# Men's Hair Loss > Hair Loss Treatments > Cutting Edge / Future Treatments >  Updated Research and Knowledge - Cutting Edge

## Chemical

Hi all, I've been doing some research on MBP for the pat few weeks and I thought I'd share some of the interesting insights on this topic, its a long read but you can skip the abstracts and articles for my conclusions. Please do not hesitate to ask questions and I strongly encourage people to dispute this research, as this will lead to a beneficial discussion on points that I havent considered.

Lets look at how DHT works to cause MBP - in depth.




> *The Anticancer Testosterone Metabolite 3β-Adiol*
> 
> Since DHT is generally agreed to be carcinogenic, the thought was that reducing the transformation of testosterone into DHT would reduce cancer risk. In the Prostate Cancer Prevention Trial (PCPT), 18,882 men over age 55 years with normal prostate examinations and a PSA below 3.0 were randomly assigned finasteride or placebo for seven years.
> Slightly Lower Cancer Risk, Much Higher Killer Cancer Risk
> The results were a surprise to the researchers, who found  as they expected  a lower rate of cancer in the finasteride group than in the placebo group (18.4% versus 24.4%). What surprised them was the considerably higher number of significantly aggressive cancers  for the technically inclined, higher Gleason scores  among those in the finasteride group with cancer versus those with cancer in the placebo group (37% versus 22.3%). Thats why you never saw a television commercial about finasteride preventing prostate cancer. To be accurate it would have to say, Take finasteride! It lowers your risk of prostate cancer, but if you do get prostate cancer, youre more likely to die of it!
> The PCPT researchers concluded, finasteride prevents or delays the appearance of prostate cancer, but this possible benefit and a reduced risk of urinary problems must be weighed against sexual side effects and the increased risk of high-grade prostate cancer. A separate meta-analysis published as a Cochrane Review found that 5a-reductase inhibitors, including finasteride, have inadequate evidence to say that these patent medicines reduce mortality, in terms of prostate cancer.
> Its true that if DHT alone is considered, elevated levels of DHT might be thought to increase your prostate cancer risk, as DHT is a procarcinogenic (for the technically inclined, dedifferentiating) meta¬bolite.
> However, thanks to evolution and Nature has developed a natural way of counter acting the procarcinogenic effects of DHT! If we take the metabolites of DHT into consideration, too, elevated DHT may or may not have this effect. One of these metabolites may actually offset or even reduce any DHT-increased risk. How does that happen?
> After testosterone is converted to DHT, DHT is in turn normally metabolized into a relatively smaller quantity of 5a-androstane-3a,17b-diol (abbreviated as 3a-Adiol), and a usually larger amount of 5a-androstane-3b,17b-diol (abbreviated as 3b-Adiol). These same researchers also report that while nearly all the 3a-Adiol is converted back to DHT (which presumably makes 3a-Adiol a pre-procarcinogen), the 3b-Adiol does not convert back to 5a-DHT. Very importantly, they report that 3b-Adiol is an anticarcinogen (for the technically inclined, a redifferentiating agent) that activates estrogen receptor beta, an anticarcinogenic estrogen receptor present in large numbers in the prostate gland.5 (Estrogen receptor beta is present in many other tissues in both sexes, but thats a topic to be explored at another time.)


 

Here are some studies in support of the DHT metabolite 3Beta diol being a potent activator of Estrogen Receptor Beta.

5α-Androstane-3β,17β-diol (3β-diol), an estrogenic metabolite of 5α-dihydrotestosterone, is a potent modulator of estrogen receptor ERβ expression in the ventral prostrate of adult rats
An endocrine pathway in the prostate, ERβ, AR, 5α-androstane-3β,17β-diol, and CYP7B1, regulates prostate growth

Back to the original article on DHTs effect on the prostate.




> In a letter to the editor of the New England Journal of Medicine, Otabek Imamov, MD, et al. state, [DHT] is the fulcrum in this balance. It suggests that finasteride, by blocking the conversion of testosterone to [DHT], inhibits the production of [3b-Adiol] thus suppressing [the anticarcinogenic activity of] ERb and preventing the [re]-differentiation of epithelium. This mechanism could account for the higher incidence of poorly differentiated tumors in the finasteride group in the Prostate Cancer Prevention Trial.6
> A review in the Biology of Reproduction Journal states, We believe that a higher incidence of low-differentiated [more aggressive] tumors in the finasteride-treated arm observed in the PCPT is caused by altering the normal differentiation of prostatic epithelium in the environment lacking the natural ERb ligand  [3b-Adiol].7
> Research has found some very specific things that 3b-Adiol does to inhibit prostate cancer growth. According to the title of a 2005 research report: The androgen derivative 5a-androstane-3b, 17b-diol [3b-Adiol] inhibits prostate cancer cell migration through activation of the estrogen receptor beta subtype.8 Other researchers reported that 3b-Adiol not only inhibits PC3-Luc cell [a specific type of prostate cancer cell] migratory properties, but also induces a broader antitumor phenotype [type of cell] by decreasing the proliferation [growth] rate, increasing cell adhesion [cancer cells dont stick as normal cells do] and reducing invasive capabilities in vitro.9 But these researchers went beyond test-tubes to living mice, writing In vivo, continuous administration of 3b-Adiol reduces growth of established tumors and counteracts metastasis formation when PC3-Luc cells are engrafted subcutaneously in nude mice or are injected into the prostate.
> The conclusion to this research article was very encouraging: Since 3b-Adiol has no androgenic activity, and cannot be converted to androgenic compounds, the effects here described entail a novel potential application of this agent against human PC.9 A novel potential application of 3b-Adiol, a totally natural human testosterone, against human prostate cancer! Where are the headlines? This article was published in 2010!
> For the really technically inclined, here are several mechanisms of action of *3b-Adiol, all of which come from stimulation of estrogen receptor beta:
> - repression of VEGF-A (vascular endothelial growth factor A) expression
> - destabilization of HIF-1a (hypoxia-inducible factor 1a)*
> - reduction of Snail1″ relocation from the cytoplasm to the nucleus of cancer cells
> 
> According to the researchers who published the above mechanisms of action,  high Gleason grade cancers  exhibit significantly more HIF-1a and VEGF-A and Snail1 nuclear localization compared to low Gleason grade cancers.


 It appears VEGF expression is crucial for hair follicle anagen induction and maintenance as described by this study:




> The murine hair follicle undergoes pronounced cyclic expansion and regression, leading to rapidly changing demands for its vascular support. Our study aimed to quantify the cyclic changes of perifollicular vascularization and to characterize the biological role of VEGF for hair growth, angiogenesis, and follicle cycling. *We found a significant increase in perifollicular vascularization during the growth phase (anagen) of the hair cycle*, followed by regression of angiogenic blood vessels during the involution (catagen) and the resting (telogen) phase. Perifollicular angiogenesis was temporally and spatially correlated with upregulation of VEGF mRNA expression by follicular keratinocytes of the outer root sheath, but not by dermal papilla cells. Transgenic *overexpression of VEGF in outer root sheath keratinocytes of hair follicles strongly induced perifollicular vascularization, resulting in accelerated hair regrowth after depilation and in increased size of hair follicles and hair shafts. Conversely, systemic treatment with a neutralizing anti-VEGF antibody led to hair growth retardation and reduced hair follicle size.* No effects of VEGF treatment or VEGF blockade were observed in mouse vibrissa organ cultures, which lack a functional vascular system. These results identify VEGF as a major mediator of hair follicle growth and cycling and provide the first direct evidence that improved follicle vascularization promotes hair growth and increases hair follicle and hair size.
> http://www.ncbi.nlm.nih.gov/pmc/articles/PMC199257/


 This shows that VEGF is crucial for hair follicle survival and a reduction in VEGF mediated angiogenesis (formation of new blood vessels to supply hair follicles) could prevent the transition from telogen to anagen.

Heres a study demonstrating the role of hif-1 in VEGF regulation:




> Transcriptional Regulation Controls Angiogenesis in Hypoxia
> One important HIF-1 function is to promote angiogenesis; HIF-1 directs migration of mature endothelial cells toward a hypoxic environment [2,5]. This is done via HIF-1 regulation of vascular endothelial growth factor (VEGF) transcription. VEGF is a major regulator of angiogenesis, which promotes endothelial cell migration toward a hypoxic area. During hypoxia, *HIF-1 binds the regulatory region of the VEGF gene, inducing its transcription and initiating its expression* [12,15,16]. Such endothelial cells ultimately help to form new blood vessels, supplying the given area with oxygenated blood [14].
> http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2140184/


 Androgens actually enhances the expression of HIF-1 via Androgen Receptor mediated signalling which explains the procarcinogenic effect of androgens in prostate cancer:
http://www.ncbi.nlm.nih.gov/pubmed/24035332



> Androgens stimulate hypoxia-inducible factor 1 activation via autocrine loop of tyrosine kinase receptor/phosphatidylinositol 3'-kinase/protein kinase B in prostate cancer cells.
> Dihydrotestosterone (DHT) activates HIF-1alpha nuclear protein expression in LNCaP cells but not in androgen receptor-negative PC-3 cells. HIF-1alpha expression is correlated with the transactivation of a hypoxia-responsive element-driven reporter gene and with the production of VEGF protein. *The effect of DHT on HIF-1 was blocked by nonsteroidal antiandrogens, flutamide and bicalutamide. DHT does not affect HIF-1alpha mRNA levels but regulates HIF-1alpha protein expression through a translation-dependent pathway*. PC-3 cells when incubated with increasing amounts of conditioned medium from LNCaP cells treated with DHT experienced a dose-dependent increase in HIF-1alpha. This induction was not seen either when LNCaP cells were treated with flutamide or conditioned medium were pretreated with antibody to the epidermal growth factor (EGF). HIF-1 activation by DHT was blocked by LY294002, a potent inhibitor of the phosphatidylinositol 3'-kinase signaling pathway, whereas HIF-1 activation by EGF, as ligand, was not inhibited by flutamide. In contrast, HIF-2alpha protein was not affected by androgens or antiandrogens.


