Updated Research and Knowledge - Cutting Edge

[tylerbose]

Minox + OL - night

hi everyone, sorry for the off topic question.

did you put the OL caps inside the minox jar ? or are t they separated ?
if so, would it be possible to use something other than ethanol as vehicle ?
thank you

[Chemical]

"the possibility that hair follicles develop de novo following wounding was raised in studies on rabbits, mice and even humans fifty years ago. Subsequently, these observations were generally discounted because definitive evidence for follicular neogenesis was not presented. Here we show that, after wounding, hair follicles form de novo in genetically normal adult mice. The regenerated hair follicles establish a stem cell population, express known molecular markers of follicle differentiation, produce a hair shaft and progress through all stages of the hair follicle cycle. Lineage analysis demonstrated that the nascent follicles arise from epithelial cells outside of the hair follicle stem cell niche, suggesting that epidermal cells in the wound assume a hair follicle stem cell phenotype. Inhibition of Wnt signalling after re-epithelialization completely abrogates this wounding-induced folliculogenesis, whereas overexpression of Wnt ligand in the epidermis increases the number of regenerated hair follicles."

I wonder if this (WnT signalling) study has been conducted in the presence of DHT and receptor diminishing regimen for such.

Hope you're still around, some excellent research you've got here.

I've read alot of studies on Wound Induced Hair regeneration in humans and most of them point towards pge2 mediated anagen induction instead of actual HF formation. The platelet derived growth factors seem like another potential candidate in increasing the probability of anagen induction. I'm more interested in new HF formation with wounding, in the hope that it might create HF's that arent susceptible to DHT.

Hair regrowth following a Wnt- and follistatin containing treatment: safety and efficacy in a first-in-man phase 1 clinical trial.
...
These results demonstrate that a single intradermal administration of HSC improved hair growth in subjects with androgenetic alopecia and is a clinical substantiation of previous preclinical research with Wnts, follistatin, and other growth factors associated with wound healing and regeneration.

Histopathological evaluation of the treatment site biopsies taken at 22 and 52 weeks post-treatment revealed no abnormal morphology, hamartomas, or other pathological responses. Trichoscan image analysis of HSC-treated sites at 12 and 52 weeks showed significant improvements in hair growth over the placebo. At the initial 12-week evaluation period, HSC-treated sites demonstrated an increase in hair shaft thickness (6.3%±2.5% vs. -0.63%±2.1%; P=0.046), thickness density (12.8%±4.5% vs. -0.2%±2.9%; P=0.028), and terminal hair density (20.6±4.9% vs. 4.4±4.9%; P=0.029). At one year, a statistically significant increase in total hair count (P=0.032) continued to be seen. These results demonstrate that a single intradermal administration of HSC improved hair growth in subjects with androgenetic alopecia and is a clinical substantiation of previous preclinical research with Wnts, follistatin, and other growth factors associated with wound healing and regeneration.

This is histogen (GK Naughton)

How is it that just one injection continues to work even after 1 year? Either this is complete bs or its a scientific breakthrough. I'm tempted to say that these growth factors created new follicles that weren't affected by DHT and since non AGA DPC secrete growth factors to nearby cells it could very well be that this started a chain reaction. Follistatin can be purchased and this has my curiosity.

I've only ready the first link SriHanuman , that is a good read and shows RB-SCE a decent alternative to minoxidil.

In terms of the stack, Linoleic Acid (LA) is there (here I'm lumping together GLA and LA) with Evening Primrose*.

The new item appears to be Gamma Oryzanol (OZ).

I wish the study went out past 4 weeks. The OZ really kicked in at week 4.

I forgot to say thanks for posting this, pointed me in the direction to discover GLA is a fairly powerful 5ar inhibitor.

chemical im going to be buying some Sodium Valproate when kane gets back from holidays, what vehicle would you use 70/30 eth/pg or is their a better one. was thinking about using 70mg a day

Water! 50mg/ml vs 30mg/ml ethanol. Distilled water preferably.

But why obfuscate the fact it inhibits PGE2 in such a detailed analysis? I can't think of a reason. And haven't cox 2 inhibitors been tried and failed already potentially for this very reason?

Been on the minox approaching 2 months so don't think its causing the shed (although can't be sure).

I couldnt find any obvious research indicating that OL directly inhibited PGE2 but as with anything that inhibits COX-2 there is bound to be a decrease in PGE2 synthesis. COX-2 is an inflammatory response so I wouldnt be surprised if an anti-inflammatory reduces COX-2.

Yo McChemical dog you have misinterpreted this study

http://www.ncbi.nlm.nih.gov/pubmed/1632630

Ketocanzole and miconazole inhibited the HYDROXYLATION of beta-diol, this means less beta-diol will be converted into it's hydroxy metabolites, which means if you use Keto there will be MORE beta-diol lying around. Food for thought dog.

The inactivation of 5 alpha-dihydrotestosterone involves reversible reduction to 5 alpha-androstane-3 beta,17 beta-diol by 3 beta-hydroxysteroid oxidoreductase followed by 6 alpha-, 7 alpha-, or 7 beta-hydroxylation. 5 alpha-Androstane-3 beta,17 beta-diol hydroxylation represents the ultimate inactivation step of dihydrotestosterone in rat prostate and is apparently catalyzed by a single, high-affinity (Km approximately 0.5 microM) microsomal cytochrome P450 enzyme. The present studies were designed to determine if 5 alpha-androstane-3 beta,17 beta-diol hydroxylation by rat prostate microsomes is inhibited by agents that are known inhibitors of androgen-metabolizing enzymes. Imidazole-type antimycotic drugs (ketoconazole, clotrimazole, and miconazole; 0.1-10 microM) all markedly inhibited 5 alpha-androstane-3 beta,17 beta-diol hydroxylation in a concentration-dependent manner, whereas triazole-type antimycotic drugs (fluconazole and itraconazole; 0.1-10 microM) had no inhibitory effect.

Hydroxylation is the process by which 5ar gets converted to 3beta-diol. Keto and mico inhibit this step thus preventing the conversion of DHT to 3Beta-diol.

The GLA and LA may have other functions, like inhibiting 5AR, but those activities are probably weak, and trumped by including something like RU in the regimen.

One caveat to all of these oils is whether they will dissolve in a vehicle. In my own experiments at home, I haven't found any oils that mix into a vehicle. One possible exception to that is Castor oil, which I read is miscible with ethanol. I don't have the patience to put several different things on my head every day.

GLA and LA's predominant function in AGA would be 5ar suppression imo. Studies show GLA to be far better than any other nutraceutical at inhibiting 5ar. There havent direct comparisons with Serenoa Repens (saw palmetto) but on paper it seems far superior. The whole point of these outlandish treatments to block 5ar is to avoid the systemic effects of HPTA overcompensation increasing T production and subsequently offsetting the effects of fin or dut. Oils are tricky in terms of absorption but I guess the whole point of oils is that they'll eventually get absorbed.

OK, I just did a pubmed search, and pulled up every paper that mentions both EGCG and PGE2. In just about every instance, EGCG causes a reduction in PGE2.

This first paper below is the most important one, it's topically applied EGCG to human skin. The second paper below shows that EGCG reduced beta-catenin activity, which is very bad if you want to grow hair. The remaining abstracts just reiterate the point that every time EGCG is put on something, PGE2 goes down, across a wide variety of animals and cell types. I highlighted the relevant parts in bold for easy skimming. In the second to last abstract listed, EGCG even downregulated the PGE2 receptors.

What you've posted are studies done on carcinogenic cell lines demonstrating the wonderful paradoxical effects of EGCG on different tissues. Its something that I was puzzled by but apparently thats how EGCG works, its a powerful anti-tumor agent in cancerous cells but acts as a mitogenic agent in normal cells.

[Chemical]

There was another study done a few years before Krajcik where they transplanted (healthy) human hair onto SCID mice. They found that the follicles soon went into something called dystrophic catagen -- large-scale apoptosis of the hair follicle -- followed by regeneration. Perhaps this process "resets" the follicle by removing senescent cells and other cells with screwy gene expression.

