Updated Research and Knowledge - Cutting Edge

[Keeper]

We have our first participant!

@Keeper

You've convinced me. They seem to be genuine, and they're offering their help free of charge, this'll benefit alot of users. And we'll have a consistent formula. Lets make this thing happen.

Can they create formulations with ingredients besides oleuropein? Like EGCG.

To be honest I don't know what ingredients they can also mix in. I could ask them but I don't want too bother them too often, so I think the best is if we collect our ideas what could be in and how to mix it with what for producing sth, and then I will ask them in one rush. (I guess sth like a cream/topiclal solution would be the best.)

They emailed me that they can produce every Oleuropein concentration we want. Would be nice if more people would think about the ideas and try to help figuring out a plan.

[potato1987]

I am not on any routine, the only thing I do at present is castor oil and derma rolling, I would be happy to document and take part, the only thing is on swisstemples he believes it may reduce PGE2, but I think castor oil increases it, so a mix Emu, Castor and Oleuropein? I would leave in over night and apply every night.

cheers,

[joshuk]

just thought i would post an update been using Oleuropein ethanol tincture of amazon for 2 weeks now, i stopped minox 2 months ago due to face/skin probs ( it was the minox my face is better not 100% but alot better now)

the small vellus hairs that minox made were about 1-2mm in length across the hairline since using the Oleuropein some of these have doubled in length to 3-4mm nothing groundbreaking but they are getting longer for sure so im hoping in the next few weeks they might keep growing.

this is not cosmetic regrowth as the hairs in question are not pigmented just thought i would share

[Chemical]

Chemical, do you think it is ok if I mix EGCG with oleuropein; same ethanol mixture?

And it would be really great to hear your opinion on this:
http://www.ncbi.nlm.nih.gov/pubmed/24676213

Yes it should be fine, but before you go ahead, which brand of EGCG are you using? I'd recommend teavigo since you can get away with a lower concentration of the extract and get the raw EGCG that you actually need. I'm ot sure how the other polyphenols affect, so for now we'll stick to pure EGCG as used in the study. EGCG also has a dose dependent effect so start off with 1 capsule, then see if it leaves a residue after applying. If not, add another. Rub the skin softly to see if you get any residue, even if its just a little, dont add anymore to the solution. Also what concentration of ethanol are you using?

Hi Chemical I am willing to try this out.
I found a tincture in my local shop that has 65% olea europea (with water and alcohol but not sure of the exact ratio yet) and I also found 500 mg capsules of green tea which contain 15% ECGC. I intend to mix them by adding a bit more alcohol and water i n a seperate bottle.
What ratio would you use?

I would encourage you to try and source the extracts online because then you know what concentration to use. It's sometimes difficult to know exactly how much oleuropein a tincture has and if its got any additional unwanted ingredients. iherb is a great place to order extracts/supplements and they deliver worldwide. Theyre very cheap and alot of people including myself can vouch for them - that is if you cant find what youre looking for on amazon. 15% EGCG isnt a high enough concentration imo, so for those reasons I'd suggest acquiring the ingredients elsewhere. I'm not 100% sure on the EGCG mixing since I'm yet to try it, but use my answer to SriHanuman as a starting point.

I read that minoxidil sulfate is the active metabolite that actually promotes hair growth. Minoxidil sulfate is made by the sulfotransferase enzyme. It is common knowledge that minoxidil doesn't work for everybody, possibly because we have different sulfotransferase enzyme activity.

So I am wondering: Is it important to have a right pH balance for minoxidil and other treatments to work, or to work better? Is temperature important? I was always applying minoxidil after a hot shower to increase permeation but is it possible that at the same time I was inactivating the sulfotransferase enzyme? I don't know but from now on I will only wash my head with cold water

http://dmd.aspetjournals.org/content/37/5/1083.full

Absolutely correct paleocapa89. Minoxidil sulfate is the active metabolite that stimulates hair follicles.