 This could explain the vellus to terminal transition of follicles all over the body in males, especially facial hair. We can conclude from this that it is indeed possible for vellus hair follicles to become terminal given the right environmental conditions (local growth promoting agents) and time. Males require a significant number of years of exposure to androgens to develop thick facial and body hair.
Evolution could be responsible for this DHT driven inhibition of androgen's procarcinogenic effects i.e the males with higher androgens were physically and mentally (androgen increases grey matter/spatial ability) stronger but were also susceptible to early cancer related deaths, so the males that ended up surviving were the ones that had the AR related edge and developed a feedback loop that canceled out the negative effects of androgens. Some of these biological feedback loops have been refined over thousands of years an as a result have become increasingly complex due to natural selection (survival of the fittest/most adaptable).
This is all interesting with regards to the prostate but what about the scalp?
First lets take a look at how exactly 3 beta diol is synthesized.



> 5α-Androstane-3β,17β-diol, also called 3β-androstanediol, and often shortened to 3β-diol, is an endogenous steroid hormone. It is a *5α-reduced and 17β-hydroxylated metabolite of dehydroepiandrosterone (DHEA)* as well as a *3β-hydroxylated metabolite of dihydrotestosterone (DHT)*. *Similarly to DHEA*, 3β-diol is a high-affinity full agonist of the ERβ, and hence, an estrogen.
> https://en.wikipedia.org/wiki/5%CE%B...,17%CE%B2-diol


 


> 3 beta diol can be synthesized from either DHEA or DHT via the respective enzymes. An important bit of information to note is that both 3 beta diol and DHEA activate ERbeta.
> 
> *Regional scalp differences of the androgenic metabolic pattern in subjects affected by male pattern baldness.*
> Regional differences in the androgen metabolism were established in alopecic and non alopecic areas of patients affected by male pattern baldness (MPB). 5-alpha-reductase (5-alpha-R) activity was measured by the formation of dihydrotestosterone (DHT), using 3H-testosterone as substrate: this activity was higher in the alopecic areas (3.4 pmol/g tissue/h) than in the non alopecic skin (1.5 pmol/g tissue/h). *3-alpha,beta-hydroxysteroid oxoreductase* (3-alpha, beta-HO) was studied using 3H-DHT as precursor and measuring the corresponding formed 3-alpha- and 3-beta-androstanediols (alpha DIOL and beta DIOL). *The beta DIOL was the predominant metabolite and total 3-alpha, beta-HO activity was higher in alopecic skin* (12.4 pmol/g tissue/h) *than in non alopecic areas* (8.4 pmol/g tissue/h). Also 17, beta-hydroxysteroid oxoreductase was measured using either testosterone or DHT as substrates: androstenedione formed from testosterone was higher in hairy skin (12 pmol/g tissue/h) than in alopecic areas (6 pmol/g tissue/h); androstanedione formed from DHT was also higher in non alopecic areas (8.1 pmol/g tissue/h) than in alopecic skin (2.8 pmol/g tissue/h). The greater formation of beta DIOL in the sebaceous glands-enriched alopecic skin supports the hypothesis for a specific role of this metabolite in the control of the sebaceous activity.


 This study is incredibly crucial to understanding the regional differences in Androgen metabolism and how the hairline is affected the most. We know that 5ar is elevated in balding regions but furthermore, the enzymes responsible for catalyzing the conversion of DHT to 3beta diol are also elevated in alopecic regions, whereas the enzymes responsible for synthesizing anrostanedione were higher in non alopecic regions.

When I read this I immediately recalled a compound that is commonly used for treating Androgenetic Alopecia: Ketoconazole (nizoral).
Ketoconazole is actually a really effective inhibitor of androgen synthesis, and a weak inhibitor of Androgen receptor.




> *In vitro inhibition by ketoconazole of human testicular steroid oxidoreductases.*
> An oral antimycotic agent, ketoconazole has been demonstrated to be an inhibitor of cytochrome P-450-dependent monooxygenases. To investigate its effect on steroid oxidoreductases, in vitro studies were carried out using subcellular fractions of human testes. *Ketoconazole competitively inhibited activities of 3 beta-hydroxy-5-ene-steroid oxidoreductase/isomerase* and NADH-linked 20 alpha-hydroxysteroid oxidoreductase for steroid substrate and the Ki values were 2.9 and 0.9 microM, respectively. In contrast, ketoconazole inhibited neither 17 beta-hydroxysteroid oxidoreductase nor NADPH-linked 20 alpha-hydroxysteroid oxidoreductase, indicating that the two 20 alpha-hydroxysteroid oxidoreductases are distinct. Further, ketoconazole inhibited non-competitively the above enzyme activities for the corresponding cofactors of NAD and NADH. From the binding mode of ketoconazole to cytochrome P-450 and the present findings, it appears likely that the agent binds to a site which is different from that of steroids or pyridine nucleotides.


 *3 beta-hydroxy-5-ene-steroid oxidoreductase = 3 beta HSD
* 


> *Hydroxylation of 5 alpha-androstane-3 beta,17 beta-diol by rat prostate microsomes: potent inhibition by imidazole-type antimycotic drugs and lack of inhibition by steroid 5 alpha-reductase inhibitors.*
> 5 alpha-Dihydrotestosterone, the principal androgen mediating prostate growth and function in the rat, is formed from testosterone by steroid 5 alpha-reductase. 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*. *Inhibitors of steroid 5 alpha-reductas*e (4-azasteroid analogs; 10 microM) or* inhibitors of 3 beta-hydroxysteroid oxidoreductase* (trilostane, azastene, and cyanoketone; 10 microM) had *no appreciable effect* on the 6 alpha-, 7 alpha-, or 7 beta-hydroxylation *of 5 alpha-androstane-3 beta,17 beta-diol* (10 microM) by rat prostate microsomes. 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. The* rank order of inhibitory potency* of the imidazole-type antimycotic drugs was *miconazole greater than clotrimazole greater than ketoconazole.* In the case of clotrimazole, the inhibition was shown to be competitive in nature, with a Ki of 0.03 microM. The imidazole-type antimycotic drugs [B]inhibited all three pathways of 5 alpha-androstane-3 beta,17 beta-diol hydroxylation to the same extent[B], which provides further evidence that, in rat prostate microsomes, a single cytochrome P450 enzyme catalyzes the 6 alpha-, 7 alpha-, and 7 beta-hydroxylation of 5 alpha-androstane-3 beta,17 beta-diol. These studies demonstrate that certain imidazole-type compounds are *potent, competitive inhibitors* of 5 alpha-androstane-3 beta,17 beta-diol *hydroxylation* by rat prostate microsomes, which is consistent with the effect of these antimycotic drugs on cytochrome P450 enzymes involved in the metabolism of other androgens and steroids.


 So for clarification:

Ketoconazole/miconzole inhibit 3 Beta HSD
3 beta HSD converts DHT to 3beta diol.
3beta diol activates ERbeta.
ERbeta destabilizes HIF-1 and results in loss of VEGF expression.
Hair follicles fail to enter anagen due to lack of bood supply.
This is a potential explanation of the mechanism behind ketoconazoles hair growth/hair loss prevention effects.

I also wanted to talk a bit about PGD2. Specifically its metabolite:




> 15-dPGJ2 and PGD2 inhibit hair growth in mouse and human hair follicles
> Given the temporal peak of PGD2 before the apoptotic catagen stage, the published ability of its metabolite 15-dPGJ2 to induce apoptosis in other cell types, we tested the effects of the prostaglandins on primary cell culture of keratinocytes isolated from neonatal foreskin. *15-dPGJ2 induces apoptosis* (fig. S2A), as evidenced by plasma membrane blebbing and cell retraction/shrinkage. *15-dPGJ2 also decreased cell density, cell division, and live-cell numbers* (fig. S2, B to D). Perhaps because the origin of these keratinocytes was not the hair follicle, *PGD2 had no such effect on the cells.* However, 15-dPGJ2 did increase sub-G1 DNA quantities and activated caspase 3 in human keratinocytes, which are features of apoptotic cell death (fig. S2, E to G). We therefore hypothesized that at least 15-dPGJ2, if not also PGD2, could directly inhibit hair growth in vivo.
> 15-dPGJ2 was applied topically to dorsal back skin of C57BL/6 mice that had been depilated to synchronize the hair follicle cycle. Starting on day 8 after depilation and continuing every other day, we applied 10 μg of 15-dPGJ2 or acetone vehicle. Hair length was measured on days 4, 12, 14, and 16 after depilation. On days 12 to 16, hair at the site of treatment was shorter than in vehicle-treated animals (Fig. 6A). To determine a minimal effective dose, we tested the application of 1 μg of both PGD2 and 15-dPGJ2 as above and measured hair length on day 20 after depilation. PGD2 inhibited hair growth, but to a lesser extent than 15-dPGJ2 (Fig. 6B). We found no evidence of changes in hair follicle cycling grossly or by histologic examination.


 

http://www.spandidos-publications.com/etm/1/2/257

Im still doing research on this by product of PGD2 as it may hold more relevance than PGD2 itself.

Another very crucial effect of DHT is the elevation Dickkopf-1 (DKK1). 



