[dr_chaos]

http://www.fasebj.or...stract/22/3/703

Quote

Methylene blue delays cellular senescence and enhances key mitochondrial biochemical pathways
Hani Atamna¹, Andy Nguyen, Carla Schultz, Kathleen Boyle, Justin Newberry, Hiroyuki Kato and Bruce N. Ames Nutrition and Metabolism Center, Children's Hospital Oakland Research Institute, Oakland, California, USA





¹Correspondence: Nutrition & Metabolism Center, Children's Hospital Oakland Research Institute, 5700 Martin Luther King Jr. Way, Oakland, CA, 94609-1673, USA. E-mail: hatamna@chori.org<script type="text/javascript">

Methylene blue (MB) has been used clinically for about a century to treat numerous ailments. We show that MB and other diaminophenothiazines extend the life span of human IMR90 fibroblasts in tissue culture by >20 population doubling (PDLs). 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 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. We propose 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. Mitochondrial dysfunction and oxidative stress are thought to be key aberrations that lead to cellular senescence and aging. MB may be useful to delay mitochondrial dysfunction with aging and the decrease in complex IV in Alzheimer disease.—Atamna, H., Nguyen, A., Schultz, C., Boyle, K., Newberry, J., Kato, H., Ames, B. N. Methylene blue delays cellular senescence and enhances key mitochondrial biochemical pathways.

So what do you think about that?

[Jonathan Rose]

Yeah I heard about this last month. They are trialing a drug of methylene blue here in the uk to treat alzheimer's disease. It is called rember. More information can be found here:

good informative movie from the bbc http://news.bbc.co.u...lth/7532180.stm


Its really cheap and easy to get hold of aswel, a common lab chemical. Any healthy volenteers want to give it ago?

[kismet]

Don't know if you regularly visit the board over at mprize, just wanted to remind, Michael (who else?) posted about rember: http://mfoundation.org/forums/showthread.p...ighlight=rember
However, I haven't seen any in vivo research on methylene blue's life extension capabilities. It may be good for AD and aging of the brain, though.

[AgeVivo]

What do you think about testing it in MPrize@ home?

[geddarkstorm]

Apparently there has been evidence of MB increasing life span in mice, but since we're taking about a much lower dose than that used for most medicinal purposes, it's been largely overlooked.

I take MB and have for about half a year. Considering it has also been shown, at lower than medicinal doses, in rats, to significantly increase learning of all types, and memory, I have to comment subjectively that that apparently has been the same for me. That is incredibly subjective, without stringent memory tests, and I did do a lot of mental exercising which recorded mental parameters (lumosity.com) a few months previously to taking MB, but none after.

This post has been edited by geddarkstorm: 03 February 2009 - 05:03 AM

[niner]

geddarkstorm, on 3-Feb 2009, 01:01 AM, said:

I take MB and have for about half a year.

Geddarkstorm, how much MB do you take, and where do you get it?

[VespeneGas]

Just want to echo niner's query. I eagerly follow your posts (very informative, thank you btw), and am interested to hear that you've noted subjective benefits from MB.

[geddarkstorm]

niner, on 2-Feb 2009, 11:05 PM, said:

geddarkstorm, on 3-Feb 2009, 01:01 AM, said:

I take MB and have for about half a year.

Geddarkstorm, how much MB do you take, and where do you get it?

The ease or difficulty of finding MB seems to vary per region in the country, I'm afraid. For instance, it's sold over the counter at Walgreen's down in Florida as a 2% solution, and I have reports of the same in Kansas. Where I live, I could only find 1% MB at a science/hobby store, so there's a good chance any place like that will have it.

I take ~100 microliters of 1% MB in tea once every morning (I have a syringe, but this is approximately two drops from any standard eye dropper). This gives about ~1mg of MB, which if factoring in an absorbance percent in the gut of about 50% (actual aqueous solution studies have shown it's about 70% absorbed orally in humans, but I like to factor it down a little for reasons I'll explain in a second), gives about 300-500 nM in my blood (assuming the standard human has 5 liters of blood). We want to keep MB around 100 nM in the blood as long as possible, so by taking around 300 or 500 nM, since we have to be aware of absorption and excretion kinetics, that should keep levels around 100 nM for as long as possible without spiking up towards 1 uM where MB's mitochondrial effects are halved. This also should allow proper dosage of MB to filter into peripheral tissues.

