Updated Research and Knowledge - Cutting Edge

[Chemical]

@Sogeking

So long as you're getting 65% actual oleuropein and the not the leaf extract at 65% of the solution, then the dosing math should be accurate. The only concern is making sure you get less than 2mg/ml HealthMonthly deliver to europe (if you're in EU?), and they stock the oleuropein that I use, maybe thats another option? http://www.healthmonthly.co.uk/swans...FasBwwodO5oO6g

[iroquoispliskin]

Found this on amazon for US people:

http://www.amazon.com/Foods-Olive-Glycerite-Oleuropein-ounce/dp/B0019LWTWQ

Its in liquid form standardized to 18% oleuropein. Would there be any problem in using this directly as a topical? The liquid is glycerol and water. From what I've read, glycerol is a penetration enhancer when used on skin (its in a lot of soaps and cosmetic products) Only thing is based on the numbers its 22mg/ml of oleuropein, which is way more than what Chemical said. Is there a reason that exceeding 2mg/ml is detrimental? Could always dilute it further if necessary.

[potato1987]

Trial

Chemical, when you say wait as somebody can have a batch mixed this would be totally without minox yes? (hoping so).

Happy to order and test as soon as someone can mix a batch!

[Sogeking]

@Sogeking

So long as you're getting 65% actual oleuropein and the not the leaf extract at 65% of the solution, then the dosing math should be accurate. The only concern is making sure you get less than 2mg/ml HealthMonthly deliver to europe (if you're in EU?), and they stock the oleuropein that I use, maybe thats another option? http://www.healthmonthly.co.uk/swans...FasBwwodO5oO6g

Ah yes. Thank you for that mate. It is the leaf extract at 65 % of the solution. I will most likely order from your link. In which case I just use 1 pill diluted in 50 ml of ethanol/pg mix.

[TubZy]

just mix the OLE powder into adenogen. That is what I do..I used to use neogenic but it doesnt contain PG and adenogen does so I switched to adenogen as my vehicle

[Chemical]

I'm sorry I forgot to answer this question. Setipiprant is a PGD2 blocker, I'm under impression PGD2 isnt the main culprit but it's metabolite 15-dPGJ2 that kills hair follicles - as stated by Cotsarelis. By reducing the expression of COX-2, PGD2 should be rate limited and subsequently result in less 15-dPGJ2 being synthesized. Its a factor, but not the only or most important one, I will investigate this matter in detail later on.

PGDTS can be induced by Estrogen (aromatased from Testosterone), but exactly which receptor mediates this effect I do not know. My guess is the ER Beta, which ketoconazole and miconazole address via 3BetaDiol suppression. Keto/Mico are like the poor mans version of Seti.

Chemical, when you say wait as somebody can have a batch mixed this would be totally without minox yes? (hoping so).
Happy to order and test as soon as someone can mix a batch!

Without minox yes, we will have to wait for Keeper and the outcome of our experiments to devise an optimal formulation with the help of Keeper's contact.


I've been doing some more research and I came across some new information on DKK-1. We know that oxidative stress (ROS) induces the expression of DKK1, but until now I didnt know how that happened. This excerpt sheds some light on this matter:

To investigate the upstream stimuli responsible for JNK pathway activation and DKK1 up-regulation by lenalidomide, we analyzed changes in gene expression that correlated with DKK1 up-regulation following lenalidomide treatment in vivo. Microarray analysis showed a clear indication of the activation of genes involved in the cellular response to oxidative stress, which is in line with previous reports indicating that thalidomide could induce oxidative injury in the rabbit model system by forming free radical-initiated reactive oxygen species. Interestingly, in the 40 most highly correlated genes, we identified genes encoding caspase 4 (CASP4, 8.7-fold increase; most up-regulated gene) and calpain 2 (CAPN2, 5.3-fold increase), which are involved in endoplasmic reticulum stress-induced apoptosis, and glutaredoxin (GLRX, 7.45-fold increase), glutathione peroxidase (GPX1, 5.7-fold increase), and glutathione S-transferase (GSTO1, 5-fold increase), which play important roles in cellular detoxification, suppressing activities of JNK and its upstream kinases through direct in,eractions.

Therefore, to determine whether oxidative stress plays a role in mediating lenalidomide-induced DKK1 up-regulation, we administered the drug along with the antioxidant agent PBN. As shown in Figure 3A, pretreating cells with PBN significantly reduced lenalidomide-induced DKK1 up-regulation (expression reduced 1.5- to 3.3-fold; mean: 2 ± 0.1; P < .01). Interestingly, preincubating cells with PBN alone could reduce the basal level DKK1 expression (data not shown). Furthermore, in line with previous reports, PBN counteracted c-Jun–binding activation (P < .01; Figure 3B) and JNK phosphorylation (data not shown). Taken together, these data demonstrate that oxidative stress is an upstream stimulus responsible for JNK pathway activation and DKK1 up-regulation induced by lenalidomide.

