59 replies to this topic

[zawy]

On december 29, 2007 a new study was published:

http://www.ncbi.nlm....Pubmed_RVDocSum

This finding may change everything about resveratrol theories. The importance is that MnSOD was 6 times more plentiful and 14 times more active. This greatly reduces cellular oxidation and increases efficiency at the core of respiration so it has a strong connection to calorie restriction independent of SIRT1. Possibly SIRT1 changes because of the MnSOD change. MnSOD is at a deeper, more fundamental level of respiration (aka aging) than SIRT1.

14 times more Active???
Any chemical reaction in the body that suddenly occurs 14 times more often should be so unnatural that it kills you. However, we already know it almost doubles lifespan in 5 wildly different species and has no relevant toxicity in any animal. If you're interested in how resveratrol works, the importance of what reservatrol does to MnSOD cannot be underestimated.

Importance of increasing MnSOD: In order to make ATP (cellular energy) there are three pumps in the membrane of mitochondria that use the energy from the chemicals from the krebs cycle to pump H+ to an outer-mitochondria area. A superoxide oxidation radical is formed in the process which is deactivated by MnSOD to less reactive peroxide H2O2. That provides a whopper of an antioxidant effect. But it also increases respiration effificiency because those H+ ions need to gather up on the outside before being pushed back into the inner mitochondria to create ATP. Remember, like charges like H+ repel each other. But those superoxide radicals are negatively charged and are attracted to the H+ and cross the membrane. Without the MnSOD, more H+ may get neutralized by the superoxides and not be allowed to create ATP energy. Superoxides outside the membrane will decrease the electrical push needed to get the H+ back inside to create ATP. Increased efficiency in the process means fewer calories are burned for the same amount of energy. Normally there is something like a 4% "leakage" of superoxides from respiration, but the hyperactive MnSOD may stop it.

Cancer: Many cancers are the result of MnSOD being altered on chromosone 6 or p53 being altered. p53 increases MnSOD to fight cancer. I believe a bad MnSOD/superoxide ratio in the presence of higher catalase allows cancer to use oxidation to breakapart collagen and spread quickly. Altering the MnSOD or p53 genes is a common way that cancer does this. They can give MnSOD to kill cancer cells, so why not increase increase it's activity by a factor of FOURTEEN. Everyone with cancer should be taking 1 to 4 g/day RESV and 2 to 4 g/day Quercetin.

The big picture
MnSOD, resveratrol, exercise, and calorie restriction all 4 reduce cancer, lower blood sugar, and increase life span. This is because they improve the energy liberation process of respiration. That is, they improve life itself, at the core.

Toxicity
I read many posts where people are skeptical that 1 g/day is safe despite the evidence of safety. Let me say that RESV is a natural compound designed and manufacturable by DNA-based systems. It is one of the most common flavonoids found in nature. We were designed to eat these DNA-manufactured compounds that are created under low-temp, low-pressure conditions (unlike the pharms). That is why there is no toxicity of ANY compound created by DNA when ingestion is kept below about 1 gram a day. Rare exceptions include purposefully-made toxins. Aspirin is one of the more dangerous of DNA compounds and shouldn't be taken at 1 g per day. The billions of DNA-based compounds also have no known interaction with each other. Compare to pharmaceuticals which have a hard time being safe at any level, and frequently have interactions with each other.

Resveratrol is a toxin to fungus which may be yeast concentrate it in making wine (for their own protection against competitors. If you have a beneficial fungus in you, you might need to avoid massive RESV.

Absorption, metabolites, half life
RESV is well-absorbed but <1% stays as resveratrol when it enters the blood. The digestive system alters with a sulfate so that it will transfer into cells more easily where it is converted back. Half-life in the blood of humans is 9 hours.

http://dmd.aspetjour.../32/12/1377.pdf Wallet, 2004 (good data, but provided doubt)
http://pubs.acs.org/.../mp700071d.html Wallet, 2007 (more positive about the metabolites)

[inawe]

Zawy,
Do you have access to the full paper? Seems that work was purely in vitro. It would be interesting to see how it translates in vivo.

[smithx]

there is no toxicity of ANY compound created by DNA when ingestion is kept below about 1 gram a day.

Uh... not quite.

In fact, not at all. There are millions of highly toxic natural compounds -- all "produced by DNA" in your way of speaking.

These include ricin (from castor beans), hydrogen cyanide (found in the pits of cherries, apricots, bitter almonds and apples), aflatoxin (from molds), urshinol (found in poison ivy, etc.)... Any of these can kill or cause severe damage in single doses of well below 1 gm.

I take about a gram a day of resveratrol, but I do it because of safety studies which have been performed on mice not because of misplaced blind faith in "natural compounds".

[edward]

The study looks interesting, if I get around to it ill try to email the authors and get a copy since I can't seem to find a way to get it online. Any one else who wants to do this go for it.

One note though, Zawny, I don't really agree with your conclusions, there are a lot of leaps and use of half information in your post above but thank you for alerting us to the study. Again did you read the whole study (if so please post a copy or a way to access it) I'll have to read it for myself and then do some additional research on MnSOD.