Now, the studies done in rats showing significant mental improvements used 1mg/kg per day dosages. When following the recommended conversion to effective human dose from a rat, that gives us about 10mg of MB, or about 3 uM in the blood. That isn't so bad, but that does put us above the best dose that was seen for human cells in the paper the OP listed, by 30x. It could be though that in vivo effective concentrations are lower than in vitro due to the influence of serum agents that may prevent MB entry into cells (and elimination kinetics of course). Yet another reason why I set my limit roughly 3x that of the optimal dose found in the OP's paper.

The absolute minimal dosing scheme for MB is also detailed nicely here, in section [037] of the patent. I fully believe, however, that their dosage is far too low, considering 10 nM of MB did nothing in human cells. Moreover, the patent did not even take into consideration absorption percentages. There's no way 160 ug will get you at a sustained level of 100 nM - you'd be immensely lucky to even spike at that level. Still, the patent gives some nice ideas about dosage schemes as well for those who want to play around with it.

As an aside, I will tell you that the dosage level I recommend may at times, depending on how hydrated you are, slightly discolor the urine with a faint blue/green tint. Going to a 10 mg MB dosage will definitely discolor it, just so no one freaks out when it happens, it is not a bad thing as we'll see below.

On a final note, the amount of MB typically used to fight methylhemoglobinemia is ~240 mg, and according to the NIH toxicological studies, the lowest dose of MB needed to see the very slightest amount of toxicity is ~670mg or so for an average human. So the dosage I'm recommending is ~600x less than the lowest toxic dose, and ~200x less than that used medicinally in humans for over a century. The LD50 of MB in humans should be around 9 grams.

VespeneGas, on 2-Feb 2009, 11:16 PM, said:

Just want to echo niner's query. I eagerly follow your posts (very informative, thank you btw), and am interested to hear that you've noted subjective benefits from MB.

It is hard for me to relay much subjective information, since I started taking quercetin and resveratrol a few months after MB. Nonetheless, I've seen marked improvements in my ability to retain and relate information in presentations and science talks; which has been very nice. There is no way I can say for certain it was MB, especially since my dose is a little lower than that seen for mental improvement in rats. The only other marked affect, which others have reported too, is improved healthy look to the skin, nails, and hair. That's highly subjective, but my skin has also been thoroughly tested by my new kitten, which has been lacerating the heck out of my fiancee, but does considerably less damage to me - my skin is apparently tougher now. But if it's due to MB or not, I cannot say for sure.

This post has been edited by geddarkstorm: 03 February 2009 - 05:26 PM

[100YearsToGo]

You can buy it here:

http://www.amazon.com/NOVALEK-KORDON-Methl...rd_i=B000PDSJ5A

2.303% water solution

This post has been edited by 100YearsToGo: 03 February 2009 - 10:58 PM

[rubegoldberg]

http://www.methylene-blue.com/

Provepharm research results in the first cGMP Pharmacopoeial grade Methylene Blue (Methylthioninium chloride Ph.Eur) obtained using

• New innovative synthetic pathway
  
• Heavy-metal-free process
  
• “Clean chemistry” process involving pharmaceutical grade reagents
  

Provepharm offers Methylene Blue with

• Metal impurity concentrations up to 50 times lower than current pharmacopoeial limits
  
• Full regulatory package in preparation including USDMF, EDMF and CEP including up to 12 months stability data under ICH conditions
  
• Stock build to ensure future reliable supply
  

Provepharm Methylene Blue enables confidence to

• Update and resume commercialisation of existing Marketing Authorisations containing methylene blue as drug substance in Europe
  
• Ensure compliance with world best practice for existing products containing methylene blue as drug substance commercialised around the world
  
• Undertake research for new therapeutic applications of Methylene Blue
  

[resvhead]

rubegoldberg, on 3-Feb 2009, 09:07 PM, said:

http://www.methylene-blue.com/

Provepharm research results in the first cGMP Pharmacopoeial grade Methylene Blue (Methylthioninium chloride Ph.Eur) obtained using

