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Aspirin Activates AMPK - But What's the Minimum Effective Dose?

aspirin ampk diabetes life extension

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#1 Bateau

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Posted 19 December 2013 - 07:42 AM


Back in April of 2012 it was discovered that salicylate, the active metabolite of aspirin, allosterically activates the master regulatory enzyme of metabolism, AMPK, in a unique manner that is only shared by the research chemical A-769662. Although salicylate is a weaker activator of AMPK than A-769662, as well as weaker than most AMPK activators in general, the manner in which it activates AMPK is unique in that salicylate attaches directly to the enzyme (AMPK) and prevents it from deactivating (dephosphorylating). This has an additive effect with other AMPK activators that activate AMPK through either upstream pathways (e.g. LKB1), the AMP+ADP/ATP ratio, ROS, osmotic stress or hormones (primarily leptin and adiponectin), since salicylate stabilizes AMPK though the beta regulatory subunit, while essentially every other way AMPK is activated is through the gamma regulatory subunit. This allows salicylate to be used in conjunction with other AMPK activators and have an additive effect, potentially magnifying and prolonging therapeutic effects.

The problem is the dosage. The initial research that revealed the discovery of the AMPK-activating effects of salicylate implied that high doses of aspirin would be necessary to achieve the effects on AMPK [1], ~4 grams a day by my amateur calculations [2]. Various experiments using salicylate and salsalate also showed that similar [3], if not higher[4][5] doses would be needed for effects on AMPK. However these studies were all using salicylate and salsalate, not aspirin. Several studies have come out using specifically aspirin that suggest that potentially much lower doses could be effective [6][7][8], with as little as 600mg a day possibly having the effect (inconclusive data IMO).

So my question to all the pharmacologists out there: According to the current evidence, what do you think is the minimum effective dose to achieve the AMPK-preserving effects of Aspirin? Please explain your reasoning on how you arrived at your conclusion.


Starting point for those unfamiliar with AMPK:

AMP-activated kinase (AMPK) is activated when the cellular (AMP+ADP)/ATP ratio rises; it therefore serves as a detector of cellular "fuel deficiency." AMPK activation is suspected to mediate some of the health-protective effects of long-term calorie restriction. Several drugs and nutraceuticals which slightly and safely impede the efficiency of mitochondrial ATP generation-most notably metformin and berberine-can be employed as clinical AMPK activators and, hence, may have potential as calorie restriction mimetics for extending healthspan. Indeed, current evidence indicates that AMPK activators may reduce risk for atherosclerosis, heart attack, and stroke; help to prevent ventricular hypertrophy and manage congestive failure; ameliorate metabolic syndrome, reduce risk for type 2 diabetes, and aid glycemic control in diabetics; reduce risk for weight gain; decrease risk for a number of common cancers while improving prognosis in cancer therapy; decrease risk for dementia and possibly other neurodegenerative disorders; help to preserve the proper structure of bone and cartilage; and possibly aid in the prevention and control of autoimmunity. While metformin and berberine appear to have the greatest utility as clinical AMPK activators-as reflected by their efficacy in diabetes management-regular ingestion of vinegar, as well as moderate alcohol consumption, may also achieve a modest degree of health-protective AMPK activation. The activation of AMPK achievable with any of these measures may be potentiated by clinical doses of the drug salicylate, which can bind to AMPK and activate it allosterically.

http://link.springer...3-9595-y#page-1


[1] http://www.ncbi.nlm....66/#!po=8.33333
[2] http://www.ncbi.nlm....1380040/?page=4
[3] http://www.ncbi.nlm....les/PMC3638122/
[4] http://www.ncbi.nlm....les/PMC3689003/
[5] http://www.sciencedi...006295213004863
[6] http://www.sciencedi...741521411023299
[7] http://www.sciencedi...53155651300051X
[8] http://www.ncbi.nlm....les/PMC3682211/

More studies on Aspirin, Salicylate and/or Salsalate and AMPK:
http://www.ncbi.nlm....pubmed/23801715
http://www.ncbi.nlm....pubmed/23721051
http://www.sciencedi...043276013001045
http://www.ncbi.nlm....pubmed/24190973
http://www.ncbi.nlm....pubmed/21514281
http://www.cell.com/...3-X?script=true

Edited by Bateau, 19 December 2013 - 08:37 AM.