[marcopolo]

An employee who works in the family business is dying of cancer, I considered telling him about resveratrol at a Christmas party my parents had for the management and he and I were both there, but I decided maybe it wasn't such a good idea because for all I know it could hasten his demise. So I feel guilty for not telling him and I would feel guilty if I did tell him and he died shortly afterward. I didn't get much of a chance to bring it up to him anyway, I mostly ate the food and listened to other people chit chat. Would you go as far as to recommend this stuff to someone who has cancer? If so, how would you bring it up to them?

[VP.]

This finding may change everything about resveratrol theories. The importance is that MnSOD was 6 times more plentiful and 14 times more active. This greatly reduces cellular oxidation and increases efficiency at the core of respiration so it has a strong connection to calorie restriction independent of SIRT1. Possibly SIRT1 changes because of the MnSOD change. MnSOD is at a deeper, more fundamental level of respiration (aka aging) than SIRT1.

You might have it backwards. Maybe MnSOD changes because because of SIRT1 activation. Check out this new study linked from Ouroburos:

Feel the burn: Exercise boosts sirtuin activity
Posted by ouroboros under Calorie restriction, Resveratrol, Sirtuins

We know that resveratrol, an activator of SIRT1, boosts exercise tolerance and performance in mice (see Resveratrol: Breakfast of champions). Now it appears that the converse is also true: exercise increases SIRT1 activity in aged rats. From Ferrara et al.:

Exercise Training Promotes SIRT1 Activity in Aged Rats

The objective of this study was to determine the effects of aging and exercise training on SIRT1 activity and to identify a pathway linking SIRT1 to antioxidant response and cell cycle regulation in rats. SIRT1 is a NAD+-dependent deacetylase involved in the oxidative stress response and aging. The effects of aging and of moderate and prolonged exercise training in rats are unknown. We measured SIRT1 activity in heart and adipose tissue of young (6 months old), sedentary old (24 months), and trained old (24 months) rats using an assay kit. … Aging significantly reduced SIRT1 activity in heart, but not in adipose tissue, increased TBARS and 4-HNE and decreased Mn-SOD and catalase expression in both heart and adipose tissue. Aging did not affect FOXO3a protein expression in the heart or FOXO3a mRNA in adipose tissue. Exercise training significantly increased FOXO3a protein in the heart and FOXO3a mRNA in adipose tissue of aged rats. It also significantly increased Mn-SOD and catalase levels in both heart and adipose tissue. … We concluded that exercise training, which significantly increases SIRT1 activity, could counteract age-related systems impairment.
Note that the effects are observed both in cardiac tissue, where SIRT1 levels decrease with age, and in adipose, where SIRT1 activity remains constant throughout the lifespan.

These findings are consistent with recent observations that resveratrol induces the heat shock response, leading some to speculate that the compound exerts some of its positive effects via hormesis. Exercise is the quintessential example of beneficial hormesis: stress the body acutely now to make it more resistant to chronic stress later. Could resveratrol and exercise (and for that matter, calorie restriction) converge on a common pathway that confers stress resistance?

For those of you who lack the hepatic fortitude to consume 1000 bottles of red wine, I’m betting that New Year’s resolution to get off your duff and hit the gym is looking a lot more appealing.

http://ouroboros.wor...rtuin-activity/

[Hedgehog]

The study looks interesting, if I get around to it ill try to email the authors and get a copy since I can't seem to find a way to get it online. Any one else who wants to do this go for it.

One note though, Zawny, I don't really agree with your conclusions, there are a lot of leaps and use of half information in your post above but thank you for alerting us to the study. Again did you read the whole study (if so please post a copy or a way to access it) I'll have to read it for myself and then do some additional research on MnSOD.

See attachment for article.

Marco, depending on what type of cancer he has I would seriously look at a compound called cyclopamine. Genentech is hopefully bringing a version of this molecule to the market.

[niner]

Here's the abstract. I note that it's an in vitro experiment, and although they mention "micromolar concentrations", they don't put a number on it, so we can't tell from the abstract if this will occur at physiological concentrations or not. A 5 gram oral dose of resveratrol produced a 2.4 micromolar plasma level in one study in humans (Boocock et al.).
edit: I see hedgehog posted the paper while I was writing this- thanx! They used 50 and 100 micromolar treatments, so hard to say what it means for humans. The effect on MnSOD was pretty large, so maybe there's something happening in vivo.

Biochem Biophys Res Commun. 2007 Dec 31 [Epub ahead of print]
Molecular mechanisms of oxidative stress resistance induced by resveratrol: Specific and progressive induction of MnSOD.Robb EL, Page MM, Wiens BE, Stuart JA.
Department of Biological Sciences, Brock University, 500 Glenridge Road, St. Catharines, Ont., Canada L2S 3A1.

trans-Resveratrol (3,4',5-trihydroxystilbene; RES), a polyphenol found in particularly high concentrations in red wine, has recently attracted intense interest for its potentially beneficial effects on human health. Here, we report the effects of long-term exposure to micromolar concentrations of RES on antioxidant and DNA repair enzyme activities in a human cell line (MRC-5). RES had either no effect on, or reduced the activities of glutathione peroxidase, catalase and CuZn superoxide dismutase (SOD), in treatments lasting up to 2 weeks. RES failed to induce activities of the DNA base excision repair enzymes apurinic/apyrimidinic endonuclease and DNA polymerase beta. However, it dramatically and progressively induced mitochondrial MnSOD expression and activity. Two weeks exposure to RES increased MnSOD protein level 6-fold and activity 14-fold. Thus, long-term exposure of human cells to RES results in a highly specific upregulation of MnSOD, and this may be an important mechanism by which it elicits its effects in human cells.

