognitive enhancement

___


Creatine— protects ATP during transport Study Shows Intelligence, Memory Improvements With the Dietary Supplement (but still hoping to clear up if this is partially a cerebral vasodilator, I personally have felt that I feel more sharp when I take creatine before working out, but did not make the connection that creatine could be synergizing)

I found this interesting as well:
Oral creatine monohydrate supplementation improves brain performance: a double-blind, placebo-controlled, cross-over trial.

Creatine supplementation is in widespread use to enhance sports-fitness performance, and has been trialled successfully in the treatment of neurological, neuromuscular and atherosclerotic disease. Creatine plays a pivotal role in brain energy homeostasis, being a temporal and spatial buffer for cytosolic and mitochondrial pools of the cellular energy currency, adenosine triphosphate and its regulator, adenosine diphosphate. In this work, we tested the hypothesis that oral creatine supplementation (5 g d(-1) for six weeks) would enhance intelligence test scores and working memory performance in 45 young adult, vegetarian subjects in a double-blind, placebo-controlled, cross-over design. Creatine supplementation had a significant positive effect (p < 0.0001) on both working memory (backward digit span) and intelligence (Raven's Advanced Progressive Matrices), both tasks that require speed of processing. These findings underline a dynamic and significant role of brain energy capacity in influencing brain performance.

L-Arginine- Vasodilator
Pyritinol HCL- Vasodilator
Vinpocetine- Vasodilator
Hydergine- Vasodilator, but a fungus, MB is an anit-fungal, so it would have to be seperately timed doses.


This sounds logically correct, could it work is the question

It is an effective treatment under the trade name "Rember" for DAT. http://pipeline.cora...es_comeback.php And I would recommend it for Dementia sufferers of all types since it increases brain energy associated with aging. Since most people with Dementia have aged quite a bit, this would be helpful in every day-to-day activities. Even though Rember is primarily for DAT, it is probably prescribed for the other types.

Methylene Blue is a vasoconstrictor by nature, but it may not be a very strong vasoconstrictor. It is a known metabolic enhancer, it also increases the metabolic capacity and function in areas of the brain that are being used during some sort of cerebral activity.
Rember is this, just very slightly different. But I am intrigued about the combination of the methylene blue and a strong vasodilator.
I envision it as, methylene blue either boosts or revives parts of the brain. It depends on which portion of the brain an activity activates, and that brain portion's past activity use (similar to a muscle). It seems during a cerebral activity, it either primes the portion of the brain for above normal functioning when this portion of the brain is used normally in the past; and it seems to revive "back to normal" the portions of the brain that have not in the past been used, but are being used in a current activity, like a metaphorical dust off of mental cobwebs.
Mixing this effect, with that of a cerebral vasodilator, could in my theory, and from what I saw exemplified in the first link I provided in the last post, could in theory work together to oxygenate the same portions of the brain that MB would be metabolically enhancing. Also, what I envision is the vasodilator will put the MB, deeper in the brain, or that it would bathe the currently being used brain region in a more thorough manner with Methylene Blue.
It would be interesting to find more information on this

Regarding Creatine:

being a temporal and spatial buffer for cytosolic and mitochondrial pools of the cellular energy currency

Methylene Blue is a Mitochondrial Optimizer, could this quote about creatine optimize the mitochondrial function even further?

Similar in the way devin said about one of ALCAR's properties, in how it shuttles fat easier to the mitochondria for use, and how this could work together with Methylene Blue which optimized mitochondrial function

[thedevinroy]

Methylene Blue is a vasoconstrictor by nature, but it may not be a very strong vasoconstrictor. It is a known metabolic enhancer, it also increases the metabolic capacity and function in areas of the brain that are being used during some sort of cerebral activity.
Rember is this, just very slightly different. But I am intrigued about the combination of the methylene blue and a strong vasodilator.
I envision it as, methylene blue either boosts or revives parts of the brain. It depends on which portion of the brain an activity activates, and that brain portion's past activity use (similar to a muscle). It seems during a cerebral activity, it either primes the portion of the brain for above normal functioning when this portion of the brain is used normally in the past; and it seems to revive "back to normal" the portions of the brain that have not in the past been used, but are being used in a current activity, like a metaphorical dust off of mental cobwebs.
Mixing this effect, with that of a cerebral vasodilator, could in my theory, and from what I saw exemplified in the first link I provided in the last post, could in theory work together to oxygenate the same portions of the brain that MB would be metabolically enhancing. Also, what I envision is the vasodilator will put the MB, deeper in the brain, or that it would bathe the currently being used brain region in a more thorough manner with Methylene Blue.
It would be interesting to find more information on this

Regarding Creatine:

being a temporal and spatial buffer for cytosolic and mitochondrial pools of the cellular energy currency

Methylene Blue is a Mitochondrial Optimizer, could this quote about creatine optimize the mitochondrial function even further?

Similar in the way devin said about one of ALCAR's properties, in how it shuttles fat easier to the mitochondria for use, and how this could work together with Methylene Blue which optimized mitochondrial function

Creatine is a known nootropic for that reason (http://www.ncbi.nlm....85/?tool=pubmed), and it also has been suggested to offset the fatigue associated with Bacopa's (indirect) down regulation of Creatine Kinase (http://www.ncbi.nlm....creatine kinase). I imagine it would be pretty good with Methylene Blue. Come to think of it, I and others have reported a headache associated with MB, and that could be a pressure headache from vasoconstriction that a vasodilator would relieve.

ALCAR is definitely nice with Methylene Blue - ALCAR gives energy to power the MB.

I have a vasodilator, Picamilon, but its GABA effects far outweigh its vasodilation effects, so I take it before bed. I didn't notice a lot that first day when I took them together, although I didn't take a lot of either - just the 60mcg/MB and 50mg/Picamilon. I'm actually looking for a better vasodilator, but none of the ones you mentioned (for each their own reasons). Ginkgo (also an NETi) is more of a blood thinner than a vasodilator, but I'd try it again knowing what I know now about extract strengths and synergy.