• New innovative synthetic pathway
  
• Heavy-metal-free process
  
• “Clean chemistry” process involving pharmaceutical grade reagents

Provepharm offers Methylene Blue with

• Metal impurity concentrations up to 50 times lower than current pharmacopoeial limits
  
• Full regulatory package in preparation including USDMF, EDMF and CEP including up to 12 months stability data under ICH conditions
  
• Stock build to ensure future reliable supply

Provepharm Methylene Blue enables confidence to

• Update and resume commercialisation of existing Marketing Authorisations containing methylene blue as drug substance in Europe
  
• Ensure compliance with world best practice for existing products containing methylene blue as drug substance commercialised around the world
  
• Undertake research for new therapeutic applications of Methylene Blue

Doesn't look like they will actually sell the stuff to an end user. Have you been able to buy from them?

[niner]

Provepharm raises the specter of heavy metal contamination. Depending on the exact nature of the metal, this may or may not be a problem at the doses being considered here. I lean toward "not", but this isn't the sort of thing that I would want to leave to chance. MB has been used as a human pharmaceutical; would that suggest a clean source? Someone with access to an analytical lab could run a sample of MB for metals, but having this done at a contract lab would run into some money. Another concern, FWIW, is

Quote

MB also induces phase-2 antioxidant enzymes in hepG2 cells.

This could impact the bioavailability of other supplements, but might not be a problem at the low doses being discussed here.

[lynx]

niner, on 3-Feb 2009, 11:37 PM, said:

Provepharm raises the specter of heavy metal contamination. Depending on the exact nature of the metal, this may or may not be a problem at the doses being considered here. I lean toward "not", but this isn't the sort of thing that I would want to leave to chance. MB has been used as a human pharmaceutical; would that suggest a clean source? Someone with access to an analytical lab could run a sample of MB for metals, but having this done at a contract lab would run into some money. Another concern, FWIW, is

Quote

MB also induces phase-2 antioxidant enzymes in hepG2 cells.

This could impact the bioavailability of other supplements, but might not be a problem at the low doses being discussed here.

If it is actually a phaseII inducer that would be good in my opinion, however, very frequently the term inducer is mistakenly used for "substrate".

[niner]

lynx, on 4-Feb 2009, 01:05 AM, said:

niner, on 3-Feb 2009, 11:37 PM, said:

Provepharm raises the specter of heavy metal contamination. Depending on the exact nature of the metal, this may or may not be a problem at the doses being considered here. I lean toward "not", but this isn't the sort of thing that I would want to leave to chance. MB has been used as a human pharmaceutical; would that suggest a clean source? Someone with access to an analytical lab could run a sample of MB for metals, but having this done at a contract lab would run into some money. Another concern, FWIW, is

Quote

MB also induces phase-2 antioxidant enzymes in hepG2 cells.

This could impact the bioavailability of other supplements, but might not be a problem at the low doses being discussed here.

If it is actually a phaseII inducer that would be good in my opinion, however, very frequently the term inducer is mistakenly used for "substrate".

A phase II inducer might be good if you are exposed to carcinogenic hydrocarbons on a regular basis, but if, like most of us, you aren't, then it might be a negative in that it would lead to quicker clearance of a lot of beneficial phytochemicals such as polyphenols, flavanols, etc. People take things like bioperine and quercetin in order to inhibit phase II enzymes. The confusion between inducers and substrates might derive from the fact that many substrates are inducers of the enzyme than metabolizes them. That is a sensible response, since if you have a lot of some xenobiotic chemical exposure, you will want more of the enzyme that gets rid of it.

[frankbuzin]

Here are 2 links for what is advertised as "UltraPure Grade":

http://www.bioexpress.com/index.html?wscde...200212000212260


http://search.anaspec.com/?keywords=methylene+blue (Will have to select country and redo the search)


Those came from the comments that follow this article:

http://pipeline.corante.com/archives/2008/...es_comeback.php

Note comment #61 geddarkstorm

[imhotep]

I tried finding Methylene blue in a Walgreens in Florida, The pharmacist went nuts, she acted like I was trying to buy LSD. Any idea where it is at?

[geddarkstorm]

imhotep, on 4-Feb 2009, 10:24 AM, said:

I tried finding Methylene blue in a Walgreens in Florida, The pharmacist went nuts, she acted like I was trying to buy LSD. Any idea where it is at?