PMID: 18167310

Edited by niner, 10 January 2008 - 06:31 AM.

[marcopolo]

Marco, depending on what type of cancer he has I would seriously look at a compound called cyclopamine. Genentech is hopefully bringing a version of this molecule to the market.

Thanks for the info. I have no idea what he does and doesn't know, the last time I really had a long conversation with him was last summer before he was diagnosed. He is a very intelligent guy and I know if I were him I would be spending every spare moment I had learning about anything and everything I could do or take to stop it before it killed me.

[zawy]

inawe, i just read the full article after hedgehog posted it.

smithx, all of the examples you mentioned as DNA-created toxins fall under the "purposefully toxic" category I mentioned as exceptions in the sentence immediately after the sound bite you extracted. But your rhetoric is a fair balance against mine.

velopismo, Does RESV act on SIRT1 to increase MnSOD or vice versa? It's neither. The article cites [19] (Oct 2007) which says RESV might alter SIRT1 downstream pathways instead of SIRT1 activity directly. Specifically, RESV moves the SIRT1 target FOXO3a to the nucleus. Another [20] reference (2004) says SIRT1 forms a complex with FOXO3a to fight oxidative stress but inhibits FOXO3a ability to cause cell death. So the answer is that RESV causes SIRT1 and FOXO3a to increase MnSOD which decreases the superoxide radical. So the finger has to point at all 3: SIRT1, FOXO3a, and MnSOD. But MnSOD/superoxide effect is the primary effect. This means RESV increases number of possible cell divisions whereas CR increases time between division (Ref 15). Unfortunately, I've seen articles where SIRT1 activity (and presumably MnSOD activity)do not increase CR effects and even reduces effect of extreme CR.

Summary of references:
1) MnSOD decreases Superoxide. (its sole purpose)
2) Superoxide increases FOXO3a creation. ref 5
3) FOXO3a increases MnSOD creation ref 3 and SIRT1 creation if p53 is present. ref 4
4) NAD+ increases SIRT1 activity at increasing FOXO3a activity in creating MnSOD. ref 6 combined with ref 4
5) But artificially ADDED SIRT1 suppresses FOXO3a creation of MnSOD. ref 5 (needed to add NAD+ too?)
6) RESV increases ability of SIRT1 to use FOXO3a to create MnSOD ([19], [20], [21])
7) Speculative: decreasing superoxide(2-) protects and increases NAD+ at complex 1
8) Increased NAD+ is a really good thing ref 7 and key to SIRT1 activity (SIRT1 does not do anything without NAD+ -- see 4) and ref 12
9) RESV causes SIRT1 to move FOXO3a to the nucleus to create MnSOD (

To summarize all this simplistic and limited view of the feedback in a sentence and ignoring the contradiction of 5):
Superoxide radical increases FOXO3a which increases SIRT1 so that SIRT1 and NAD+ can activate FOXO3a to create more MnSOD to reduce the superoxide that triggered the FOXO3a. RESV shifts the balance towards more SIRT1/FOXO3a activity and MnSOD and therefore less superoxide radical and therefore less FOXO3a. A better ratio of NAD+/superoxide(-) ratio from MnSOD activity may make SIRT1 more capable of activating the dimished FOXO3a to keep making MnSOD. This implies there's less SIRT1 (because of less FOXO3a) but that it's more active (more NAD+). If RESV activating SIRT1 causes the increased MnSOD, then it could follow this chain: RES increases SIRT1 which increases FOXO3a which increases MnSOD. The feedback from the MnSOD/NAD+ would limit the SIRT1. Another possibility is

SIRT, CR, Resveratrol, and Lifespan - research in past 4 years.
Ref 8 says RESV activates SIRT1 only in the right subtrate. Ref 9 and ref 13 say calorie restriction works even better without SIRT1. ref 14 says CR works without any of the SIR family. Ref 10 says multiple members of mammilian SIRT family may regulatae lifespan. Ref 11 shows RESV directly increases rate of reaction of SIRT1 "for both the acetylated substrate and NAD(+)". Ref 15 says yeast sirtuins increase longevity indepedent of CR by increasing number of viable cell divisions, not by delaying cell divisions (CR method). It also says "elevated respiration capacity is an important determinant of chronological longevity" (indepedent of calories consumed) which seemes to imply exercise works similarly to CR.

Dosage, Plasma Levels
They also stated no affect above 50 uM, and possibly a bit lower is just as effective. For 70 kg, that would be 0.8 grams. This is just a guide as to what oral minimum is needed to get their maximum. An older reference says half life is 9 hours which would be 2 g/day. The 3 hour half life in boocock et al would mean 6 g/day. Blood plasma was shown to be 2.4 uM in Boocock as niner mentioned, but don't forget the two links at the bootom of my first post (and another ref i saw) show the sulfated metabolites are probably more important than reservatrol itself for getting into the cell and being active. The metabolites were 23 times higher. 2.4 x 23 = 55 uM peak RESV metabolite blood concentration with a 5 g dose after 1.5 hour.