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#2 Darryl

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Posted 19 December 2013 - 04:36 PM

I think 2300 mg / day (~7 * 325 mg tablets) could achieve the 1 mM salicylate concentration for significant AMPK induction in Hawley et al 2012.

In Keystone et al 1982, 3.9 g/d aspirin (given in different dosing schedules), achieved serum salicylate levels of 18-24 mg / dL. Hence 2.3-3.0 g/d would seem the right neighborhood for achieving 1 mM in serum
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#3 hav

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Posted 21 December 2013 - 11:29 PM

Here's a related study increasing the oral availability of berberine cloride using hydroxypropyl-β-cyclodextrin with the objective being to get an effective 100 mg dosage for treatment of gastroenteritis:

Physicochemical Characterization of Berberine Chloride

Phase solubility studies demonstrated the formation of berberine chloride–HPβCD inclusion complex with 1:1 stoichiometry, and the aqueous solubility of the drug improved almost 4.5-fold in the presence of 20% HPβCD.


They also tried various surfactants like tween 80 but they didn't do the trick.

Howard

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#4 joelcairo

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Posted 22 December 2013 - 12:00 AM

There are numerous studies showing anticancer effects of aspirin use in humans at normal therapeutic doses, which is at least partly attributed to AMPK activation.

For example, here's a meta-analysis of 11 studies investigating the protective effect of low-dose aspirin against cancer.
http://www.amjmed.co...0094-0/abstract

Personally I don't see why aspirin wouldn't be having a longterm effect at all dosage levels, although it might be hard to measure at doses like 80 mg - 325 mg.

#5 Bateau

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Posted 22 December 2013 - 12:42 AM

Personally I don't see why aspirin wouldn't be having a longterm effect at all dosage levels, although it might be hard to measure at doses like 80 mg - 325 mg.



The anti-cancer effects of aspirin could be attributed entirely to the inhibition of mTOR signaling that aspirin causes, which seems to be at least partially independent of its effects on AMPK. It's entirely possible that the typical dose of aspirin prevents cancer through mTOR signaling inhibition without ever activating AMPK. The reason this thread was made was so we can figure out the dose that aspirin activates AMPK, not just inhibits mTOR signaling.

Aspirin inhibits mTOR signaling, activates AMP-activated protein kinase, and induces autophagy in colorectal cancer cells.

BACKGROUND & AIMS: Aspirin reduces the incidence of and mortality from colorectal cancer (CRC) by unknown mechanisms. Cancer cells have defects in signaling via the mechanistic target of rapamycin (mTOR), which regulates proliferation. We investigated whether aspirin affects adenosine monophosphate-activated protein kinase (AMPK) and mTOR signaling in CRC cells.
METHODS: The effects of aspirin on mTOR signaling, the ribosomal protein S6, S6 kinase 1 (S6K1), and eukaryotic translation initiation factor 4E binding protein 1 (4E-BP1) were examined in CRC cells by immunoblotting. Phosphorylation of AMPK was measured; the effects of loss of AMPKα on the aspirin-induced effects of mTOR were determined using small interfering RNA (siRNA) in CRC cells and in AMPK(α1/α2-/-) mouse embryonic fibroblasts. LC3 and ULK1 were used as markers of autophagy. We analyzed rectal mucosa samples from patients given 600 mg aspirin, once daily for 1 week.
RESULTS: Aspirin reduced mTOR signaling in CRC cells by inhibiting the mTOR effectors S6K1 and 4E-BP1. Aspirin changed nucleotide ratios and activated AMPK in CRC cells. mTOR was still inhibited by aspirin in CRC cells after siRNA knockdown of AMPKα, indicating AMPK-dependent and AMPK-independent mechanisms of aspirin-induced inhibition of mTOR. Aspirin induced autophagy, a feature of mTOR inhibition. Aspirin and metformin (an activator of AMPK) increased inhibition of mTOR and Akt, as well as autophagy in CRC cells. Rectal mucosal samples from patients given aspirin had reduced phosphorylation of S6K1 and S6.
CONCLUSIONS: Aspirin is an inhibitor of mTOR and an activator of AMPK, targeting regulators of intracellular energy homeostasis and metabolism. These could contribute to its protective effects against development of CRC.