I missed this sorry. (whats up with the sci-hub.io links? I never get to see the damn article, it shows me some russian page and I feel left out of your scientific research)

I wonder why the cells go into catagen only to regenerate? I was doing some in depth reading on the DPC and how even in AGA telogen the DPC remains intact - although senescent. Which means you'll always have the dysfunction no matter how many times you go through anagen and its only the DPC that are the faulty cells. If you permanently destroy the DPC however, the dermal sheath can regenerate a new DPC! (would recommend reading whole article, very interesting)

Review of hair follicle dermal cells

DS has been considered as a cellular reservoir of DP cells during the hair follicle cycle[9], and it was hypothesized that stem cells might reside in the DS similar to its epithelial counterpart, the outer root sheath [10]. DS cells share similar characteristics with the DP and can regenerate a new DP after loss of the DP [11–13]. In classic transection studies of rat vibrissae, Oliver and colleagues demonstrated that removal of the lower follicle resulted in regeneration of the DP, apparently from the DS, while removal of the follicle at the level of the bulge did not allow for DP regeneration [14].

My question is do the DS cells contain the AGA code? Doing some more digging around I found out that the DS cells are continually renewed from the stem cell bulge, they move all around the HF from the bulge so my hope is that stem cells do not contain the code.

Also Androgens cause senescence of stem cells via WNT inhibition/GSK3b upregulation (study), I suspect this is due to insufficient WNT feedback from DPC which keeps the stem cell pool pluripotent.

But in the stains, it looks like very little AR or beta-catenin is in the nucleus after Wnt is added in AGA40M?

Yeah I noticed that too but I wasn't sure if that was just a staining thing. your explanation below is probably my best bet, something to do with epigenetics.

Anyway, I'm wondering if AR binding to beta-catenin could maybe alter the way in which beta-catenin interacts with Wnt response elements -- for instance by recruiting HATs and HDACs to remodel chromatin structure, or by other interactions with certain transcription factors -- and thereby downregulate some Wnt target genes and upregulate others. DKK1 for example is actually a Wnt target gene -- and indeed DKK1 is upregulated over 2x in response to Wnt in DPCs -- and as you've pointed out DKK1 is (further) upregulated in balding DP cells in response to DHT. The new gene transcription "profile" of DPCs in response to Wnt could then fail to induce epithelial stem cell activation. Highly speculative, but could it be possible?

Increased sensitivity to androgens could happen by

- Upregulation of AR
- An increase in androgen-independent activation of AR. Let's say z is the threshold in AR activation that causes hair loss, x is the androgen-independent level of activation, and y is activation caused by androgens. And let's say androgen-independent activation is very low in early stages of balding, but rises somehow over time as a result of DHT-bound AR transcribing its target genes. Left untreated, x rises over time and it takes less y to push x + y over the threshold (z). Eventually x > z and no anti-androgen or even castration could push AR activation below z -- the DPC's inductive ability is lost.

In this model castration > dut > fin. A prediction of this model is that someone who is maintaining on fin who goes on dut, regrows some on dut, but then switches back to fin would lose the hair he gained on dut (is this how it is in reality?). This would be expected based on a model where AR is upregulated or androgen-independent activation of AR increases over time with exposure to DHT, or both. At least, I think it would fit such a model better than a model where DP cells in recently lost hair are "repaired" in response to androgen removal.

I'm just speculating here, of course.

What you're describing reminds me of prostate cancer how it starts off as androgen dependent then becomes independent during Androgen depletion. The model is eerily similar to PC and the reality of it further points me to believe this is true. Even when people start RU they dont see complete regrowth after being on it for a long time, which completely goes against intuition. The DPC arent cancerous so its not mutation based adaption, so it has to be something subtle like you described above.

Sulforaphane? Don't know how good it would be in practice though. Seems to have a half-life of only a couple hours too, so even if it did work you might have to use it 20 times a day or so, including at night.

I'm a bit reluctant about messing with HSP90 because it's used by quite a few pathways notably PI3K/AKT.

Here's a cool diagram I found:



MDV-3100 would've answered our problems but its not exactly a viable solution.

Yes, but dystrophic catagen could remove senescent cells and other malfunctioning cells from the cell cycle. Then dermal stem cells regenerate the dermal papilla. Sure, over time it will malfunction again, but you would just need maintenance treatments then to stop that from happening. And then eventually, once the technology is available, you could get the AR gene knocked down permanently in your scalp and never have to worry about AGA again.

I dont think the malfunction would happen at all if the cells were renewed. The problem is only the DPC, they control the stem cell pool pluripotency and the whole cycle is orchestrated by the DPC themselves so if they're replaced it could very well be the cure.

The more I look into it, the more I think this is probably correct. It's probably not so much that AR binds beta-catenin and prevents it from acting (as FGF11 thought). I think it's more likely that AR hijacks the Wnt pathway for its own nefarious purposes by binding to beta-catenin and altering the way that beta-catenin/TCF4 interacts with Wnt response elements, either enhancing them or (relatively) repressing them. From this model you can actually derive, for example, why PGD2 is upregulated in AGA with reference to both theory and experimental results. You can also imagine how a similar mechanism could actually account for why androgens stimulate facial hair growth while causing hair loss on your head. However, I'd like to develop this idea further before really getting into that (maybe I'll create a new thread sometime).

This is not exactly a novel idea, as something similar is known to happen in prostate cells, for example.

The androgen receptor can signal through Wnt/β-Catenin in prostate cancer cells as an adaptation mechanism to castration levels of androgens

Transient transfections of several human prostate cancer cell lines with the AR and multiple components of the Wnt signaling pathway demonstrate that the AR overexpression can potentiate the transcriptional activities of Wnt/β-Catenin signaling. In addition, the simultaneous activation of the Wnt signaling pathway and overexpression of the AR promote prostate cancer cell growth and transformation at castration levels of androgens. Interestingly, the presence of physiological levels of androgen or other AR agonists inhibits these effects. These observations are consistent with the nuclear co-localization of the AR and β-Catenin shown by immunohistochemistry in human prostate cancer samples. Furthermore, chromatin immunoprecipitation assays showed that Wnt3A can recruit the AR to the promoter regions of Myc and Cyclin D1, which are well-characterized downstream targets of the Wnt signalling pathway. The same assays demonstrated that the AR and β-Catenin can be recruited to the promoter and enhancer regions of a known AR target gene PSA upon Wnt signaling. These results suggest that the AR is promoting Wnt signaling at the chromatin level.

You are correct, the immediate effect of AR isnt to inhibit WNT, but merely a side effect of AGA mediated AR gene expression in DPC.

Its important not to forget this crucial bit of information;

Wnt3a-dependent keratinocyte growth was suppressed by the addition of dihydrotestosterone in coculture with DP cells that were derived from AGA patients, but growth was not suppressed in coculture with DP cells from non-AGA males. Whereas DP cells from both scalp regions expressed AR protein, the expression levels of AR and cotranslocation with β-catenin, a downstream Wnt signaling molecule, were higher in DP cells of AGA patients than in DP cells from non-AGA males. In addition, significant suppression of Wnt signal-mediated transcription in response to dihydrotestosterone treatment was observed only in DP cells from AGA patients.

DPC do not respond anabolically in single cell cultures to WNT or DHT which tells us its all paracrine/autocrine signalling. The Keratinocytes (HaCaT) do respond directly to WNTs by hypertrophying/growing. But we know its the DPC that are in charge of the entire cycle.

Wnt3a treatment significantly accelerated the growth of HaCaT cells in the single cell culture, but the addition of DHT did not affect growth (Fig 1D1D).). On the other hand, the growth of DP cells was not affected by Wnt3a in single cell culture conditions (data not shown). It has been reported that androgen does not influence growth of DP cells in a single cell culture condition (26,27).

