11 replies to this topic

[rwac]

Methylene Blue

XMRV in MCF-7 (Breast Cancer Cell Line)

EC50 = 0.42 uM
EC90 = 2.7 uM
CC50 = 7.2 uM

HIV-1 in PBM (Peripheral Blood Mononuclear cells)

EC50 = 0.024 uM
EC90 = 0.5 uM
CC50 = 6.1 uM

I'm not sure exactly why someone would ignore great results like this.
Well, it looks great on the surface at least.

Methylene Blue even has a high EC50/CC50 ratio.

Raltegravir Is a Potent Inhibitor of XMRV, a Virus Implicated in Prostate Cancer and Chronic Fatigue Syndrome

http://www.plosone.o...al.pone.0009948
(look in figure 1)

Edited by rwac, 04 January 2011 - 03:03 PM.

[Rational Madman]

Methylene Blue

XMRV in MCF-7 (Breast Cancer Cell Line)

EC50 = 0.42 uM
EC90 = 2.7 uM
CC50 = 7.2 uM

HIV-1 in PBM (Peripheral Blood Mononuclear cells)

EC50 = 0.024 uM
EC90 = 0.5 uM
CC50 = 6.1 uM

I'm not sure exactly why someone would ignore great results like this.
Well, it looks great on the surface at least.

Methylene Blue even has a high EC50/CC50 ratio.

Raltegravir Is a Potent Inhibitor of XMRV, a Virus Implicated in Prostate Cancer and Chronic Fatigue Syndrome

http://www.plosone.o...al.pone.0009948
(look in figure 1)

I'm still open to the notion of using Methylene Blue, but I'm waiting for an attainable pharmaceutical grade that meets my standards for potency and purity. The best candidate, Rember, is still undergoing clinical trials, but if it continues to perform in accordance with its past record and expectations, then I imagine that it will be released into the marketplace within the next few years.

[maxwatt]

Kingherbs, a former sponsor of the Product Discussion forum, has a laboratory grade methylene blue though it's not on their website. You have to ask about it at this point.

[Rational Madman]

Kingherbs, a former sponsor of the Product Discussion forum, has a laboratory grade methylene blue though it's not on their website. You have to ask about it at this point.

That's right, I remember you mentioning this source before. This time, I'll make an inquiry, and see what I find.

[AgeVivo]

urolene blue is an approved drug against some urinary tract issues. It is medical grade methylene blue, used in a weak antiseptic manner. this page describes it along with side effect warnings: http://www.medicinen...ral/article.htm

[Rational Madman]

After making an inquiry to KingHerbs yesterday, I received their thoughtful reply in my inbox. For interested parties, here is the body of the message:

We'll have it up on the Kingherbs.com website shortly. We have 30 ml bottles such that 4 drops from the included dropper., = 0.2 ml provide 60 micrograms of MB. This is the amount calculated to achieve the optimum serum levels as found in Ames' paper for a 70 kg primate, and makes it easier to dose rodent drinking water for testing. Besides the dropper, it comes with a more precise micro-pipette. Price $30 plus $5 for shipping. I can send an invoice now if you want it sooner.

We can also provide methylene blue tri-hydroxide, safe to handle than pure MB. It is laboratory grade. Technically weit cannot be called pharmaceutical grade once the bulk package is opened, unless performed in an FDA registered pharmaceutical facility. It is sterile, though, and of sufficient purity. Price is to be determined, but I think 10 grams of the powder will be $50.

Where are you located?

Richard

Calculating the mg equivalent/per bottle in my head, there should be approximately 9 mg of Methylene Blue for each unit. For the time being, though, I don't want to dive into PubMed, since my need isn't that pressing. But does anyone know of any research that concurs with Ames' conclusions? With the limited information that I have, I would prefer to defer to Maxwatt and rwac on the subject. However, the suggested dosage does strike me as being somewhat low, even though I'm not suffering from any of the features that warranted larger doses in the Rember trials. If my more modest goal is human enhancement and longevity, then I think a daily dosage somewhere in the middle might be more apt, though I don't really have much of a basis for making this conclusion. So there is a question of approach, because I'm wondering if this agent might be able to replace a number of my other mitochondrial supplements, whose prices are predatorilly inflated. Because of this potential, I'm inclined to embark on a one week trial of using much higher than recommended doses, since I can't think of any plausible reason of potentially adverse effects of any significant duration, which anyway still seem to be somewhat in the theoretical plain. But methodological soundness matters, and since I haven't been as dilligent with my health diary and blood tests as of late, I'm wondering if I should monitor health markers for a month with my current regimen, and then next month, drop many of my mitochondrial supplements in place of Methylene Blue? Because of the price, though, a 30 mg/day dose is clearly prohibitive, and ill-advised due to the state of the research. So I think a 1-4 mg/day dose might be more in the ballpark. Alternatively, I could request an assay on the bulk powder, and just prepare the solution myself, which wouldn't be that complicated, and probably not risky in any serious way. Anyway, I'm becoming increasingly intrigued with this agent, and I would have to credit the enthusiasm of forum members for melting my initial skepticism. To save me the time, I'd appreciate .pdfs of any relevant papers, but only if anyone has them on their hard drive, of course. Otherwise, any suggestions would be welcome, and if I decide to begin a trial, I'll certainly keep everyone apprised.

