46 replies to this topic

[timothymburke]

I was researching today the possibility of buying some older 18 month old mice or shrews to begin feeding the them a diet infused with a telomerase activator on/off regimen every other day. I'd like to know if anyone else is doing similar experimentation on these shorter lived mammals? I've done this on plants and insects, the fruit fly, with good results, but not on a mammal yet, except for my cat, only 5 years old now.

I'm thinking of creating two feed mixes, one Astragalus-Astragaloside, apricot seed, melatonin feed mix then alternating it each day a Clove, Green tea, Goji Berry, feed mix while keeping them in a Schumann Resonance caged area. Such experiments should be easy to do and further elaborate upon. Results would be obvious within less than 2 years since a shrew's average lifespan while in captivity in only 30 months.

I know there's probably only a few thousand people in the whole world on Astragaloside and that only a handful of those are scientists, but hasn't anyone in here tried this experiment yet? I'd like to set it up with a website & 24-hour streaming webcam, the whole 9 yards, if nobody has done it yet.

Leave your comments please. Thanks!

Edited by timothymburke, 09 December 2010 - 06:23 PM.

[timothymburke]

** UPDATE **

After a month and over 100 views, still no takers on this project, so I've gone ahead and ordered the supplies needed to start the experiment myself beginning in January of 2011.

I've decided to start with 2 Russian Dwarf Hamsters; average life span of 2-3 years.
http://www.petsmart....oductId=3893799

I ordered RevGenetics Astral Fruit Telemere DNA support supplement and Manna Bread
http://www.revgeneti...supplement.aspx
http://www.mannaorga...log&categoria=3

I already have apricot seed, melatonin, Clove, Green tea, & Goji Berry.

I also plan to purchase a Geoclense Orgone-Schumann resonance device just for kicks.
http://www.orgoneene...ium-harmonizers

Initial startup costs will amount to at least $500.

It would be nice if I could find a webcam site that pays me for hits to the streaming video or if I could find a sponsor for this research project. If you know of any please post them here.
For now I will probably use livestream with Procaster for the streaming webcam.
http://www.livestrea...tform/procaster

I will keep you updated on my progress...

Edited by timothymburke, 05 January 2011 - 10:23 PM.

[xEva]

That's interesting. But how will you assess the success/failure of your experiment? Will you be able to distinguish Astragalus-Astragaloside influence from other antioxidants/vitamins in the mix? Also, I've read a number of times that rodents (mice for sure) have rather long telomeres to begin with and their great length has no relevance to their usual death of cancer in the end of their rather short lives.

[timothymburke]

That's interesting. But how will you assess the success/failure of your experiment? Will you be able to distinguish Astragalus-Astragaloside influence from other antioxidants/vitamins in the mix? Also, I've read a number of times that rodents (mice for sure) have rather long telomeres to begin with and their great length has no relevance to their usual death of cancer in the end of their rather short lives.

No I won't be able to distinguish the difference with just 2 subjects in the same cage; however, if a year from now I have the time, money, and space to expand my research (and they are still alive LOL) I can always break down each control by adding new test subjects, though I don't really see the point. I believe the problem most labs have is they refuse to test multiple controls at once trying to isolate one single item that will make all mammals live forever and/or kill them faster. No results from testing show that any single element for longevity kills them much faster, yet still no single change makes them live longer than double their normal lifespan, so clearly this is the wrong approach to search for an "unlimited" lifespan. Mammals have very complex needs. You have to provide lots of changing broad input variables to their environment in order for them to live long healthy normal lives.

Such a variable experiment with all possible life extension methods combined more closely resembles the lifestyle I lead, and possibly many of us here in this forum. We don't know which ones to take so we take several each time a new fad says we should add this or that. If the animals die early, then this tells us taking a combination of supplements is bad, though that is extremely unlikely since our body requires a broad range of differentiating supplements to remain healthy.

For example, lets say we feed 1 lab rat pure Goji and another was a control with generic cheap pet food. Clearly the Gojo rat would die from cancer faster even though it might have a diet higher in anti-oxidants simply because it would be lacking the broad range of vitamins it needs to sustain healthy DNA replication, or might even have excessive diarrhea. Another example, lets say we expose 1 lab rat to cell phone waves and the other to Schumann, maybe the Schumann will live longer, but not much longer than any average control because I seriously doubt frequencies alone have that much positive prevention to cancer, plus there would be little difference for the average life span because it's natural lifespan will kick in without any other major change in their diet, so we definitely won't achieve a 300% or better lifespan with any single simple change in control.

Now let's combine them. There is no need for a control because that has been done. There is also no need for tests on individual supplements either, because we know those all yield negligible results. The only thing left is the ultimate combination of them all but not to any dangerous toxic extreme level, similar to that of our typical diverse lives, but using exclusively healthy and diverse life extending inputs. The Schumann provides healthy frequencies, the supplements also provide ultimate nutrition, the diverse diet and lack of excessive extremes prevents dangerous and harmful effects other lab animals wouldn't withstand. It stands to reason that this is the only relatively true test that can be done to attempt longevity with the information we have today since we know these things aren't harmful individually, but we don't know their effects when they are all combined since any good scientist is taught from the beginning to put as many controls on an experiment as possible thus making it almost completely irrelevant to the average human lifestyle, which is quite chaotic/variable to say the least...

I have already been taking all these supplements and many more for over 5 years now, so this a test to see how my life (our lives) might turn out if I (we) were to continue this pattern for an entire lifetime to an utmost extreme. If the animals die early, then we are screwed. If they live over 3 years, then I have no reason to stop. If they live close to 6 years, then I shouldn't stop; anything over 6 years, then we are approaching an "unlimited" lifespan pattern to follow and further testing should continue with thousands of animals to verify.