The DPC transform their internal response to WNTs/Androgens into a positive or negative extracellular response that the adjacent DPC can respond to physiologically. A successful canonical response to WNT/β-catenin pathway leads to enhanced VEGF, IGF-1R, PDGF which kicks off and maintains anagen but the AGA response is DKK1, TGF-Beta, and none of the pro-cytokines.

AGA changes the way androgen interacts with β-catenin like you've pointed out, and the response becomes a destructive one.

By using image analysis software, we examined the degree of nuclear colocalization of AR and β-catenin semiquantitatively using the ratio of merged dots in the nucleus/AR dots in cells treated with both Wnt3a and DHT (Fig. 3A3A).). The ratios in DP cells from AGA males was higher by 2.5-fold than in DP cells of non-AGA males. These results indicate that AR and β-catenin in the nucleus was more highly colocalized in DP cells of AGA40M and other AGA males than in that of non-AGA males.

AR and β-catenin bind regardless of AGA/non-AGA but the binding is enhanced in AGA!? wtf

A direct interaction of AR and β-catenin was detected even in both untreated AGA and non-AGA DP cells. In addition, the amount of AR-β-catenin complex was increased by the treatment with Wnt3a in AGA40M cells but not the non-AGA male cells. This result is consistent with the result of the subcellular localization analysis.

The more β-catenin you have, the worse it becomes...

Lef/Tcf-mediated transcriptional activity was induced by treatment with Wnt3a in AGA40M cells, but this induction was significantly suppressed by the addition of DHT (Fig. 4A4A).). The same result was observed in DP cells of other AGA male. In contrast, in DP cells of non-AGA males, the transcriptional activity was not suppressed by DHT. These results indicate that the Wnt-β-catenin signaling pathway is negatively influenced by ligand-activated AR in DP cells of AGA but not DP cells of non-AGA males.

And here the LEF/TCF response is fine in non-AGA (typical IGF-1, IGF-1R, VEGF, PDGF upregulation) but in AGA, you get none of that but instead we get the awesome (!) destructive DKK1 + TGFBeta.

I am convinced the polymerization of AR and β-catenin causes a different gene expression in complete contrast to the canonical TCF/LEF. In non-AGA there is some binding but not nearly as much as AGA, and the canonical pathway is still functional. WHY the frick is the AR/β-catenin binding enhanced - is it AGA itself or an indirect effect we can fix? I can't believe I didnt read this properly all this time, the answer is right here.

You should definitely make a new thread specifically for this topic. I'd like to investigate further.

What about your progress ( Oleuropein, etc....)

Thanks

Give me some time to post pictures. And please dont post in the read only thread.

[Chemical]

I think some people on here like chemical should be researching an actual treatment for MPB. Some of the guys here know as much as the researchers almost. Can you use those supplements orally instead of having to mix a topical vehicle? how do you know using that stuff causes any hair growth verses it being from the minox which is obviously one of the best regrowth agents.
What is the best and safest way to stop the most androgens?

Unfortunately I dont have a degree in biochem and hate chemistry with a passion (not biology) so I wouldn't be appropriate to work on an actual cure. I'm just trying to understand how this works and if we're lucky someone will be able to use this info to come up with a cure.

As for whether my results were all minox, I dont know. Frankly theres no way to find out since the treatments wont have any effect when used by themselves and so my best guess is they have small additive effect when stacked. If I could do some lab experiments with biopsies and stuff I'd know for sure but right now its all calculated guesswork based on theoretical models. Orally? I'd say oleuropein but it would be very hard to notice any results using oral growth promoters and even if they did work systemically you'd see growth everywhere. To stop most androgens safely (only on your scalp) I'd say... GLA, Rosemary, Serenoa Repens, EGCG, RU, CB. This is all still in experimental stages and dont feel you should jump the gun and try them all.

hi everyone, sorry for the off topic question.

did you put the OL caps inside the minox jar ? or are t they separated ?
if so, would it be possible to use something other than ethanol as vehicle ?
thank you

Yes, in the minox bottle. I personally havent tried any other vehicle since the study used Ethanol + water (I recommend PG because it keeps ethanol from drying up too fast).

[machi]

chemical , I have no knowledge of alopecia. I suffer aga for 16 years . I was deceived by a surgeon in Spain twice and had to go to Dr. Hasson to solve the problem .
I took finasteride for 7 years with very good results but I gave it up because of side effects . 8 years ago I left finasteride.
currently I do not take drugs for hair loss and I 'm sentenced because I have several capillaries operations.
I read your posts but I do not understand anything . I'm depressed , bitter. please chemical , you seem to have much knowledge. I think we are many users who believe in you. Do you think you come to any conclusion on your research and get a possible future treatment ?

[Benis23]

My current progress

I stopped everything around 2 months ago. Before I stopped I was using the following for two weeks:

Minox by itself - morning
Minox + EGCG - night
Minox + OL - night
Water + VPA - night
Keto + Mico - morning & night
Evening primrose - night

After stopping, a week later I saw a ridiculous amount of vellus hairs all along my nw0 hairline, so much that it looked like I'd cured my recession. I strongly suspect this a residual effect of all the treatments finally showing results - or it could be because I stopped. I really really regret not taking pictures which means we're going to have to write it off as not happening but I have hope that I'll be able to recreate the results again and this time will take pictures. I didnt have my treatments with me so I couldnt continue but I had good density and I was quite happy. 6 weeks later I started receding again and lost all the vellus hairs but surprisingly I didnt lose the hairs on the side I was concentrating all my treatments (pics soon). I've got more density than I've had in the past year and I'm back on minox, keto, mico, EPO and borage oil. The EGCG is quite irritating so I've dropped it and I've ordered some more OL. I've managed to regrow hair using only minox and OL in the past - without an AR blocker, so the EPO and Borage oil should definitely help. I might use the VPA again and I'm considering creating a topical out of my fin tablets.

Chemical,

I've followed this thread with interest and appreciate all your hard work and research. I'd like to replicate your treatment on myself, but I don't have a chemistry background and want to make sure I'm using the right products and dosages. Is it possible to go into the details of your current regimen so that the followers of this thread can easily replicate what you're doing? If you can provide exact dosages and even brand names you've been using to buy some of these products, that would be really helpful. Thanks again!

[InBeforeTheCure]

I missed this sorry. (whats up with the sci-hub.io links? I never get to see the damn article, it shows me some russian page and I feel left out of your scientific research)

https://en.wikipedia.org/wiki/Sci-Hub

I post the Sci-Hub links when the full articles are behind a paywall. It should link to a PDF. Sometimes it makes you copy a word into the textbox to proceed to the PDF (to make sure you're human and not a bot).

I wonder why the cells go into catagen only to regenerate? I was doing some in depth reading on the DPC and how even in AGA telogen the DPC remains intact - although senescent. Which means you'll always have the dysfunction no matter how many times you go through anagen and its only the DPC that are the faulty cells. If you permanently destroy the DPC however, the dermal sheath can regenerate a new DPC! (would recommend reading whole article, very interesting)

Review of hair follicle dermal cells

Yeah, I've read that. That and the Krajcik study gives one confidence that the DP can regenerate under the right circumstances. It's just a question of how to do it.

My question is do the DS cells contain the AGA code? Doing some more digging around I found out that the DS cells are continually renewed from the stem cell bulge, they move all around the HF from the bulge so my hope is that stem cells do not contain the code.

I think they probably do actually, and over time after becoming DP cells will develop the AGA phenotype without treatment. I don't see why they wouldn't, since they come from the same source as the other DPCs.

Also Androgens cause senescence of stem cells via WNT inhibition/GSK3b upregulation (study), I suspect this is due to insufficient WNT feedback from DPC which keeps the stem cell pool pluripotent.

I think you mean quiescent, not senescent?