Misc:
Decreased superoxide radical also decreases H2O2:
http://books.google....HxakXg#PPA28,M1

2008 article quote: "MnSOD overexpression has been associated with extension of lifespan [16,17], resistance to ischemia–reperfusion injury and resistance to irradiation-induced damage [18]. In addition, overexpression of MnSOD has been reported to inhibit cancer cell growth. Interestingly, most of these same observations have been made for RES (Table 3). We therefore propose that the RES-mediated induction of MnSOD may be an important mechanism by which this polyphenol confers its beneficial effects."

I kind of like the idea of mixing the other natural gunk in with RESV at 50%. For all we know the mixture increases absorption and may have complex matrix effects inside the cells. It's a little closer to the natural state. I'm going from 200 mg/day to 1 g/day using 50% purity. These two post may be the result of extra mental activity.

Edited by zawy, 11 January 2008 - 03:44 PM.

[missminni]

An employee who works in the family business is dying of cancer, I considered telling him about resveratrol at a Christmas party my parents had for the management and he and I were both there, but I decided maybe it wasn't such a good idea because for all I know it could hasten his demise. So I feel guilty for not telling him and I would feel guilty if I did tell him and he died shortly afterward. I didn't get much of a chance to bring it up to him anyway, I mostly ate the food and listened to other people chit chat. Would you go as far as to recommend this stuff to someone who has cancer? If so, how would you bring it up to them?

I would tell him about it, that it might be helpful, and encourage him to do his own research on it. Let him know that if he
has any questions to feel free to ask you, but he should really pursue it himself.

[zawy]

I've made many changes to my second post as my knowledge and number of references increased. The short story is that RESV primary action is to use MnSOD to decrease free radicals which allows a larger number of cell divisions instead of longer time between divisions. So it's different from CR, but one article says RESV decreased lifespan from the maximum when used in extreme CR. SIRT1/NAD+/FOXO3a are just the point of RESV action that causes the MnSOD increase. SIRT1 is not directly affected. Only it's affect on FOXO3a is important as the step before increased MnSOD. I also provided the link that indicates exercise may operate fundmentally different from CR and RESV. It seems there are enough published articles out there to show the difference between the 3.

[Hedgehog]

Absorption, metabolites, half life
RESV is well-absorbed but <1% stays as resveratrol when it enters the blood. The digestive system alters with a sulfate so that it will transfer into cells more easily where it is converted back. Half-life in the blood of humans is 9 hours.

Has anybody added up the numbers to see if any of the resveratrol is hiding somewhere else in the body besides plasma such as cell walls? I guess we could look at studies that look at resveratrol in blood plasma + urine, + "poop". See if all the known metabolites + Resveratrol add up to close to 100% of the original dose?

[zawy]

The 2nd to last link in the first post in this thread shows that after 12 hours 17% remains after subtracting feces and urine for 25 mg oral dose and 26% remains for 0.2 mg I.V. So orally it appears to be 65% as effective (absorbed) as I.V. Let's say we take 50 mg every hour, so that we've take 600 mg in 12 hours. That will result in an absolute minimum of 17% of the 600 mg still in the body (non-feces, non-urine). After another 12 hours, at least 17% of that initial 17% (3%) should still be in the body. So we have at least 20% of 600 mg after 24 hour and subsequent days will not add much except to keep up this steady-state level of 120 mg in the body. For 50 kg of non-bone tissue, that's 10 um/L. The new article under discussion said 50 um/L was apparently way more than needed to see the 14-fold effect on MnSOD.

[zawy]

The yeast studies used 2 to 22 mg/L media for growing, and showed up to 100% increase in life-span as measured by number of viable cell divisions. The fish study increased lifespan 30% on 120 ug/g resveratrol in food and 56% increase on 600 ug/g. No increase was observed with 24 ug/g. The treatment began at 4 weeks of age which is 30% of the maximum recorded lifespan for the species. 50% of the 600 ug/g group lived longer than had ever been observed before in this species (without calorie restriction). 90% of the high-dose group were still sexually active after 90% of the control group were dead. Using the simple dose per day per kg body-weight, the fish dosages of 24, 120, and 600 ug/g at 50 mg/g comsumption/day is 84, 420, 2100 mg/day for a 70 kg person. This daily dose per body-weight is suspect since the metabolic rate of humans is much slower. The dose per calorie consumed is 10, 50, and 250 mg/day for a 2000 calorie/day diet. The effects of any compound on different species varies widely. This species of fish is the shortest-lived vertabrete known with a high metabolic rate. Humans are one of the longest-lived vertabretes with a much slower metabolic rate.