I'm still waiting on the supplies I ordered to arrive. I doubt I will get running until after the 15th of January. Also I have cold, probably bird flu, so I don't want to bring the animals in until I get well, otherwise I might make them sick from the beginning, then again, we get sick, so maybe I should let them catch it too, to create the best chaotic control environment possible. So you see, the more restrictions/controls you put on a test like human longevity, the less relevant are your results, so contrary to what you might think, more variables are better in this case quite the contrary to black and white "by the book" suggestion for less broad experimental testing. Such extreme environments don't exist in real life and are clearly dangerous pointless situations you will never see outside the lab, we can't live by them and survive, so there is no need to bother.



I'm thinking of USTREAM for the steaming night vision webcam. I hear it pays and is easy to use.

Edited by timothymburke, 09 January 2011 - 06:05 AM.

[xEva]

Well, Timothy, good luck in your experiments and keep us posted on your progress

[timothymburke]

Thanks Xev! I will keep this thread up-to-date so that hopefully soon we'll be like the Brunnen-G and live forever!

[timothymburke]

Thanks Xev! I will keep this thread up-to-date so that hopefully soon we'll be like the Brunnen-G and live forever!

Still waiting on supplies to arrive. They are coming in from the UK, no tracking, it may be next month before they get here... Spent $750 already...

[AgeVivo]

hi Timothy
similarly to you, I started Mprize at home: http://www.imminst.o...474-mprizehome/

a few persons showed their interestt, but the project was somewhat abandonned (i remained the only one to test, a little like you). one reason is that we need to be quite a lot for it to have an interest, another one is that it is not legal to raise "lab mice" like C57BL6 so we end up using petshop mice whose lifespan may differ from one petshop to another, another one is that caliban felt particularly concerned about longecity organizing such a thing (without that last one I think it would have worked). on the mprize at home thread you can see that my cost is much smaller than what you have (i had even overestimated the long term costs in the thread), so even students should generally afford it.

I had chosen to test methylene blue; the Intervention Testing Program decided to test methylene blue so my test became definitely useless. I kept only one mouse. My mouse alternates drinks: tap water, dilluted methylene blue, and some water-cleaning solution for fish. it is now still alive and healthy after more than 2 years, still very active and doesn't look old. I don't think it means much right now, if it is still alive in 6 months or even one year it will mean something to me, despite the n=1.

don't hesitate to PM me if you wish. So far I'm not convinced by the probability that the compounds and treatment you indicate shall extend lifespan, but if you convince me to test your thing by having 2 cages and 2 mice per cage, i may accompany you, and perhaps others as well. but i don't think we will be a lot.

in theory if every one in this forum was participating with care and in a coordinated way then we would get quite a lot of results every year. the main trick to do that is to choose a webplace where people send picture of their animals each time they change the cage. but i am really not convinced anymore that a lot of people would follow, because having an animal at home is "borderline" in terms of acceptance, especially if testing things on it (even food & drink), so with such a webplace perhaps some animal defensors could make some noise. stupid noise i think because the animals are well care as the goal is to extend their life and as the webplace allows some pretty good check that cages are replaced often enough and that animals are well, but if only one person makes such noise i don't think there is much to argue against. yes, non-scientific fear sometimes destroys innovation, that's a shame...

Edited by AgeVivo, 29 January 2011 - 09:30 PM.

[DeadMeat]

I had chosen to test methylene blue; the Intervention Testing Program decided to test methylene blue so my test became definitely useless. I kept only one mouse. My mouse alternates drinks: tap water, dilluted methylene blue, and some water-cleaning solution for fish. it is now still alive and healthy after more than 2 years, still very active and doesn't look old. I don't think it means much right now, if it is still alive in 6 months or even one year it will mean something to me, despite the n=1.

What's the water cleaning solution for fish and what's it for? Or do you mean that you use that as a different source for the methylene blue?

And I don't think the NIA test makes your test useles. If I didn't mess up the calculations, they seem to be using way to high dosages of methylene blue to be relevant for lifespan. Or at least not relevant for the 100 nM target.

http://en.wikipedia.org/wiki/Mouse

Food intake is approximately 15 g (0.53 oz) per 100 g (3.5 oz) of body weight per day; water intake is approximately 15 ml (0.53 imp fl oz; 0.51 US fl oz) per 100 g of body weight per day.[4]

http://www.nia.nih.g...dsInTesting.htm

They are using 28 ppm methylene blue in food. So 28 mg/kg food. And methylene blue is 319.85 g/mol so this is 87.5 µmol/kg food. And this should then be sort of similar to 87.5 µM in drinking water. And 4.2 mg/kg body weight.

[timothymburke]

"they seem to be using way to high dosages of methylene blue to be relevant for lifespan"

They always seem to give way too much to the mice when testing, but I get caught up in the calculations myself.

Hamsters weigh roughly 365 times less than us but die 35 times faster. Does that mean dosages should be 10 times our daily dosage divided by 365?
We don't eat or drink our weight in food and water everyday like they do. Something here doesn't add up. HOW DO laboratories calculate their dosage?

[timothymburke]

hi Timothy
similarly to you, I started Mprize at home: http://www.imminst.o...474-mprizehome/

I had chosen to test methylene blue...

... to test your thing by having 2 cages and 2 mice per cage, i may accompany you, and perhaps others as well. but i don't think we will be a lot.

...having an animal at home is "borderline" in terms of acceptance

Ok, I got caught up around the house and found some time toget back to this experiment…

Looking at the average weight of a hamster, from 90 to 120grams, and the average weight of a human, from 140 to 180 pounds, I have calculated the daily dosage dilution needed by body weight for a typical hamster.