What you're describing reminds me of prostate cancer how it starts off as androgen dependent then becomes independent during Androgen depletion. The model is eerily similar to PC and the reality of it further points me to believe this is true. Even when people start RU they dont see complete regrowth after being on it for a long time, which completely goes against intuition. The DPC arent cancerous so its not mutation based adaption, so it has to be something subtle like you described above.

Yeah, I doubt AGA is actually a selected phenotype. I suspect it's probably either random or an off-target effect of some other adaptation(s), and the selection pressure against it was very low, so it wasn't weeded out.

I'm a bit reluctant about messing with HSP90 because it's used by quite a few pathways notably PI3K/AKT.

Here's a cool diagram I found:



MDV-3100 would've answered our problems but its not exactly a viable solution.

Ideally, you would want to knock out AR. Gene therapy would obviously be the best option for this, but sadly that won't be available to us for who knows how many years. Maybe something will come out of prostate cancer research, since they have basically the same goal. I think that's much more likely than anything miraculous coming from the small and pitiful hair loss field. Besides cell-based treatments, that is.

I dont think the malfunction would happen at all if the cells were renewed. The problem is only the DPC, they control the stem cell pool pluripotency and the whole cycle is orchestrated by the DPC themselves so if they're replaced it could very well be the cure.

I think replacing them would be a temporary, but not permanent, cure. They would eventually malfunction just like their cousins did after they became DP cells. I don't see why they wouldn't?

The androgen receptor can signal through Wnt/β-Catenin in prostate cancer cells as an adaptation mechanism to castration levels of androgens

Thank you. Very interesting study.



Hmm...Very interesting shape to that graph. Have we seen anything like that before?



Ayyy. Add DHT and you enhance the nuclear localization of AR/beta-catenin. Keep adding more and more DHT and AR/beta-catenin is ever more inclined to go transcribe AR target genes rather than Wnt target genes -- Wnt target genes like DKK1. That's probably the main reason why DHT inhibited Lef/Tcf luciferase activity in AGA DPCs in the Japanese study also.

So either beta-catenin is over at AR target genes, or it's at Wnt target genes but transcribing them differentially because it's bound to AR. Either way, when AR is bound to beta-catenin, you're screwed.

You are correct, the immediate effect of AR isnt to inhibit WNT, but merely a side effect of AGA mediated AR gene expression in DPC.

Its important not to forget this crucial bit of information;



DPC do not respond anabolically in single cell cultures to WNT or DHT which tells us its all paracrine/autocrine signalling. The Keratinocytes (HaCaT) do respond directly to WNTs by hypertrophying/growing. But we know its the DPC that are in charge of the entire cycle.

Yes, you can also manipulate the growth factors yourself to achieve regrowth as well (as many have done), but totally mimicking the paracrine growth factors is very hard to do manually. It's true what they say about the "MPB hydra". The argument then comes down to which factors are most important to address.

The DPC transform their internal response to WNTs/Androgens into a positive or negative extracellular response that the adjacent DPC can respond to physiologically. A successful canonical response to WNT/β-catenin pathway leads to enhanced VEGF, IGF-1R, PDGF which kicks off and maintains anagen but the AGA response is DKK1, TGF-Beta, and none of the pro-cytokines.

AGA changes the way androgen interacts with β-catenin like you've pointed out, and the response becomes a destructive one.

Yeah, I honestly think we may have stumbled upon the core etiology of AGA here. I think the theory works and can explain the details of AGA in a coherent way, but of course it would have to be tested experimentally to really know. As Richard Feynman said, "If it disagrees with experiment, it’s wrong. In that simple statement is the key to science. It doesn’t make any difference how beautiful your guess is, it doesn’t matter how smart you are or who made the guess, or what his name is… If it disagrees with experiment, it’s wrong. That’s all there is to it."

AR and β-catenin bind regardless of AGA/non-AGA but the binding is enhanced in AGA!? wtf



The more β-catenin you have, the worse it becomes...



And here the LEF/TCF response is fine in non-AGA (typical IGF-1, IGF-1R, VEGF, PDGF upregulation) but in AGA, you get none of that but instead we get the awesome (!) destructive DKK1 + TGFBeta.

True, true. I think TGFbeta is probably an AR target gene though and not a Wnt target gene like DKK1.

I am convinced the polymerization of AR and β-catenin causes a different gene expression in complete contrast to the canonical TCF/LEF. In non-AGA there is some binding but not nearly as much as AGA, and the canonical pathway is still functional. WHY the frick is the AR/β-catenin binding enhanced - is it AGA itself or an indirect effect we can fix? I can't believe I didnt read this properly all this time, the answer is right here.

There's probably some post-translational modification to AR in AGA, but I'm not sure what that would be exactly.

You should definitely make a new thread specifically for this topic. I'd like to investigate further.

I'll post a thread on this...in 5-10 years.

Seriously though, I want to really develop some of the details as I've only scratched the stuface, and then I'll post a thread. That could be as much as a few weeks from now, but we'll see.

[InBeforeTheCure]

True, true. I think TGFbeta is probably an AR target gene though and not a Wnt target gene like DKK1.

I should rephrase that and say: TGFbeta is probably a downstream effect of AR transcription of AR target genes, possibly as a response to oxidative stress.

[burtandernie]

Could it be for people using fin/dut that when MPB resumes after years of use that its because the AR use the wnt pathway instead of the lower levels of androgens? So maybe they dont upregulate just use a different pathway

[InBeforeTheCure]

Could it be for people using fin/dut that when MPB resumes after years of use that its because the AR use the wnt pathway instead of the lower levels of androgens? So maybe they dont upregulate just use a different pathway

There are still some androgens present -- ~30% of original DHT with fin and ~10% with dut, or something like that, plus testosterone. And AFAIK castration stops progression of AGA permanently in 100% of cases, so I would say that progression of AGA is androgen-dependent (but maintenance of AGA is androgen-independent except for recent loss). In those cases the remaining low levels of androgens probably (slowly) drive progression of AGA.

[Chemical]

https://en.wikipedia.org/wiki/Sci-Hub

I post the Sci-Hub links when the full articles are behind a paywall. It should link to a PDF. Sometimes it makes you copy a word into the textbox to proceed to the PDF (to make sure you're human and not a bot).

I noticed I could access the pdfs on my phone then realised sci-hub was like a piratebay but for research papers lol. This is better than frickin christmas! Thanks to you I've made some discoveries on some potential treatments.

Yeah, I've read that. That and the Krajcik study gives one confidence that the DP can regenerate under the right circumstances. It's just a question of how to do it.

It might be unfeasible to try and destroy the DPC without surgical means and right now it might not even be possible. But if the DPC is dead due to AGA and the DS is intact, then I dont see why it shouldnt be possible without forced intervention. Perhaps in the distant future this will be commercially viable, its a possibility.

I think they probably do actually, and over time after becoming DP cells will develop the AGA phenotype without treatment. I don't see why they wouldn't, since they come from the same source as the other DPCs.

I'm trying to stay ignorantly optimistic. Please dont hit me with a dose a reality.

I think you mean quiescent, not senescent?

Quiescent yes, the stem cells do not lose their proliferative capacity or reach their telomerase limit, thanks for correcting me. Its the DPC that become scenescent (study) (study).

Yeah, I doubt AGA is actually a selected phenotype. I suspect it's probably either random or an off-target effect of some other adaptation(s), and the selection pressure against it was very low, so it wasn't weeded out.

My theory is that the frontal DPC's sensitivity to sex steroids has been unintentionally conserved by men when selecting women with thick hairlines, perceiving them to be more youthful and attractive. And this regional difference in DPC response becomes the complete opposite in males, but since women might have given less preference to looks and more on dominance, the genes ended up being conserved.

Ideally, you would want to knock out AR. Gene therapy would obviously be the best option for this, but sadly that won't be available to us for who knows how many years. Maybe something will come out of prostate cancer research, since they have basically the same goal. I think that's much more likely than anything miraculous coming from the small and pitiful hair loss field. Besides cell-based treatments, that is.