Edited by Bateau, 22 December 2013 - 01:01 AM.


#6 joelcairo

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Posted 22 December 2013 - 01:45 AM

"Could be", "seems to be", "entirely possible". I don't see why longterm aspirin use wouldn't have a beneficial effect on AMPK activation, so that's my two cents.

If you're trying to increase activation by some enormous amount, like by 100% or whatever, than that's a separate matter.
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#7 Darryl

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Posted 22 December 2013 - 03:30 AM

At this moment, we know nothing about AMPK induction with salicylate concentrations lower than 1 mM, which achieved a 50% increase in AMPK activity in Hawley et al. 2012. 100% increases required 3 mM, which I estimate would require about twice (7000-9000 mg) the label's maximum daily dosage (12 x 365 mg = 3900 mg).

There's no question smaller doses would be effective for AMPK induction in the intestines, and demonstrably are for colorectal cancer prevention.

My hope is that by combining moderate dose aspirin (perhaps 2-3 325 mg tablets/day), I might achieve perhaps a 25-30% independent contribution to AMPK induction, which as Bateau suggests, might have synergistic effects with other AMPK activators like fasting, exercise, and mild respiratory complex I poisons like berberine or metformin.

Its a pity salsalate isn't a an inexpensive over-the counter drug here, as for this purpose ASA is a prodrug (only the salicylate has AMPK activity), and I'd rather not have the platelet inhibiting "side effects" of ASA on intracranial and gastric bleeding. I've also looked into sodium salicylate, which sold for animal agriculture and is remarkably cheap, and white willow bark, which would increase pill volume by a factor of 10. Perhaps in a decade we'll see more experimental compounds like ZLN024 (EC50 around 1 µM, several thousand times as potent as salicylate) find approval for diabetes, and make off-label use of them.

Edited by Darryl, 22 December 2013 - 03:45 AM.

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#8 8bitmore

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Posted 22 December 2013 - 07:06 PM

If anyone is thinking of taking larger doses of aspirin it might well pay off to include piracetam in the equation since it seems to protect from the ulcer issues that aspirin carries with it: http://www.ncbi.nlm..../pubmed/2351418
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#9 hav

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Posted 22 December 2013 - 09:53 PM

Berberine cloride might be the best choice because aspirin would amplify its inherent ampk effects while berberine's gastroenteritis mitigating side effect might protect against aspirins ulcer promoting effect. And could bring the amount of aspirin needed down to a typical dosage. In fact, if you complex the berberine with a little cyclodextrin, it could all probably fit into a single nice little potent capsule.

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#10 Darryl

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Posted 17 January 2014 - 05:28 PM

Another alternative for those considering adding a salicylate to their regimens is magnesium salicylate, sold over the counter in the U.S. as a backache remedy (Doan's, Percogesic, generics). Unlike acetylsalicylate, it doesn't cause irreversible COX inhibition (with its gastric and anti-thrombotic effects), but its bioequivalent to aspirin in increasing plasma salicylate concentrations.

580 mg magnesium salicylate tetrahydrate tablets have 397 mg salicylate + 70 mg magnesium, and cost about $0.11 each (48 ct., store brand, on sale). The magnesium may be considered a useful supplement with anti-inflammatory and anti-sudden cardiac death effects (1, 2, 3).