Dose-response studies showed that minoxidil sulfate is 14 times more potent than minoxidil in stimulating cysteine incorporation in cultured follicles. Three drugs that block production of intrafollicular minoxidil sulfate were tested for their effects on drug-induced hair growth. Diethylcarbamazine proved to be a noncompetitive inhibitor of sulfotransferase and prevented hair growth stimulation by minoxidil but not by minoxidil sulfate.

Biochemical evidence for minoxidil sulphation by two phenol sulphotransferases has been found in human scalp skin[22] and Dooley[21] reported finding mRNA expression for four sulphotransferases in human epidermal keratinocytes. There are interindividual variations in scalp sulphotransferase activity and this correlates with the level in platelets.[22] In a clinical setting, scalp sulphotransferase activity was higher in men who responded to minoxidil compared with those who did not respond [23]
http://www.medscape.com/viewarticle/470297_3

The variation in sulphotransferases enzymes impacts how people respond to minoxidil. You are also correct that PH and temperature affect the sulphotransferase enzymes. Enzymes in general have an optimal environmental state that allows them to catalyse reactions. I wouldnt worry to much about PH and temperature, your body's natural homeostasis should take care of that for you:

Normal human body temperature of 37C provides a good internal environment for enzymes to work efficiently. Enzymes in the stomach, such as pepsin ( which digests protein ), work best in very acid conditions ( pH 1 - 2 ), but most enzymes in the body work best close to pH 7.

I don't want too bother them too often, so I think the best is if we collect our ideas what could be in and how to mix it with what for producing sth, and then I will ask them in one rush. Would be nice if more people would think about the ideas and try to help figuring out a plan.

Good idea, I agree that we should wait a little till I've got some more experimental outcomes to go on before bothering them with additional requests.

The only thing I do at present is castor oil and derma rolling, I would be happy to document and take part, the only thing is on swisstemples he believes it may reduce PGE2, but I think castor oil increases it, so a mix Emu, Castor and Oleuropein? I would leave in over night and apply every night.

cheers,

Thats an interesting site you linked, I'm going to go over the stuff wen I'm free to see if theres anything of interest. As for PGE2, my next post will be an in depth analysis on how PGE2 actually works to promote hair growth. In short, Ricinoleic acid (found in castor oil) activates the EP3 receptors in women. It should in theory do the same in men and exert it effects when applied topically. EP3/EP2 agonists increase Beta Catenin, and thats why they grow hair. Not sure about Castor oil inhibiting PGE2 synthesis - negative feedback? Anyways, I dont think castor oil is bad. My recent findings indicate oleuropein works faster if you use an ethanol vehicle, and using it in an oil based solution will yield sub par results. You want to aim for 1-2mg per ml and use the oils after the ethanol has dried up.

A caution for people making ethanol vehicles, you can only use ethyl alcohol/190 proof alcohol found in drinks, NOT denatured ethanol or Isopropyl/Isopropanol. This study on topical ethanol/PG formulations of minox shows that anything above 90% Ethanol/10 % PG results in absorption less than that of 50% ethanol/50% PG. With 50/50 to 90/10 having optimal absorption. PG seems to keep the minox from forming crystals/residue on the skin and improves absorption if my understanding is correct, whereas a 100% ethanol vehicle just doesnt work. This probably applies to oleuropein as well and possibly EGCG.

[Chemical]

The small vellus hairs that minox made were about 1-2mm in length across the hairline since using the Oleuropein some of these have doubled in length to 3-4mm nothing groundbreaking but they are getting longer for sure so im hoping in the next few weeks they might keep growing.

This is promising! Which brand are you using?