160lbs/105g = 72,575g/105g = 691

So 1 day's recommended dosage for us lasts 691 days for a hamster.

I will have 2 in a cage, meaning it will last roughly 346 days. (close to 365)

A 105g hamster drinks roughly 15 grams/ml of water a day.

2 hamsters make it 30 grams a day for 346 days.

30g X 346days = 10,380g or 10.38 liters.

So I can dilute 1 daily dosage across 3 three liter bottles.(1.38 liters more concentrate won't make that much of a difference (~12%) since we can assume some will be lost to the cage floor.) The issue here is that some supplements shouldn't be taken with others since they cancel each other out, such as Anti-Oxidants stopping Cycloastragenol Telomerase Activators, so I will need to split these 2 apart into separate days. The end result would be 1.5 three litter bottles with Telomerase non-conflicting supplement dilutions and 1.5 three little bottles with the other supplement dilutions, then changing their water every other day. I have ½ oz (15 gram) dropper bottles that I will use to draw out 30 grams of water to fill and monitor their water usage eachday.)

Does this sound about right? Should I multiply dosage by 10 since they die 10 times faster than us? (I wish I had the space for 2 cages too. Then I could test both!)

I'm still gathering more items to add to the supplement mix. I'm considering buying the Regeneration Bundle from zptech.net to get some M-state elements added to the regimen for validation. That's another $230 though to add to what I've already spent bumping costs to over $1000. I've already looked into methylene blue, my 4 inch goldfish lived 10 years on that alone, but it can be toxic, I'll look back at it again.

These rats better appreciate what I'm doing for them, living the high life! (I won't be giving them anything I'm not takingmyself at the same time.) That's ok I'm broke though, I'll save up money for a while because I need to hold off until I can build some special shelving into my room to place their cage upon and supplies without being in my workspace but still at a place where I can mount the webcam to see around them but you won't have to see me in my boxers. (yes, we will share the same living area too.)

Remember my test is only for the combined effect of all the best known elements for extended lifespan when taken together, quite different from individual testing of any 1 single supplement. I don't consider this borderline since I'm taking these supplements myself. They are all already approved for consumption. You can get most my supplements off of my makeshift website: neverdie.weebly.com. It consists of over 100 supplements and their daily amount. I only add/change depending on how tight money gets. Roughly $12 a day is a lot to spend on supplements!

I'll keep updating when I get the time.

Edited by timothymburke, 22 February 2011 - 07:58 PM.

[timothymburke]

My dosage calculations for a hamster appear to be right according to the FDA but according to common sense I should multiply it by 6.6!


FDA Dosage Calculator for Humans vs. Animals: http://www.accessdat...animalquery.cfm
Hamster Weight : 105g
Human Weight : 70kg
FDA at 1000mg (14.28mg per kg) for a 70kg daily human dosage yields 1.5mg for a hamster giving a divisor of 666. (very close to my estimate of 691)
Divide by 2 for 2 hamsters and get 1/333 dosage in the water each day, so 1 day is nearly a full year's supply, the same results I had.

The problem here is metabolism and lifespan!

We eat 5% of our body weight per day; they eat 33% (multiplier of 6.6).

We live 70 years; they live 2 years (multiplier of 35).

They weigh 105g; we weigh 70kg (multiplier of 666).

I'm thinking we could increase their dosage by 33% / 5% or by a multiple of 6.6 to account for the high metabolism? (but this ignores the relational life span multiplier.)

But if we allow food intake to remain stay high, organ breakdown will be faster than our own, as it is already 10 times faster, so slowing their metabolism or calorie restriction makes sense too. I may have to meet in the middle - calorie restriction - hoping they eat half as much food since they won't need it with all those supplements then also double the dosage? (I'm assuming they won't be fat, so in the end they may end up lower than average rate, yielding a relative 3.3 multiplier (double dose, 66% weight, 33% less intake). This would potentially double the equivalent of the physical limitations of their bodies already, simply because of the lower stress put on their organs over the years from calorie restriction combined with their extremely healthy diet & weight.

Does this sound reasonable?

I couldn't find any human relational dosage recommendations in any of the forums for mice, shrews, or hamsters....
It could possibly be 1/666th, 1/66th, 1/35th, 1/100th and/or any combination of these....
I don't trust the FDA on this! It makes no sense at all.

[maxwatt]

There are two methods used; one scales by inverse cube of body weight to attempt to compensate for the metabolic difference in energy consumption; a larger body size retains heat more efficiently. Another method attempts to scale by surface area. But in either case, the "equivalent" is a first approximation, used for initial human doses to avoid toxicity. Difference in biochemistry can overwhelm this factor. But the FDA recommendation is a good starting point.

[timothymburke]

OK, I saw something I missed, it looks like they meet in the middle as well, at less than 6.6 with a factor of 4.1 for hamsters and 3 for mice. This may be why most people consider the test dosages to the way too high but the FDA does not and neither do I when you consider the small animals are dying a lot faster than we are because of their high metabolism due to their small size.

FDA Recommended Human to Animal Dosage Conversion Website
"the FDA recommends the following conversion factors: Mouse = 3, Hamster = 4.1, Rat = 6, Guinea Pig = 7.7.... ... Multiply the conversion factor by the animal dose in mg/kg to obtain the dose in mg/m2 for human dose equivalent."

This may not work though, by the time I dilute all these supplements into the water they may not even want to drink it.