Edited by Rol82, 17 January 2011 - 03:09 AM.

[maxwatt]

A 60 microgram dose is optimum according to Ame's paper (full text free) if we've calculated correctly based on his stated in vitro concentration and our estimated blood volume for a "70 kg primate"; more than twice that amount was said to negate the effects, which were much the same as those claimed for acetyl-l-carnitine (ALCAR): reduced ROS generation. It's cheaper too. Also, more than twice that dose turns ones urine faintly but definitely green. The dose ROL82 is considering would result in intensely green or blue urine. I believe it's been estimated that the low dose is gone from one's system in four hours, so it might be desirable to take it three times a day. I take it in the morning, and sometimes before afternoon workouts and again before bed.

Does it work? I think so, at least as well as ALCAR for exertion at one's anaerobic threshold. But then it could be all placebo effect for both of them. I also hae a number of confounding factors, including other supplements. There is noting I've found in the literature that supports Ames' conclusions at these low doses, and neither is there anything I've found for his earlier findings on ALCAR and R-lipoic acid. I dropped ALCAR when I started using methylene blue.

FWIW, my current supplements are the Imminst designed multi (#3 in the sidebar at the left of this screen) plus additional vitamin D to bring up my levels of D hydroxy 25 toward 40, some extra D2 as MK7, though not consistently. Due to a genetic polymorphism I also take 800 to 1000 mcg of methyl-tetra-hydrofolate.* Other than vitamins, resveratrol (400 mg) in the morning, with a small amount of luteolin (50mg) and recently 500 mg of curcumin - which has a synergistic effect with resveratrol according to many studies. If nothing else, it might prevent cancer of the colon. With this combination, I need less resveratrol to control arthritis symptoms.

I'd give up the methylene blue before giving up resveratrol, but continue to use MB as it seems to help stamina in a different way than resveratrol. Resveratrol improves endurance, MB seems to improve muscle contractile strength with short anaerobic efforts, or for longer periods at one's limit. Again, these are only my impressions. YMMV

Disclaimer: I nagged Kingherbs to formulate this, so I'd have a ready source without having to mix my own from fish-store solutions. I don't think they consider it will be a profit center.

* this form of folate is an effective anti-depressant, at least in those with one or two copies of the MTHFR polymorphism.

[Rational Madman]

A 60 microgram dose is optimum according to Ame's paper (full text free) if we've calculated correctly based on his stated in vitro concentration and our estimated blood volume for a "70 kg primate"; more than twice that amount was said to negate the effects, which were much the same as those claimed for acetyl-l-carnitine (ALCAR): reduced ROS generation. It's cheaper too. Also, more than twice that dose turns ones urine faintly but definitely green. The dose ROL82 is considering would result in intensely green or blue urine. I believe it's been estimated that the low dose is gone from one's system in four hours, so it might be desirable to take it three times a day. I take it in the morning, and sometimes before afternoon workouts and again before bed.

Does it work? I think so, at least as well as ALCAR for exertion at one's anaerobic threshold. But then it could be all placebo effect for both of them. I also hae a number of confounding factors, including other supplements. There is noting I've found in the literature that supports Ames' conclusions at these low doses, and neither is there anything I've found for his earlier findings on ALCAR and R-lipoic acid. I dropped ALCAR when I started using methylene blue.

FWIW, my current supplements are the Imminst designed multi (#3 in the sidebar at the left of this screen) plus additional vitamin D to bring up my levels of D hydroxy 25 toward 40, some extra D2 as MK7, though not consistently. Due to a genetic polymorphism I also take 800 to 1000 mcg of methyl-tetra-hydrofolate.* Other than vitamins, resveratrol (400 mg) in the morning, with a small amount of luteolin (50mg) and recently 500 mg of curcumin - which has a synergistic effect with resveratrol according to many studies. If nothing else, it might prevent cancer of the colon. With this combination, I need less resveratrol to control arthritis symptoms.