[Hedgehog]

The yeast studies used 2 to 22 mg/L media for growing, and showed up to 100% increase in life-span as measured by number of viable cell divisions. The fish study increased lifespan 30% on 120 ug/g resveratrol in food and 56% increase on 600 ug/g. No increase was observed with 24 ug/g. The treatment began at 4 weeks of age which is 30% of the maximum recorded lifespan for the species. 50% of the 600 ug/g group lived longer than had ever been observed before in this species (without calorie restriction). 90% of the high-dose group were still sexually active after 90% of the control group were dead. Using the simple dose per day per kg body-weight, the fish dosages of 24, 120, and 600 ug/g at 50 mg/g comsumption/day is 84, 420, 2100 mg/day for a 70 kg person. This daily dose per body-weight is suspect since the metabolic rate of humans is much slower. The dose per calorie consumed is 10, 50, and 250 mg/day for a 2000 calorie/day diet. The effects of any compound on different species varies widely. This species of fish is the shortest-lived vertabrete known with a high metabolic rate. Humans are one of the longest-lived vertabretes with a much slower metabolic rate.

Common formula for dose translation studies

HED (mg/kg) = animal dose mg/kg * (animal km/ human km)

(HED) human equivalent dose
km factor for adult human = 37 for 60kg person
km factor for rat = 6 0.15kg rat
km factor for mouse = 3 .02kg mouse

Also see Du Bois formula

[Hedgehog]

HED (mg/kg) = animal dose mg/kg * (animal km/ human km)

(HED) human equivalent dose
km factor for adult human = 37 for 60kg person
km factor for rat = 6 0.15kg rat
km factor for mouse = 3 .02kg mouse

Also see Du Bois formula

Source of formula.
Center for Drug Evaluation and Research, Center for Biologics

Evaluation and Research. (2002) Estimating the safe starting dose in

clinical trials for therapeutics in adult healthy volunteers, U.S. Food

and Drug Administration, Rockville, Maryland, USA


So if biotech companies start with this guideline or formula based on animal studies and conduct clinical trials to narrow the proper dosage down.

If we use this formula and we want what a 200mg/kg study used in a mouse it would come out to approx 1 gram of resveratrol for a human. However, the blood plasma [concentrations] could tell a different story and one could assume you would need to take more or less.

[zawy]

My method of "dose/calories consumed" is supposed to adjust for any difference in metabolic rate. I believe this is the most accurate method for our purposes. The RESV action is at the core of respiration, this may be an enormously accurate method. When i was calculating the doses i was constantly wondering why they chose 24, 120, and 600 ug RESV per gram food for the fish. They eat 50 mg food per g body weight per day. The calorie content of typical food is 2000 calories in 417 grams (30% calories from fat, 70% protien-carbs). When i converted I got EXACTLY 10, 50, and 250 mg/day for a 2000 calorie diet and then I knew exactly what they were thinking...and that my conversion was legitimate. Those little research devils had human consumption in mind right from the beginning.

So those of us taking 1,000 mg/day should be doing better then the fish.

Let's assume that RESV is similar to B vitamins when trying to compare utilization in different species. Why B vitamin? They're in every species, going all the way back to yeast, and my intuition just says they do things somewhat like RESV. I found fortified mice/rat food that averaged 6.8 mg B-complex/1000 calories for the 5 B vitamins (B1,B2,B3,B6, pantethine) in my B-50 complex supplement that contains 50 mg each of these. I believe one B-50 per day is good enough for good health (not RDA!) for 2000-calories/day (25 mg/1000 calories) in a manner close to how the B vitamins in the fortified mice/rat is good for them. So by this reasoning, we need 25/6.8 = 3.7 times more supplements/calorie than a mouse/rat to get the same benefit from B-vitamins. If we take the same approach with RESV and the fish, we can use the 3.7 times RESV/calorie used for the fish. The fish weighs 20 times less than a mouse/rat of 30 grams and has a lifespan 10 times less (both weight and lifespan are related to metabolic rate), so the factor might need to be a little bigger. Not a lot bigger because we weigh 3500 times more than a mouse/rat and have 70 times the lifespan. So I'll just use 4 as the factor for going from fish to human. Calorie based method said we should take 250 mg/day to have the same RESV/calorie. Times 4 equals 1,000 mg/day. My B-complex probably provides a better margin of safety so the 3.7 factor should be high.

As a check, I found channel catfish fingerlings require at least 14 mg niacin per kg food to achieve maximal growth with. Actually, it was 10 mg/kg but they used curve fitting to get a margin of safety. I'll assume 14 mg/kg for max growth is equivalent to the purposes of the RDA. In humans the RDA is 16 mg / 2000 calories (0.417 kg) = 38 mg/kg food. This gives a factor of 38/14 = 2.7 which comes out close to the 3.7 factor for the mice/rat food. 2.7 x 250 mg/day=675 mg RESV/day is needed to get the same results as the high-dose RESV fish.

Exploring other methods of calculating HED:

HED (mg/kg) = animal dose mg/kg * (human kg/ animal kg)^0.33 if it's a "direct acting" (concentration dependent?)
HED (mg/kg) = animal dose mg/kg * (animal kg/ human kg)^0.33 if it's "bioactivated" (respiration dependent?)

I don't know if we should use the first or second formula, but it makes a 1000-fold difference as shown at the bottom of this post. RESV is intimately tied to respiration so that would make it appear the second one is right which gives 61 mg/day which is a lot lower than my calorie methods.

After a fair amount of looking, I can't find the weight of this fish. Length is from 4 to 10 cm. Average is 5 cm. Looking at the pics, I get a surface area of 5cm x 1 cm tall x 2 sides = 10 cm^2 = 0.001 m^2.
Human man is 1.9 m^2. A ratio of 2,000.