> Dihydrotestosterone-inducible dickkopf 1 from balding dermal papilla cells causes apoptosis in follicular keratinocytes.
> Abstract
> Recent studies suggest that androgen-driven alteration to the autocrine and paracrine factors produced by scalp dermal papilla (DP) cells may be a key to androgen-potentiated balding. Here, we screened dihydrotestosterone (DHT)-regulated genes in balding DP cells and found that dickkopf 1 (DKK-1) is one of the most upregulated genes. 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. A co-culture system using outer root sheath (ORS) keratinocytes and DP cells showed that DHT inhibits the growth of ORS cells, and neutralizing antibody against DKK-1 significantly reversed the growth inhibition of ORS cells. Analysis of co-cultured ORS cells showed a significant increment of sub-G1 apoptotic cells in response to DHT. Also, recombinant human DKK-1 inhibited the growth of ORS cells and triggered apoptotic cell death. In addition, DHT-induced epithelial cell death in cultured hair follicles was reversed by neutralizing DKK-1 antibody. Moreover, immunoblotting showed that the DKK-1 level is up in the bald scalp compared with the haired scalp of patients with androgenetic alopecia. Altogether, our data strongly suggest that DHT-inducible DKK-1 is involved in DHT-driven balding.


 So how exactly does dkk1 work?



> DKK1 is a high affinity antagonistic ligand for LRP6, which is a Wnt coreceptor that acts together with the Frizzled serpentine receptor to initiate Wnt signal transduction. Two different models have been proposed to account for the mechanism by which DKK1 antagonizes LRP6 function. One model suggests that DKK1 binding to LRP6 disrupts Wnt-induced Frizzled-LRP6 complex formation, whereas the other model proposes that DKK1 interaction with LRP6 promotes LRP6 internalization and degradation, thereby reducing the cell surface LRP6 level.


 Lets take a look at the importance of LRP6.






> WNT - Canonical pathway
> The canonical Wnt pathway (or Wnt/β-catenin pathway) is the Wnt pathway that causes an accumulation of β-catenin in the cytoplasm and its eventual translocation into the nucleus to act as a transcriptional coactivator of transcription factors that belong to the TCF/LEF family. *Without Wnt signaling, the β-catenin would not accumulate in the cytoplasm since a destruction complex would normally degrade it.* This destruction complex includes the following proteins: Axin, adenomatosis polyposis coli (APC), protein phosphatase 2A (PP2A),* glycogen synthase kinase 3 (GSK3)* and casein kinase 1α (CK1α).[14] It degrades β-catenin by targeting it forubiquitination, which subsequently sends it to the proteasome to be digested.[11][15] However, as soon as *Wnt binds Fz and LRP5/6, the destruction complex function becomes disrupted.* This is due to Wnt causing the translocation of the negative Wnt regulator, Axin, and the destruction complex to the plasma membrane.Phosphorylation by other proteins in the destruction complex subsequently binds Axin to the cytoplasmic tail of LRP5/6. Axin becomes de-phosphorylated and its stability and levels are decreased. Dsh then becomes activated via phosphorylation and its DIX and PDZ domains inhibit the GSK3 activity of the destruction complex. This allows β-catenin to accumulate and localize to the nucleus and subsequently induce a cellular response via gene transduction alongside the TCF/LEF (T-cell factor/lymphoid enhancing factor)[16]transcription factors.[15]


 So without LRP6, the canonical WNT pathway cannot be activated. This means Beta Catenin will be degraded by GSK3. 
Its very clear that without Beta Catenin hair will not form:




> Further analysis demonstrates that β-catenin is essential for fate decisions of skin stem cells: in the absence of β-catenin, stem cells fail to differentiate into follicular keratinocytes, but instead adopt an epidermal fate.


 The real question is how does DHT actually increase DKK1? This is something that Ive been struggling to figure out. Finding an effective and feasible way to prevent the increase of DKK1 as a result of DHT may allow us to focus our attention away from DHT inhibitors, and also allow other hair promoting agents to work better (although even without agonists the bodys natural wnt proteins can have a chance to bind to the LRP6 and do their job).

Ive come across mainly dead ends and no studies to date actually document how Dkk1 is regulated.

http://www.nature.com/jid/journal/v1.../5700999a.html

We know that L-ascorbic acid 2-phosphate and  L-threonate, an ascorbate metabolite inhibits DHT induced DKK-1.
Dkk-1 is also induced by p53: http://www.ncbi.nlm.nih.gov/pubmed/10777218

DHT increases p53 and p21 significantly: http://www.ncbi.nlm.nih.gov/pubmed/22859066
This is a weak connection at best. 

Other possible links include:
http://carcin.oxfordjournals.org/con.../1877.full.pdf




> The Wnt antagonist DICKKOPF-1 gene is induced by 1a,25-dihydroxyvitamin D3
> The Wntb-catenin pathway is aberrantly activated in most colon cancers. DICKKOPF-1 (DKK-1) gene encodes an extracellular Wnt inhibitor that blocks the formation of signalling receptor complexes at the plasma membrane. We report that 1a,25- dihydroxyvitamin D3 [1,25(OH)2D3], the most active vitamin D metabolite, increases the level of DKK-1 RNA and protein in human SW480-ADH colon cancer cells. This effect is *dose dependent, slow and depends on the presence of a transcription-competent nuclear vitamin D receptor (VDR)*. Accordingly, 1,25(OH)2D3 activates a 2300 bp fragment of the human DKK-1 gene promoter. Chromatin immunoprecipitation assays revealed that 1,25(OH)2D3 treatment induced a pattern of histone modifications which is compatible with transcriptionally active chromatin.  Exogenous expression of E-cadherin into SW480-ADH cells results in a strong adhesive phenotype and a 17-fold increase in DKK-1 RNA. In contrast, an E-cadherin blocking antibody inhibits 1,25(OH)2D3-induced differentiation of SW480-ADH cells and DKK-1 gene expression. Remarkably, in vivo treatment with the vitamin D analogue EB1089 induced DKK-1 protein expression in SW480-ADH cells xenografted in immunodeficient mice, and a correlation was observed in the expression of VDR and DKK-1 RNA in a series of 32 human colorectal tumours. These data indicate that *1,25(OH)2D3 activates the transcription of the DKK-1 gene, probably in an indirect way* that is associated to the promotion of a differentiated phenotype. DKK-1 gene induction constitutes a novel mechanism of inhibition of Wnt signalling and antitumour action by 1,25(OH)2D3.


 It appears VDR (Vitamin D receptor) mediates DKK-1 protein expression in the presence of E cadherin by binding to a promoter region on Dkk1 gene.
Furthermore, DHT has been shown to upregulate VDR via - wait for it  ERBeta!




> Sex steroids induced up-regulation of 1,25-(OH)2 vitamin D3 receptors in T 47D breast cancer cells.
> 
> Abstract
> There is evidence indicating that 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] through binding to its specific receptor (VDR) exerts an antiproliferative effect on breast cancer cells. Considering the importance of receptor regulation in modulating the target cell responsiveness to hormones, the effect of dihydrotestosterone (DHT) and estradiol-17 beta (E2) on the regulation of VDR number was investigated in T 47D human breast cancer cells that also express androgen and estrogen (ER) receptors. Exposure to 10(-7) M DHT for 72 h resulted in a significant increase in VDR levels. Similar results were obtained with 10(-7) M E2. *DHT- and E2-induced up-regulation was completely suppressed by 10(-6) M tamoxifen (TAM) addition but unaffected by 10(-6) M flutamide*. TAM treatment alone produced a significant dose-dependent increase in VDR content, that was maximal at 10(-6) M. Our data strongly suggest, for the first time, an up-regulation of VDR by DHT and E2 via an ER-mediated mechanism.


 Although this studys focus is on breast cancer cells, its still points us in the right direction.

Vitamin D has been shown to have a biphasic effect on hair growth:




> http://www.ncbi.nlm.nih.gov/pubmed/8077696
> At higher concentrations of 1,25(OH)2D3, there was a dose-dependent inhibition of both follicle and fiber growth (IC50 values of 100 nM), in part due to reduction in the growth periods. There was a *marked delay between the onset of 1,25(OH)2D-induced hair follicle and hair fiber growth inhibition.*


 This exactly inline with the oxford study:




> Time-course and dose-curve experiments showed that 1,25(OH)2D3 (107 M) caused a *slow 3- to 5-fold induction of DKK-1 RNA at 2448 h* upon treatment. The effect of 1,25(OH)2D3 was specific, as several hormones (dexamethasone, retinoic acid, progesterone and oestradiol) acting through members of the superfamily of nuclear receptors similar to VDR did not induce DKK-1. The induction of DKK-1 was confirmed at the protein level and in another colon cancer cell line. Immunofluorescence studies confirmed the increase in DKK-1 protein expression following 1,25(OH)2D3 exposure and showed its preferential localization in the cell periphery, Golgi apparatus and vesicles of the exocytic route. These results confirmed that 1,25(OH)2D3 induces DKK-1 expression with slow kinetics, which precluded the use of translation inhibitors such as cycloheximide to investigate whether the induction is direct or indirect.


 So to conclude, its quite apparent there are a multitude of factors involved in the prognosis of MBP, but certain pathways have been overlooked which I believe has stagnated potential progress.

I strongly believe that inhibiting DKK-1 or reducing it will enable other treatments to work far more effectively, along with mitigating the detrimental effects of downstream DHT pathways i.e the actual effect instead of the overall mediator. The answer lies in the details, we must fully understand the mechanisms  the how and why before trying to exploit the pathway for our own gain. This is the key to hacking and efficient problem solving, you find the weakest link and plan your attack on that. 
In the next post I will discuss therapeutic applications of these findings and some experiments Ive been conducting (with interesting results), after all, what good is theory without application?