Wouldn't Arginine be a bit contradictory in the mitochondrial-antiaging sense for Methylene Blue? MB is an antioxidant, and Arginine increases NOS.

[aaron43]

Creatine is a known nootropic for that reason (http://www.ncbi.nlm....85/?tool=pubmed), and it also has been suggested to offset the fatigue associated with Bacopa's (indirect) down regulation of Creatine Kinase (http://www.ncbi.nlm....creatine kinase). I imagine it would be pretty good with Methylene Blue. Come to think of it, I and others have reported a headache associated with MB, and that could be a pressure headache from vasoconstriction that a vasodilator would relieve.

ALCAR is definitely nice with Methylene Blue - ALCAR gives energy to power the MB.

I have a vasodilator, Picamilon, but its GABA effects far outweigh its vasodilation effects, so I take it before bed. I didn't notice a lot that first day when I took them together, although I didn't take a lot of either - just the 60mcg/MB and 50mg/Picamilon. I'm actually looking for a better vasodilator, but none of the ones you mentioned (for each their own reasons). Ginkgo (also an NETi) is more of a blood thinner than a vasodilator, but I'd try it again knowing what I know now about extract strengths and synergy.

Wouldn't Arginine be a bit contradictory in the mitochondrial-antiaging sense for Methylene Blue? MB is an antioxidant, and Arginine increases NOS.

For the arginine:

With increased Arginine/NOS to the mitochondria, it has been shown to reduce Ca2+ uptake by 67% and 85%, and the rate of mitochondrial Ca2+ release decreased by 11% and 8%. Less is being taken up, and less is being let out, I still cant figure out the dosage used in the study: Effects of mitochondrial L-arginine/nitric oxide system on mitochondrial Ca2+ transport in rat myocardium L-Arg/NO systems take part in the regulation Ca2+ transportation as well, so L-Arg/NO are key players with Ca2+.
But:
Ca2+ participates in cell activation and the enhancement of aerobic metabolism in the mitochondria, and is ultimately the positive modulator of mitochondrial ATP synthesis.
But this same Ca2+ , if there is a major increase, is known that it causes cell death. It is also established that mitochondria are a major target of Ca2+-dependent necrosis.
Over accumulation of Ca2+ in the mitochondria leads to cell death, but from what I have found l-arginine/NOS, reduce the uptake of Ca2+ by ~74%

Found this interesting as well, and still relates:

The link between cytochrome c release, apoptosis and mitochondrial Ca2+ was immediately clear: overaccumulation of Ca2+ by mitochondria was already known to activate the PTP (in a cyclosporine A sensitive mechanisms, see below) and this large membrane pore in the inner mitochondrial membrane can lead to matrix swelling, causing the rupture of the outer mitochondrial membrane and release of cytochrome c.

L-arginine inhibits uptake of Ca2+ which reduces pre-mature cell death
For a cell's death cycle to be activated, , cytochrome c and Apaf-1 form together to initiate the death of the cell
But, Methylene Blue reduces cytochrome C (which leads to increase in cytochrome oxidase activity),
Explified from a MB study:

Brain cytochrome oxidase activity in the MB-treated group was approximately 70% higher

So it seems that an l-arginine/MB could actually be beneficial in reducing pre-mature deaths of cells

Cytochrome oxidase is the mitochondrial enzyme that catalyzes the utilization of oxygen for the electron transport chain during cellular respiration. (This brings up the vasodilator aspect as well, as they increase oxygen available for use)
In this same way, the memory effects of MB are shown, Methylene blue, which has been shown to increase oxygen consumption in vitro, was used to restore mitochondrial electron transport in order to facilitate memory consolidation.
So the main aspect, of MB and it's nootropic properties is that it increases oxygen consumption to restore the mitochondrial electron transport chain, in turn increasing cytochrome oxidase activity, which in turn leads to improved memory consolidation. and cytochrome oxidase is described below,

Oxidase Activity: It is the last enzyme in the respiratory electron transport chain of mitochondria (or bacteria) located in the mitochondrial (or bacterial) membrane. It receives an electron from each of four cytochrome c molecules, and transfers them to one oxygen molecule, converting molecular oxygen to two molecules of water. In the process, it binds four protons from the inner aqueous phase to make water, and in addition translocates four protons across the membrane, helping to establish a transmembrane difference of proton electrochemical potential that the ATP synthase then uses to synthesize ATP.

I am still going to look more into how creatine could possibly work together with all this

Niner: I can't post the link, because I go to a university, and if I did, all you guys would just get the abstract because your not logged in. And even if it was already posted in the thread, snuff asked for it, so Im giving it to her/him/it, as well as the other study that is not provided in this forum. Please stop trying to tell me what I can and cannot do, I won't listen because I don't care, the only thing I'm interested in is the academic information. It will be a waste of threadspace to try and critique my posts, and is weird since your all about not wasting threadspace. And this nitpicking needs to stop because again this person asked for the full study, something that I have access too, and I gave it to them. I don't need to know your personal opinion on the way I post, but would much rather here about what you think about the information that is included in the posts. If you have nothing to say on the subject that is in the post, just like you tried to tell me, then you don't need to say anything

[thedevinroy]

For the arginine:

With increased Arginine/NOS to the mitochondria, it has been shown to reduce Ca2+ uptake by 67% and 85%, and the rate of mitochondrial Ca2+ release decreased by 11% and 8%. Less is being taken up, and less is being let out, I still cant figure out the dosage used in the study: Effects of mitochondrial L-arginine/nitric oxide system on mitochondrial Ca2+ transport in rat myocardium L-Arg/NO systems take part in the regulation Ca2+ transportation as well, so L-Arg/NO are key players with Ca2+.
But:
Ca2+ participates in cell activation and the enhancement of aerobic metabolism in the mitochondria, and is ultimately the positive modulator of mitochondrial ATP synthesis.
But this same Ca2+ , if there is a major increase, is known that it causes cell death. It is also established that mitochondria are a major target of Ca2+-dependent necrosis.
Over accumulation of Ca2+ in the mitochondria leads to cell death, but from what I have found l-arginine/NOS, reduce the uptake of Ca2+ by ~74%

Found this interesting as well, and still relates:

The link between cytochrome c release, apoptosis and mitochondrial Ca2+ was immediately clear: overaccumulation of Ca2+ by mitochondria was already known to activate the PTP (in a cyclosporine A sensitive mechanisms, see below) and this large membrane pore in the inner mitochondrial membrane can lead to matrix swelling, causing the rupture of the outer mitochondrial membrane and release of cytochrome c.