[timothymburke]

After careful consideration I will be adding methylene blue to the combination. Apparently it "can be" toxic according to the FDA but only because it "may in some cases" make cyanide poisoning worse and increase cyanide in the blood when it is being used to treat cyanide poisoning. Plus now I recall some pranksters squirting in quite a bit in a drink to turn my buddies pee blue back in college and don't recall any side effects, except for a few laughs.

Are you taking it orally already? If so, is your pee blue?

Bitter Almonds (or Apricot Seed) are said to contain high levels of a cyanogenic glycoside called amygdalin.
Could laetrile (or vitamin B17) containing items combined with methylene blue induce cyanide poisoning?
(I have been eating them for years and haven't died any times that I can recall. LOL)

Laetrile was also once used as a cancer curing agent but once the FDA found the a potential relation to cyanide the sale of apricot seeds were banned in the USA. (I think it has more to do with the fact that eating small numbers of apricot seeds, 8 a day, were a cheap & easy way to avoid buying all those expensive cancer treatments that drive our economy today.) Cherries and regular almonds also have amygdalin, they taste good, and we eat large numbers of them often but they didn't make those illegal when they found out. The law was just silly and is ignored today in the USA, so apricots as well as their seeds are pretty easy to find for sale.

The cancer preventative properties of laetrile and methylene blue may be related.

Edited by timothymburke, 28 February 2011 - 07:27 PM.

[AgeVivo]

here is a post about the methylene blue dosage i'm using: http://www.longecity...post__p__362529

[AgeVivo]

What's the water cleaning solution for fish and what's it for? Or do you mean that you use that as a different source for the methylene blue?

To get methylene blue I had first bought solution for fish containing methylene blue (in fact it contains other things as well and as a result is not only blue but also greenish). When I saw that the MPrize at home test wasn't going on I decided to take care of my mouse in a more flexible manner and I thought it was worth using that solution from time to time (2 drops in the big water bottle during one week every other month approximately; by eye I'd say it is slightly more dilluted than the methylene blue solution I use) to slightly "clean the mouse" just like it does for fish. Btw also I sometimes now give some cheese and my little son plays with the mouse (just saying in case my n=1 experiment finally becomes a great finding, but i really don't think ;-)

[DeadMeat]

What's the water cleaning solution for fish and what's it for? Or do you mean that you use that as a different source for the methylene blue?

To get methylene blue I had first bought solution for fish containing methylene blue (in fact it contains other things as well and as a result is not only blue but also greenish). When I saw that the MPrize at home test wasn't going on I decided to take care of my mouse in a more flexible manner and I thought it was worth using that solution from time to time (2 drops in the big water bottle during one week every other month approximately; by eye I'd say it is slightly more dilluted than the methylene blue solution I use) to slightly "clean the mouse" just like it does for fish. Btw also I sometimes now give some cheese and my little son plays with the mouse (just saying in case my n=1 experiment finally becomes a great finding, but i really don't think ;-)

Thanks AgeVivo. My two pet rats have been getting methylene blue for about three weeks now. Also at 100 nM in their drinking water.

Made by adding 4 drops(drop size: 17 drops in a ml) from a 1% MB solution to 44 ml demineralized water to make a stock solution. And from that stock solution, 2 drops added daily to their 200 ml water bowl to make it a MB concentration of 100 nM.

[timothymburke]

What's the water cleaning solution for fish and what's it for? Or do you mean that you use that as a different source for the methylene blue?

To get methylene blue I had first bought solution for fish containing methylene blue (in fact it contains other things as well and as a result is not only blue but also greenish). When I saw that the MPrize at home test wasn't going on I decided to take care of my mouse in a more flexible manner and I thought it was worth using that solution from time to time (2 drops in the big water bottle during one week every other month approximately; by eye I'd say it is slightly more dilluted than the methylene blue solution I use) to slightly "clean the mouse" just like it does for fish. Btw also I sometimes now give some cheese and my little son plays with the mouse (just saying in case my n=1 experiment finally becomes a great finding, but i really don't think ;-)

Thanks AgeVivo. My two pet rats have been getting methylene blue for about three weeks now. Also at 100 nM in their drinking water.

Made by adding 4 drops(drop size: 17 drops in a ml) from a 1% MB solution to 44 ml demineralized water to make a stock solution. And from that stock solution, 2 drops added daily to their 200 ml water bowl to make it a MB concentration of 100 nM.

So 4 drops of 1% into 44ml where 17 drops is 1 ml:

17 drops X 44 ml = 748 drops
4 drops / (748 drops + 4 drops) = 0.0053191489361702
0.0053191489361702 X 1% = 0.0053191489361702%

Then you add 2 drops into 200 ml:
200 ml X 17 drops = 3400 drops
2 drops / (3400 drops + 2 drops) = 0.0005878894767783657
0.0053191489361702% X 0.0005878894767783657 = 0.00000312707168499129940992157304214%

1% / 0.00000312707168499129940992157304214% = 319788
That's 1 drop of 1% solution added to 319,787 drops of water.

17 drops are 1 ml so...
319,787 drops / 17 drops = 18,811 ml (18.811 liters)

You end up with 1 drop of 1% solution for every 18.811 liters.

My fish solution is 2.303% so I need 1 drop per every 43.321733 liters to match the dosage you stated.

But 1 drop of 1% solution added to 43.321733 liters doesn't equal 100 nm....
0.01 part of MB per 43.321733 liters is 1 part per 4,332.1733 liters.
1 nanomole is 10e-9 molar or 1 part per 10,000,000,000 liters.
100 nanomole would be 1e-6 or 1 part per 1,000,000 liters.
1,000,000 / 4,332.1733 = 230.8310242344183
You dosage appears to actually be 231 times higher than 100 nm?!!