Even if we did have gene therapy, to me it seems like it would be very difficult to localise it to just the epidermis of the frontal scalp. And the cost might not justify not getting a transplant instead either from healthy regions or cultured DPC if they get that far in the future.

I think replacing them would be a temporary, but not permanent, cure. They would eventually malfunction just like their cousins did after they became DP cells. I don't see why they wouldn't?

My belief is that the DPC become conditioned during embryogenesis or at some point in the womb and that this is only specific to the DPC. It could very well be that the entire frontal scalp contains the AGA code, but that just seems unintuitive. The stem cells should have the same code regardless of the location (I tell myself). I'm just trying to be optimistic with my delusions.

One thing I find strange is how it takes quite a while for the initial recession to become noticeable but happens quite fast when stopping treatments. I remember reading a study on Hic5/ara55 (Androgen co-activator) and how it starts to get upregulated after puberty. And also reports of Testosterone/DHT being able to upregulate the expression of AR. Perhaps continued activation of AR causes permanent upregulation or enhanced stability of AR. Its also peculiar that RU users stop responding after a while - I mean its not like theres a negative feedback loop here? What do you think?

Looking at CB and RU they seem like the realistic and most ideal AR antagonists currently on the market, they have virtually no systemic effects on the HPTA or GnRH release and only work peripherally. Its just the price and lack of reputable sources (I'm a little cautious of chinese manufacturers) that put me off.

Hmm...Very interesting shape to that graph. Have we seen anything like that before?

Ayyy. Add DHT and you enhance the nuclear localization of AR/β-catenin. Keep adding more and more DHT and AR/β-catenin is ever more inclined to go transcribe AR target genes rather than Wnt target genes -- Wnt target genes like DKK1. That's probably the main reason why DHT inhibited Lef/Tcf luciferase activity in AGA DPCs in the Japanese study also.

So either β-catenin is over at AR target genes, or it's at Wnt target genes but transcribing them differentially because it's bound to AR. Either way, when AR is bound to β-catenin, you're screwed.

Functional localization and competition between the androgen receptor and T-cell factor for nuclear beta-catenin: a means for inhibition of the Tcf signaling axis

Also AR is upregulated significantly so that might play a part in increasing the receptor saturation ceiling. It also doesnt make the conclusion robust given that non-AGA DPC have less AR. I want to know what would happen if AR was boosted to AGA levels in non AGA DPC.

We know that DHT/T that causes the binding of β-catenin to AR, and the study showed that without DHT, WNT3a did activate the TCF/LEF genes. Something that worries me is that maybe even an antagonist that binds to AR can recruit β-catenin. I'm thinking if we can prevent the binding of β-catenin to AR, we might not have to worry about Androgens at all. Something that fits into the pocket in place of β-catenin! What about SARMS and steroids? I know there are steroids that have significantly higher AR binding affinities that could easily saturate the receptors, and most of them have altered anabolic:androgenic ratios. Perhaps its the androgenic part that causes this negative response of AGA? Edit: looks like R1881 elicits the same response as T/DHT

Yes, you can also manipulate the growth factors yourself to achieve regrowth as well (as many have done), but totally mimicking the paracrine growth factors is very hard to do manually. It's true what they say about the "MPB hydra". The argument then comes down to which factors are most important to address.

Most treatments themselves do not contain the growth factors, they merely stimulate the release indirectly and I'm wondering if all epidermal cells have the ability to release paracrine growth factors. Furthermore, there is only so much you can proliferate the HF shaft with exogenous growth factors, which explains why people only see vellus hairs getting longer. A functioning anagen DPC with an adequate blood supply thickens the diameter of the hair shaft significantly and continually releases growth factors in comparison the infrequent applications of treatments that may or may not reach target tissues. I'm thinking if theres another way to activate TCF/LEF genes without β-catenin, that would be even better than using indirect growth factors.

Yeah, I honestly think we may have stumbled upon the core etiology of AGA here. I think the theory works and can explain the details of AGA in a coherent way, but of course it would have to be tested experimentally to really know. As Richard Feynman said, "If it disagrees with experiment, it’s wrong. In that simple statement is the key to science. It doesn’t make any difference how beautiful your guess is, it doesn’t matter how smart you are or who made the guess, or what his name is… If it disagrees with experiment, it’s wrong. That’s all there is to it."

All the research points to this theory, the AGA DPC inhibiting growth of keratinocytes but normal DPC promoting growth of HaCat. We already know its androgens, and we kind of knew that AR inhibited β-catenin somehow. I think this is the definitive answer, its the binding of AR to our precious β-catenin that causes AGA.

I should rephrase that and say: TGFbeta is probably a downstream effect of AR transcription of AR target genes, possibly as a response to oxidative stress.

I remember making a post about tgf-beta and dug this up:

The involuntary tonic contraction of occipitofrontalis muscle is related to psychological stress conditions,[23] facial expression,[24] the maintenance of visual field,[19] and an aponeurotic tension model of human craniofacial growth,[25] so the galea aponeurotica supports a continuous stress which is transmitted to ECM and cells of each tissue, dermal papilla, and dermal sheath cells included. The deformation energy does not cause apparent damage to scalp skin, but its interplay with androgens could be fatal in organ remodeling of hair follicles. This androgen-mediated molecular response to mechanical stimulation can play the anabolic role instead of biological virilization role, as it has been extensively studied in tissues whose function is closely linked to the physical force support.[26] Furthermore, it has been reported that TGFβ-1 increases the expression of Hic-5 in hypertrophic scars fibroblasts[27] and it potentiates AR transactivity in balding dermal papilla cells[28] by autocrine loop [Figure 3]. Hence, the long-lasting cyclic strain would cause a slow, chronic, and progressive environmental adaptation process in balding hair follicles since puberty.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4639964/

original post

So TGF-beta can potentiate AR via autocrine loop? the f***, it just keeps getting worse

Androgen receptor transactivity is potentiated by TGF-b1 through Smad3 but checked by its coactivator Hic-5/ARA55 in balding dermal papilla cells

(was behind paywall but thats not going to stop me now)

Our data presented here suggest that TGF-b1 can enhance androgen sensitivity through Smad3 in the dermal papilla of AGA in an autocrine manner. Because TGF-b1 from bald DPCs inhibits hair follicle epithelial cell growth in a paracrine manner [1], TGFb1 exerts its pathogenic roles with dual secretion, autocrine and paracrine, between epithelium and dermal papilla in AGA. On the other hand, although Hic-5/ARA55 upregulates androgen sensitivity via coactivation for AR in DPCs [6], the data obtained in our current study indicated that this molecule impedes the AR stimulation by TGF-b1. This may be due to crosstalk between Hic-5/ARA55 and Smad3 [8] or possibly the attenuated effect of TGF-b1 on the high expression of Hic-5/ARA55, which is reportedly increased by TGF-b1 [9]. Given that Hic-5/ARA55 is highly expressed in the androgen-sensitive DPCs from AGA [6], a complex compensatory mechanism through reciprocal interaction must be in place between TGF-b-Smad and androgen-AR signaling pathways in the hair follicles of AGA.

So hic5/ARA55 is already upregulated in AGA when it starts and that is known to enhance AR. Then AR -> ROS -> TGF-beta -> more hic5/ARA55 ¬ (TGF-beta -> smad3 -> AR)

So it looks like hic5/ARA55 is bad, but is keeping check on the additional AR effect of TGF-beta. A powerful or mild anti-oxidant is probably all we need to fix this part but I think we should both look into hic5/ara55 and other co-activators that could be making AGA worse over time.

Induction of transforming growth factor-beta 1 by androgen is mediated by reactive oxygen species in hair follicle dermal papilla cells

Another thing I just noticed is that rat DPC overexpressing AR has the same phenotype as AGA:

During sub-cultivation of DPCs, sensitivity to androgens may be low because of the reduced expression level of AR (11). AR has been detected in the DPCs of human skin (5), and the DPCs of bald frontal scalps express higher levels of AR than those of non-balding occipital scalps (4). Therefore, to confirm our hypothesis, we used rat DPCs that over-express AR.