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## bananana

Very interesting, I found some wiki pages that confirm your theory...

I got ONE QUESTION:

*DKK-1 - How do we kill it?
*

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## Chemical

> Very interesting, I found some wiki pages that confirm your theory...
> 
> I got ONE QUESTION:
> 
> *DKK-1 - How do we kill it?*


 This is the million dollar question. 

One way is to inhibit DHT, but that is a difficult task especially using topical agents.

Another is l-threonate/ascorbate derivatives.




> We recently reported that DHT inhibits the growth of co-cultured ORS keratinocytes using this in vitro co-culture system and demonstrated that the growth suppression is largely due to DKK-1 (11). Consistent with this, the growth of cocultured ORS cells was significantly suppressed in the presence of 100 nM DHT. Since L-threonate repressed DHT-induced DKK-1 expression, we investigated whether or not L-threonate attenuates DHT-induced growth suppression of co-cultured keratinocytes. We indeed observed that L-threonate reversed the DHT-induced growth inhibition of co-cultured keratinocytes. In summary, our data demonstrates that* L-threonate repressed DHT-induced DKK-1 expression* in cultured DPCs and attenuated DHT-induced growth inhibition of co-cultured keratinocytes. Although further investigation is needed to elucidate the mechanism of L-threonate-mediated repression of DHT induced DKK-1 expression, our data in this study strongly suggest that L-threonate has an inhibitory effect on androgen-driven balding (Fig. 4).


 L threonate or Ascorbate derivatives but they do not last long or work effectively as topical treatments. (I also havent seen any supporting evidence)

Another way could be completely inhibiting Estrogen Receptor Beta to prevent the DHT -> VDR -> DKK-1 pathway with topical tamoxifen which has a moderately long tissue half life (7 days when applied topically to breast tissue), there currently isnt any topical available but it shouldnt be too hard to make a formulation. The study used off the shelf inregredients to make their own topical - I'll have to dig it out if anyones interested as it could combat all the negative effects of DHT if it works, including:

-stabilizing HIF-1 alpha, disinhibiting VEGF, and inhibiting DKK1. 

Here's some more evidence on why DKK1 is of the main culprits:




> The neutralizing antibody against DKK-1 reverses the DHT-induced cell death in cultured hair follicles
> To further investigate the role of DHT-inducible DKK-1 in hair growth, we employed a hair-follicle organ culture system (Philpott et al., 1994). Significant cell death was observed in epithelial cells surrounding DP in response to 100 nm DHT compared with control follicles (Figure 6a and b). *The neutralizing antibody against DKK-1 reversed the DHT-induced cell death in epithelial cells* (Figure 6c), demonstrating that the cell death is due largely to DKK-1.
> 
> DKK-1 level is up in the balding scalp compared with the haired scalp of patients with AGA
> To strengthen the argument that our in vitro observations translate to a significant role for DKK-1 in AGA, we examined DKK-1 expression in balding and non-balding scalp of patients. Immunoblots showed a strong band around 35 kDa from balding scalp, whereas we observed a weak signal from non-balding scalp (Figure 7). *This result clearly shows that DKK-1 is upregulated* in the balding scalp compared with the haired scalp of patients with AGA.


 However, a working, and readily available supplement that moderately inhibits DKK1 and enhances a myriad of other growth factors is: oleuropein. I'll be making an in-depth post on oleuropein this weekend, how it works, supporting evidence, and results of my own experiments with this compound in a topical formulation  (very very exciting results).

----------


## BaldingEagle

Quality post. 

Thanks for taking the time. Very interesting theory.

----------


## bananana

Sounds good, oleuropein is basically extract of* olive leaf*?

Seems a bit farfetched because I see bunch of ORAL supplements selling on ebay, so probably a lot of people using them already,
and no remarks of hair growth... Or you think it should be applied topically? 

regards

----------


## tiktok

Solid work so far!
I'm partial the idea that heightened estrogen levels are a likely cause of propecia not working for some people (couples sometimes with a burning/flushed scalp, another symptom of elevated estrogen), and your research seems to back this up.

Regarding topical Tamoxifen, Afimoxifene (4-hydroxytamoxifen) looks like a great candidate as it works topically and doesn't seem to increase much in serum (http://clincancerres.aacrjournals.or.../3672.abstract).

----------


## burtandernie

Yeah good work with piecing all that together but its a bit over my head. How do you put this to practical use? How do you stop DKK-1?
The complexity of all this is what makes me not want to try propecia. I really never liked the idea of screwing this big complicated system up, but what choice do you have. Nothing else seems to work

----------


## jamesst11

> Yeah good work with piecing all that together but its a bit over my head. How do you put this to practical use? How do you stop DKK-1?
> The complexity of all this is what makes me not want to try propecia. I really never liked the idea of screwing this big complicated system up, but what choice do you have. Nothing else seems to work


 You don't... this is a nice regurgitation, and a lot of effort and that's awesome but molecular biologists have been trying to sort this out for years.  when we are 60, perhaps they will have a solution.

----------


## Magster

Looking forward to your experiments with topical oleuropein, Chemical. I am interested in doing similar.

----------


## Chemical

In this post Ill be describing the effects of Oleuropein on hair follicles and just how awesome this herb is.
Ill start off with the main study which luckily is a free pubmed article:

*Topical Application of Oleuropein Induces Anagen Hair Growth in Telogen Mouse Skin*

Im not going to bother with the cliffs, were going to look at the findings in depth because thats where the devil lives.




> Recently, we observed that oleuropein reduced body weight gain and visceral adiposity in high-fat diet (HFD)-fed mice. The protective effect of oleuropein against HFD-induced adiposity in mice appeared to be mediated through the *upregulation of genes involved in Wnt10b-mediated signaling cascades* [11]. In the present study we investigated whether oleuropein could induce anagenic hair growth in C57BL/6N mice and explored the underlying mechanism.


 What does WNT10b do?




> Here, we showed that in response to *prolonged ectopic Wnt10b-mediated β-catenin activation, regenerating anagen hair follicles grew larger in size.* In particular, the hair bulb, dermal papilla and hair shaft became enlarged, while the formation of different hair types was unaffected. Interestingly, we found that the effect of exogenous WNT10b was mainly on Zigzag and less on the other kinds of hairs. We observed dramatically enhanced proliferation within the matrix, DP and hair shaft of the enlarged AdWnt10b-treated hair follicles compared with those of normal hair follicles at P98. Furthermore, expression of CD34, a specific hair stem cell marker, was increased in its number to the bulge region after AdWnt10b treatment. Ectopic expression of CD34 throughout the ORS region was also observed. Many CD34-positive hair stem cells were *actively proliferating in AdWnt10b-induced hair follicles*. Importantly, subsequent co-treatment with the Wnt inhibitor, DKK1, reduced hair follicle enlargement and decreased proliferation and ectopic localization of hair stem cells. Moreover, injection of DKK1 during early anagen significantly reduced the width of prospective hairs. Together, these findings strongly suggest that Wnt10b/DKK1 can modulate hair follicle size during hair regeneration.
> 
> http://www.ncbi.nlm.nih.gov/pubmed/24750467


 We have our first evidence that oleuropein has the potential to significantly stimulate hair growth in the absence of DKK1.




> We investigated the effect of oleuropein on the proliferation of DPCs. DPCs were treated with various concentrations of oleuropein (10, 20, and 50 μM), and cell proliferation after day 5 was assessed by using an MTT assay and trypan blue exclusion assay. Outcomes from trypan blue assay were highly consistent with that from MTT assay (Fig 1B). *Treatment with oleuropein significantly increased DPC proliferation relative to untreated controls* (Fig 1). The highest degree of proliferation was achieved using 20 μM oleuropein. *The effect of 20 μM oleuropein on increasing cell proliferation was significantly higher than that of minoxidil at 100 μM*.


 

Strangely the optimal concentration that achieved the highest percentage of cells with increased proliferation was 20um, instead of the 50um, perhaps theres a biphasic effect? Or maybe a statistical/extraneous anomaly because the two concentrations have similar efficacy. Ill come back to this.




> To investigate whether oleuropein modulates the Wnt/β-catenin pathway in DPCs, we examined the expression of nuclear β-catenin by western blotting analysis. As shown in Fig 2A, the protein level of *nuclear β-catenin was significantly increased in oleuropein-treated DPCs (a 212% increase)* compared with control cells treated with dimethyl sulfoxide (DMSO). Cells treated with oleuropein for 24 h were evaluated for expression of Wnt/β-catenin signaling target genes by using RT-PCR. We found that compared with control cells treated with DMSO, cells treated with oleuropein showed significantly increased expression of LEF1 (P = 0.0027) and Cyc-D1 (P = 0.0034) (Fig 2C). In addition, cells treated with oleuropein showed significantly increased nuclear accumulation of β-catenin and *mRNA expression of LEF1 and Cyc-D1 relative to those treated with minoxidil*.


 (Cyc d1 is an indicator of cell cycle progression and correlates with proliferation)
Oleuropein increases BetaCatenin which as we know is a very good thing.