The instructions on the bottle say "add approximately 1/3 teaspoon (1.64 ml) per 10 gallons for each required 1 ppm increase."
1/3 teaspoon (1.64 ml) is 27.88 drops per 37.8541178 L to get 1 part per 10e-5 (1ppm). (1 drop per 1.357751714490674 liters)
We would have to multiply liters by 10,000 to reach1 nanomole or 10e-9(0.0001ppm).
So that amounts to only 1 drop of solution per 13,577.5 liters.


Something has to be wrong otherwise we couldn't even create these kinds of dosages in our own homes.


100 nM is probably way too little, even your dosage at 231 times that is small when compared to what we give to a little fish at 2.788 drops per gallon.
According to the bottle I need to put 2 drops per each 3 liter bottle to reach the recommended dosage if they drink 20 mL per day.
Roughly 6 drops a year. For a human that weighs 365 times more:
6 drops / 4.2 FDA hamster ~ 1.43 drops X 365 mass ~ 521.43 / 365 days ~ 1.42 drops per day for a 170 pound person (assuming a fish has the same multiplier as a hamster.)

I guess though we are assuming that we treat the water, not the fish, so the dosage is only relevant in proportion to how much space a fish would take up in a 10 gallon tank and how much MB the fish would actually absorb from the water. The average goldfish weighs 1.5 grams. 10 gallons of water is 37,854.118 grams. The goldfish is 1/25,236th of the water weight thus it would get a relative dosage?
The fish would "drink/fill" the equivalent of 1 drop per 37,854 liters. Roughly 1/3 of the dosage at 1 drop of solution per 13,577.5 liters or 100 nM.

Can you provide the calculations that you used to determine the proper dosage or is this all just a shot in the dark?

Edited by timothymburke, 08 March 2011 - 06:17 PM.

[timothymburke]

What's the water cleaning solution for fish and what's it for? Or do you mean that you use that as a different source for the methylene blue?

To get methylene blue I had first bought solution for fish containing methylene blue (in fact it contains other things as well and as a result is not only blue but also greenish). When I saw that the MPrize at home test wasn't going on I decided to take care of my mouse in a more flexible manner and I thought it was worth using that solution from time to time (2 drops in the big water bottle during one week every other month approximately; by eye I'd say it is slightly more dilluted than the methylene blue solution I use) to slightly "clean the mouse" just like it does for fish. Btw also I sometimes now give some cheese and my little son plays with the mouse (just saying in case my n=1 experiment finally becomes a great finding, but i really don't think ;-)

Yes, 2 drops per 3 liters every 4 months sounds about right if your fish solution is 2.303% Methylene Blue and you want 1 mouse to intake 100 nM per day, according to my calculations.

Where did you get your dosage calculation from? I'd like to double check my figures.

[DeadMeat]

Can you provide the calculations that you used to determine the proper dosage or is this all just a shot in the dark?

The 1% is a mass percentage(not a mole percentage or a volume percentage). My starting solution is 1% so 10 gram MB/1000 gram water = 10 gram/l. The molar mass of the stuff is 319.85 gram/mol. So the concentration in mole is 10/319.85 = 0.03179 mol/l = 0.03179 M = 31790000 nM

I added 4 drops of this starting solution to the stock solution, so I added (4/17)*10^-3 = 0.000235 l of the starting solution. And in nano mole that's 0.000235*31790000 = 7480 nmol.

The volume of the stock solution is 44 ml = 0.044 l (plus 4 drops but who cares). So the concentration of the stock solution becomes 7480/0.044 = 170000 nmol/l.

I add 2 drops of this stock solution to the final solution, so I add (2/17)*10^-3 = 0.000118 l of the stock solution. And in nano mole that's 0.000118*170000 = 20 nmol.

The volume of the final solution is 200 ml = 0.200 l. So the final concentration becomes 20/0.2 = 100 nmol/l = 100 nM.

Or a bit shorter.
c = 31790000*(4/(44*17))*(2/(200*17)) = 100 nM

Edited by DeadMeat, 08 March 2011 - 10:46 PM.

[timothymburke]

Can you provide the calculations that you used to determine the proper dosage or is this all just a shot in the dark?

The 1% is a mass percentage(not a mole percentage or a volume percentage). My starting solution is 1%
so 10 gram MB/1000 gram water = 10 gram/l.
The molar mass of the stuff is 319.85 gram/mol.
So the concentration in mole is 10/319.85 = 0.03179 mol/l = 0.03179 M = 31790000 nM
I added 4 drops of this starting solution to the stock solution, so I added (4/17)*10^-3 = 0.000235 l of the starting solution. And in nano mole that's 0.000235*31790000 = 7480 nmol.
The volume of the stock solution is 44 ml = 0.044 l (plus 4 drops but who cares). So the concentration of the stock solution becomes 7480/0.044 = 170000 nmol/l.
I add 2 drops of this stock solution to the final solution, so I add (2/17)*10^-3 = 0.000118 l of the stock solution. And in nano mole that's 0.000118*170000 = 20 nmol.
The volume of the final solution is 200 ml = 0.200 l. So the final concentration becomes 20/0.2 = 100 nmol/l = 100 nM.
Or a bit shorter.
c = 31790000*(4/(44*17))*(2/(200*17)) = 100 nM

2.303% solution "add approximately 1/3 teaspoon (1.64 ml) per 10 gallons for each required 1 ppm increase."
My starting solution is 2.303% so
23.03 gram MB/1000 gram water = 23.03 gram/l
"molar mass of the stuff is 319.85 gram/mol. So the concentration in mole is"
23.03/319.85 = 0.0720025011724246 mol/l = 0.0720025012724246 M = 72002501 nM per liter
20 drops = 1mL, 1 liter = 20000 drops, 72002501 nM / 20000 drops = 3600.125 nM per drop
3600.125 nM divided into [(35 water drops) + (1 MB drop)] = 100nM per drop
So to get 100 nM per drop I need to add 35 more drops of water then add 1 drop of that to the water each day.
20000 / 35 = 571 drops added to 1 liter for a mouse.