And they only used DHT which leads me to believe AR will increase ROS regardless of β-catenin being present (or maybe there is some β-catenin being released intracellularly). Maybe in AGA the binding of β-catenin to AR doesnt affect the transcription of AR genes but binds - just because it can and by the time it reaches the nucleus it cant activate TCF/LEF. I was incorrect about β-catenin making the DPC response worse, it just cant target its canonical genes, thats all.

We also know that DKK-1 is induced by reactive species, specifically JNK mediated. And looking back over the research it looks like the DPC themselves produce DKK-1:

DKK-1 messenger RNA is upregulated in 3-6 hours after 50-100 nM DHT treatment and ELISA showed that DKK-1 is secreted from DP cells in response to DHT.
http://www.ncbi.nlm.nih.gov/pubmed/17657240

But I dont understand why ROS is induced in AGA DPC when in PC it represses it. The androgen receptor represses transforming growth factor-beta signaling through interaction with Smad3.

So although there are quite a few similarities its not quite apples to apples which irritates me.

My current understanding:

Increasing β-catenin will saturate the AR in the presence of DHT/T and will not successfully activate TCF/LEF

However without DHT/T, β-catenin will be able to activate TCF/LEF which is good because we can just try to reduce AR. I'm unsure if AR antagonists will still bind β-catenin.

Increasing β-catenin to suraphysical levels could in theory bypass the AR recruitment

AR will increase ROS and subsequent TGF-β1 + DKK-1 via DPC regardless of β-catenin being present. This can be counteracted with an anti-oxidant

AR can increase GSK3β thereby reducing available β-catenin

Next post will be new treatments and an approach plan along with my current progress.

[Chemical]

Did some more reading on prostate cancer and AR beta-catenin interaction.

Functional localization and competition between the androgen receptor and T-cell factor for nuclear beta-catenin: a means for inhibition of the Tcf signaling axis

Reports have shown a role for the beta-catenin/Tcf pathway in prostate cancer including nuclear localization and beta-catenin mutations in primary prostate tumor samples (Chesire and Isaacs, 2002; Chesire et al., 2002). beta-Catenin has also been described as a ligand-dependent coactivator of the AR (Truica et al., 2000; Mulholland et al., 2002; Pawlowski et al., 2002; Yang et al., 2002). Previously, we have shown that translocating AR can provide a means of nuclear entry and accumulation of beta-catenin (Mulholland et al., 2002). This mode of beta-catenin trafficking has also been shown to hold true in neuronal cells (Pawlowski et al., 2002). Cotrafficking of AR and beta-catenin to the nucleus likely has important implications both for AR and Wnt signaling. While nuclear accumulation of beta-catenin has been shown to correlate with increased AR transcriptional activity, the effects on Tcf signaling have only begun to be explored. Recently, it has been shown that the AR has the ability to inhibit the beta-catenin/Tcf signaling pathway, ligand-dependently (Chesire and Isaacs, 2002). By way of transcriptional reporter assay, this report showed reduced luciferase activity for the Tcf reporter in a ligand-dependent manner in several prostate and colon cancer cell lines. This study also observed reduced transcriptional activity with the use of an AR deletion mutant (for beta-catenin binding), suggesting the possibility of a reciprocal balance of nuclear beta-catenin between the AR and Tcf. In the present study, we corroborate these results but provide mechanistic data to support the hypothesis that repression of the beta-catenin/Tcf signaling is mediated by ligand-occupied AR that is in competition with Tcf for nuclear beta-catenin. Specifically, using transcriptional reporter assays, we show that overexpression of WT Tcf reduced the activity of an AR (ARR3-Luc)-responsive reporter, while overexpression of a DeltaNt Tcf mutant did not have this effect.

Results

TOPFLASH activity = beta-catenin TCF/LEF activity

To evaluate the repressive effect that AR and its physiological ligand had on Tcf signaling, several titrations were performed in PC3 cells. Specifically, we observed decreased TOPFLASH activity with increased amounts of transfected AR (mug/three wells) with no detectable changes observed in either beta-catenin or Tcf4 protein levels (Figure 1a). Basal levels of BCT (TOPFLASH) in untreated PC3 cells were low, while in SW480 cells luciferase counts were 6–8-fold higher. In either cell line, control FOPFLASH values were about 10% of basal TOPFLASH levels. To confirm the ligand dependency of Tcf inhibition, we used AR deletion mutants (Figure 1b), including those coding the amino- and DNA-binding domain regions (Nt/DBD), as well as the DNA-binding region plus ligand-binding region (DBD/LBD). While the Nt/DBD mutant showed little ability to repress TOPFLASH, the DBD/LBD mutant was capable of a 3.5–4-fold repression in the presence of DHT. This suggests that in an AR overexpressed state, the ARNt is dispensable for Tcf repression. Further verification that the LBD is vital for repression is shown by increased relief of AR-mediated repression in cells treated with the pure AR antagonist Casodex (1 and 10 mum), which was able to efficiently relieve AR (DBD/LBD)-mediated repression of TOPFLASH. Having shown that repression is both AR and ligand dependent, we next evaluated the effect of increasing the concentration of DHT.

So bicalutamide reduced beta-catenin recruitment! Woohoo!

As a prerequisite, we verified that we could, in fact, detect an AR/beta-catenin complex upon transfection with AR (Figure 6b, arrow). Consistent with our previous studies (Mulholland et al., 2002), we detected more AR/beta-catenin complex from cells treated with DHT, suggesting a ligand-sensitive interaction. We also detected an association with beta-catenin and Tcf-HIS, but did not detect this association in the DeltaNt Tcf-myc deletion mutant. Importantly, using this assay, interactions between AR and Tcf were very weak as compared to our detected AR/beta-catenin and beta-catenin/Tcf complexes both in prostate and colon cancer cells. Nonimmune, control precipitations were only slightly less than those of the AR/Tcf complex, suggesting that only a small fraction of AR associates with Tcf. We wanted to know whether Casodex, which abrogated both HcRed-Tcf focal accumulation (data not shown) and TOPFLASH activity, could alter the binding of AR and beta-catenin. By treating cells with 5 mum Casodex (dissolved in EtOH) with and without 5 nm DHT, we observed reduced physical interaction between AR and beta-catenin (arrowhead), while also relieving AR-mediated repression of the beta-catenin/Tcf-HIS complex (Figure 6b, star and open arrow heads). To evaluate whether a decreased beta-catenin/TcfHIS complex could be directly affected by the AR TcfHIS and ARS35, in vitro translated products were used in series of binding assays with HIS-tagged recombinant beta-catenin (Figure 6c). Using Ni-NTA beads to precipitate in vitro binding reactions, we observed that upon increased levels of ARS35 (a–d) there was a corresponding decrease in TcfS35 (a–e) precipitated with the HIS-tagged recombinant beta-catenin. Although ARS35 levels up to 10 mul were added per binding reaction, we observed no further increase in detectable AR/beta-catenin-HIS binding greater than 6 mul (point d) or 60% of input by volume.

There is a saturation limit which we can exploit to get beta-catenin to activate TCF/LEF.

Some closing analysis:

The reduction of AR transcription (in the presence of DHT) upon overexpression of Tcf provides evidence of competition. These data are consistent with the notion that modulation of the beta-catenin-Tcf/Lef assembly may be the mechanism by which AR exerts its repression on Tcf/Lef signaling. To test this further, we evaluated the amount of beta-catenin associated with the Tcf/Lef complex with and without AR+androgen, both in vivo and in vitro. Data achieved with these data corroborate morphological data and our overall hypothesis.

Could it be that AR requires beta-catenin to exert its effects?