> To determine whether oleuropein promoted hair growth, we measured the length of 10 hairs plucked from the dorsal skin center area of each mouse at 0, 7, 14, 21, and 28 days. As shown in Fig 3B, *the hairs in the oleuropein-treated group were significantly longer than those in the control group. Moreover, the hairs in the oleuropein-treated group were observed to be longer than those in the minoxidil-treated group.* 
> ---
> In the representative longitudinal sections, the number and size of hair follicles were significantly increased in the oleuropein-treated group compared to the control group (Fig 4). In addition, the *number and size of hair follicles in the oleuropein-treated group were significantly greater than those of minoxidil-treated group.*


 The hairs were physically longer than the tried and proven minoxidil! And there were more follicles in anagen (up from telogen). Im curious as to what concentration of minoxidil they used and if using  higher concentration wouldve yielded better results.




> Mice treated with oleuropein exhibited a *significant increase in the mRNA levels of IGF-1* (P = 0.0008), hepatocyte growth factor (HGF, P = 0.0021), *vascular endothelial growth factor* (VEGF, P = 0.0062), and *keratocyte growth factor* (KGF, P = 0.0052) in their skin tissues compared with mice treated with vehicle (Fig 5A). The effect of oleuropein on increasing mRNA levels of these growth factors was significantly greater than that of minoxidil. ELISA analysis revealed that *dermal IGF-1 levels were significantly increased in mice treated with oleuropein (71% increase)* compared with mice treated with vehicle only (Fig 5B). The oleuropein-treated mice demonstrated significant increases in dermal IGF-1 level and immunohistochemical expression compared with control mice (Fig 5B and 5C).


 

Oleuropein somehow manages to increase IGF1 (by a lot) and VEGF too. This study shows that oleuropein can increase VEGF via WNT10b, however Im not sure how IGF-1 was elevated. The figure clearly shows just how much these growth factors are elevated, but Im surprised minoxidil didnt come anywhere near close enough to oleuropein in increasing VEGF.




> DPC incubated with increasing minoxidil concentrations* (0.2, 2, 6, 12 and 24 mumol/L) induced a dose-dependent expression of VEGF mRNA.* Quantification of transcripts showed that *DPC stimulated with 24 mumol/L minoxidil express six times more VEGF mRNA than controls*. Similarly, VEGF protein production increases in cell extracts and conditioned media following minoxidil stimulation. These studies strongly support the likely involvement of minoxidil in the development of dermal papilla vascularization via a stimulation of VEGF expression, and support the hypothesis that minoxidil has a physiological role in maintaining a good vascularization of hair follicles in androgenetic alopecia.


 The oleuropein study used 3mg of minoxidil  not sure howthat translates to mumol, maybe someone an help?




> and the minoxidil group received vehicle containing 3 mg of minoxidil.


 Anyways, now comes the best part:




> Mice treated with oleuropein exhibited a *significant increase in the mRNA levels of WNT10b* (P = 0.0045),* LRP5* (P = 0.0004),* FZDR1* (P = 0.0123), and the Wnt-responsive transcription factor LEF1 (P = 0.0142), along with its target genes, such as Cyc-D1 (P = 0.009), in their skin tissues compared with mice treated with the control vehicle (Fig 6A). In addition, oleuropein-treated mice showed a *significant decrease in the mRNA level of DKK1* (P = 0.0003) in their skin tissues compared with control vehicle-treated mice. The oleuropein-treated mice showed significant increases in the mRNA levels of Wnt signaling-related genes (Wnt10b, LRP5, FZDR1, LEF1, and Cyc-D1) relative to minoxidil-treated mice. This effect of oleuropein on increasing protein levels of β-catenin in hair follicles was greater than that of minoxidil.


 

THIS IS THE ****ING HOLY GRAIL. The rare SYNERGY. When you decrease an inhibitory protein and boost an agonist protein you achieve synergy, where 2 + 2 does not equal 4 (additive) but rather 7. You can see that DKK1 was reduced from control levels. They did not elucidate the actual pathway but if oleuropein targets the gene expression or transcription then this could mean DHT can no longer exert its effects on the hair follicle via DKK1. I am still trying to figure out how this inhibition works because I dont like believing studies that do not have supporting evidence from other sources. If someone can assist with research Id be very very grateful.
Lets talk about the dosages and how it translates to human doses.




> Based on our preliminary study involving different oleuropein dosages (0.4 to 3.0 mg/mous), *0.4 mg of oleuropein per mouse was found to be the minimal effective dose* for anagen hair induction in C57BL/6N mice.


 0.4mg as in 10um? Or 20um? probably 20 as the minimum EFFECTIVE dose. 3mg might have resulted in diminishing returns since they didnt see dose dependant growth. I believe theres a plateau of diminishing returns and oleuropein isnt biphasic but thats still open to debate. 




> Each mouse received 200 μL of the reagent applied topically with a plastic spatula to the shaved dorsal skin daily for 28 days. The control group received vehicle alone, the oleuropein group received vehicle containing 0.4 mg of oleuropein, and the minoxidil group received vehicle containing 3 mg of minoxidil. *All reagents used for the hair growth test were dissolved in a vehicle composed of 50% (v/v) ethanol, 30% water, and 20% propylene glycol.*


 Nothing fancy here, just your standard 50% ethanol, water and propylene glycol. Which means anyone can make an oleuropein vehicle very cheaply. Heck, you could even dissolve it in minoxidil (which is what Ive been doing).
To conclude the study:




> Li et al. reported that Wnt10b could induce the biological switch of hair follicles from the telogenic phase to the anagenic phase in mouse skin [31]. In addition, they showed that *siRNA suppression of β-catenin inhibited hair follicle regeneration, even when Wnt10b was overexpressed* [31]. Recently, Choi et al. reported that β-catenin-deleted and DKK1-expressing hair follicles showed greatly diminished expression of Cyc-D1, a direct Wnt/β-catenin target gene that helps initiate the transition from late G1 to S phase of the cell cycle, likely contributing to decreased hair follicle matrix proliferation [32]. In the present study,* in parallel to the significant upregulation of Wnt10b, LRP5, FZDR1, β-catenin, LEF1, and Cyc-D1, a prominent decrease of DKK1 levels* was observed in skin tissue of oleuropein-treated mice compared with the control group. On the basis of these results, we propose that oleuropein may cause premature entry of telogenic follicles into the anagen phase via activation of the Wnt10b/β-catenin signaling pathway.


 ------

Imagine what would happen if we combined Minoxidil with this. 
A decrease in DKK1 -> more LRP6 available to bind -> the body's natural WNTs can finally work.
Oleuropeins WNT10b upregulation -> increased IGF-1, VEGF, KGF -> BetaCatenin
An increase in VEGF both from oleuropein AND Minoxidil -> more blood vessels to hair follicle, which as Ive discussed in my first post, leads to increased hair follicle size and growth.
A ridiculous amount of beta catenin from oleuropein and minoxidil combined  does anyone see where Im going with this?

Thats the agonist side covered, and not to forget:

ketoconazole and miconazole nitrate for 3 beta HSD suppression -> less DHT converted to 3 Beta diol  which is a potent ERBeta agonist -> disinhibition of VEGF pathways and stabilization of HIF-1.
Throw in topical saw palmetto just for safe measure and youve got yourself a serious stack.

Bringing me to the part that is most important.
My experiments. First a little bit about myself:

My hairline started receding when I was 17 because I stupidly decided to mess with anabolic steroids (which is where I got my biology knowledge). I used emu oil, and nizoral but didnt touch minoxidil or fin, saw a slight improvement but nothing significant. My father started receding at 30, so Ive definitely got the genes and wouldve receded eventually even if I hadnt messed around with my androgens.

I became a NW2 at 18, after using letrozole to boost testosterone to suraphysical levels not realizing it would **** up my hairline. I started using seakelp bioferment, microneedling, emu oil, folligen, and some other supplements. Saw slight improvements but nothing too great.

Then I became a NW3 at 19 when I had the worst shed of my life after a car accident. I was in icu for 2 months and the stress really destroyed my hair. I used emu oil, folligen and microneedling but still wasnt seeing any significant growth. 
Fast forward this year april. I was desperate. So I took the plunge. I decided to go on fin, minoxidil (everyday), and ketoconazole cream (everyday). I used emu oil religiously to help with absorption. 2 months in and I saw great regrowth, increased density and I became a NW2.2 ish.
I took the fin every 3 days because I didnt want it to affect my pregnenolone levels too much (Ive got anxiety and fin ED made it worse). I stopped after the 2 month mark. I incorporated iodine into te minoxidil thinking it would help. didnt do shit. Kept using minoxidil mixed with emu oil with minor improvements in hair density and decreased shedding.

August 2015, I started putting the pieces together and I had a few breakthroughs in terms of research.

Bought myself Oleuropein capsules from amazon (extra strength), and mixed it with minoxidil. I had one bottle left so I dissolved like 5 capsules into the liquid and gave it a good shake. I also added a few saw palmetto capsules which I regret because it did not dissolve at all. I let it sit for 24 hour then shook it again.
I then used the dropper to mix around 10ml into my emu oil dispenser which was half full (25ml). And that was my treatment. I used it most nights but it was a real pain to keep looking at the ceiling to prevent it from pouring down my forehead. In the morning it would all be absorbed thanks to the minoxidil but thered be a few remnants of the undissolved saw palmetto and the oleuropein. 

Every week I kept adding more drops of the minoxidil (with the dissolved oleuropein and saw) because I wanted to see what would happen. I also used ketoconazole cream 2% (Janssen pharma) twice a day non stop because it was easy to apply.  