L-arginine inhibits uptake of Ca2+ which reduces pre-mature cell death
For a cell's death cycle to be activated, , cytochrome c and Apaf-1 form together to initiate the death of the cell
But, Methylene Blue reduces cytochrome C (which leads to increase in cytochrome oxidase activity),
Explified from a MB study:

Brain cytochrome oxidase activity in the MB-treated group was approximately 70% higher

So it seems that an l-arginine/MB could actually be beneficial in reducing pre-mature deaths of cells

Cytochrome oxidase is the mitochondrial enzyme that catalyzes the utilization of oxygen for the electron transport chain during cellular respiration. (This brings up the vasodilator aspect as well, as they increase oxygen available for use)
In this same way, the memory effects of MB are shown, Methylene blue, which has been shown to increase oxygen consumption in vitro, was used to restore mitochondrial electron transport in order to facilitate memory consolidation.
So the main aspect, of MB and it's nootropic properties is that it increases oxygen consumption to restore the mitochondrial electron transport chain, in turn increasing cytochrome oxidase activity, which in turn leads to improved memory consolidation. and cytochrome oxidase is described below,

Oxidase Activity: It is the last enzyme in the respiratory electron transport chain of mitochondria (or bacteria) located in the mitochondrial (or bacterial) membrane. It receives an electron from each of four cytochrome c molecules, and transfers them to one oxygen molecule, converting molecular oxygen to two molecules of water. In the process, it binds four protons from the inner aqueous phase to make water, and in addition translocates four protons across the membrane, helping to establish a transmembrane difference of proton electrochemical potential that the ATP synthase then uses to synthesize ATP.

I am still going to look more into how creatine could possibly work together with all this

Great info! Arginine reduces Ca uptake in the Mitochondria, thereby preventing apoptosis from overswelling... and here I though it caused free radical damage without the presence of antioxidants. Since MB increases oxygen uptake, I bet the extra NOS from arginine would be particularly beneficial. Sounds like a good combo.

Also, I read the study you posted, and the quote was taken out of context. Here is the correct context: "After L-Arg or SNP treatment, mitochondrial Ca2+ uptake were decreased by 67% and 85%, respectively..." The compound SNP is sodium nitroprusside, a potent vasodilator with an iron core used to treat severe hypertension. Despite it's use as a drug, it is pretty toxic and was probably used just as a validation tool for the arginine's relative effectiveness.

Let me know if you uncover any potential creatine synergy effects.

[Elus]

I have a question regarding methylene blue, vitamin B12, and cyanide. I read on wikipedia (Top portion of the Vitamin B12 page) that Vitamin B12 "is converted to the physiological forms,methylcobalamin and adenosylcobalamin, leaving behind the cyanide, albeit in minimal concentration."


I also read on wikipedia (Methylene Blue article, cyanide poisoning section) that "large doses of methylene blue are sometimes used as an antidote to potassium cyanidepoisoning."


My question is: Could there be a relationship between the beneficial effects of methylene blue be via its functionality in treating residual cyanide left over from Vitamin B12 conversion? Perhaps I'm not clear on some details, so if anyone has any ideas let me know.

[thedevinroy]

I have a question regarding methylene blue, vitamin B12, and cyanide. I read on wikipedia (Top portion of the Vitamin B12 page) that Vitamin B12 "is converted to the physiological forms,methylcobalamin and adenosylcobalamin, leaving behind the cyanide, albeit in minimal concentration."

I also read on wikipedia (Methylene Blue article, cyanide poisoning section) that "large doses of methylene blue are sometimes used as an antidote to potassium cyanidepoisoning."

My question is: Could there be a relationship between the beneficial effects of methylene blue be via its functionality in treating residual cyanide left over from Vitamin B12 conversion? Perhaps I'm not clear on some details, so if anyone has any ideas let me know.

Here's the full text: http://www.ncbi.nlm....00433-0082b.pdf cited from Wikipedia. I can't conclude the article... it's pretty two-faced.

[niner]

I have a question regarding methylene blue, vitamin B12, and cyanide. I read on wikipedia (Top portion of the Vitamin B12 page) that Vitamin B12 "is converted to the physiological forms,methylcobalamin and adenosylcobalamin, leaving behind the cyanide, albeit in minimal concentration."


I also read on wikipedia (Methylene Blue article, cyanide poisoning section) that "large doses of methylene blue are sometimes used as an antidote to potassium cyanidepoisoning."


My question is: Could there be a relationship between the beneficial effects of methylene blue be via its functionality in treating residual cyanide left over from Vitamin B12 conversion? Perhaps I'm not clear on some details, so if anyone has any ideas let me know.

No. Dose, guys. Dose. It's all about dose.