I've been doing some more research on PGE2 and thought I'd share my findings here. We'll start off with this study that shows PGE2's effects on β-catenin:

Prostaglandin E2 promotes colon cancer cell growth through a Gs-axin-beta-catenin signaling axis.
Abstract

We show that PGE2 stimulates colon cancer cell growth through its heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptor, EP2, by a signaling route that involves the activation of phosphoinositide 3-kinase and the protein kinase Akt by free G protein betagamma subunits and the direct association of the G protein alphas subunit with the regulator of G protein signaling (RGS) domain of axin. This leads to the inactivation and release of glycogen synthase kinase 3beta from its complex with axin, thereby relieving the inhibitory phosphorylation of beta-catenin and activating its signaling pathway. These findings may provide a molecular framework for the future evaluation of chemopreventive strategies for colorectal cancer.

PGE2 inhibits axin mediated formation of the BetaCatenin destruction complex involving GSK3Beta. It does this via the EP2 receptor and also the EP3/4 receptors:

Prostaglandin E2 promotes human cholangiocarcinoma cell proliferation, migration and invasion through the upregulation of β-catenin expression via EP3-4 receptor.

To understand what the Axin/APC destruction complex is, heres a graphical representation of what it looks like:



Wnt Signaling through Inhibition of β-Catenin Degradation in an Intact Axin1 Complex

The proteins surrounding β-catenin are involved in the destruction of β-catenin. First CKI prepares β-catenin for GSK3Beta. Once GSK3Beta has phosphorylated β-catenin, Axin and APC form a scaffold that that enables the β-catenin to be degraded. If any of these proteins are inactivated then β-catenin will not be destroyed and translocate to the nucleus and exert its effects with the help of TCF4. WNTs like WNT10b - which oleuropein upregulates - protect β-catenin from destruction:

The removal of β-catenin from the destruction complex is accomplished simply by direct degradation by the proteasome. This step recycles the destruction complex for another round of β-catenin degradation. We show that Wnt receptor-ligand interaction leaves the destruction complex compositionally unchanged and does not affect the activity of its kinases. The only change that we observe is the association of Axin1 with phosphorylated Lrp6 and the dissociation of β-TrCP. Indeed, phosphorylated β-catenin—still bound to the Axin1 complex—is no longer ubiquitinated and degraded. It saturates and thus effectively inactivates the Axin1 complex. We have previously reported that only β-catenin that is newly synthesized after initiation of the Wnt signal is signaling competent (Staal et al., 2002). This notion is in agreement with our current model, which predicts that newly synthesized, nonphosphorylated β-catenin will be stable in a free cytosolic form once the destruction complex is saturated. It can then translocate to the nucleus to associate with TCF and activate the Wnt transcriptional program.

Anything that blocks Axin or GSK3 or any of the proteins required to destroy β-catenin will lead to increased β-catenin. Thats why GSK3 inibitors like Valproic acid work topically. PGE2 -> ( EP2/3 ¬ Axin ) -> β-catenin. This is one possible explanation for castor oils hair growth promoting effects.

PGE2 is quite difficult to increase and analogues are very expensive. Castor oil is a viable alternative however. If there was a different molecule that could inhibit Axin then we could use that to directly increase β-catenin bypassing WNT. Thats where EGCG comes in:

The anti-adipogenic effects of (-)epigallocatechin gallate are dependent on the WNT/β-catenin pathway.