6 weeks in and I could feel stubble, small prickly hairs. I was ****ing ecstatic. My existing hairs became thicker especially the vertex area, and shedding stopped  (almost completely) but I was still skeptical, I thought it mustve been a delayed reaction to minoxidil and ketoconazole. My frontal hairline grew a few hairs initially because thats where I was using it the most, but I wanted to see if this solution actually did anything so I started using it only on my left side and tilting my head to the right to make it pour down to the other side and the backwards to reach the rest of my scalp.
I regret not taking enough pictures before starting although I wasnt really planning to share this until I genuinely saw an improvement. Ive got a few shitty pictures taken with a potato before the regimen:



( 6 weeks into the regimen):





The left side was the same as the first pic before the regimen:



 

The past few weeks Ive been really busy with work and have only been using the solution heavily on the weekends (3-5 times a day) to make up for not using it on the weekdays. I've been feeling more stubble, mainly on the left side, and thicker vertex hair. I also got carried away with the oleuropein and now the solution leaves a hard residue after drying so Ive had to buy more minoxidil and emu oil which should be here this week. I ran out of ketoconazole also so Ill be starting a proper log with decent pictures now that Ive got a better phone.

I'll be posting more recent pics tomorrow.

I would strongly recommend people get some ketoconazole in cream form and maybe miconazole too to apply everyday, as often as possible. The reason being, testosterone is always present in the blood, which means DHT is being converted all the time by the hair follicles. You need to keep the hair follicles saturated with antagonists and agonists to keep them from being slowly killed by DKK1/DHT. Agonists like minoxidil and your body's own WNT proteins WILL NOT work if DKK1 has internalized the LRP6 receptors. 

I'd also suggest people not to take finasteride systematically because the body will naturally increase testosterone to offset the negative feedback loop, and since finasteride only inhibits ~60%, that increase in testosterone will give 5AR additional substrate to convert into DHT, defeating the whole purpose of fin. I'd like to see people try this with RU as an augmentation, since theoretically there should be less DKK1 as a result of impaired AR signalling.

Sorry for the long read.

----------


## Left High Kick

Hi guys, 

I'm currently doing dermarolling once a week and i want to try oleuropein as a daily hair loss care. 

I have some questions about oleuropein. If someone has knowledge about chemistry and would be willing to help me it would be cool. 

1. Oleuropein molar mass is 540 dalton. Which means that according to the 500 dalton rule, it won't penetrate the skin. 
Would vehicule made of 50% of ethanol 30% of water and 20% of PPG will allow some penetration of oleuropein despite its initial molar mass ? 

2. Can i replace ethanol with vodka ? Will Vodka have effectivness in this vehicule ? (I don't want to use DMSO btw).

Thank you very much for you help. 

Take care !

----------


## TheKingofFighters

inbeforethecure, please read the below:

http://www.educatedtherapists.com/fo...-play-in-acne/

----------


## TheKingofFighters

> inbeforethecure, please read the below:
> 
> http://www.educatedtherapists.com/fo...-play-in-acne/


 The *main activator of mTOR* is a *variety of amino acids* and the hormone insulin.  Testosterone also is capable of activating mTOR.

Protein, especially leucine
Excess calories
Excess carbs
Exercise  activated in brain, muscle and heart.Inhibited in liver and fat cells.
Orexin
*IGF-1*
Insulin
*Testosterone*
Ghrelin  in hypothalamus
Leptin  in the hypothalamus
Thyroid hormone  in the hypothalamusand other cells
Oxygen
*Ketamine.  (In the brain  produces antidepressant effect.)*
IL-6   in muscle and fat
Natural Inhibitors of mTOR

*Protein restriction
Leucine restriction
Glutamine restriction
Methionine restriction
Lysine restriction
Arginine restriction
Threonine restriction
Isoleucine restriction*<== weight gainers and whey protein formulations gives me an uber itchy scalp within half an hr after consumption.
Calorie restriction
*Ketogenic Diets*
Cortisol/Glucocorticoids
*Metformin*
NAC
Resveratrol
Aspirin
Cod liver/ Omega-3
Extra Virgin Olive Oil
EGCG/Tea
Curcumin
R-Lipoic Acid
Caffeine
Apigenin
Quercetin
*Genistein*
DIM (R)
Ursolic acid
Emodin (found in Fo-Ti, Resveratrol, Rhubarb, Aloe,)
Andrographis/Andrographolide
Pomegranate/Ellagic acid
Reishi
Milk thistle/Silymarin
Oleanolic acid
Anthocyanins/Grape Seed Extract
*Rhodiola*
Carnosine


What is FOX01
FOX01 stands for forkhead box class O1 transcription factor.  FoxO1 is an important transcription factor that modulates the expression of genes involved in cell cycle control, DNA damage repair, apoptosis, oxidative stress management, cell differentiation, glucose and lipid metabolism, inflammation, and innate and adaptive immune functions. FoxO1 is expressed in all mammalian tissues including human and plays an important role in the regulation of metabolism. FoxO1 has been proposed to function as a key regulator in the pathogenesis of acne as FoxO1 senses external nutrient and internal growth factor signals and relays these to FoxO1-dependent gene regulation.

High glycemic diet, dairy products and animal protein has been recognized as affecting this expression through activation of IGF-1/insulin, which, in turn, influence mTORC1. Transcriptor factor sterol regulatory element binding protein-1 (SREBP-1) is influenced by mTORC. This pathway leads to pathogenesis in sebaceous glands and more synthesis of free fatty acids, with well known role in acne vulgaris pathogenesis.

I have frequently heard Aestheticians say that if a client is having hormonal acne that they should get their hormone levels checked.  Often the therapist is shocked that the client comes back reporting that her levels of hormones are normal and within healthy limits.  *The hormone levels of acne clients and non-acne clients are usually normal.  It is not the level of these hormones that is the issue it is how acne-prone skin reacts to them.   Acne skins have increased conversion of testosterone to DHT.(just like AGA- not circualting serum levels of androgens, but how the local tissue like the balding scalp react to the circulating androgen)
*
And this is where genes come into play. Because of genetic factors acne patients are deficient in nuclear transcription factor FoxO1. Sebum production and skin cell growth are out of control in acne patients. FoxO1 acts like a break to these processes, and its malfunctioning. It reduces sensitivity to androgens by suppressing androgen receptors and regulates cell growth and inflammation. Thus theres a good reason to believe that the less FoxO1 is present in the skin the more prone to acne it is.

*Insulin and IGF-1 can make the situation even worse by further reducing FoxO1 levels.*(the last fiure in the 106 study states that IGF-1 is not expressed in both balding and non-balding scalp DPCs)

So how does this work and how does it affect Acne

What you eat can show up on your skin, and one way this happens is through hormones. Studies link acne to Western-style diets (high in sugar and calories), and given what we know this is not a surprise. Eating sugar and refined carbohydrates causes the pancreas to release large amounts of insulin and IGF-1. Over time this type of diet leads to insulin resistance and chronically high levels acne-causing hormones.

Eating minimally processed low glycemic index foods can reverse the situation, and this has been now demonstrated in several studies.  A low glycemic load diet has been shown to improve acne symptoms, and decrease IGF-1 and skin oil production in several studies(1-3).(This is agreeable becos I find that weight gainers and protein shakes gave me noticeably increased hair loss and itchy scalp)

FOXO1 INHIBITS LIPOGENESIS

FoxO1 not only suppresses protein synthesis and cell growth, but also lipid metabolism. FoxO1 regulates the key transcription factor of lipid synthesis SREBP-1c.  IGF-1 induced SREBP-1 expression and enhanced lipogenesis in SEB-1 sebocytes via activation of the PI3K/Akt pathway, whereas FoxO1 antagonized the expression of SREBP-1c. Thus, reduced expression of SREBP-1 should be expected from a low glycaemic load diet associated with attenuated IIS. In fact, a 10-week low glycaemic load diet reduced SREBP-1 expression in the skin of acne patients, reduced the size of sebaceous glands, mitigated cutaneous inflammation and improved acne. Furthermore, FoxO1 suppresses the activity of peroxisome proliferatoractivated receptor-γ (PPARγ) and LXRα that both costimulate sebaceous gland lipogenesis.

Isotretinoins sebum-suppressive effect has recently been associated with upregulated FoxO1 expression. Reported reductions in IGF-1 serum levels during isotretinoin treatment.

FOXO1 SUPPRESSES ANDROGEN SIGNALLING

Sebaceous gland growth and acne are androgen dependent. The growth of androgen-responsive tissues is coordinated with general somatic growth. IGF-1 stimulates gonadal and adrenal androgen synthesis as well as intracutaneous intracrine conversion of testosterone to tenfold more active dihydrotestosterone, the most potent androgen receptor (AR) ligand. Enhanced hepatic IGF-1 synthesis by Western Diet may thus increase the availability of potent androgens in the skin.

*Intriguingly, FoxO1 functions as an androgen receptor cosuppressor.* Nuclear extrusion of FoxO1 by high IIS relieves FoxO1-mediated repression of androgen receptor transactivation. Thus the western diet stimulates androgen receptor-mediated signalling, which explains enhanced peripheral androgen responsiveness.  Both androgen receptors and IIS synergistically increase SREBP-1-mediated lipogenesis and upregulate lipogenic pathways.

*FOXO1 REDUCES OXIDATIVE STRESS*

Overnutrition and anabolic states with enhanced mTORC1 activity are associated with increased oxidative stress, which has been observed in acne vulgaris. FoxOs upregulate defense mechanisms against reactive oxygen species (ROS). FoxO1 induces the expression of haeme oxygenase 1 *(this is a vasculature-related gene downregulated in balding scalp DPCs)* and thereby reduces mitochondrial ROS formation. FoxO1 and FoxO3 mediate the expression of the ROS scavenger sestrin. FoxO3 stimulates the expression of ROS-degrading enzymes manganese superoxide dismutase and catalase. Hence, FoxOs are key players of redox signalling and link western diet to enhanced metabolic oxidative stress in acne vulgaris.