[niner]

Here is the MB paper from Bruce Ames' lab:

Atamna H, Nguyen A, Schultz C, Boyle K, Newberry J, Kato H, Ames BN (2008) Methylene Blue Delays Cellular Senescence and Enhances Key Mitochondrial Biochemical Pathways Relevant to Aging. FASEB J. 22:703-12. FREE Full Text

Methylene Blue (MB) and other diaminophenothiazines extend the life span of human IMR90 fibroblasts in tissue culture by >20 population doubling (PDLs) (527). MB has been used clinically for about a century to treat numerous ailments. MB delays senescence at nM levels in IMR90 by enhancing mitochondrial function. MB increases mitochondrial complex IV by 30%, enhances cellular oxygen consumption by 37–70%, increases heme synthesis, and reverses premature senescence caused by H2O2 or cadmium. MB also induces ~250 phase-2 antioxidant enzymes in hepG2 cells. Flavin-dependent enzymes are known to use NAD(P)H to reduce MB to leucomethylene blue (MBH2), whereas cytochrome c reoxidizes MBH2 to MB. Experiments on lysates from rat liver mitochondria suggest the ratio MB/cytochrome c is important for the protective actions of MB. It is proposed that the cellular senescence delay caused by MB is due to cycling between MB and MBH2 in mitochondria, which may partly explain the increase in specific mitochondrial activities. Cycling of MB between oxidized and reduced forms may block oxidant production by mitochondria. MB may be useful to delay mitochondrial dysfunction and oxidative stress, which are thought to be key aberrations that lead to cellular senescence and aging and the decrease in complex I in Parkinson’s disease and complex IV in Alzheimer’s disease (527).

Two points I draw from this: There is an important dosage, determined by the ratio of MB/cytochrome c. Dosing on the basis of psychotropic effects may or may not match this protective level. Secondly, because of the redox cycling that appears to be part of the protective mechanism, it doesn't seem to me that ascorbic acid would be needed; that's just a way of creating leuco-MB, which the body will do anyway.

[rwac]

Secondly, because of the redox cycling that appears to be part of the protective mechanism, it doesn't seem to me that ascorbic acid would be needed; that's just a way of creating leuco-MB, which the body will do anyway.

Its probably a good way to avoid blue stains though.

[MrHappy]

Here is the MB paper from Bruce Ames' lab:

Atamna H, Nguyen A, Schultz C, Boyle K, Newberry J, Kato H, Ames BN (2008) Methylene Blue Delays Cellular Senescence and Enhances Key Mitochondrial Biochemical Pathways Relevant to Aging. FASEB J. 22:703-12. FREE Full Text

Methylene Blue (MB) and other diaminophenothiazines extend the life span of human IMR90 fibroblasts in tissue culture by >20 population doubling (PDLs) (527). MB has been used clinically for about a century to treat numerous ailments. MB delays senescence at nM levels in IMR90 by enhancing mitochondrial function. MB increases mitochondrial complex IV by 30%, enhances cellular oxygen consumption by 37–70%, increases heme synthesis, and reverses premature senescence caused by H2O2 or cadmium. MB also induces ~250 phase-2 antioxidant enzymes in hepG2 cells. Flavin-dependent enzymes are known to use NAD(P)H to reduce MB to leucomethylene blue (MBH2), whereas cytochrome c reoxidizes MBH2 to MB. Experiments on lysates from rat liver mitochondria suggest the ratio MB/cytochrome c is important for the protective actions of MB. It is proposed that the cellular senescence delay caused by MB is due to cycling between MB and MBH2 in mitochondria, which may partly explain the increase in specific mitochondrial activities. Cycling of MB between oxidized and reduced forms may block oxidant production by mitochondria. MB may be useful to delay mitochondrial dysfunction and oxidative stress, which are thought to be key aberrations that lead to cellular senescence and aging and the decrease in complex I in Parkinson’s disease and complex IV in Alzheimer’s disease (527).

Two points I draw from this: There is an important dosage, determined by the ratio of MB/cytochrome c. Dosing on the basis of psychotropic effects may or may not match this protective level. Secondly, because of the redox cycling that appears to be part of the protective mechanism, it doesn't seem to me that ascorbic acid would be needed; that's just a way of creating leuco-MB, which the body will do anyway.

Very valid point.. I noticed from the graphs that the senescence delay dropped significantly in the 1000nM dosage, compared to 100nM. I wonder how that scales up to humans. Good link!

[MrHappy]

UPDATE: Have been taking Modafinil. Ran out of Acetyl-L-Carnitine. The switch took place around the same time. Ran out of vitamins, too. The effects of Methylene Blue are still apparent: increased word-find & similar connections, increased quality decision making, lack of self-defeat, etc. However, the effects SUCKED without the aid of a multivitamin. When I ran out of vitamins, I had the worst day. So I stole my sisters Women's multivitamin this morning and was back to normal. ALCAR boosted the effects of MB more than Modafinil. I still haven't determined if I like Modafinil as an ADHD aid... seems to work sometimes and not others... and has a terrible interaction with caffeine for me. Regardless, I began taking more MB per dose, and interestingly, with the same logic as Isochroma: to gauge absorption better. I take 8 drops of the 2.303% solution 4x a day.

Other research I have read indicated that calcium or sodium ascorbate was required as part of the process. Maybe worth experimenting with that?

[MrHappy]

UPDATE: Have been taking Modafinil. Ran out of Acetyl-L-Carnitine. The switch took place around the same time. Ran out of vitamins, too. The effects of Methylene Blue are still apparent: increased word-find & similar connections, increased quality decision making, lack of self-defeat, etc. However, the effects SUCKED without the aid of a multivitamin. When I ran out of vitamins, I had the worst day. So I stole my sisters Women's multivitamin this morning and was back to normal. ALCAR boosted the effects of MB more than Modafinil. I still haven't determined if I like Modafinil as an ADHD aid... seems to work sometimes and not others... and has a terrible interaction with caffeine for me. Regardless, I began taking more MB per dose, and interestingly, with the same logic as Isochroma: to gauge absorption better. I take 8 drops of the 2.303% solution 4x a day.

Other research I have read indicated that calcium or sodium ascorbate was required as part of the process. Maybe worth experimenting with that?

Ahh found it! MB + Vitamin C = leucomethylene blue!

