• Log in with Facebook Log in with Twitter Log In with Google      Sign In    
  • Create Account
  LongeCity
              Advocacy & Research for Unlimited Lifespans

Photo
- - - - -

Autophagy Induction with Liposomal Trehalose

autophagy liposomal trehalose trehalose tau protein osteoarthritis arthritis

  • Please log in to reply
20 replies to this topic
⌛⇒ support MITOMOUSE via LongeCity!

#1 Fafner55

  • Member
  • 228 posts
  • 220
  • Location:USA
  • NO

Posted 29 September 2018 - 04:14 PM


Summary: Liposomal trehalose is prepared and a therapeutic oral dose is estimated.
 
Background: Research indicates that the disaccharide trehalose initiates autophagy by blocking a glucose transporter. Cells are thus nutrient deprived, so it's something like a CR mimetic. In mice and humans, trehalose has been shown to reduce tau pathology, induce mitophagy, improve osteoarthritic cartilage and artery tissues.
  1. “Trehalose ameliorates dopaminergic and tau pathology in parkin deleted/tau overexpressing mice through autophagy activation” (2015) https://drive.google...kPqNZqGFIWdZR  
  2. “Trehalose ameliorates oxidative stress-mediated mitochondrial dysfunction and ER stress via selective autophagy stimulation and autophagic flux restoration in osteoarthritis development” (2017) https://www.nature.c...es/cddis2017453
  3. “Oral trehalose supplementation improves resistance artery endothelial function in healthy middle‐aged and older adults” (2016) https://www.ncbi.nlm...ing-08-1167.pdf
  4. Trehalose works by blocking glucose transport and mimicking calorie restriction, thus activating AMPK.  “Mystery solved: Trehalose kickstarts autophagy by blocking glucose transport” (2016) https://drive.google...ufL3iPcKgJeh9  
 
Preparation: Trehalose is water soluble and easily prepared in liposomal form. Instructions are attached.
 
Starting dose: In mice, a 2% oral dose of trehalose induces autophagy. Assuming an average 25 g mouse drinks 4 ml water daily, a 2% solution is 3200 mg/kg. The human equivalent dose is 260 mg/kg, or for a 70 kg individual, this dose is 18.2 gm. For 0.5% bioavailability, only 0.091 gm of 18.2 gm reaches the bloodstream. 
 
Assuming liposomal formulations encapsulate at a 50% rate (my guess), 30% phosphatidylcholine content, and 175 gm trehalose/1000 gm solution, then the target equivalent liposomal dose of trehalose is 3.46 gm.
Then, 1 tsp (5 gm) should be a good starting point for self-experimentation.
 
Results: 
  • 1 tsp/day for 10 days had no apparent effect. 
  • 2 tsp, 1x/day for 7 days lead to fading of lipofuscin accumulation in spots on my hands and face beginning within a couple of days. This fading appeared to slow on days 6 and 7.
  • 2 tsp, 2x/day for 3 days resulted in dramatic increase in spot fading but minor muscle aches and fatigue on day 2 and slight fever on day 3. My doctor suggested that the aches were a symptom of rhabdomyolysis. Comprehensive blood panel tests 2 weeks later showed no issues.
Oral dose: At least for me, 2 tsp, 1x/day for 5 or 6 days seems safe and effective. 2 tsp, 2x/day is harmful.
 
References: Background references for manufacturing liposomal trehalose and research on the effects of trehalose on mice and humans is included in the attachment.
 
 

  • Informative x 3
  • WellResearched x 2
  • like x 2

#2 John250

  • Guest
  • 1,451 posts
  • 103
  • Location:Temecula
  • NO

Posted 29 September 2018 - 06:23 PM

Can this still induce autophagy if your not fasted?

sponsored ad

  • Advert
Click HERE to rent this advertising spot for SUPPLEMENTS (in thread) to support LongeCity (this will replace the google ad above).