FOXO1 LINKS NUTRITIONAL STATUS TO INNATE AND ADAPTIVE IMMUNITY

FoxO family members suppress the highly substrate- and energy-dependent process of T-cell activation, whereas FoxO1 deficiency in vivo resulted in spontaneous T-cell activation and effector differentiation. Increased CD4+ T-cell infiltration and enhanced IL-1 activity have been detected in acne-prone skin areas prior to comedo formation.  Thus, FoxO1 links nutrient availability and metabolic conditions to T-cell homoeostasis.
FoxOs control antimicrobial peptide synthesis.

Downregulated FoxO signalling by western diet may thus favour an AMP-deficient follicular microenvironment, which may allow overgrowth of P. acnes.   Western Diet would not only overstimulate sebum production favouring P. acnes growth but may diminish AMP-controlled host responses against P. acnes, which may ultimately stimulate inflammatory TLR-mediated innate immune responses against hypercolonized P. acnes. Upregulated TLR-driven innate immune responses against P. acnes with overexpression of TNF-α may further enhance sebaceous gland lipogenesis via activated proinflammatory mTORC1 signalling.

MTORC1: CONVERGENCE POINT OF NUTRIENT SIGNALLING IN ACNE

Western diet overactivates mTORC1 by providing an abundance of dairy- and meat-derived essential amino acids, increased IIS induced by dairy protein consumption and high glycaemic load and suppressed AMPK activity by calorie excess. As protein and lipid biosynthesis, cell growth and proliferation are coordinated by mTORC1, it is obvious that mTORC1 plays a key role in acne pathogenesis, characterized by increased proliferation of acroinfundibular keratinocytes, SG hyperplasia and increased SG lipogenesis.

ACNE AND MTORC1-DRIVEN INSULIN RESISTANCE

Nutrient signalling of western diet results in increased activation of downstream substrates of mTORC1. S6K1-mediated phosphorylation of insulin receptor substrate 1 (IRS-1) downregulates IIS and thus induces insulin resistance. Dietary fatty acids directly activate S6K1 independent of mTORC1. Insulin resistance is considered to be a physiological feature of increased growth during puberty. However, pathologically persistent insulin resistance is associated with the metabolic syndrome as well as acne-associated syndromes. Thus, increased mTORC1/S6K1 signalling explains the reported associations between western diet, acne, increased body mass index (BMI) and insulin resistance.

MTORC1 REGULATES LIPID SYNTHESIS

Increased sebaceous gland lipid biosynthesis is responsible for seborrhoea and sebaceous gland hyperplasia. Importantly, the key transcription factor of lipid biosynthesis SREBP-1 depends on mTORC1 activation. mTORC1 phosphorylates lipin-1, which controls the access of SREBP-1 to the promoter region of SREBP-1-dependent lipogenic genes in the nucleus.

FOXO1: THE RHEOSTAT REGULATING MTORC1

As both mTORC1 and FoxO1 integrate nutrient and growth factor signals, it is conceivable that they interact with each other to coordinate cellular responses to nutrient availability. FoxOs are pivotal inhibitors of mTORC1 and have emerged as important rheostats that modulate the activity of mTORC1. FoxO1, FoxO3 and FoxO4 induce the expression of sestrin3 that activates AMPK, which inhibits mTORC1.  Furthermore, Akt-phosphorylated cytoplasmic FoxO1 binds to TSC2 and thereby dissociates the TSC1/TSC2 complex, which activates mTORC1. Thus, activated Akt inhibits FoxO1, FoxO3 and FoxO4 through direct phosphorylation and indirectly activates mTORC1, which in turn increases protein and lipid synthesis and induces insulin resistance.  In summary, FoxO transcription factors, especially FoxO1, inhibit the activity of mTORC1 at multiple levels of cellular regulation.

----------


## InBeforeTheCure

Well guys, here's a quick look at the network "reverse engineered" from differentially expressed genes in balding DPCs (> 1.5 or < 0.67 fold change) using X2K. The algorithm infers differentially regulated transcription factors from the gene expression data, then from that builds a protein network that regulates those TFs, and then from that infers upstream kinases that act on those proteins. This gives a nice hierarchical model.



You can right click and select "view image in another tab" or whatever to see a larger version. Red = transcription factors, yellow = proteins, green = kinases.

The image shows just the connections between the top 10 most differentially regulated TFs, and the top 10 most differentially regulated kinases. I'll have more on that later.

@TheKingofFighters

Rapamycin, which inhibits mTORC1, has been shown to extend lifespan in a variety of species:




> *Inhibition of the TOR signalling pathway by genetic or pharmacological intervention extends lifespan in invertebrates, including yeast, nematodes and fruitflies*1, 2, 3, 4, 5; however, whether inhibition of mTOR signalling can extend lifespan in a mammalian species was unknown. *Here we report that rapamycin, an inhibitor of the mTOR pathway, extends median and maximal lifespan of both male and female mice when fed beginning at 600 days of age. On the basis of age at 90% mortality, rapamycin led to an increase of 14% for females and 9% for males.* The effect was seen at three independent test sites in genetically heterogeneous mice, chosen to avoid genotype-specific effects on disease susceptibility. Disease patterns of rapamycin-treated mice did not differ from those of control mice. In a separate study, rapamycin fed to mice beginning at 270 days of age also increased survival in both males and females, based on an interim analysis conducted near the median survival point. *Rapamycin may extend lifespan by postponing death from cancer, by retarding mechanisms of ageing, or both.* To our knowledge, these are the first results to demonstrate a role for mTOR signalling in the regulation of mammalian lifespan, as well as pharmacological extension of lifespan in both genders. These findings have implications for further development of interventions targeting mTOR for the treatment and prevention of age-related diseases.


 (link)

This is probably thanks to reversing mTOR inhibition of autophagy.




> Organismal lifespan can be extended by genetic manipulation of cellular processes such as histone acetylation, the insulin/IGF-1 (insulin-like growth factor 1) pathway or the p53 system. *Longevity-promoting regimens, including caloric restriction and inhibition of TOR with rapamycin, resveratrol or the natural polyamine spermidine, have been associated with autophagy (a cytoprotective self-digestive process) and in some cases were reported to require autophagy for their effects.* We summarize recent developments that outline these links and *hypothesize that clearing cellular damage by autophagy is a common denominator of many lifespan-extending manipulations.*


 (link)

FOXOs also play a role in longevity, and in fact SNPs around FOXO3 have been linked to a higher chance of living to 95+ years old.

----------


## TheKingofFighters

> Well guys, here's a quick look at the network "reverse engineered" from differentially expressed genes in balding DPCs (> 1.5 or < 0.67 fold change) using X2K. The algorithm infers differentially regulated transcription factors from the gene expression data, then from that builds a protein network that regulates those TFs, and then from that infers upstream kinases that act on those proteins. This gives a nice hierarchical model.
> 
> 
> 
> You can right click and select "view image in another tab" or whatever to see a larger version. Red = transcription factors, yellow = proteins, green = kinases.
> 
> The image shows just the connections between the top 10 most differentially regulated TFs, and the top 10 most differentially regulated kinases. I'll have more on that later.
> 
> @TheKingofFighters
> ...


 hi ur hardwork is appreciated- but can u explain what's the correlation in differences with the sizes of the circles? Downregulated or upregulated??

----------


## InBeforeTheCure

> hi ur hardwork is appreciated-


 Thank you. Lots of work left to do though.  :Smile: 




> but can u explain what's the correlation in differences with the sizes of the circles? Downregulated or upregulated??


 Size of circle (nodes) = number of other circles (nodes) connected to it

Anyway, the results are definitely interesting. I'll go over some quick observations before studying these things more deeply...

First, it seems that the computer likes FOXA2 rather than PAX1 as the causative gene at 20p11. Despite it not being expressed in dermal papilla cells, we see its "shadow", as well as the shadow of HNF4A (which is also not expressed in DPCs). Why? Is it a fictitious result? Some sort of statistical artifact? Here's a crazy idea: What if instead of dermal papilla cells, FOXA2 and HNF4A are expressed transiently in the _precursors_ to dermal papilla cells (neural crest cells?). FOXA2 and HNF4A are pioneer factors -- they modify chromatin so that transcription factors bind particular sites and not others in particular cell types. FOXA2 and HNF4A are known to cooperate in differentiation of liver cells (link). Notice also that FOXA2 binds to the homeobox HOXA5, which in the oPossum results was one of the most enriched transcription factors in balding DPCs.

But can such an effect on chromatin persist even after FOXA2 and HNF4A are no longer expressed? Possibly. Something like this has been observed for another pioneer factor called NeuroD1:




> Our study also addressed for the first time how transcription factors that are induced for a brief period during development have potential to orchestrate long-term transcriptional program by inducing epigenetic memory. NeuroD1 is highly induced very briefly during the onset of neurogenesis, while many of its target genes are kept active for much longer period despite its later absence. We find that a transient expression of NeuroD1 was sufficient to trigger changes at its target sites that ensured a long-term maintained loss of repressive transcription factor landscape as well as heterochromatin and consequently, a transcriptionally induced state of its targets. These striking findings demonstrate that the brief period of NeuroD1 action can generate epigenetic memory at its target sites that lasts longer than the presence of NeuroD1 itself and contributes to the maintained expression profile and phenotypic state. In combination with our findings of a strong sequence specificity in NeuroD1 recruitment, these observations provide new concepts of how genetic determinants may guide epigenetic memory to ensure stability of distinct transcription programs during development.