Patent 7,335,505 Reduced methylene blue (leucomethylene blue) is more effective and crosses the blood brain barrier better. One recipe for reduction is 2 - 2.5 mg vitamin C per mg MB for three hours. IC50 for inhibiting tau filament formation is 4 microM.

[niner]

Ahh found it! MB + Vitamin C = leucomethylene blue!

Patent 7,335,505 Reduced methylene blue (leucomethylene blue) is more effective and crosses the blood brain barrier better. One recipe for reduction is 2 - 2.5 mg vitamin C per mg MB for three hours. IC50 for inhibiting tau filament formation is 4 microM.

Thank you for that, MrHappy! That patent is a treasure trove. In a nutshell, oxidized MB, the blue form, is a charged molecule and does not cross membranes well. If you reduce it to the MBH2 (colorless) form known as leucomethylene blue, it is a neutral molecule and crosses the blood brain barrier, and membranes in general a lot better. It's known from pharmacokinetic studies done in the early 70's that MB is converted to leucoMB in the body:

Methylene blue was administered orally to seven normal human subjects at a dose of 10 mg. in capsule form. Total urinary recovery ranged from 53 to 97% of the dose, with an average of 74%. Of the material recovered, an average of 78% was excreted as Ieucomethylene blue (stabilized in some salt, complex, or combination form) and the remainder as methylene blue. Some excretion rate-time plots of both methylene blue and leucomethylene blue showed evidence of a circadian rhythm. In a male dog and a female dog, administered 15 mg./kg. methylene blue orally, no drug was detected in blood. The female dog was catheterized and urine was collected for 10 hr. postdosing; recovery was 2.4% of the dose. The female dog was also administered a 10-mg. dose of methylene blue orally, and urine was collected by catheter over 14 hr. Recovery was 3.8% of the dose. It was concluded that methylene blue is well absorbed in man and poorly absorbed in the dog after oral administration.

This suggests to me that MB will work without ascorbate, but that ascorbate will be a small to medium-sized improvement. In the patent, it says:

It is highly likely that variability in oral absorbtion is determined largely by the efficiency of initial reduction in the GI tract. One way to achieve more reliable absorbtion is therefore be to pre-reduce methylene blue with ascorbic acid. We have shown from in vitro studies that this conversion is rather slow, so that it takes 3 hours to achieve 90% reduction of methylene blue in water in the presence of 2× mg ratio of ascorbic acid. Therefore, the dosage of methylene blue which is most likely to ensure reliable absorbtion will be 3.5 mg/kg/day of methylene blue pre-reduced for at least 3 hours in the presence of 7 mg/kg/day of ascorbic acid.

This means that (according to them, anyway) if you want whatever improvement ascorbate can give you, you can't just swallow a vitamin C tablet at the same time you take the MB. You need to pre-incubate the two compounds in solution for several hours first. Considering the small doses that we're using, we could probably use a large excess of ascorbate to accelerate the reaction. To be honest, I'm kind of baffled that this should take any time at all; this sort of redox reaction is usually very fast. Also, I would think that if you see the color change, then it's done. Note that these guys are trying to get a patent on the use of leucomethylene blue as a therapeutic agent, so they are probably inclined to make it sound tricky in order to flummox the hapless patent examiner. If someone happens to have some MB and some ascorbate handy, mix a little together and let us know what happens.

[rwac]

If someone happens to have some MB and some ascorbate handy, mix a little together and let us know what happens.

I've done it before, the MB becomes colorless in a few minutes. However I was using 12g of ascorbate/ascorbic acid with ~10mg of MB.

[MrHappy]

This morning I woke up hungover and dead tired and thought to myself I should call in sick to work. I went into the kitchen to take my 60mcg dose of MB and thought eff it I should just take one drop right from the Kordon bottle in a glass of water and maybe it will push me through the work day. So I did.

A dramatic difference, within an hour or two, the hungover tired feeling was gone and for lack of a better analogy I began feeling like I was on mushrooms minus the hallucinations. Serene , calm , centered, colors are brighter, everything is brighter, a general feeling of well being.

I guess if it really is safe, I think I might move to this much higher dose. It feels a bit surreal after all these years of pissing out expensive garbage that did nothing, that a $5 bottle of blue dye would give me what I was looking for. There is a nagging voice somewhere in the distance... saying nothing like this ever comes without a cost or side effect, you should know better, and that a year of dosing from now I'll find out MB causes ear wax cancer or something. Maybe the Blue Man Group are all MethB addicts and eventually I'll have to join them too, I best start practicing my drumming now.

I was going to post a reminder about "NTP Toxicology and Carcinogenesis Studies of Methylene Blue Trihydrate (CAS No. 7220-79-3) in F344/N Rats and B6C3F1 Mice (Gavage Studies)." .. when I realised something was nagging me about grouping these results with the others. http://www.ncbi.nlm....pubmed/18685714

These results were obtained using MB Trihydrate (CAS No. 7220-79-3). http://www.chemicalb...60_EN.htm#MSDSA

We are using vanilla MB - CAS No. 61-73-4. http://www.chemicalb...B2748858_EN.htm

I know they are nearly identical, but worth mentioning in case the dosage is different.

[thedevinroy]

This morning I woke up hungover and dead tired and thought to myself I should call in sick to work. I went into the kitchen to take my 60mcg dose of MB and thought eff it I should just take one drop right from the Kordon bottle in a glass of water and maybe it will push me through the work day. So I did.

A dramatic difference, within an hour or two, the hungover tired feeling was gone and for lack of a better analogy I began feeling like I was on mushrooms minus the hallucinations. Serene , calm , centered, colors are brighter, everything is brighter, a general feeling of well being.

I guess if it really is safe, I think I might move to this much higher dose. It feels a bit surreal after all these years of pissing out expensive garbage that did nothing, that a $5 bottle of blue dye would give me what I was looking for. There is a nagging voice somewhere in the distance... saying nothing like this ever comes without a cost or side effect, you should know better, and that a year of dosing from now I'll find out MB causes ear wax cancer or something. Maybe the Blue Man Group are all MethB addicts and eventually I'll have to join them too, I best start practicing my drumming now.