#3 Fafner55

  • Topic Starter
  • Member
  • 228 posts
  • 220
  • Location:USA
  • NO

Posted 29 September 2018 - 06:31 PM

Can this still induce autophagy if your not fasted?

 

I did not fast during this test. Liposomal trehalose induces autophagy independently of fasting. Trehalose works by blocking glucose transport and mimicking calorie restriction, thus activating AMPK.

“Mystery solved: Trehalose kickstarts autophagy by blocking glucose transport” (2016) https://drive.google...qmufL3iPcKgJeh9



⌛⇒ support MITOMOUSE via LongeCity!

#4 Benko

  • Guest
  • 210 posts
  • 324
  • Location:US

Posted 30 September 2018 - 12:16 AM

“At least for me, 2 tsp, 1x/day for 5 or 6 days seems safe and effective. 2 tsp, 2x/day is harmful.”

What harm did you experience from 2 tsp 2x/day?

#5 Fafner55

  • Topic Starter
  • Member
  • 228 posts
  • 220
  • Location:USA
  • NO

Posted 30 September 2018 - 12:40 AM

“At least for me, 2 tsp, 1x/day for 5 or 6 days seems safe and effective. 2 tsp, 2x/day is harmful.”

What harm did you experience from 2 tsp 2x/day?

 

Minor muscle aches and fatigue on day 2 and slight fever on day 3. These are symptoms of rhabdomyolysis.


Edited by Fafner55, 30 September 2018 - 12:42 AM.

  • Needs references x 1

#6 Fafner55

  • Topic Starter
  • Member
  • 228 posts
  • 220
  • Location:USA
  • NO

Posted 02 October 2018 - 02:05 AM

I should add that my 3 trials (1 tsp/day, 2 tsp/day, 2tsp, 2x/day) were consecutive, one after the other without recovery time in between.


  • Needs references x 1

#7 Ducky-001

  • Member
  • 30 posts
  • 23
  • Location:Europe
  • NO

Posted 31 October 2018 - 12:42 PM

Did the spots fade permanently? Any other "permanent" changes?

 



#8 Fafner55

  • Topic Starter
  • Member
  • 228 posts
  • 220
  • Location:USA
  • NO

Posted 31 October 2018 - 01:35 PM

I did a second round of liposomal trehalose (2 tsp/day for 5 days) about 3 weeks after the initial round. The spots on the back of my hands still appeared faded at the beginning of that round, and are still faded today (it has been less than 3 weeks), so I can only know for sure that those spots remain faded for 3 weeks.



#9 able

  • Guest
  • 731 posts
  • 333
  • Location:austin texas
  • NO

Posted 31 October 2018 - 01:57 PM

Do you follow any particular diet?

 

I'm curious how this would work for someone following a keto diet like me.  

 

For those keto adapted to burn fat and not be dependent on glucose, it might not have as much effect.  

 

Or, it might allow higher dosage and work even better.

 

Yet another test I'll have to try - thanks for the info.



⌛⇒ support MITOMOUSE via LongeCity!

#10 Fafner55

  • Topic Starter
  • Member
  • 228 posts
  • 220
  • Location:USA
  • NO

Posted 31 October 2018 - 08:55 PM

Do you follow any particular diet?

 

I'm curious how this would work for someone following a keto diet like me.  

 

For those keto adapted to burn fat and not be dependent on glucose, it might not have as much effect.  

 

Or, it might allow higher dosage and work even better.

 

Yet another test I'll have to try - thanks for the info.

 

I considered fasting (by a fasting mimicking diet) but decided against it for the reasons:

1) I wanted to determine the efficacy of lipo trehalose in activating autophagy without complicating factors

2) I presumed that if lipo trehalose was truly effective, I would not need to further promote autophagy. Indeed it was, since taking to much for too long apparently caused rhabdomyolysis.

 

In my second trial, I experienced slight muscle aches on the 4th and 5th days, and stopped after day 5. I didn't want to push it any further.