Surface area is supposed to be scaled by (weight ratio)^0.66
weight^0.66 = 2000 which gives weight ratio = 51,000. 70kg/51,000 = 1.6 grams for fish. As a check, i found minnows are 1.6 g at 4.5 cm. Good.

This gives:
HED g/kg = 50 mg food/g bdy wt/day*600 ug RESV/g food * 1000 g/kg * (0.0016/70)^0.33 = 0.00088 g/kg
times 70 kg = 62 mg/day for "bioactivated" method.

HED g/kg = 50 mg/g/day*600 ug/g * 1000 g/kg * (70/0.0016)^0.33 = 1.02 g/kg
times 70 kg = 70 g/day for "direct acting" method

Body weight only (another saturation method):
50 mg/g/day*600 ug/g * 1000 g/kg = 0.03 g/kg
70 kg x 0.03 = 2.1 g/day

Edited by zawy, 12 January 2008 - 02:53 PM.

[drmz]

maybe posted already, bumped into it when searching on pubmed

The effect of resveratrol on a cell model of human aging.Stefani M, Markus MA, Lin RC, Pinese M, Dawes IW, Morris BJ.
Basic & Clinical Genomics Laboratory, School of Medical Sciences and Bosch Institute, The University of Sydney, Sydney, New South Wales 2006, Australia.

The natural polyphenol resveratrol stimulates sirtuins and extends lifespan. Here resveratrol inhibited expression of replicative senescence marker INK4a in human dermal fibroblasts, and 47 of 19,000 genes from microarray experiments were differentially expressed. These included genes for growth, cell division, cell signaling, apoptosis, and transcription. Genes involved in Ras and ubiquitin pathways, Ras-GRF1, RAC3, and UBE2D3, were downregulated. The changes suggest resveratrol might alter sirtuin-regulated downstream pathways, rather than sirtuin activity. Serum deprivation and high confluency caused nuclear translocation of the SIRT1-regulated transcription factor FOXO3a. Our data indicate resveratrol's actions might cause FOXO recruitment to the nucleus.

[zawy]

This is a correction to my previous post that tries to convert the high-dose fish group to human consumption. This lowers what appears to be the human-equivalent dose. I calculated the fish RESV consumption to be 250 mg / 2000 calories. But this was based on ideal dry weight food, 30% fat, of 4.8 calories/gram. They were fed drip-dried frozen bloodworm larvae. I don't know the calorie content of larvae, but let's assume it's about like dry and soft dog food, 3.1 cals/gram. It could be less, but probably not more. Canned dog food is 1.3 cals/gram. So fish RESV consumption was 250 *3.1/4.8= 135 mg /2000 calories or LESS. My comparison of our b-vitamin RDAs to the B vitamins in rats/mice and fingerling fish contains the same error, which means the 2.7 and 3.7 factors should be lower: 1.75 and 2.4. This means our equivalent dose should be between 236 and 324 mg RESV/ 2000 calories.

500 mg/day in humans should be plenty for an equivlant dose that increased fish lifespan by 56%.

[niner]

This is a correction to my previous post that tries to convert the high-dose fish group to human consumption. This lowers what appears to be the human-equivalent dose. I calculated the fish RESV consumption to be 250 mg / 2000 calories. But this was based on ideal dry weight food, 30% fat, of 4.8 calories/gram. They were fed drip-dried frozen bloodworm larvae. I don't know the calorie content of larvae, but let's assume it's about like dry and soft dog food, 3.1 cals/gram. It could be less, but probably not more. Canned dog food is 1.3 cals/gram. So fish RESV consumption was 250 *3.1/4.8= 135 mg /2000 calories or LESS. My comparison of our b-vitamin RDAs to the B vitamins in rats/mice and fingerling fish contains the same error, which means the 2.7 and 3.7 factors should be lower: 1.75 and 2.4. This means our equivalent dose should be between 236 and 324 mg RESV/ 2000 calories.

500 mg/day in humans should be plenty for an equivlant dose that increased fish lifespan by 56%.

Except that you aren't accounting for differential metabolism between fish and humans, not to mention other differences related to our wildly different lifespans, so you may be off by an order of magnitude or two.

[zawy]

Dosage per calorie consumed is the DEFINITION of accounting for the difference in metabolism. There is not likely to be a better measure. My post concerning the B-vitamins was a check as much as it was an adjustment. It's an excuse to have a safety factor just as the RDA has a safety factor when trying to prevent disease, except I'm shooting for optimum health. I then almost doubled that to come up with 500 mg RESV as a gross overestimate of what would be needed to achieve the same dose as the high-dose fish.

Another way to look at dosage per calorie consumed is that it's a percentage of incoming amino acids that make up SIRT1 (Sirs), FOXO3a, NAD+, and MnSOD. Therefore you can see dosage/calorie consumed is enormously accurate compared to any other method until relative absorption and utilization are known. The objective is to have the same mg RESV/body weight. Since our protein turnover rate is slower with our slower metabolism, you don't use mg/kg/d which everyone knows is a gross overestimate ( 2100 mg/day ) at least as far as trying to get the same dose as the fish.