Earlier this month, I discovered a high-aspirin regimen allowed me to donate a pint of blood in under 5 minutes. Not such a good thing in the event of accidental trauma. So I've decided to opt for 2 magnesium salicylate tablets daily instead, each taken with 500 mg nicotinic acid for anticipated synergies in NAD+ levels/sirtuin activation, slower clearance, and reduced flushing.
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#11 Logic

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Posted 17 January 2014 - 06:13 PM

I seem to remember Vit C being very good at countering the ulcer effects of Aspirin?

#12 Nate-2004

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Posted 28 June 2016 - 12:42 PM

I seem to remember Vit C being very good at countering the ulcer effects of Aspirin?

 

http://www.huffingto...g_b_529058.html

 

That article has some references or at least an explanation. 

 

I've been under the impression that aspirin is good for you because of its anti-inflammatory properties. That article paints a terrible picture about what it can do to your stomach and intestines. So in that case maybe it's one of those things you have to cycle in and out?

 

Any updates on this AMPK activation?



#13 ironfistx

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Posted 28 June 2016 - 02:42 PM

I'm pretty sure adding aspirin to alcohol is a bad way to get AMPK activation.


Will you give more information about irreversable COX inhibition?  Could that refer to something that never gets back to how it was?



#14 Skyguy2005

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Posted 28 June 2016 - 03:17 PM

 

I seem to remember Vit C being very good at countering the ulcer effects of Aspirin?

 

http://www.huffingto...g_b_529058.html

 

That article has some references or at least an explanation. 

 

I've been under the impression that aspirin is good for you because of its anti-inflammatory properties. That article paints a terrible picture about what it can do to your stomach and intestines. So in that case maybe it's one of those things you have to cycle in and out?

 

Any updates on this AMPK activation?

 

 

Aspirin and Willow Bark are unbelievably potent, IMO. Ginkgo Biloba is second only to Willow Bark in terms of the feeling I get (milder, less potent but still powerful and noticeable).

 

In the Concordia study they had dose-response curves for lifespan extension: http://www.impactjou...or-preview=5wx 

 

Search "supplementary files" to see the dose details and the graphs, for Plant Extracts 1-37. The Red ones indicate lifespan extension. 

 

Willow Bark peaked at 0.1 dose (same as celery seed), Ginkgo Biloba peaked at 0.3 dose, Black Cohosh and Valerian root peaked at 0.5, and they all declined after (U-shaped curve). Except True passionflower that was a little less potent, peaking at 1.0, and never actually declined probably because the dose was not high enough. 

 

The point is, Willow Bark and probably it's derivatives are potent but also the most finely balanced in terms of dose, just look at the supplemental figures in this study. Compared to Ginkgo Biloba and Valerian Root that don't reach quite the same peak, but possess a smoother, gentler curve.  

 

I don't know how this aspect would tie in to the AMPK activation. 


Edited by Skyguy2005, 28 June 2016 - 03:27 PM.

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#15 sthira

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Posted 03 January 2017 - 05:45 PM

Another alternative for those considering adding a salicylate to their regimens is magnesium salicylate, sold over the counter in the U.S. as a backache remedy (Doan's, Percogesic, generics). Unlike acetylsalicylate, it doesn't cause irreversible COX inhibition (with its gastric and anti-thrombotic effects), but its bioequivalent to aspirin in increasing plasma salicylate concentrations.

580 mg magnesium salicylate tetrahydrate tablets have 397 mg salicylate + 70 mg magnesium, and cost about $0.11 each (48 ct., store brand, on sale). The magnesium may be considered a useful supplement with anti-inflammatory and anti-sudden cardiac death effects (1, 2, 3).

Earlier this month, I discovered a high-aspirin regimen allowed me to donate a pint of blood in under 5 minutes. Not such a good thing in the event of accidental trauma. So I've decided to opt for 2 magnesium salicylate tablets daily instead, each taken with 500 mg nicotinic acid for anticipated synergies in NAD+ levels/sirtuin activation, slower clearance, and reduced flushing.