571 drops is 28.57142857142857 ml
1 US fluid ounce = 29.5735296 ml
4 ounces are in the bottle
So if I pour 2 ounces of 2.303 solutions into a 2 liter bottle (and the other 2 ounces in another 2 liter bottle)
Then you have your solution for a mouse.

Your figure is very roughly 5/34 but at 1% instead of 2.303% so 5/2.303 = 2.17/34 or ~ 200nM per drop
A mouse drinks from 20 ml(g) to 100ml(g) a day so adding 200 ml to your 2ml (37drops) provides enough water for a mouse to last 2 days yielding no less than 100nM per day.

This strong a concentration would surely turn us both myself and the poor mouse BLUE! They are blue right?!
A human would have to drink like 8 liters a day of this blue stuff or increase the concentration by 4 times to only drink 2 liters.
That's way too much, it would look like blueberry cool-aid LOL
1 drop a day in 2 liters of water is already giving me a bad headache so there is no way this can be right.

[DeadMeat]

2.303% solution "add approximately 1/3 teaspoon (1.64 ml) per 10 gallons for each required 1 ppm increase."
My starting solution is 2.303% so
23.03 gram MB/1000 gram water = 23.03 gram/l
"molar mass of the stuff is 319.85 gram/mol. So the concentration in mole is"
23.03/319.85 = 0.0720025011724246 mol/l = 0.0720025012724246 M = 72002501 nM per liter
20 drops = 1mL, 1 liter = 20000 drops, 72002501 nM / 20000 drops = 3600.125 nM per drop
3600.125 nM divided into [(35 water drops) + (1 MB drop)] = 100nM per drop
So to get 100 nM per drop I need to add 35 more drops of water then add 1 drop of that to the water each day.

You are confusing 100 nM with 100 nmol. The M stands for the unit of concentration: mol per liter(mol/l). Saying that something is 100 nM per liter is nonsense because that would mean it would be 100 nmol per liter per liter.

There is a tiny little factor of 20000 between saying a drop contains 100 nmol and saying that the drop has a concentration of 100nM. It should be:

My starting solution is 2.303% so
23.03 gram MB/1000 gram water = 23.03 gram/l
"molar mass of the stuff is 319.85 gram/mol. So the concentration in mole is"
23.03/319.85 = 0.0720025011724246 mol/l = 0.0720025012724246 M = 72002501 nmol per liter
20 drops = 1mL, 1 liter = 20000 drops, 72002501 nmol / 20000 drops = 3600.125 nmol per drop
3600.125 nmol divided into [(35 water drops) + (1 MB drop)] = 100 nmol per drop
So per drop I need to add 35 more drops of water then add 1 drop of that to 1 liter of water each day to get a concentration of 100 nmol per liter which is 100 nM.

And then you have 1 liter water with a concentration of 100 nM. Which is as good as colorless. And my rats get 100 nM as their drinking water(they don't get 100 nmol through their drinking water if thats what you were thinking).

[timothymburke]

Yes, that's what I was thinking exactly! Now I see what you ware saying. Basically you have 1 liter of water. That water is 100nM of blue and you give them that water each day.

The measurements would be easier for us to make if they were worked out into drops per liter instead of nM. I just want to take an empty 2 or 3 liter bottle of cola, fill it with water, then add the proper number of drops and refrigerate to have their water for a whole 3 months prepared. I won't have the time, patience, or accuracy to keep measuring out drop per drop like you are every day or couple of days just to get their water right.

ok so...
~0.0720025012 moles is a liter of ~72,002,501.2 nanomole.
1 liter = ~20000 drops.
~72,002,501.2 / ~20000 = ~3600.12506 nM.
If I add 1 drop of 2.303% solution to 1 liter of water I get a solution of ~3600.12506 nM.
but I can't add 36 liters of water to 1 liter of 2.303% solution since I don't have that much solution or space.
so I'll add 36 drops to 1 drop of the 2.303% solution instead...
My dropper bottles are 1/2 oz.
1/2 fluid ounce = 14.7867648 mL
4.7867648 mL = 295.735296 drops in my medicine dropper bottle.
Nice! That will last a while.
I will count out 36 drops of water then add 1 drop of 2.303% solution.
If I do that 8 times the 1/2 oz medicine dropper will be full 8 X 37 drops = 296 drops or 1/2 oz.
Now I just have to add 1 drop from this medicine dropper to every 1 liter of water to get a 100nM solution.
I have 3 liter bottles, so with 3 drops, I've got their water prepped for months.

I've calculated that a 70 kg human needs about 365 times more than an average mouse.
Mice have a conversion factor of 4.2 times more than normal for their weight when compared to humans because of a higher metabolism.

The mice drink from 20mL to 100 mL each day.
20 mL * 365 / 4.2 = 1.738 liters
so easy, I can add 1 drop from the dropper mix to my 2 liter bottle of coke, drink it all day, and i will definitely get higher concentration than the mouse.
100 mL * 365 / 4.2 = 8.69 liters
I'm not gonna drink that much water in a day, but maybe a really huge sumo person can, or a really huge sumo mouse could do the equivalent. The FDA conversion factors and estimated mouse drinking rates create a huge margin of error as much as 500% too much when getting human dosages from that of mouse experimentation results.