It was reported in the hispanic/ethnic remedy section. I'm not sure where in Florida that was, and since I don't live there, I can't say for sure. MB is used in a prescription drug combination for fighting urinary track infections, from my understanding, which might be why she freaked out, haha.

@niner: I've seen analytical reports on MB purity for the kind sold to consumers, and all the heavy metal levels where well below the strictest limits. Even if some metals like mercury or lead, for the sake of argument, were in the low ppb range, when you are taking just 100 microliters of that solution, you'll only be getting the equivalent of 100 parts per quadrillion amounts; well below detection, and you probably get an order of magnitude or two more than that in your drinking water. Furthermore, MB is used to treat fish of fungal diseases and other pests, and fish are more sensitive to heavy metals than us, from what I know.

My area of research is directly on a mouse/human metal sensor protein (for zinc), which also responds to heavy metals. Our bodies have a lot of heavy metal defenses. Metals just aren't quite as dangerous (nor is radiation) as we treat them, but better safe than sorry!

Still, I completely believe that company is creating a "false dilemma" to get people to buy their radically overpriced product - especially since they claim they are the new, and best process, and heavily imply all other processes are "inferior" and let in a lot of heavy metals. All I can say is, what? I highly, highly doubt that is the case since it's used in medicine, and I'm sure the FDA would throw a fit if everyone else were letting in a substantial amount of metals. After I saw their site, I worried quite a bit about this and went looked around everywhere to investigate, but no where could I, at least, find any evidence what so ever that there are substantial heavy metals in MB synthesis - except maybe the version used as an industrial fabric dye, but then that also has strict heavy metal limits (though not as strict) as you aren't allowed to put lead or mercury in consumer clothing. Getting back to it being used in people for 100 years without an apparent problem, all the way back when we didn't even have real heavy metal standards, and certainly not anywhere near as strict as now, I personally don't worry about this issue unless some reputable source showed otherwise.

Nonetheless, it is something to think about, and everyone needs to be aware.

This post has been edited by geddarkstorm: 05 February 2009 - 05:47 PM

[tham]

You can find methylene blue at aquarium shops.
Not sure of the concentration though.

I used quite a bit during my fish rearing days as a kid.
Dripped in till the aquarium turned all bluish, whenever
the fish fell sick. Never really worked anyway.

[tham]

Looks like 2.3 %.

http://www.americana.../Quickcure.html

http://www.everythingforpets.com/interpet_...6253.dept.1131/

http://www.aquariumg...hyleneblue.html

This post has been edited by tham: 05 February 2009 - 06:35 PM

[lynx]

niner, on 4-Feb 2009, 12:31 AM, said:

lynx, on 4-Feb 2009, 01:05 AM, said:

niner, on 3-Feb 2009, 11:37 PM, said:

Provepharm raises the specter of heavy metal contamination. Depending on the exact nature of the metal, this may or may not be a problem at the doses being considered here. I lean toward "not", but this isn't the sort of thing that I would want to leave to chance. MB has been used as a human pharmaceutical; would that suggest a clean source? Someone with access to an analytical lab could run a sample of MB for metals, but having this done at a contract lab would run into some money. Another concern, FWIW, is

Quote

MB also induces phase-2 antioxidant enzymes in hepG2 cells.

This could impact the bioavailability of other supplements, but might not be a problem at the low doses being discussed here.

If it is actually a phaseII inducer that would be good in my opinion, however, very frequently the term inducer is mistakenly used for "substrate".

A phase II inducer might be good if you are exposed to carcinogenic hydrocarbons on a regular basis, but if, like most of us, you aren't, then it might be a negative in that it would lead to quicker clearance of a lot of beneficial phytochemicals such as polyphenols, flavanols, etc. People take things like bioperine and quercetin in order to inhibit phase II enzymes. The confusion between inducers and substrates might derive from the fact that many substrates are inducers of the enzyme than metabolizes them. That is a sensible response, since if you have a lot of some xenobiotic chemical exposure, you will want more of the enzyme that gets rid of it.

Since fertility rates, sperm motility, testosterone levels have all been dropping like crazy, I assume that everyone has significant exposure.

This post has been edited by lynx: 06 February 2009 - 06:54 PM