Is this combo (Doan's 2x + 500 niacin 2x) for possible AMPK activation still your routine?

#16 Darryl

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Posted 03 January 2017 - 08:28 PM

Yes, I'm still using OTC magnesium salicylate. The only alternatives for high dose salicylate without aspirin are magnesium salicylate (Doan's), veterinary sodium salicylate, or Rx salsalate.

 

Salsalate is preferable due to extended half-life (larger areas under the curve) for those who can obtain a prescription and who have good drug insurance, but as a single manufacturer generic its expensive. I hope salsalate comes into favor as an adjunct in diabetes treatment, as that might bring more manufacturers and reasonable pricing for off-label use in AMPK activation.



#17 Darryl

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Posted 04 January 2017 - 12:02 AM

Correction, there are at least three salsalate manufacturers. I've no idea why its still > $2.70 / g (Rx retail) given its a rather well-known synthesis.

 

Salsalate_synthesis.png



#18 zorba990

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Posted 22 January 2017 - 06:41 PM

Bumping this. I see some other salicylate sources as with Schweitzer formula (tin foil hat warning)
http://www.schweitzerformula.com/

And also copper salicylate which claims non irritation and actually ulcer healing
http://www.strideintohealth.com/


Ref
http://www.healing-y...alicylates.html
Willow bark v aspirin
https://www.ncbi.nlm...les/PMC3399766/

Bulk magnesium salicylate
http://www.citychemi...salicylate.html

Edited by zorba990, 22 January 2017 - 07:16 PM.

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#19 Dorian Grey

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Posted 22 January 2017 - 08:36 PM

For what it's worth, regular intake of aspirin (and ibuprofen & acetaminophen) have recently been associated with permanent hearing loss, particularly in males and often at a young age.  

 

https://www.ncbi.nlm...les/PMC2831770/

 

"During 369,079 person-years of follow-up, 3488 incident cases of hearing loss were reported. Regular use of each analgesic was independently associated with an increased risk of hearing loss. Multivariate-adjusted hazard ratios of hearing loss in regular users (2+ times/week) compared with men who used the specified analgesic <2 times/week were 1.12 (95% CI, 1.04–1.20) for aspirin, 1.21 (95% CI, 1.11–1.33) for NSAIDs, and 1.22 (95% CI, 1.07–1.39) for acetaminophen. For NSAIDs and acetaminophen, the risk increased with longer duration of regular use. The magnitude of the association was substantially higher in younger men. For men younger than age 50, the hazard ratio for hearing loss was 1.33 for regular aspirin use, 1.61 for NSAIDs, and 1.99 for acetaminophen.

Conclusions

Regular use of aspirin, NSAIDs, or acetaminophen increases the risk of hearing loss in men and the impact is larger on younger individuals.

---------------------------------

 

My girlfriend has taken an aspirin/acetaminophen/caffeine pain reliever fairly regularly for the last 30 years and now has substantial hearing loss that has seriously affected her life.  I sometimes tease her about how many times she'll say "huh, what" during normal conversation, even in a quiet room.  She also startles quite easily.  She can't hear me moving around the house and about jumps out of her shoes when she comes around a corner and sees me in a room she isn't expecting to find me.  She no longer appreciates music, & can't even hear something boiling over on the stove if she's not in the kitchen.  

 

I've always shunned pain meds as they wreck my gut, and my hearing is so acute I can tell where the cat is sleeping at night as I can hear it breathing.  This after attending dozens of rock concerts throughout the 70s, 80s & 90s, & smoking for over 40 years.  

 

You never realize just how much mild to moderate hearing loss can truly affect quality of life unless you've lived with someone who can no longer hear even normal conversation. Something to consider when thinking about long term / regular use of ototoxic substances.  

 


Edited by synesthesia, 22 January 2017 - 08:44 PM.