The antioxidant PBN could prevent the oxidative stress induced DKK-1. Just like Ascorbic Acid. Oleuropein is also a free radical scavenger. But the interesting point here is that, antioxidants could reduce DKK-1 below basal levels without a stress response stimulus. This explains why oleuropein reduced DKK-1 in the mice study. In the presence of testosterone, anti-oxidants will probably only keep DKK-1 at basal levels, but if AR is suppressed significantly, DKK-1 can be further reduced thus allowing WNT to freely bind whenever and however it wants! It seems DKK-1 is a solved case, its now time to find agents that can activate the canonical WNT pathway.

Baicalin activates the WNT pathway
Baicalin inhibits AR and grows hair

The only problem is its not very water soluble. Heat can increase the solubility of baicalin in ethanol. As can a surfactant like tween 80/polysorbate 80.

This excellent study goes over some of the effects of the different flavanoids on WNT/BetaCatenin. Flavanoids have differential effects depending on cell type, exerting pro-apoptotic factors in tumors and promoting survival in others. Does anyone else have any knowledge on herbs/extracts that can increase BetaCatenin and has good solubility properties?

[joshuk]

chemical about to make my first mix of EGCG/OLE do you think a mixture of 50ml ethanol 10ml of PG is ok?? its about 80/20 split.

also i was thinking of applying this all over my scalp as i shaved my head for this experiment lmao looks wierd but im gonna give it a good go.im gonna use 1/2 cap of OLE and 1 cap of EGCG. so its about 1.35mg/ml OLE in the mix.

i have 50grams of seti that i have not started do you think it will help benifit the WNT/BetaCatenin pathways?? i just dont have any capsules to put it into and dont want to use it topically as im already using a fair amount

[Chemical]

That should be perfect. The Seti might augment this protocol, but skip if for a few weeks since you've already got a heavy stack. If you dont see any change, then you might want to throw it in. I wish you all the best!

[dunester01]

Hi Chemical - First, add me to the growing number of posters (and lurkers) in these forums grateful for your willingness to wade through the scientific lit related to AGA and share your insights. While the prospect of a standardized formula that incorporates oleuropein or ECGC (or both) is exciting (I'm referring to Keeper's efforts), I want to experiment on my own. I've had decent results using 5% Minoxidil so I want to continue to use it. If I understand your recent posts correctly, to replicate the formula that you'd currently recommend, I'd simply add 1/2 capsule of the Swanson's Superior Herbs Olive Leaf Extract Super Strength and 1 capsule of either the Swanson's Superior Herbs Teavigo EGCG or the Healthy Origins Teavego EGCG to a single 60 mL bottle of 5% Minox. Please correct me if I've misunderstood, and thanks again for your consideration.

[InBeforeTheCure]

Ironically many people have also tried olive oil with little success = oleuropein doesnt work?

People at the other hairloss forum have tried: http://www.*****************/interac...r-growth-study

Some guy claims to have seen results:

http://www.regrowth.com/hair-loss-forums/topic/my-results-with-egcg-and-ode-to-my-methodology/

Someone has been using it on their face to reduce facial hair too:



This patent by Gillette on syrian hamsters that have AR dependent hair, showed that EGCG significantly reduced hair size and growth at concentrations up to 30%.

I think alot of people that try different stuff probably try them in a vaccuum, and if it works then only very small minority will post their findings on the internet whereas the vast majority will say that it didnt work. The theory is there, and on paper/animals there is convincing evidence of its efficacy. But we dont know for sure if using it in a proper vehicle or along side other treatments will yield results. Like Josh has said, this is still an experiment, I'll never know until I try.

It does say in the abstract of the mouse hair EGCG study:

Topical EGCG down-regulated the T-induced expression of androgen receptor but did not down-regulate 17β-hydroxysteroid dehydrogenase (HSD) and three β-HSD expression.

Perhaps the continued expression of β-HSD could limit results despite AR inhibition? So using EGCG alongside ketoconazole or miconazole (or maybe setipiprant?) to downregulate the ER-β pathway could have a nice synergistic effect.

By the way, the following protocol is the one I'm (tentatively) considering.