I'd give up the methylene blue before giving up resveratrol, but continue to use MB as it seems to help stamina in a different way than resveratrol. Resveratrol improves endurance, MB seems to improve muscle contractile strength with short anaerobic efforts, or for longer periods at one's limit. Again, these are only my impressions. YMMV

Disclaimer: I nagged Kingherbs to formulate this, so I'd have a ready source without having to mix my own from fish-store solutions. I don't think they consider it will be a profit center.

* this form of folate is an effective anti-depressant, at least in those with one or two copies of the MTHFR polymorphism.

I was aware that one of the possible effects of administering a larger dose was a change in the hue of my urine, but I was under the impression that this effect was pretty innocuous. But anyway, there still seems to be quite a wide gap between doses being administered. So before I go forward, I want to get some notion of the optimal dose and plasma level for Methylene Blue, because there's tremendous potential with this agent---given its effects on a number of important correlates of ageing, disease, and individual well-being. If I find confidence raising findings that will permit me to move forward with my experiment, then I'll take the leap----although sometimes I feel like one of the infamous mad scientists, whom subject themselves to a wide range of chemical torture:
http://www.youtube.com/watch?v=U2D-T6O7myQ

[rwac]

Here's the pharmacokinetics of a 100mg oral dose of MB in human test subjects.
 blood-conc.png   56.12KB   45 downloads

a 100mg dose results in a 100nM concentration in blood, which dissipates over a few hrs.
It appears that the intestine releases the methylene blue into the blood slowly, since there's a high conc in the intestine, and fairly low conc in the blood.

Now in rats, the distribution of MB 1 hr after oral administration is as follows, note the gray bars.
 brain-conc.png   98.39KB   45 downloads

Assuming this works similarly in humans brain conc can reach about 10xBlood conc (or more, our brains are more metabolically active than rats' brains)= maybe more than 1000nM for a dose of 100mg, which is too high for our purposes anyway.

I'm trying to get a hold of PMID 5044807 which hopefully will tell us the pharmacokinetics of a 10mg dose.

I really can't remember how we calculated the 60mcg dose anymore ...

I take 1.5 mg right now split into 5(!) doses. My urine is only very lightly green.

Attached Files

•  Pharmacokinetics and organ distribution of intravenous and oral methylene blue.PDF   123.79KB   63 downloads

Edited by rwac, 17 January 2011 - 02:02 AM.

[FunkOdyssey]

Here's the pharmacokinetics of a 100mg oral dose of MB in human test subjects.
 blood-conc.png   56.12KB   45 downloads

a 100mg dose results in a 100nM concentration in blood, which dissipates over a few hrs.
It appears that the intestine releases the methylene blue into the blood slowly, since there's a high conc in the intestine, and fairly low conc in the blood.

Now in rats, the distribution of MB 1 hr after oral administration is as follows, note the gray bars.
 brain-conc.png   98.39KB   45 downloads

Assuming this works similarly in humans brain conc can reach about 10xBlood conc (or more, our brains are more metabolically active than rats' brains)= maybe more than 1000nM for a dose of 100mg, which is too high for our purposes anyway.

What purpose are you using it for? You got me all excited with the first post demonstrating XMRV inhibition, but as I read further it seems like those are impossibly high concentrations. The EC50 is 420nM and you can only reach 100nM in the blood with a 100mg dose? That would probably make the doses you're actually talking about and using inconsequentially small from the XMRV perspective (sad face).

[rwac]

What purpose are you using it for? You got me all excited with the first post demonstrating XMRV inhibition, but as I read further it seems like those are impossibly high concentrations. The EC50 is 420nM and you can only reach 100nM in the blood with a 100mg dose? That would probably make the doses you're actually talking about and using inconsequentially small from the XMRV perspective (sad face).

I'm going more by the HIV numbers. The XMRV numbers are in a cancer cell line(MCF-7) so you can't directly translate them to human numbers, or indeed compare them to the HIV numbers which are in Peripheral Blood Mononuclear cells.

Numbers in other papers disagree, for instance tenofovir was found to inhibit XMRV at comparable concentrations to HIV in this paper(table 1).

Anecdotally, these guys seem to be getting some benefit out of tenofovir at similar doses to HIV treatment (on AZT/raltegravir/tenofovir).

Additionally some of the drugs these guys dismiss as having high EC50s are effective against other MLVs.

In any case, I consider the potential downside to Methylene Blue low.

[stephen_b]

* this form of folate is an effective anti-depressant, at least in those with one or two copies of the MTHFR polymorphism.

Thanks for that. I have one copy myself.