In order to make sure the mouse gets at least as much we will then no less than 1 drop and up to 5 drops from the dropper can be added to every 1 liter of water. When you are filling the dropper, this would mean a minimum of 1 drop of blue with 36 drops of water or up to 5 drops of blue with 36 drops of water until the dropper becomes full. After this dilution step you can add 1 drop from the blue/water dropper mix to 1 liter of water to get the mouse dosage, but if you add 5 drops of blue to 36 drops of water then DO NOT use this to create the drinking water for yourself. 1 drop of blue with 36 drops of water, then this added to 1.738 liters of water to be drank throughout the day is already the maximum dosage for a human to definitely obtain the same 100nM proportional dosage equivalent throughout a day that the mice will be getting.

So that's why when I added 1 drop of blue 2.303% to 2 liters of drink I got a headache because my mind ran in circles, it was because I was taking 36-37 times more blue than was recommended, but even diluted across 36 drops of water, it could still be 5 times more than what a smaller mouse gets, but will be fine, I can just stretch the 2 liter out over a day every now and then.
I will try the new watered down dosage and see how it goes.
If it doesn't bother me then I will add 5 drops to 36 drops of water, label as FOR MOUSE ONLY, then dilute in 1 liter to be sure the mouse gets exactly the same relevant amount as me according to the FDA website for human to hamster conversion dosages.

Edited by timothymburke, 18 March 2011 - 10:47 PM.

[timothymburke]

The easy way:

1. Put 8 drops of 2.303% methylene blue into a 1/2 ounce dropper bottle using the medicine dropper.
2. Fill the rest of the dropper bottle with water to the 1/2 ounce line on the side.
3. Add 1 drop of the mixed solution you made in the dropper bottle to 1 liter of water to get a 100 nM solution.

If a mouse drinks 100g from this 1 liter of water they may get a maximum of 100 nM for the day.
Rodent daily water requirement > 20 ml < 100g per day.

If a human drinks 1 liter of this 1 liter of water they will get a minimum of 100 nM for the day.
Human daily water requirement ~ 8 glasses (2L) per day.

I've been doing this for 3 days now. So far no headaches from excessive thinking this time...
I started at the onset of a sinus infection and sore throat from all this pollen.
The pollen and sneezing is still here but the infection and sore throat is gone.
More testing is still needed...

[maxwatt]

Note: I changed the name of this topic as after the initial posts, the topic changed to Methylene Blue. @Agevivo, keep us posted on you n=1 methylene blue-fed mouse. If it is still alive after six more months, it will mean something to me too. When it does die, you might want to autopsy and count the tumors, or not unusual organ size, lesions or other factors contributing to cause of death.

FWIW, I've been taking 60 mcg MB AM and PM.

[Ark]

I started Methylene Blue last week, I have noticed a few things.

Deeper sleep
super increase in cardio during kickboxing(never had wind like this before)

blue piss (*shrug)

[timothymburke]

I started Methylene Blue last week, I have noticed a few things.

Deeper sleep
super increase in cardio during kickboxing(never had wind like this before)

blue piss (*shrug)

You are joking right?
put 8 drops of 2.303% solution into 1/2 oz of water then only put 1 drop of mixture that into a 2 liter bottle of water to drink throughout 1 day.
No blue pee and no headaches this way.

After a couple of weeks I can take 2-3 drops a day of the watered down mix, but never have blue pee.
Yes, I have noticed I do get winded easier, I wake up sore everyday though (probably since I'm off the micronized creatine) but weight gains are up.
I'll get on my electric shock body mass index scales tonight to see if it is muscle mass, water, or fat I'm gaining.
I would think M-Blue delivers more energy to the brain than the muscles, and the brain burns a lot of energy, so less physical endurance makes sense.
I had very good dream state sleep when taking way too much M-Blue as well. I'm really missing that now...

[timothymburke]

Note: I changed the name of this topic as after the initial posts, the topic changed to Methylene Blue. @Agevivo, keep us posted on you n=1 methylene blue-fed mouse. If it is still alive after six more months, it will mean something to me too. When it does die, you might want to autopsy and count the tumors, or not unusual organ size, lesions or other factors contributing to cause of death.

FWIW, I've been taking 60 mcg MB AM and PM.

I was planning on getting an autopsy, the vet said they don't normally do small animals but a university nearby does it for free plus they will even send it off for me for free so I won't have to travel far.

[timothymburke]

I'm not only giving them M-Blue though. Here is a short list of supplements I will be providing them in their daily water.