- Oleuropein
- EGCG
- Setipiprant
- PGE2
- Valproic acid

Is there anything critical missing? Something like ketoconazole or miconazole for example, or does seti take care of all the downstream effects of ER-β?

[JDW]

Total respect to all those on this topic who are pushing the boundaries and testing out these new potential solutions. You're doing a top job

[lukey]

How come no more than 2mg/ml? Is it unsafe or is the excess OL just not doing anything?

[Chemical]

I want to experiment on my own. I've had decent results using 5% Minoxidil so I want to continue to use it. I'd simply add 1/2 capsule of the Swanson's Superior Herbs Olive Leaf Extract Super Strength and 1 capsule of either the Swanson's Superior Herbs Teavigo EGCG or the Healthy Origins Teavego EGCG to a single 60 mL bottle of 5% Minox.

Exactly like you've said. I hope you will keep us updated?

Perhaps the continued expression of β-HSD could limit results despite AR inhibition? So using EGCG alongside ketoconazole or miconazole (or maybe setipiprant?) to downregulate the ER-β pathway could have a nice synergistic effect.
Is there anything critical missing? Something like ketoconazole or miconazole for example, or does seti take care of all the downstream effects of ER-β?

Minoxidil increases 17 beta-hydroxysteroid dehydrogenase and 5 alpha-reductase activity of cultured human dermal papilla cells from balding scalp.

40% increase in 17βHSD from minoxidil. This might be the reason why Keto is included in the big three. HIF-1 stabilization probably maintains sustained VEGF expression, and a decrease cuts off the blood supply, I'm unsure if it makes a big difference given that β-Catenin can induce VEGF independently, albeit for unknown amount of time. Seti would eliminate all probability of PGD2 and its metabolite killing hair cells, thats the only guarantee you get.

Keto and mico quite potently reduce 3BetaDiol, although no numbers. This study shows bifonazole, keto and mico to be very potent at reducing only 17 alpha-hydroxylase in the testes. Bifonazole managed to bring the Ki down from 1090 to 86 +/- 3.3, and keto managed: 160 +/- 4.92. Mico wasn't as strong: 599 +/- 7.22. My assumption is that Keto and mico are additive hence my recommendation for both, but if you must choose one, get keto. They noticed a dose dependent inhibition, and thats why I use it twice a day (dont exceed 3x/week with shampoo).

Your choice of valproic is surprising, its quite harsh on skin according to anecdotal reports. Even at ~8% this study shows there was some irritation. If you must satisfy your curiosity, use no more than 2% and with an anti-inflammatory like emu oil. I will look into dosing a little more because GSK3b inhibitors are showing more and more promise. I am tempted to suggest LiCl but not until I've done testing first.

Now for some more research findings!

It turns out activating β-Catenin pathway triggers a feedback loop increasing Axin2 (minoxidil study) (another). Axin2 can substitute axin1 and I'm not sure if the prostaglandin receptors or even EGCG can reduce Axin2 expression. Without Axin2, you'd have sustained β-Catenin level which could become cancerous in sensitive cell types. For hair this would translate to ridiculous hair regrowth rates. (More reading)

Axin1 is the rate limiting factor for β-Catenin degradation, so if its substituted with Axin2, you're back at square one - β-Catenin will be destroyed. But if you reduce GSK3b, regardless of Axin1/2, β-Catenin will stay elevated.

Indirubin from Angelica sinensis extract - Dong Quai can inhibit GSK3b. Another study.

Androgens increase GSK3b:

Androgen treatment revealed a significant decrease in the cytoplasmic/total β-catenin protein ratio and upregulation of the activity of glycogen synthase kinase-3β in DPC, indicative of canonical Wnt pathway inhibition.

http://www.ncbi.nlm.nih.gov/pubmed/22283397

I suspect this is due to AR's negative feedback loop found in the prostate cells, wherein GSK3b represses AR activity (controversial atm).

By reducing AR this additional increase in GSK3b will be negated. Its becoming very complex again so I will make another diagram soon.

I also realise PGE2 is a beast:

Here, we show that PGE2 activated the E-catenin/TCFdependent transcription in colon cancer cells through the cAMP/PKA pathway. Expression of cyclin D1 and vascular endothelial growth factor (VEGF) was induced by PGE2 in LS-174T cells. Moreover, PGE2 and mutated Ecatenin stimulated the transcription of cyclin D1 and VEGF in a synergistic fashion. Mechanistically, PGE2 increased the phosphorylation of GSK-3 and consequently accumulated E-catenin.

PGE2 -> EP2/4 -> PKA ¬ GSK3b
PGE2 -> EP2/4 ¬ Axin1

Castor oil anyone?