#11 Fafner55

  • Topic Starter
  • Member
  • 228 posts
  • 220
  • Location:USA
  • NO

Posted 15 December 2018 - 05:28 PM

Phosphatidylcholine Content of Soybean versus Sunflower Lecithin
 
Background
Previously I made liposomal trehalose using an expensive soybean lecithin product that advertised 40% PC content. To determine if other sources of phosphatidylcholine might be used, and to better gauge how the dose of liposomal trehalose might be affected, I extracted the alcohol soluble content of different lecithin products. 
 
From lecithins, alcohol extractions yield mostly phosphatidylcholine (PC) since other  phospholipids (such as phosphatidylinositol (PI) and phosphatidylethanolamine (PE)) have lower alcohol solubility. After evaporating the alcohol, I found the remaining residue to be entirely water soluble, demonstrating that the oil/fat/fatty acid content of the extraction was low.
 
Result
By my experiments,
  • Soybean lecithin (Now brand) has 26.3% alcohol soluble content (isopropyl). This is close to product labels.
  • Sunflower lecithin (Swanson brand) has 19.5% alcohol soluble content.(isopropyl). This is lower than product labels.
  • A brand that advertises a high (40%) phosphatidylcholine content (Happy Bodies, HB-PC Phosphatidyl Choline-40%) had a 28% alcohol soluble content (isopropyl).
 
Conclusion
Inexpensive soybean lecithin is a good starting material for homemade liposomal products.
 
 

Attached Files


  • WellResearched x 3
  • Informative x 1
  • like x 1

#12 Munin

  • Member
  • 4 posts
  • 3
  • Location:EU
  • NO

Posted 15 January 2019 - 11:00 AM

Quality control of the Phosphatidylcholine content of Soybean Lecithin

 

I have followed the example of Fafner55 and made a control of the PC content in Soy Lecithin.

 

Soy Lecithin brand:  NEMKUR (made in Germany)

 

Process:        40 g Soy Lecithin was dissolved in 200 g Isopropyl (Isopropanol) 99.5%.

                      The blend (in a jar with lid) was mixed for two days by magnetic stirring.

 

                      The mixture settled for 12 h and a yellow transparent liquid was formed over a sludge on the bottom.                        

                       I poured of the alcohol liquid and was careful of not getting any sludge into the new beaker. This repeated several times.

 

                       I had now 135 g of isopropyl plus alcohol soluble substances.

                       The new beaker was placed in a water bath which was heated to 95 C. After two h had all the isopropyl evaporated and left was 4.5 g of a oily, yellowish                                   transparent liquid with a viscosity somewhere between oliveoil and syrup.

 

                       Assuming that all the alcohol in the sludge has the same concentration of soluble substances (PC and others) then this would give a total 6.9 g of soluble                                 content, i.e. 17 % of the used 40 g Soy Lecithin.

 

 

Discussion:     The Nemkur label states more then 20 % PC which might have been extracted if one had a more optimized process. But if I want to do my own Liposomal                                 mixture (following the Fafner55 process) then this is probably the concentration of PC I can expect using Nemkur Soy Lecithin.

 

 

 

 


  • Good Point x 2

#13 Daniel Cooper

  • Guest
  • 1,062 posts
  • 174
  • Location:USA

Posted 15 January 2019 - 03:44 PM

Did you sonicate or use a high speed mixer in making your preparation?

 

 

 



#14 Munin

  • Member
  • 4 posts
  • 3
  • Location:EU
  • NO

Posted 15 January 2019 - 05:44 PM

I only used a magnetic stirrer.

 

No mixer(blender) or ultrasound equipment was used. This could perhaps increase yield.



#15 Fafner55

  • Topic Starter
  • Member
  • 228 posts
  • 220
  • Location:USA
  • NO

Posted 15 January 2019 - 06:03 PM

Nice work Munin. It points to a large range of phosphatidylcholine (PC) content in commercially available lecithins. This isn't surprising given differences in soybean seed and growing conditions.

 

Knowing the PC content will help adjust a liposomal trehalose dose. When you get to that point, if you take a dose that compares to one I took (such as 2 tsp, 1x/day), please let us know if and when you experience muscle pain.