|[ 001 ]| Alpha Lipoic Acid 230 mg
|[ 002 ]| Amino Acid Blend: (L-Taurine, L-Glutamine, L-Arginine, L-Valine, L-Cysteine, L-Leucine, N-Acetyl Carnitine Hydrochloride, L-Isoleucine, LCarnitine, L-Methionine) 150 mg
|[ 003 ]| Antioxidant Cardio Blend: (Garlic (Allium sativum) (bulb), Citrus Bioflavonoids Citrus limon) (fruit), Bilberry (Vaccinium myrtillus) (leaf), Elderberry Sambucusnigra) (fruit), Black Currant (Ribes nigrum) (fruit), Grape Seed Extract (Vitis vinifera), Brussels Sprouts Extract, Kale Extract, Cauliflower Extract ,Celery Seed (Apium graveolens), Carrot Extract, Broccoli Extract, Cabbage Extract, Astaxanthin Complex, Lutein, Zeaxanthin, Super Oxide Dismutase, Coenzyme Q-10, Pycnogenol® Pinus pinaster) (Maritime Pine Bark Extract)) 120 mg
|[ 004 ]| Apple Cider Vinegar 600 mg
|[ 005 ]| Apricot Seed, Raw Green 5 a day
|[ 006 ]| Apsirin, Low Dose 81 mg
|[ 007 ]| Astragalus 470 mg
|[ 008 ]| B-12, sublingual 5000 mcg
|[ 009 ]| Biotin 1502 mcg
|[ 010 ]| Calcium(as Calcium Citrate, Calcium Carbonate and Dicalcium Phosphate) 200 mg
|[ 011 ]| Chloride (as Potassium Chloride) 26 mg
|[ 012 ]| Choline Bitartrate 10 mg
|[ 013 ]| Chromium (as Chromium Picolinate) 200 mcg
|[ 014 ]| Cinnamon 1000 mg
|[ 015 ]| Cloves, Fresh Ground (Szygium aromaticum) 450 mg
|[ 016 ]| Coenzyme Q-10 100 mg
|[ 017 ]| Copper (as Cupric Oxide) 2 mg
|[ 018 ]| Creatine 700 mg
|[ 019 ]| DHEA (Dehydroepiandrosterone) 12.5 mg
|[ 020 ]| Energy Blend: (Ginkgo Biloba (Ginkgo biloba) (leaf), Green Tea Extract (Camellia sinensis) (leaf), Cayenne Pepper (Capsicum annuum) (fruit), Spirulina (Spirulina platensis) (plant), Korean Ginseng (Panax ginseng) (root)) 100 mg
|[ 021 ]| Fenugreek (Trigonella foenum-graecum) (seed) 610 mg
|[ 022 ]| Flaxseed Oil (Omega 3-6-9) 800mg-382mg-307mg
|[ 023 ]| Folic Acid 1200 mcg
|[ 024 ]| Garlic, Odorless (Allium sativum) 500 mg
|[ 025 ]| Green Tea Extract (Camellia sinensis) (leaf) 1000 mg
|[ 026 ]| Inositol 10 mg
|[ 026 ]| Niacin (as Inositol Hexanicontinate) 100 mg
|[ 027 ]| Iodine (as Potassium Iodide) 150 mcg
|[ 028 ]| Joint Health Complex: (Methylsulfonylmethane (MSM), Silica(as Silicon Dioxide), Collagen (Hydrolyzed Gelatin), Glucosamine Hydrochloride, Chondroitin Sulfate) 100 mg
|[ 029 ]| Lactobacillus Acidophilus 10 mg
|[ 030 ]| L-Arginine 1000 mg
|[ 031 ]| L-Carnitine 250 mg
|[ 032 ]| L-Carnosine 500 mg
|[ 033 ]| Lecithin, Mega Soy 1325 mg
|[ 034 ]| L-Glutamine 500 mg
|[ 035 ]| Magnesium (as Magnesium Oxide) 100 mg
|[ 036 ]| Manganese (as Manganese Gluconate) 2 mg
|[ 037 ]| Melatonin (plus 25 mg Theanine) 5 mg
|[ 038 ]| Men's Bioactive Blend: (Pumpkin Seed Cucurbita pepo), Pygeum (Pygeum africanum) bark), Oyster Extract, Saw Palmetto Extract (Serenoa repens) (fruit), Lycopene) 120 mg
|[ 039 ]| Molybdenum (As Sodium Molybdate) 75 mcg
|[ 040 ]| Niacin (as Nicotinic Acid) 50 mg
|[ 040 ]| Niacin (as Inositol Hexanicontinate) 400 mg
|[ 041 ]| Pantothenic Acid (as d-Calcium Pantothenate) 50 mg
|[ 042 ]| Parsley (Petroselinum crispum) 100 mg
|[ 043 ]| Potassium (as Potassium Chloride) 30 mg
|[ 043 ]| Potassium (from 595mg Potassium Gluconate) 99 mg
|[ 044 ]| Resveratrol, Natural Mega Potency 200 mg
|[ 045 ]| Riboflavin (Vitamin B-2) 50 mg
|[ 046 ]| Saw Palmetto 450 mg
|[ 047 ]| Selenium (as Sodium Selenate) 200 mcg
|[ 048 ]| Simethicone 80 mg
|[ 049 ]| Thiamin (Vitamin B-1) (as Thiamin Mononitrate) 50mg
|[ 050 ]| Turmeric Curcumin (50mg at 95% Curcuminoids) 500 mg
|[ 051 ]| Vanadium(as Sodium Metavanadate) 10 mcg
|[ 052 ]| Vitamin A (as Beta-Carotene) 5000 IU
|[ 053 ]| Vitamin B-12 (as Cyanocobalamin) 50 mcg
|[ 054 ]| Vitamin B-6 (as Pyridoxine Hydrochloride) 30 mg
|[ 055 ]| Vitamin C (as Ascorbic Acid) 300 mg
|[ 056 ]| Vitamin D (as Cholecalciferol) 400 IU
|[ 057 ]| Vitamin E (as d-Alpha Tocopheryl Acetate) 100 IU
|[ 058 ]| Vitamin K (as Phytonadione) 80 mcg
|[ 059 ]| Zinc (as Zinc Oxide) 25 mg

There have been a few changes to the list though.
I added M-Blue and switched out the Apricot Seed with Laetrile made in Mexico plus ran out of creatine and protein drink mix a month ago.
I also bought some Astral Fruit-NF RGTA™ Complex and added it to the mouse mix but I ran out of that only yesterday. The mice have enough to last 6 months.

I've been experimenting with Tesla coils and slow charge spark gap homemade battery capacitors as well as homemade fuel cells and hydrogen storage techniques so this project is falling behind a bit until I can find more room to work.

Edited by timothymburke, 30 March 2011 - 09:36 PM.