[maxwatt]

Dosage per calorie consumed is the DEFINITION of accounting for the difference in metabolism. There is not likely to be a better measure. My post concerning the B-vitamins was a check as much as it was an adjustment. It's an excuse to have a safety factor just as the RDA has a safety factor when trying to prevent disease, except I'm shooting for optimum health. I then almost doubled that to come up with 500 mg RESV as a gross overestimate of what would be needed to achieve the same dose as the high-dose fish.

Another way to look at dosage per calorie consumed is that it's a percentage of incoming amino acids that make up SIRT1 (Sirs), FOXO3a, NAD+, and MnSOD. Therefore you can see dosage/calorie consumed is enormously accurate compared to any other method until relative absorption and utilization are known. The objective is to have the same mg RESV/body weight. Since our protein turnover rate is slower with our slower metabolism, you don't use mg/kg/d which everyone knows is a gross overestimate ( 2100 mg/day ) at least as far as trying to get the same dose as the fish.

A good check on that is to take the ration of calories consumed, and compae it to the ratios determined by another method(surface area.) here-to-fore considered the gold standard.

[zawy]

Judging from the pictures and the data i found that the fish averages 5 cm long, it looks to average 1 cm tall. The tail is about 1 cm, so let's say body length is 4 cm. That's 8 cm^2 for each side. Let's say it's 10 cm^2 (0.001 m^2) since i didn't account for surface curvature. Weight seems to be about 1.6 grams (see previous post above). This gives a km factor of 1.6 kg/m^2. Human km is 37. A human is about 1.8 m^2. From a previous post above, this gives:
HED = 30 mg RESV/ kg fish/day * 1.6/37 = 1.3 mg/kg
times 70 kg = 91 mg RESV / 70 kg person. This is a little less than the 135 mg / 2000 calorie diet I had calculated. The error in the numbers above could be the difference. A previous post above based on the HED using weight only came out to 62 mg/ 70 kg based on 1.6 gram fish. I believe these HED methods are estimations of the dosage/calorie consumed method, but i couldn't find a reference. It's not possible to beat my previous arguments.

Edited by zawy, 15 January 2008 - 12:25 AM.

[niner]

Dosage per calorie consumed is the DEFINITION of accounting for the difference in metabolism. There is not likely to be a better measure. My post concerning the B-vitamins was a check as much as it was an adjustment. It's an excuse to have a safety factor just as the RDA has a safety factor when trying to prevent disease, except I'm shooting for optimum health. I then almost doubled that to come up with 500 mg RESV as a gross overestimate of what would be needed to achieve the same dose as the high-dose fish.

Another way to look at dosage per calorie consumed is that it's a percentage of incoming amino acids that make up SIRT1 (Sirs), FOXO3a, NAD+, and MnSOD. Therefore you can see dosage/calorie consumed is enormously accurate compared to any other method until relative absorption and utilization are known. The objective is to have the same mg RESV/body weight. Since our protein turnover rate is slower with our slower metabolism, you don't use mg/kg/d which everyone knows is a gross overestimate ( 2100 mg/day ) at least as far as trying to get the same dose as the fish.

Zawy, when I said "metabolism" I was talking about xenobiotic metabolism; what the body does with chemicals that are not food. Humans have very efficient mechanisms for getting rid of molecules like resveratrol. We sulfate and glucuronidate the hell out of them. I don't think fish do that to anywhere near the extent that humans do.

We already know from pharmacokinetic studies in mice and in human volunteers that in order to get the same plasma levels of free resveratrol, humans need to take a lot more mg/kg than mice do, not a lot less as the usual interspecies scaling guidelines would predict. I would expect humans would need a lot LOT more mg/kg than fish to get the same effects.

[zawy]

Can you refer me to the studies that show resveratrol in mice plasma? As i pointed out before, there's one or two new articles that are say the sulfated form is more useful than the free form. One of the authors is the one who first publsished the low level of free resv in plasma.

[zawy]

I finally found calorie content on this order of fly larvae: 70% moisture, 15% fat = 1.4 Cal/g. They consumed an estimated 50 mg food/g body weight/day. This is 0.07 Cal/g body weight/day. However, this size animal should consume between 0.6 and 0.8 Calories/gram body weight/day. Therefore the paper has a serious error that makes it impossible to check how much resveratrol the fish really received. The only explanation i can think of is that the larvae which where "frozen and drip dried" where somehow actually previously dried which would be about 3.8 Cal/g. But that's still only 0.2 Cal/gram.

They stated in parenthesis 2.5 uM for the 600 ug Resv/g food when it should be 2500 uM. Also they stated "1.2 mg/ul resveratrol stock was prepared in 5% ethanol". What does that mean?

Judging from the pictures here, these fish have a volume of 4/3 *pi * r1 * r2 * r3 (ellipsoid). gives between 1.8 and 3 ml volume, 1.8 to 3 g. 35 gram mice need 10 calories/day. Using the weight adjustment for metabolism and 3 gram fish: 10/35*(35/3)^(.33) = 0.64 Calories/gram needed at least. 0.52 if the ^0.25 factor is used in place of 0.33. Using 5 g fish and the 0.25 to see how low is possible: 10/35*(35/5)^0.25 = 0.46

Edit: I'm told the lavae may have as much as 6 Cal/g (would have to be dry weight and 50% fat, contrary to info above). This gracious view would be 0.3 Cal food / g bdy wt / day at the estimated 50 mg food/g body weight/day. This still would qualify as calorie restriction since the most twisted view can get their needs down to no less than 0.46 Cal/g (see above). The control fish were fed in the same twice a day manner, but uneaten food was collected from each tank, but not measured to see if control fish got fewer calories. It is implied that they did not notice a difference and were well aware of CR and presumably prevented it.