I was going to post a reminder about "NTP Toxicology and Carcinogenesis Studies of Methylene Blue Trihydrate (CAS No. 7220-79-3) in F344/N Rats and B6C3F1 Mice (Gavage Studies)." .. when I realised something was nagging me about grouping these results with the others. http://www.ncbi.nlm....pubmed/18685714

These results were obtained using MB Trihydrate (CAS No. 7220-79-3). http://www.chemicalb...60_EN.htm#MSDSA

We are using vanilla MB - CAS No. 61-73-4. http://www.chemicalb...B2748858_EN.htm

I know they are nearly identical, but worth mentioning in case the dosage is different.

Those forms are identical as soon as they hit water and become aqueous.

I find that study interesting primarily because it says that rats developed Methemoglobinemia even though Methylene Blue is used as a treatment for Methemoglobinemia. Plus those rats are given some crazy doses. The only thing that scares me is the mutagenic study at the end. Ow, my DNA.

[niner]

You are assuming that the "2.303%" is a measurement of volume/volume, which is unusual and usually avoided outside of textbook use because when two liquids are combined, their volumes are not always additive. A "percent" solution is typically mass/mass (Mass fraction), mole/mole (Mole fraction, or amount fraction), or mass/volume (Mass Concentration, not really dimensionless but still used). Thus, your statement, "1 ml 2.303% MB solution contains 0.02303 ml of pure MB" needs sources, otherwise we will have to assume that Kordon uses the typical interpretation of % solutions.

However, if Kordon uses Mass Concentration, then 1mL of 2.303% solution does in fact yield 23.03mg of Methylene Blue. This is what I and others have concluded with little discussion. I believe it to be this way not only because the math makes chemistry simple (which is why this method is used: http://wetlab.izzid....rcent_Solution/) and also because it costs less to manufacture a 2.303% Mass Concentration than any other form of percent solution.

Thanks for this correction, Devin, you nailed it. It's a near certainty that Kordon is using mass concentration. That's the standard, and when it isn't used, the unit will get a label, like m/v.

I was going to post a reminder about "NTP Toxicology and Carcinogenesis Studies of Methylene Blue Trihydrate (CAS No. 7220-79-3) in F344/N Rats and B6C3F1 Mice (Gavage Studies)." .. when I realised something was nagging me about grouping these results with the others. http://www.ncbi.nlm....pubmed/18685714

[trihydrate vs. unhydrated]

I know they are nearly identical, but worth mentioning in case the dosage is different.

Those forms are identical as soon as they hit water and become aqueous.

I find that study interesting primarily because it says that rats developed Methemoglobinemia even though Methylene Blue is used as a treatment for Methemoglobinemia. Plus those rats are given some crazy doses. The only thing that scares me is the mutagenic study at the end. Ow, my DNA.

Exactly what I was going to write, but you beat me to it. Also, despite the methemoglobinemia and some increase in cancers, the rats lived as long as the controls, but the mice lived longer than controls in a dose-dependent manner. The fact that it was dose-dependent is a good indicator that the drug is responsible. If only mice were miniature humans, despite the number of people I've met in the corporate world who are rats... But that's a really interesting outcome. Since the Ames redox-cycling mitochondrial assist mechanism is supposed to only work at low concentration, this mouse result is presumably due to some other mechanism, unless their mitochondrial concentration is very low for some reason.

You really don't need to worry about the in vitro mutagenicity experiments at the end of the abstract. They are trumped by the in vivo work. OTOH, you might want to worry at least a little about the increased cancers they saw in the rodents. The in vitro assays are indicators that you might have something to worry about in vivo. Rodents are cancer-prone, so they are indicators that you might have something to worry about in humans. MB is the right size and shape to be a DNA intercalator, but note the crazy high dosages being used. Don't forget the mice lived longer.

[MrHappy]

You are assuming that the "2.303%" is a measurement of volume/volume, which is unusual and usually avoided outside of textbook use because when two liquids are combined, their volumes are not always additive. A "percent" solution is typically mass/mass (Mass fraction), mole/mole (Mole fraction, or amount fraction), or mass/volume (Mass Concentration, not really dimensionless but still used). Thus, your statement, "1 ml 2.303% MB solution contains 0.02303 ml of pure MB" needs sources, otherwise we will have to assume that Kordon uses the typical interpretation of % solutions.

However, if Kordon uses Mass Concentration, then 1mL of 2.303% solution does in fact yield 23.03mg of Methylene Blue. This is what I and others have concluded with little discussion. I believe it to be this way not only because the math makes chemistry simple (which is why this method is used: http://wetlab.izzid....rcent_Solution/) and also because it costs less to manufacture a 2.303% Mass Concentration than any other form of percent solution.

Thanks for this correction, Devin, you nailed it. It's a near certainty that Kordon is using mass concentration. That's the standard, and when it isn't used, the unit will get a label, like m/v.

I was going to post a reminder about "NTP Toxicology and Carcinogenesis Studies of Methylene Blue Trihydrate (CAS No. 7220-79-3) in F344/N Rats and B6C3F1 Mice (Gavage Studies)." .. when I realised something was nagging me about grouping these results with the others. http://www.ncbi.nlm....pubmed/18685714

[trihydrate vs. unhydrated]

I know they are nearly identical, but worth mentioning in case the dosage is different.

Those forms are identical as soon as they hit water and become aqueous.

I find that study interesting primarily because it says that rats developed Methemoglobinemia even though Methylene Blue is used as a treatment for Methemoglobinemia. Plus those rats are given some crazy doses. The only thing that scares me is the mutagenic study at the end. Ow, my DNA.