Other nuclear receptors and Tcf/Lef signaling

It is interesting how two stimulators of cell proliferation (AR/DHT and Wnt) can interact in a repressive manner. One possible interpretation is that androgens may not increase proliferation by, rather, promoting growth and differentiation in prostate epithelia. Prostate cancer cells have cell cycle deregulation compounded by a cell survival response to androgens. The contributions of Wnt signaling in prostate cancer are likely complex, although it is clear that nuclear beta-catenin can serve as a potent AR coactivator. We suggest a scenario whereby beta-catenin could be shuttling between Tcf/Lef-binding sites and AR elements. In the absence of androgen, AR resides mainly in the cytosol, while nuclear beta-catenin associates with Tcf (Figure 8a). In the presence of androgen, beta-catenin could be shuttled by translocating nuclear receptors to both Tcf- and AR-associated response elements to promote coactivation (Figure 8b). Consequently, less beta-catenin would be associated with Tcf and more with AR. This would simultaneously lower Tcf activity and augment AR transactivation.

Very very interesting stuff.

[Seuxin]

Hmm, so you think using bicalutamide could be good ?

[Chemical]

For those of you that havent been following the science, our current understanding is that for as long as Testosterone and DHT is around, any treatment that tries to increase beta-catenin will be significantly blunted. The AR will just suck up the beta-catenin and prevent it from working. So we need to reduce AR and 5ar using suppressors and increase aromatase within hair follicles to further divert Testosterone to convert to Estrogen which can enhance the growth of frontal hair follicles. Also since there will be some DHT/T floating around you will need to boost beta-catenin so that even after AR has bound beta-catenin until saturation, there will still be enough left to activate the good TCF/LEF genes that induce and maintain anagen.

Existing Androgen Receptor inhibitors

Ketoconazole

Ketoconazole binds to the human androgen receptor.

Ketoconazole competition with [3H]methyltrienolone (R1881) for androgen binding sites in dispersed, intact cultured human skin fibroblasts was determined at 22 degrees C. Fifty percent displacement of [3H]R1881 binding to AR was achieved by 6.4 +/- 1.8 (SE) x 10(-5) M ketoconazole. Additional binding studies performed with ketoconazole in the presence of increasing amounts of [3H]R1881 showed that the interaction of ketoconazole with AR was competitive when the data were analyzed by the Scatchard method. It should be noted, however, that the dose of ketoconazole required for 50% occupancy of the androgen receptor is not likely to be achieved in vivo, at least in plasma. Finally, androgen binding studies performed with other imidazoles, such as clotrimazole, miconazole, and fluconozole, revealed that in this class of compounds only ketoconazole appears to interact with the androgen receptor. Ketoconazole appears to be the first example of a non-steroidal compound which binds competitively to both SSBG and multiple steroid hormone receptors, suggesting that the ligand binding sites of these proteins share some features in common.

Ketoconazole as an adjunct to finasteride in the treatment of androgenetic alopecia in men.

Dihydrotestosterone (DHT) binding to androgen receptors (AR) in hair follicles is commonly accepted as the first step leading to the miniaturizing of follicles associated with androgenetic alopecia (AGA). Testosterone is converted to DHT by the enzyme 5alpha-reductase. Finasateride a 5alpha-reducase inhibitor blocks the production of DHT and is currently used to treat AGA. The inhibition is not complete but a reduction of DHT systemically and in the scalp is accomplished. Ketoconazole has been clinically shown to be effective in the treatment of AGA. In this paper, evidence is presented to support the hypothesis that ketoconazole 2% shampoo has a local disruption of the DHT pathway. It is proposed that using ketoconazole 2% shampoo as an adjunct to finasteride treatment could lead to a more complete inhibition of DHT and thus better treat AGA.

Ketoconazole has a plasma half life of 8 hours and I'm not too sure if that relates to skin retention too. The current recommended dosing is 3x a week using the shampoo but I personally encourage the use of the cream 2-3x a day. Its easier to apply, more convenient and you dont need to worry about the shampoo causing irritation from excessive use. Even though keto is a mild AR blocker people still see results using it so its not entirely useless - remember that this is using it 3x a week so imagine if you used it 2-3x a day.

Minoxidil

Minoxidil may suppress androgen receptor-related functions

Minoxidil concentrations were chosen based on most common reports in the literature: 0.1 to 10 μM for oral intake and about 1 mM for topical application of 5% minoxidil (100 mM) in skin tissue (Regaine 5% minoxidil topical solution; monograph, February 2013), assuming 1.7% absorption. In in vitro organ culture or animal studies, high concentrations, ranging from 1-100 mM, have been reported in skin tissue [31, 32]. After treating with different concentrations of minoxidil (1 to 100 μM), cells were harvested and cellular extracts were assayed for luciferase activity. As shown in Fig. Fig.1A,1A, minoxidil suppressed AR reporter activity at the concentrations tested.

To further test minoxidil suppression of AR-related function, we tested the effects of minoxidil on the growth of LNCaP cells, an androgen-sensitive prostate cancer cell line that, as noted above, endogenously expresses AR. As shown in Fig. Fig.1D,1D, minoxidil suppressed DHT-dependent LNCaP cell growth in concentration-dependent manner. Notably, at the highest concentration, cell growth was suppressed to a level comparable to that observed in the absence of DHT. These data provide the first demonstration that minoxidil can suppress AR-related functions, including AR transcription and AR-related cell growth.

To assess possible nonspecific effects of minoxidil at high concentrations, we performed control studies, testing different concentrations of minoxidil (1-100 μM) on glucocorticoid receptor (GR) transcriptional activity. As shown in Fig. Fig.1E,1E, the highest concentration minoxidil (100 μM) did affect GR transcriptional activity in reporter assays in PC-3 cells, whereas lower concentrations (1-10 μM) did not. These data suggest that minoxidil, a small hydrophobic molecule, may have multiple targets in the cell when used at high concentrations.

You can see that minoxidil can suppress AR activity in a dose dependent and time dependent manner providing more evidence that minoxidil works on two levels necessary to treat AGA. This explains why alot of people can achieve regrowth solely using minox seen both in studies and anecdotal reports.

Its important to note that minoxidil does not work if the follicles do not contain adequate sulfotransferase enzymes necessary to convert minoxidil to minoxidil sulphate which is reported to be 14 times more potent.

Minoxidil sulfate is the active metabolite that stimulates hair follicles.

An important step in understanding minoxidil's mechanism of action on hair follicles was to determine the drug's active form. We used organ-cultured vibrissa follicles to test whether it is minoxidil or its sulfated metabolite, minoxidil sulfate, that stimulates hair growth. Follicles from neonatal mice were cultured with or without drugs and effects were assessed by measuring incorporation of radiolabeled cysteine in hair shafts of the treated follicles. Assays of minoxidil sulfotransferase activity indicated that vibrissae follicles metabolize minoxidil to minoxidil sulfate. 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. Inhibiting the formation of intracellular PAPS with chlorate also blocked the action of minoxidil but not of minoxidil sulfate. Acetaminophen, a potent sulfate scavenger blocked cysteine incorporation by minoxidil. It also blocked follicular stimulation by minoxidil sulfate apparently by directly removing the sulfate from the drug. Experiments with U-51,607, a potent minoxidil analog that also forms a sulfated metabolite, showed that its activity was inhibited by both chlorate and diethylcarbamazine. These studies show that sulfation is a critical step for hair-growth effects of minoxidil and that it is the sulfated metabolite that directly affects hair follicles.

Sulfotransferase levels in scalp vary alot between individuals and that can account for the lack of response in some individuals

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

I'm currently looking into ways of boosting scalp sulfotransferase levels to improve the efficacy of minoxidil.

Valproic Acid - Sodium Valproate

I was quite surprised when I found out VA was a serious Androgen Receptor suppressor. Before we look at that I'd like to show you the study they did on men with AGA with a 8.3% Sodium Valproate spray in a 27% EtOH solution over 6 months, twice a day applications of 1ml.

Topical valproic acid increases the hair count in male patients with androgenetic alopecia: a randomized, comparative, clinical feasibility study using phototrichogram analysis.