Abstract
(-)Epigallocatechin gallate (EGCG) is the most abundant catechin in green tea and reportedly has anti-obesity and anti-adipogenic effects. In this study, we determined that the up-regulation of the WNT/β-catenin pathway is the anti-adipogenic mechanisms of EGCG in 3T3-L1 cells. EGCG treatment down-regulates the expression of major genes involved in the adipogenesis pathway including peroxisome proliferator-activated receptor (PPAR)γ, CCAAT/enhancer binding protein (C/EBP)α, fatty acid binding protein (FABP)4 and fatty acid synthase (FASN), while up-regulating the nuclear level of β-catenin. Knockdown of β-catenin using small interfering (si) RNA attenuated the inhibitory effects of EGCG on intracellular lipid accumulation. β-catenin siRNA transfection also recovered terminal adipocyte markers such as FABP4, FASN, lipoprotein lipase and adiponectin, which were down-regulated by EGCG. The DNA binding activities as well as the expression levels of PPARγ and C/EBPα, which were down-regulated by EGCG, were significantly restored by β-catenin siRNA transfection. In addition, we found that EGCG efficiently up-regulates the WNT/β-catenin pathway. Among the members of the WNT/β-catenin pathway, the expressions of low density lipoprotein receptor-related protein LRP5, LRP6, disheveled DVL2 and DVL3 were significantly up-regulated, while AXIN expression was down-regulated by EGCG, and the phosphorylation of glycogen synthase kinase 3β was increased. These results suggest that EGCG activates the WNT/β-catenin pathway, resulting in the up-regulation of β-catenin, which down-regulates the major genes of the adipogenesis pathway. Taken together, our findings clearly show that the anti-adipogenic effects of EGCG are, at least partially, dependent on the WNT/β-catenin pathway.

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

EGCG, in addition to inihibiting AR expression, downregulates Axin and reduces the amount of GSK3 available to destroy β-catenin.

So this is what the current treatment stack looks like:

(Oleuropein ¬ DDK1) -> natural WNTs can bind
Oleuropein -> WNT10b -> β-catenin

Minoxidil -> Adenosine Receptor -> β-catenin
(Minoxidil ¬ AR) -> β-catenin + TCF4 activity

(EGCG ¬ AR) -> β-catenin + TCF4 activity
(EGCG ¬ Axin) -> β-catenin

((Mico/Keto ¬ 3β-Diol) ¬ ERβ ) -> VEGF

And if you want to use castor oil:

(Ricinoleic Acid -> EP3 ¬ Axin) -> β-catenin

I think we should all try and find cost effective herbs/supplements that can downregulate AR or boost WNTs/β-catenin signalling. The more readily vailiable options we have, the greater the chance people can use what works best for them. We might even have a ridiculously powerful stack to reverse AGA or even cure it - and by cure I mean you only need to use the treatments infrequently to maintain density. I have hopes that next year we will have some interesting breakthroughs/discoveries, and we may no longer need to rely on or wait for big pharmaceutical companies for new treatments or only rely on expensive treatments to grow hair.

[joshuk]

using this ethanol tincture http://www.amazon.co.uk/gp/product/B...ilpage_o02_s00

do you think if i put a cap of EGCG into it, it would not mess up the Oleuropein im also using CB-03-01 (AR blocker??) would their be any point with EGCG if im using an AA or do you think it could compliment it

[joshuk]

chemical im going to make a new topical with EGCG and Oleuropein in it

gonna use 60/40 mix of ethanol/pg so if i make a 60ml solution being 40ml ethanol 20ml PG

Oleuropein caps are = http://www.amazon.co.uk/gp/product/B...ilpage_o03_s00
EGCG caps are = http://www.amazon.co.uk/gp/product/B...ilpage_o00_s00

how many caps of each would you add?? just 1 im thinking as a trial or would you add more

also for people wanting to try this in europe i got my ethanol from https://www.thewhiskyexchange.com/P-15105.aspx

and proplyene glycol from http://www.cremedevape.com/epages/yx...jectID=4164040

im also using CB-03-01 and setipirant/ neogenic and adenonise castor oil 1ml orally and nizoral 3 times a week.
so hoping this stacked with these will bring results ( cant use minox otherwise i would )

[Chemical]

http://www.amazon.co.uk/gp/product/B...ilpage_o02_s00

Looking at that product again it seems it may be not 100% effective as kirkland minox vehicle seeing as its got only 36% ethanol derived from cane sugar(?):

Raw Olive Leaf, Olive Leaf Extract (20% oleuropein standardised), ethanol 36% (derived from cane sugar), purified water.