Exactly what I was going to write, but you beat me to it. Also, despite the methemoglobinemia and some increase in cancers, the rats lived as long as the controls, but the mice lived longer than controls in a dose-dependent manner. The fact that it was dose-dependent is a good indicator that the drug is responsible. If only mice were miniature humans, despite the number of people I've met in the corporate world who are rats... But that's a really interesting outcome. Since the Ames redox-cycling mitochondrial assist mechanism is supposed to only work at low concentration, this mouse result is presumably due to some other mechanism, unless their mitochondrial concentration is very low for some reason.

You really don't need to worry about the in vitro mutagenicity experiments at the end of the abstract. They are trumped by the in vivo work. OTOH, you might want to worry at least a little about the increased cancers they saw in the rodents. The in vitro assays are indicators that you might have something to worry about in vivo. Rodents are cancer-prone, so they are indicators that you might have something to worry about in humans. MB is the right size and shape to be a DNA intercalator, but note the crazy high dosages being used. Don't forget the mice lived longer.

<snip>
Survival of dosed male and female groups exceeded that of the vehicle controls in a generally dose-related manner. Mean body weights of dosed female mice began to increase after weeks 29, 61, and 85, reaching final values that were 113%, 111%, and 106% of vehicle controls for the 2.5, 12.5, and 25 mg/kg groups, respectively. Dosed mice developed methemoglobinemia and a regenerative Heinz body anemia. The incidences of carcinoma and of adenoma or carcinoma (combined) of the small intestine occurred with a positive trend in males. The incidences of malignant lymphoma occurred with a positive trend in females, and the incidence in 25 mg/kg males exceeded the historical control range. The incidences of hematopoietic cell proliferation of the spleen were significantly increased in 12.5 and 25 mg/kg males and in 25 mg/kg females. The incidences of inflammation of the nose were significantly increased in 12.5 and 25 mg/kg females. GENETIC TOXICOLOGY: Methylene blue trihydrate was mutagenic in Salmonella typhimurium strains TA98 and TA100 with and without rat or hamster liver S9 activation enzymes; mutagenicity was also observed in Escherichia coli strain WP2 uvrA/pKM101 with and without rat liver S9. In cytogenetic tests with cultured Chinese hamster ovary cells, methylene blue trihydrate induced sister chromatid exchanges and chromosomal aberrations with and without S9.
</snip>

*EDIT: I'm retarded. Those doses are insanely high. I was looking at 25mg/kg and skipping over the '/kg' part.

Edited by MrHappy, 13 August 2011 - 02:55 PM.

[thedevinroy]

Their lowest dose, 2.5mg/kg ends up being 20.268mg for humans after allometric scaling (using 0.02kg mouse, 60kg human, 0.75 exponent). http://home.fuse.net.../allometry.html

Heh, I might cut back a little bit. I take more than that a day, and the mice were only fed it for 5 days a week, not 7. Since I got a good effect from only around a drop, I figure 8 drops in the morning will last me all day.

[rwac]

Their lowest dose, 2.5mg/kg ends up being 20.268mg for humans after allometric scaling (using 0.02kg mouse, 60kg human, 0.75 exponent). http://home.fuse.net.../allometry.html

Well, humans certainly don't develop Methemoglobinemia at ~20 mg, so I would argue against its being equal in effect to 2.5mg/kg in mice. In fact humans are at risk for worsening methemoglobinemia only at 5 mg/kg http://books.google...... blue&f=false

Although it is possible that humans have additional enzymes in blood cells mitigating that particular issue.

Edited by rwac, 14 August 2011 - 12:55 AM.

[MrHappy]

Their lowest dose, 2.5mg/kg ends up being 20.268mg for humans after allometric scaling (using 0.02kg mouse, 60kg human, 0.75 exponent). http://home.fuse.net.../allometry.html

Heh, I might cut back a little bit. I take more than that a day, and the mice were only fed it for 5 days a week, not 7. Since I got a good effect from only around a drop, I figure 8 drops in the morning will last me all day.

Since the half-life it 4-5.5 hours, 8 drops in the morning will only last you to lunchtime, I'd think?


The only other thing is that they were mixing MB with methyl cellulose as a delivery vehicle for the rats/mice. Methyl cellulose is made using methyl chloride (hmmm, mighty similar to MB). http://en.wikipedia....ethyl_cellulose
I wonder if there may have been another process occuring between the MB and MC, possibly altering/increasing the dosage.. or changing the composition adversely.

[niner]

Is the "Nootropic Dose" bunk? I realized recently that I didn't know where the 50-60ug dose came from, so I looked into deriving an appropriate dosage. The paper from the Ames lab (Atamna et al.) finds 100 nM to be the optimal concentration. In order to find the mass of drug we should take, we need to know the 'volume of distribution' of methylene blue. This is the hypothetical volume that, if a given mass of drug were dissolved in it, would provide the concentration that we wish to see in plasma. This takes into account all the various compartments in the body that the drug might diffuse into. I wasn't able to find a measured Vd for MB in humans, but I did find a value for dogs, from DiSanto et al. This was 0.876 L kg(-1) If we use this value, which is probably not wildly off, and assume a 65 kg patient, we get 0.876 L/kg * 65kg = 56.94 L. The MB doesn't just go into 5 L of blood, but rather it partitions into all body water, the organs, muscle, and adipose tissue.

Now we compute the moles of MB needed to make a 100nM solution in that volume, and convert to grams:

56.94 L * (100e-9 m/L) * (320g/m) = 0.0018g = 1.8mg.

This is pretty consistent with what people have been discovering on their own. The 50ug dose wasn't working for a lot of people, and they have gravitated toward a milligram or more. This might be the explanation for their observations.