#16 Daniel Cooper

  • Guest
  • 1,062 posts
  • 174
  • Location:USA

Posted 15 January 2019 - 06:06 PM

I only used a magnetic stirrer.

 

No mixer(blender) or ultrasound equipment was used. This could perhaps increase yield.

 

 

I do not believe you will get any significant amount of liposome encapsulation from magnetic stirring only.  I might be wrong about that but I've never heard of anyone making liposome encapsulations by that method.

 

This highlights the fact we really need some way to check these attempts at liposomal encapsulation to see how successful they were.


Edited by Daniel Cooper, 15 January 2019 - 06:09 PM.


#17 Munin

  • Member
  • 4 posts
  • 3
  • Location:EU
  • NO

Posted 15 January 2019 - 06:24 PM

Well, this experiment was not about making "Liposome formation". 

 

The purpose was only to establish the amount of PC in my Soy Lecithin to be used later on for Liposomal Trehalose.

 

I just followed the above described process in Phosphatidylcholine Content of Soybean versus Sunflower Lecithin.pdf

 

But please do some expreiments according to your own ideas and report back your findings. Thats the only way bring the common knowledge forward. 

 

OK?

pdf.gif  Phosphatidylcholine Content of Soybean versus Sunflower Lecithin.pdf   65.11KB   11 d


  • Agree x 1

#18 Daniel Cooper

  • Guest
  • 1,062 posts
  • 174
  • Location:USA

Posted 15 January 2019 - 06:41 PM

Ah, I misunderstood.  That's what I get for not reading carefully.

 

 

 



#19 Munin

  • Member
  • 4 posts
  • 3
  • Location:EU
  • NO

Posted 15 January 2019 - 06:43 PM

Thats easy to do. There is so much information on this site so you have to read fast  :) .



#20 granmasutensil

  • Guest
  • 44 posts
  • 9
  • Location:USA

Posted 22 January 2019 - 09:35 AM

I do not believe you will get any significant amount of liposome encapsulation from magnetic stirring only.  I might be wrong about that but I've never heard of anyone making liposome encapsulations by that method.

 

This highlights the fact we really need some way to check these attempts at liposomal encapsulation to see how successful they were.

 

This is somewhat not completely relevant yet I think it is worth mentioning. You never know what other people might know and what brainstorming might bring here. There is actually a product that has been developed called "pro-lipo™ neo" that all you have to do is stir in your lipid or water soluble compounds and it instantly makes 200-250nm liposomes. It's a fairly new technology and is now being sold to the cosmetic industry for extremely easy to make liposomes. Maybe the compounds they use are safe for ingestion? Maybe this can be replicated ourselves, or with other food safe compounds? Just thought I should throw it out there. It's a pretty amazing new concept.


  • like x 3
  • Needs references x 1
  • Informative x 1

sponsored ad

  • Advert
Click HERE to rent this advertising spot for SUPPLEMENTS (in thread) to support LongeCity (this will replace the google ad above).

#21 QuestforLife

  • Member
  • 724 posts
  • 328
  • Location:UK

Posted 22 January 2019 - 01:27 PM

Do you follow any particular diet?

 

I'm curious how this would work for someone following a keto diet like me.  

 

For those keto adapted to burn fat and not be dependent on glucose, it might not have as much effect.  

 

Or, it might allow higher dosage and work even better.

 

Yet another test I'll have to try - thanks for the info.

 

I would guess autophagy already upregulated on keto, as glucose transporters will not be active.

 

There is some evidence for upregulation of autophagy and mitophagy on a KD:

 

https://www.ncbi.nlm...pubmed/29056525

 

https://www.ncbi.nlm...les/PMC5186794/


  • Informative x 2





Also tagged with one or more of these keywords: autophagy, liposomal trehalose, trehalose, tau protein, osteoarthritis, arthritis

0 user(s) are reading this topic

0 members, 0 guests, 0 anonymous users