PKA inactivates GSK3b expression. Since PGE2 is so hard to effectively upregulate, other agents that increase cAMP mediated PKA can also inhibit GSK3b:

Effects of the cAMP-elevating agents cilostamide, cilostazol and forskolin on the phosphorylation of Akt and GSK-3beta in platelets.

PTH/cAMP/PKA signaling facilitates canonical Wnt signaling via inactivation of glycogen synthase kinase-3beta in osteoblastic Saos-2 cells.

Western blotting demonstrated that GSK-3beta was rapidly phosphorylated at Ser(9) on treatment with PTH or forskolin, leading to its inactivation. Moreover, overexpression of a constitutively active mutant of GSK-3beta abolished the TCF-dependent transactivation induced by forskolin. On the other hand, overexpression of the Wnt antagonist Dickkopf-1 (DKK1) failed to cancel the effects of forskolin on the canonical Wnt pathway

So why hasnt anyone tried forskolin yet?

Evidence that activation of protein kinase A inhibits human hair follicle growth and hair fibre production in organ culture and DNA synthesis in human and mouse hair follicle organ culture.

Human hair follicles were isolated from facial skin by microdissection...Human hair follicle growth was inhibited by the phosphodiesterase inhibitors 3-isobutyl-1-methylxanthine and Ro 20-1724, and by the adenylate cyclase activator, forskolin.

Interpret that however you wish. Apparently PGE2 should inhibit facial hair growth too.

This study shows procyanidins (found in apple skin) paired with forskolin increases hair growth rate.

http://www.ncbi.nlm.nih.gov/pubmed/10859531

Forskolin, an adenylate cyclase activator, promotes hair epithelial cell growth and boosts the growth-promoting effect of procyanidin B-2. It is speculated that the hair-growing activity of procyanidins is related to their protein kinase C-inhibiting activity.

Furthermore: Pi3K/AKT ¬ GSK3b.

I know puerarin activates Pi3K quite strongly, so I googled, and whaddayaknow!

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3765709/

These observations indicated that puerarin induced the phosphorylation of AKT at serine 473 and subsequently activated the phosphorylation of GSK-3β at serine 9, leading to GSK-3β inhibition and Wnt/β-catenin signaling.

http://www.healthmonthly.co.uk/swanson_kudzu_root?af=gaw&gclid=CKDe7fbDhsoCFYMIwwodQfgN3Q

Puerarin;s solubility in water can be increased with higher temperatures. Adding in a starch and using steel balls to mix the solution ridiculously enhances bioavailability lol: this.

Puerariae Flos (the flowers of Pueraria thomsonii) was found to inihibit 5ar and grow hair in mice: http://www.ncbi.nlm.nih.gov/pubmed/21822606

Not sure where to get this specfic extract since kudzu is broad and we need that flower specifically to inhibit AR, but the standard swanson kudzu I linked should inhibit GSK3b - in theory.

Theres conflicting literature on PKC regulating GSK3b, I'm looking into it. I will be experimenting with various GSK3b inhibitors over the course of next few weeks, including lithium chloride and forskolin to see if they hold any value for us.

@InBeforetheCure
Your stack is ideal, but I'm curious about the PGE2 - where are you getting it from?

@Lukey
OL appears to have a biphasic effect, meaning too much and it loses its effectiveness. In the original study OL wasnt more effective higher than 50um, and produced slightly worse results. Same was seen in OL/Androgen study. I've noticed some strange things too, most of my growth with high OL was in peripheral regions where little OL was penetrating. I've been using a reduced concentration and I'm seeing remarkable things which unfortuneately my shitty camera cant capture. As soon as the area has filled in I will post pics. Its not conclusive but for now less OL is more.

[joshuk]

chemical you say you are seeing good results, do you think you will see a cosmetic diffrence,as in new terminal regrowth or is it just thickening of exsisting hairs.

i have just applied my first EGCG/OLE mix so i will just report back if i see any results worth mentioning. taken pictures and shaved head for this haha. hopefully have something positive to come back with.

on another note if this does bring results... im not using minox so potential for us poor souls that cant use it

[Chemical]

It's new years eve, fvck it. You be the judge.

http://imgur.com/a/usmLd

Today I saw two terminal hairs growing right in the centre of that region, where my nw0 hairline was. You cant see it but it was more than 2 inches long. I pulled one of them and it wasnt coming out, so I pulled it until it broke halfway - thats a terminal follicle. The hair felt weird though, really really tough. I've got soft fine hair so somethings definitely going on.