[MrHappy]

Is the "Nootropic Dose" bunk? I realized recently that I didn't know where the 50-60ug dose came from, so I looked into deriving an appropriate dosage. The paper from the Ames lab (Atamna et al.) finds 100 nM to be the optimal concentration. In order to find the mass of drug we should take, we need to know the 'volume of distribution' of methylene blue. This is the hypothetical volume that, if a given mass of drug were dissolved in it, would provide the concentration that we wish to see in plasma. This takes into account all the various compartments in the body that the drug might diffuse into. I wasn't able to find a measured Vd for MB in humans, but I did find a value for dogs, from DiSanto et al. This was 0.876 L kg(-1) If we use this value, which is probably not wildly off, and assume a 65 kg patient, we get 0.876 L/kg * 65kg = 56.94 L. The MB doesn't just go into 5 L of blood, but rather it partitions into all body water, the organs, muscle, and adipose tissue.

Now we compute the moles of MB needed to make a 100nM solution in that volume, and convert to grams:

56.94 L * (100e-9 m/L) * (320g/m) = 0.0018g = 1.8mg.

This is pretty consistent with what people have been discovering on their own. The 50ug dose wasn't working for a lot of people, and they have gravitated toward a milligram or more. This might be the explanation for their observations.

That's pretty similar to the calculations I have read on a few other papers and threads. 1.15mg should be around 130nM, which allowing for absorption rates should be pretty close. I want to check on leucomethylene blue..

[maxwatt]

Is the "Nootropic Dose" bunk? I realized recently that I didn't know where the 50-60ug dose came from, so I looked into deriving an appropriate dosage. The paper from the Ames lab (Atamna et al.) finds 100 nM to be the optimal concentration. In order to find the mass of drug we should take, we need to know the 'volume of distribution' of methylene blue. This is the hypothetical volume that, if a given mass of drug were dissolved in it, would provide the concentration that we wish to see in plasma. This takes into account all the various compartments in the body that the drug might diffuse into. I wasn't able to find a measured Vd for MB in humans, but I did find a value for dogs, from DiSanto et al. This was 0.876 L kg(-1) If we use this value, which is probably not wildly off, and assume a 65 kg patient, we get 0.876 L/kg * 65kg = 56.94 L. The MB doesn't just go into 5 L of blood, but rather it partitions into all body water, the organs, muscle, and adipose tissue.

Now we compute the moles of MB needed to make a 100nM solution in that volume, and convert to grams:

56.94 L * (100e-9 m/L) * (320g/m) = 0.0018g = 1.8mg.

This is pretty consistent with what people have been discovering on their own. The 50ug dose wasn't working for a lot of people, and they have gravitated toward a milligram or more. This might be the explanation for their observations.

But does MB diffuse evenly through all tissues? I.e, if more concentrated in brain, we could be overdosing our neurons at 1.8mg.

[MrHappy]

Is the "Nootropic Dose" bunk? I realized recently that I didn't know where the 50-60ug dose came from, so I looked into deriving an appropriate dosage. The paper from the Ames lab (Atamna et al.) finds 100 nM to be the optimal concentration. In order to find the mass of drug we should take, we need to know the 'volume of distribution' of methylene blue. This is the hypothetical volume that, if a given mass of drug were dissolved in it, would provide the concentration that we wish to see in plasma. This takes into account all the various compartments in the body that the drug might diffuse into. I wasn't able to find a measured Vd for MB in humans, but I did find a value for dogs, from DiSanto et al. This was 0.876 L kg(-1) If we use this value, which is probably not wildly off, and assume a 65 kg patient, we get 0.876 L/kg * 65kg = 56.94 L. The MB doesn't just go into 5 L of blood, but rather it partitions into all body water, the organs, muscle, and adipose tissue.

Now we compute the moles of MB needed to make a 100nM solution in that volume, and convert to grams:

56.94 L * (100e-9 m/L) * (320g/m) = 0.0018g = 1.8mg.

This is pretty consistent with what people have been discovering on their own. The 50ug dose wasn't working for a lot of people, and they have gravitated toward a milligram or more. This might be the explanation for their observations.

But does MB diffuse evenly through all tissues? I.e, if more concentrated in brain, we could be overdosing our neurons at 1.8mg.

From what I've read so far, different health benefits were received from different dosages. The blood/brain barrier should mean that less is absorbed in the brain than other organs. I'm going to test the calcium ascorbate + MB (=leucomethylthionine chloride) and see if there is a perceived change in effects.

[rwac]

It is highly likely that variability in oral absorbtion is determined largely by the efficiency of initial reduction in the GI tract. One way to achieve more reliable absorbtion is therefore be to pre-reduce methylene blue with ascorbic acid. We have shown from in vitro studies that this conversion is rather slow, so that it takes 3 hours to achieve 90% reduction of methylene blue in water in the presence of 2× mg ratio of ascorbic acid. Therefore, the dosage of methylene blue which is most likely to ensure reliable absorbtion will be 3.5 mg/kg/day of methylene blue pre-reduced for at least 3 hours in the presence of 7 mg/kg/day of ascorbic acid.

This means that (according to them, anyway) if you want whatever improvement ascorbate can give you, you can't just swallow a vitamin C tablet at the same time you take the MB. You need to pre-incubate the two compounds in solution for several hours first. Considering the small doses that we're using, we could probably use a large excess of ascorbate to accelerate the reaction. To be honest, I'm kind of baffled that this should take any time at all; this sort of redox reaction is usually very fast. Also, I would think that if you see the color change, then it's done. Note that these guys are trying to get a patent on the use of leucomethylene blue as a therapeutic agent, so they are probably inclined to make it sound tricky in order to flummox the hapless patent examiner. If someone happens to have some MB and some ascorbate handy, mix a little together and let us know what happens.

There's another catch though. Leucomethylene blue is only soluble in acidic solutions (pH below 4.5ish?). So you may need to use ascorbic acid as opposed to ascorbate.
I wonder if putting a thin layer of ascorbic acid at the bottom of the diluted methylene blue solution would keep the pH sufficiently low.

http://jcs.biologist...26/113.full.pdf
page 5

Edited by rwac, 16 August 2011 - 07:08 PM.