Edited by zawy, 16 January 2008 - 04:39 PM.

[SirtuinInvestor]

New poll on resveratrol usage at http://sirtuininvestor.blogspot.com/

Will be interesting to see who much people are taking.

I finally found calorie content on this order of fly larvae: 70% moisture, 15% fat = 1.4 Cal/g. They consumed only 50 mg food/g body weight/day. This is 0.07 Cal/g body weight/day. However, this size animal should consume between 0.6 and 0.8 Calories/gram body weight/day. Therefore the paper has a serious error that makes it impossible to check how much resveratrol the fish really received. The only explanation i can think of is that the larvae which where "frozen and drip dried" where somehow actually previously dried which would be about 3.8 Cal/g. But that's still only 0.2 Cal/gram.

They stated in parenthesis 2.5 uM for the 600 ug Resv/g food when it should be 2500 uM. Also they stated "1.2 mg/ul resveratrol stock was prepared in 5% ethanol". What does that mean?

Judging from the pictures here, these fish have a volume of 4/3 *pi * r1 * r2 * r3 (ellipsoid). gives between 1.8 and 3 ml volume, 1.8 to 3 g. 35 gram mice need 10 calories/day. Using the weight adjustment for metabolism and 3 gram fish: 10/35*(35/3)^(.33) = 0.64 Calories/gram needed at least.

[niner]

Can you refer me to the studies that show resveratrol in mice plasma? As i pointed out before, there's one or two new articles that are say the sulfated form is more useful than the free form. One of the authors is the one who first publsished the low level of free resv in plasma.

Here's a link to a post that goes into some detail on rodent vs human RSV levels: http://www.imminst.o...&...st&p=197228
There's one abstract, but the rest of the refs only have title and date. (I grabbed them from another posting) so you'll have to look those up in medline. Are you talking about the Walle papers that you mentioned above as being the refs for the sulfate being more useful than free? I looked at them and only found a bit of speculation about that. Is there more to it, or something I missed there?

[zawy]

Yes, Walle was speculative. However, the two rat studies you referenced may have only been discussing metabolites and not resveratrol. The first says "resveratrol" but then mention it is metabolized to aglycone and glucuronid. The second one definitely says it only measured the sulphate or glucuronate conjugates metabolites. So we don't know what the free resveratrol concentration is after oral ingestion by rats or mice. A newer study. Another None of them seem to mention "free resveratrol" in the blood of rats.

Back in 2002:
"Incubation with beta-glucuronidase and sulfatase to release free resveratrol was used to confirm the structures of these conjugates." ref 4 Makes me wonder if mice and men do the same "de-gluconation" and "de-sulfation", and if this is a well-known method of delivering compounds to tissue. The most abundant metabolites in humans, rats, and mice are trans-resveratrol-3-O-glucuronide and trans-resveratrol-3-sulfate. I suspect the body uses the modifications to determine friend from foe and to get them transferred to different locations in the body, through different membranes. Once there, there might be a mechanism for turning it back into free resveratrol. Suppose free resveratrol does not get inside cells as well. Then these studies with direct immersion in resveratrol may not be able to get as high a dose as we can get orally. I'm just saying...it's possible.

"... the disposition of flavonoid glucuronides has also been illustrated. Despite low oral bioavailabilities of the parent compounds, [modified?] flavonoids and some of their bioactive phase II conjugates may accumulate adequate amount in the body to produce their pharmacological activities. Further investigation on the correlation between the accumulated concentrations of flavonoids and their pharmacological activities after their repeated oral administration is warranted." ref 3

Note on comparative dosing: Here's an article saying the RESV dose/kg reported in media was very unfortunate http://www.ncbi.nlm....pubmed/17942826
The most common method to adjust for the difference in metabolism is the formula: (human dose/kg) = (animal dose mg/kg) x (animal kg/human kg)^(1-P) where P=2/3 by convention, but P=3/4 is more accurate ( http://www.fda.gov/cber/gdlns/dose.htm ) and "Air Toxics and Risk Assessment" By Edward J. Calabrese, Elaina M. Kenyon, 1991 CRC Press. The P=3/4 is based on trying to estimate calories consumed from the body weight ( http://books.nap.edu...i...634&page=56 ) The formula is supposed to be used only when you don't know calories consumed for the animals being compared. It's just usually easier to measure body weight than to figure out calorie consumption. This "surface area" formula method is based on trying to adjust for less calories/kg being lost in larger bodies (volume raises as r^3 while surface raises as r^2). This is where P=2/3 comes from. But in practice, they found P=3/4 was a more accurate way of estimating calories/kg. The FDA and EPA are sticking with P=2/3 because it gives a larger margin of safety for toxicology work.

Edited by zawy, 17 January 2008 - 01:14 AM.