And although its a prepared solution, it doesnt gaurantee it'll be as effective as a custom vehicle. The oleuropein content is around 31mg/ml which is ridiculously high. for its low ethanol percentage. If you really want to avoid minox and you're under a budget, then this will have to do. Be sure to use only 1 drop, no more.

Approx 130 servings of 20 drops. Approx. 25mg oleuropein per serving.
100/130 = 0.78ml per 20 drops.
25mg/0.78ml = 31mg/ml.

chemical im going to make a new topical with EGCG and Oleuropein in it. Gonna use 60/40 mix of ethanol/pg so if i make a 60ml solution being 40ml ethanol 20ml PG.

Oleuropein caps are = http://www.amazon.co.uk/gp/product/B...ilpage_o03_s00
EGCG caps are = http://www.amazon.co.uk/gp/product/B...ilpage_o00_s00

how many caps of each would you add?? just 1 im thinking as a trial or would you add more?

I'm also using CB-03-01 and setipirant/ neogenic, adenonise, castor oil 1ml orally and nizoral 3 times a week. So hoping this stacked with these will bring results ( cant use minox otherwise i would )

It seems you've beaten me to it. A purer ethanol vehicle with a bit of PG will most likely yield better results even if only increases the chances/probability of regrowth by a small amount. 80% ethanol, 20% PG. Thats what I would do. 1 cap of that oluropein you linked will give around 1.5mg/ml if you're going with 100ml. 1 cap of EGCG too. The purpose of the ethanol is to break down the barrier of the skin, and the PG supposedly helps the molecules stay dissolved long enough to slowly be absorbed by the skin. Beware of irritation and itching from PG, you want to avoid that.

I realise getting ethanol in the UK is not an easy task, you cant even get everclear here. I was going to suggest spirytus but you've already beaten me to it lol. Its quite steep but it'll last you ages at least. For those of you that can use minox, it works out cheaper to use the kirkland solution as its preformulated and available cheaply in the UK.

You're already on quite a stack Josh, do you think those 2mm of growth could be attributed to CB or any of the other treatments?

I've been using ~1.8mg/ml oleuropein with minox for a little over 20 days now and I'm seeing some remarkable things on the area I normally apply. I'm convinced the oleuropein is making the biggest difference. Since I'm on two weeks break I've been using 0.5ml of the minox solution in the morning, then again at 6pm, then at 10pm I finish off with the emu oil (+ minox + oleuropein solution) which prevents the itching from just using minox + oleuropein alone. I'm going to wait for the area to completely fill in before I post pics, which is very very soon at this rate! I also cant wait to add in EGCG!

[joshuk]

wow i have been applying about 1ml of that amazon tincture so thats way to much, im glad ive decided to make my own. not only that it stains the scalp brown so not ideal really to use during the day.

so how much Oleuropein per ml would be in 60ml of my custom solution, im guessing 1.8mg/ml as the same as a minox solution just without the minox...

i thought it might have been the Oleuropein that was getting my results becasue RU never really grew much for me so i did not think CB would either so unless CB is way better than RU or the Oleuropein tincture i was using had some effect. either way i will try using this custom mix then hopefully if a few of us start seeing some results we can work out a base formula.

[Chemical]

I'm using the same oleuropein caps you linked. 150mg per capsule. 150mg/100ml = 1.5mg, which is ideal. For 60ml, use half a cap. Twice a day should be more than enough, less is more in the case of oleuropein. Vellus hairs can grow quite long but you need to be looking out for actual stubble. Hair that feels prickly. I'd expect that around the 30 day mark.

I hope you're not using DMSO as a vehicle for any of your treatments? because DMSO isnt too friendly to hair according to some anecdotal reports. The CB or any AA for that matter wont regrow hair that quickly but the longer your hairs are free from Androgen, the higher the chance other treatments will work faster. CB should definitely potentiate the oleurpein and EGCG, but even without those two you should notice regrowth over time.

And 1ml is way too much. I should'nt have recommended it so I apologise.