Full Study

The representative clinical photographs and macrophotographs are presented at baseline and after 24 weeks of treatment (Fig. 2). The total hair count increased after 24 weeks of treatment in the VPA group; the median hair counts were 181/cm2 (range, 125–241) at baseline and 192/cm2 (range, 153–271) at 24 weeks. However, the total hair count did not change in the placebo group; the median hair counts were 194/cm2 (range, 155–244) at baseline and 197/cm2 (range, 132–253) at 24 weeks (Fig. 3a). The median change in total hair count from the baseline was 23/cm2 (range, 17 to 39) in the VPA group and 1/cm2 (range, 68 to 70) in the placebo group, and the difference between groups was statistically significant (P = 0.047; Fig. 3b).

In the full study you can see a photo of the results, and I can definitely see an increase in hair count both in the closeup and wide angle. You have to understand that there is a very high probability the subjects might not have been too diligent with the treatment and the ages ranged from 27-45. The AGA has probably become alot worse and its very unlikely that 6 months of monotherapy will completely regrow hair. For those of you that expect complete regrowth from one treatment please reevalute your expectations, we do not live in the world of movies and fiction where scientists come up single drugs that cure diseases completely. A small but consistent positive effect of a treatment on a reasonable sample size is a good predictor that a treatment has potential to have additive or even synergistic effect when combined with other treatments. Most of the placebo individuals saw a decline in hair count but a vast majority of VA subjects saw no reduction in hair count or an increase - which means it definitely did something.

The serum levels achieved by topical VA dont seem too bad but its something to keep in mind

The serum VPA concentration was measured in 20 subjects in the VPA group and 19 subjects in the placebo group. The serum VPA level was detectable in only seven subjects in the VPA group (range, 0.4–2.3 lg/mL), whereas it was too low (<0.4 lg/mL) to be detected in the other 13 subjects in the VPA group and all of the subjects in the placebo group.

Valproic acid and its derivatives enhanced estrogenic activity but not androgenic activity in a structure dependent manner.

Full Study

Steroid hormones affect metabolic pathways and cellular functions. Valproic acid (VPA), used as antiepileptic drug, inhibits histone deacetylases and interacts with intracellular receptors. We analyzed the impact of VPA and VPA derivatives on activation of estrogen and androgen receptors (ER and AR) using reporter gene assays. VPA and its long-chain derivatives (long name 1), (long name 2) and (long name 3) enhanced 17β-estradiol-induced ERα and ERβ activation partly synergistically with a structure-activity correlation. The extent of this effect regarding to ERα activation increased with prolongation of the aliphatic side chain. Regarding AR activation, VPA, S-pentyl-4-yn- and heptyl-4-yn-VPA slightly induced AR activity when tested alone. In combination with the AR agonist 5α-dihydrotestosterone, VPA, S-pentyl-4-yn- and heptyl-4-yn-VPA showed anti-androgenic effects without an apparent structural relation. Our results indicate that VPA and its derivatives affect estrogen signaling with a structural specificity, while the (anti-)androgenic effects of these compounds are not structurally correlated.

So here we have our first indication that VPA can indeed suppress AR. No mention of how much so lets delve into the study

For VPA, concentrations between 0.001 and 3 mM and for its derivatives between 0.01 and 1 mM were investigated. The therapeutic range of VPA comprises 0.35–0.7 mM in serum [42], whereas concentrations of 3 mM can be attained in the serum of patients after an acute VPA ingestion [43].

In combination with 10−9 M E2, VPA enhanced the E2-induced ER activation synergistically in a concentration-dependent manner with a first significant effect at 0.001 mM. For instance, 2 mM VPA caused a 3.5-fold increase of ER activity compared to 10−9 M E2 alone (E2: 100% vs VPA plus E2: 348% of maximum E2- activation; Fig. 4A). Similarly for ER activation, the co-incubation of VPA (0.001–3 mM) with 10−9 M E2 caused synergistic effects (e.g. E2: 100% vs. 1 mM VPA plus E2: 408% of maximum E2-activation; Fig. 4B). Thus, VPA enhanced the E2-induced activation of both ER types in a dose-dependent manner with a slightly greater effect on ER activation.

VPA is quite powerful at potentiating the ER pathways which is exactly what we want for frontal DPC.

Co-administration of VPA (0.01–4 mM) with 10−9 M DHT reduced AR activity in a dose-dependent manner with a first significant effect at 1 mM (Fig. 7). Butyl-4-yn-VPA did not alter DHT-induced AR activation. S-pentyl-4-yn- and heptyl-4-yn-VPA (≥0.2 mM) also decreased DHT-mediated AR activation. At the highest concentration, VPA and heptyl-4-yn-VPA inhibited the AR activity induced by DHT to control values of untreated cells (Fig. 7).

Just look at what 1mM could do to DHT's ability to bind to AR. Just look at it! And look at what 2mM could do, its identical to frickin flutamide.

The anti-androgenic effects of VPA are also characterized through a reduced AR expression and cell growth inhibition revealed in cell cultures of prostate cancer cells [64,65]. For example, Iacopino et al. [66] have shown that 0.45 mM VPA suppressed DHT-stimulated proliferation of human androgen-sensitive LNCaP prostate cancer cells. Epigenetic regulations byVPA might be involved in the modulation ofAR activity. HDAC inhibition leads to AR acetylation [67] causing reduced cell growth, apoptosis induction and differentiation [64,68]. However in our study, VPA seems to have both anti-androgenic and marginal androgenic effects dependent on the VPA concentration and the presence or absence of DHT. A structural dependency of the VPA derivatives could not be attributed to the AR activity in the presence of DHT.

That was Valproic Acid, what about Sodium Valproate?

Valproate is an anti-androgen and anti-progestin.

We examined the widely used anti-convulsants valproate (VPA) and carbamazepine (CBZ) for steroidal bioactivity using a yeast-based steroid receptor-beta-galactosidase reporter assay for the androgen receptor (AR), progesterone receptor (PR) or estrogen receptor (ER). Bioassays were performed (a) to detect agonist activity by exposing yeast to 100 microM CBZ or VPA or (b) to detect antagonist activity by exposing yeast stimulated with testosterone (5 x 10(-9) M, AR), progesterone (1.6 x 10(-9) M, PR) or estradiol (2.6 x 10(-11) M, ER) together with either VPA or CBZ for 4 (PR) or 16 (AR, ER) hours. VPA showed dose-dependent (1-800 microM) inhibition of progesterone-induced PR- and testosterone-induced AR activity but had no ER antagonist bioactivity and no significant PR, AR or ER agonist bioactivity. VPA also showed a dose-dependent (1-200 microM) blockade of DHT's suppression of AR-mediated NF-kappaB activation in human mammalian cells. By contrast, CBZ had no significant PR, AR or ER agonist or AR and ER antagonist bioactivity but at the highest concentration tested (800 microM) it did antagonize PR activity. We conclude that VPA is a non-steroidal antagonist for human AR and PR but not ER. VPA's androgen and progesterone antagonism at concentrations within therapeutic blood levels (350-700 microM) seems likely to contribute to the frequency of reproductive endocrine disturbances among patients treated with VPA.

Since Sodium Valproate is the more readily available form and has excellent water solubility at 50mg/ml, in comparison to 1.3mg/ml for Valproic Acid (source) it looks like Sodium Valproate is a strong candidate to be used in our protocols. I use distilled water to dissolve the tablet after crushing them and removing the orange coating but I'm looking for capsules which should make my life easier. I would suggest starting at 5mg/ml then increasing until you get irritation or notice side effects. You can just dilute to bring the concentration down again.

This post is quite lengthy so I'll continue with the other treatments in the next post.

[Rog]

Chemical, have you seen:

http://www.dannyroddy.com/weblog/antihistaminebaldnessstress

which if correct indicates that Estrogen plays a negative role (causes terminal hair - vellus, is involved in fibrosis of scalp around follicles, etc) rather than a positive role?