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#20 zorba990

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Posted 22 January 2017 - 09:58 PM

For what it's worth, regular intake of aspirin (and ibuprofen & acetaminophen) have recently been associated with permanent hearing loss, particularly in males and often at a young age.

https://www.ncbi.nlm...les/PMC2831770/

"During 369,079 person-years of follow-up, 3488 incident cases of hearing loss were reported. Regular use of each analgesic was independently associated with an increased risk of hearing loss. Multivariate-adjusted hazard ratios of hearing loss in regular users (2+ times/week) compared with men who used the specified analgesic <2 times/week were 1.12 (95% CI, 1.04–1.20) for aspirin, 1.21 (95% CI, 1.11–1.33) for NSAIDs, and 1.22 (95% CI, 1.07–1.39) for acetaminophen. For NSAIDs and acetaminophen, the risk increased with longer duration of regular use. The magnitude of the association was substantially higher in younger men. For men younger than age 50, the hazard ratio for hearing loss was 1.33 for regular aspirin use, 1.61 for NSAIDs, and 1.99 for acetaminophen.
Conclusions
Regular use of aspirin, NSAIDs, or acetaminophen increases the risk of hearing loss in men and the impact is larger on younger individuals.

---------------------------------

My girlfriend has taken an aspirin/acetaminophen/caffeine pain reliever fairly regularly for the last 30 years and now has substantial hearing loss that has seriously affected her life. I sometimes tease her about how many times she'll say "huh, what" during normal conversation, even in a quiet room. She also startles quite easily. She can't hear me moving around the house and about jumps out of her shoes when she comes around a corner and sees me in a room she isn't expecting to find me. She no longer appreciates music, & can't even hear something boiling over on the stove if she's not in the kitchen.

I've always shunned pain meds as they wreck my gut, and my hearing is so acute I can tell where the cat is sleeping at night as I can hear it breathing. This after attending dozens of rock concerts throughout the 70s, 80s & 90s, & smoking for over 40 years.

You never realize just how much mild to moderate hearing loss can truly affect quality of life unless you've lived with someone who can no longer hear even normal conversation. Something to consider when thinking about long term / regular use of ototoxic substances.

Interesting. Wikipedia also has this to say about salicylic acid:
"When ingested, salicylic acid has a possible ototoxic effect by inhibiting prestin.[19] It can induce transient hearing loss in zinc-deficient rats. An injection of salicylic acid induced hearing loss, while an injection of zinc reversed the hearing loss. An injection of magnesium in the zinc-deficient rats did not reverse the induced hearing loss.[citation needed]"

Also from
https://www.ncbi.nlm...les/PMC3395201/
"N-acetylcysteine (NAC) NAC has been shown to be otoprotective in both noise induced, aminoglycoside as well as cisplatin ototoxicity in the animals in several studies. Some examples of the studies: Feldman et al. [49] showed otoprotection by NAC against gentamicin induced ototoxicity in hemodialysis patients."

And confusingly:
https://www.research...Threshold_Shift

"The permanent threshold shift (PTS) following exposure to intense noise may be due to the noise-induced excessive vibrations in the cochlea or to the generation of elevated levels of reactive oxygen species. Thus, it is possible that the resulting PTS may be reduced if the cochlear amplifier could be temporarily depressed beginningjust before the onset of the noise and continuing during the noise exposure or if antioxidant drugs were administered. These possibilities were assessed in mice by administering a single injection of salicylic acid (an antioxidant drug which also reversibly depresses the motor protein prestin of the cochlear amplifier) just before, and in other mice, just after, 3.5 h of 113-dB SPL broadband noise exposure. The PTS in the mice injected with salicylic acid just before the noise exposure was significantly smaller than that in mice exposed to the same noise without salicylic acid. The PTS in the latter was not significantly different from that in mice who received the drug just after the noise. Thus a single injection of salicylic acid, just before a noise exposure, can protect the ear from a noise-induced hearing loss"

Edited by zorba990, 22 January 2017 - 10:06 PM.

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