 (link)

As for the other stuff, MAPK1 (ERK2) and MAPK 3 (ERK1) are the two most altered kinases. CSNK2A1 (Casein Kinase 2) interacts with that pathway. MAPK14 is p38-alpha, a stress-activated pathway that is probably the major upstream regulator of senescence in AGA. CDK2 regulates G1/S cell cycle progression. GSK3beta inhibits beta-catenin of course, but phosphorylates a lot of other things as well. ATM is DNA damage response, and maybe HIPK2, although the latter is also involved in the ERK pathway and in regulating HIF1A. AKT1 we've talked about.

SOX2, a DP signature gene, is the most differentially regulated transcription factor. This plays a role in signaling to epithelial progenitor cells. Rendl et. al* showed that Sox2 knockout mice have higher BMP expression and slower migration of progenitor cells. Since BMPs are shown to be downregulated in balding DPCs, perhaps Sox2 is more active in AGA -- then the migration of progenitor cells would be faster? EGR1 is downstream of the ERK pathway. MITF is involved in DNA replication, DNA repair, mitosis, miRNA production, and mitochondrial metabolism. E2F1, SUZ12, PHF8, and MTF2 are involved in cell cycle regulation and senescence. POU5F1 (Oct-4) binds to Sox2...

*Avi Ma'ayan, an author of this paper, was also involved in writing the X2K program.

That's just rushing through of course. We'll get deeper into this stuff later on.

----------


## TheKingofFighters

atually- the clue is something that u've have already read before: 

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4365504/

SNP
marker
Position
(hg19)
Alleles IS
Genotype
IS Genotype
Frequency
1000G CEU
Genotype
Frequency
Disrupted TF binding site
rs11699227 21,961,920 C/T CC 1.00 0.259 TATA
rs6036003 21,961,964 A/G AA 1.00 0.259 4 altered motifs:*HNF4*, RAR, RXRA,
STAT
rs169311 21,962,333 A/C AA 1.00 0.259 4 altered motifs: BATF; COMP1, Irf, VDR
rs201545 21,962,422 A/C CC 1.00 1.000 N/A
rs5840940 21,962,533 -/T -/T 1.00 0.329 N/A
rs2424421 21,963,058 C/T CC .98* 0.259 10 altered motifs: Cart1, *Foxa*, Foxp1,
GATA, HDAC2, Hmx_2, Irf, Pax-5, RXRA,
*P300*

----------


## TheKingofFighters

BTW, on the BMPs part, not all BMPs are downregulated in balding DPCs. accoridng to a diagram in the study, BMP4's expression is only present in non-balding scalp DPCs where else BMP2's expression is only present in balding scalp DPCs.

SOX2 is mainly invovled with the shape of the individual hair follicle:

Summary

The dermal papilla comprises the specialised mesenchymal cells at the base of the hair follicle. Communication between dermal papilla cells and the overlying epithelium is essential for differentiation of the hair follicle lineages. We report that Sox2 is expressed in all dermal papillae at E16.5, but from E18.5 onwards expression is confined to a subset of dermal papillae. In postnatal skin, Sox2 is only expressed in the dermal papillae of guard/awl/auchene follicles, whereas CD133 is expressed both in guard/awl/auchene and in zigzag dermal papillae. Using transgenic mice that express GFP under the control of the Sox2 promoter, we isolated Sox2+ (GFP+) CD133+ cells and compared them with Sox2- (GFP-) CD133+ dermal papilla cells. In addition to the `core' dermal papilla gene signature, each subpopulation expressed distinct sets of genes. GFP+ CD133+ cells had upregulated Wnt, FGF and BMP pathways and expressed neural crest markers. In GFP- CD133+ cells, the hedgehog, IGF, Notch and integrin pathways were prominent. In skin reconstitution assays, hair follicles failed to form when dermis was depleted of both GFP+ CD133+ and GFP- CD133+ cells. In the absence of GFP+ CD133+ cells, awl/auchene hairs failed to form and only zigzag hairs were found. We have thus demonstrated a previously unrecognised heterogeneity in dermal papilla cells and shown that Sox2-positive cells specify particular hair follicle types.


http://dev.biologists.org/content/136/16/2815

Sox2 positive hair follicles have guard/awl/auchenne shapes while SOX2 negative 1s have zigzag(kinky) shapes. It also states that the formation of guard(straight and long) hairs have high levels of WNT expression, correlating with high levels of SOX2's expression.

http://onlinelibrary.wiley.com/doi/1...6.00933.x/full :

Valproic Acid–induced Hair-texture Changes in a White Woman

The woman in the photos had straight hair. She gradually developed curly hair during the course of VPA treatment for epilepsy.

*Valproic Acid results in decreased expression of SOX2 mRNA*
Genistein results in decreased expression of SOX2 mRNA
resveratrol results in decreased expression of SOX2 mRNA
resveratrol	resveratrol results in decreased expression of SOX2 protein

Ascorbic Acid results in increased expression of SOX2 mRNA
[Estradiol binds to ESR2 protein] which results in increased expression of SOX2 mRNA <=== Estrogen receptor Beta ups SOX2
butyraldehyde results in decreased expression of SOX2 mRNA
Curcumin results in increased expression of SOX2 mRNA
Curcumin	Curcumin results in increased expression of SOX2 protein

----------


## TheKingofFighters

> BTW, on the BMPs part, not all BMPs are downregulated in balding DPCs. accoridng to a diagram in the study, BMP4's expression is only present in non-balding scalp DPCs where else BMP2's expression is only present in balding scalp DPCs.
> 
> SOX2 is mainly invovled with the shape of the individual hair follicle:
> 
> Summary
> 
> The dermal papilla comprises the specialised mesenchymal cells at the base of the hair follicle. Communication between dermal papilla cells and the overlying epithelium is essential for differentiation of the hair follicle lineages. We report that Sox2 is expressed in all dermal papillae at E16.5, but from E18.5 onwards expression is confined to a subset of dermal papillae. In postnatal skin, Sox2 is only expressed in the dermal papillae of guard/awl/auchene follicles, whereas CD133 is expressed both in guard/awl/auchene and in zigzag dermal papillae. Using transgenic mice that express GFP under the control of the Sox2 promoter, we isolated Sox2+ (GFP+) CD133+ cells and compared them with Sox2- (GFP-) CD133+ dermal papilla cells. In addition to the `core' dermal papilla gene signature, each subpopulation expressed distinct sets of genes. GFP+ CD133+ cells had upregulated Wnt, FGF and BMP pathways and expressed neural crest markers. In GFP- CD133+ cells, the hedgehog, IGF, Notch and integrin pathways were prominent. In skin reconstitution assays, hair follicles failed to form when dermis was depleted of both GFP+ CD133+ and GFP- CD133+ cells. In the absence of GFP+ CD133+ cells, awl/auchene hairs failed to form and only zigzag hairs were found. We have thus demonstrated a previously unrecognised heterogeneity in dermal papilla cells and shown that Sox2-positive cells specify particular hair follicle types.
> 
> 
> ...


 (1)Estradiol + (2)? + (3)? = Hair

----------


## TheKingofFighters

East asians typically have straight + thick hair texture

South asians/Middle easterners typically have (1)wavy + thick or (2)curly + thick hair texture

Caucasians typically have either (1)straight + thin or (2)wavy + thin hair texture

Africans typically have curly + thin hair texture



*Balding hair textures typically turn kinky and short .* <==== this is what i can see and feel on my own head.

----------


## Seuxin

Hello,

I use VPA at 12% once a day since december.
Absolutely no changes :s

----------


## TheKingofFighters

> Thank you. Lots of work left to do though. 
> 
> 
> 
> Size of circle (nodes) = number of other circles (nodes) connected to it
> 
> Anyway, the results are definitely interesting. I'll go over some quick observations before studying these things more deeply...
> 
> First, it seems that the computer likes FOXA2 rather than PAX1 as the causative gene at 20p11. Despite it not being expressed in dermal papilla cells, we see its "shadow", as well as the shadow of HNF4A (which is also not expressed in DPCs). Why? Is it a fictitious result? Some sort of statistical artifact? Here's a crazy idea: What if instead of dermal papilla cells, FOXA2 and HNF4A are expressed transiently in the _precursors_ to dermal papilla cells (neural crest cells?). FOXA2 and HNF4A are pioneer factors -- they modify chromatin so that transcription factors bind particular sites and not others in particular cell types. FOXA2 and HNF4A are known to cooperate in differentiation of liver cells (link). Notice also that FOXA2 binds to the homeobox HOXA5, which in the oPossum results was one of the most enriched transcription factors in balding DPCs.
> ...


 How about u list down each of the 10 most differentially-regulated genes for proteins, kinases and transcription factors

----------


## SuicidalTraveler

summer - 他正在找你。

----------


## mlamber5

> Thank you. Lots of work left to do though. 
> 
> 
> 
> Size of circle (nodes) = number of other circles (nodes) connected to it
> 
> Anyway, the results are definitely interesting. I'll go over some quick observations before studying these things more deeply...
> 
> First, it seems that the computer likes FOXA2 rather than PAX1 as the causative gene at 20p11. Despite it not being expressed in dermal papilla cells, we see its "shadow", as well as the shadow of HNF4A (which is also not expressed in DPCs). Why? Is it a fictitious result? Some sort of statistical artifact? Here's a crazy idea: What if instead of dermal papilla cells, FOXA2 and HNF4A are expressed transiently in the _precursors_ to dermal papilla cells (neural crest cells?). FOXA2 and HNF4A are pioneer factors -- they modify chromatin so that transcription factors bind particular sites and not others in particular cell types. FOXA2 and HNF4A are known to cooperate in differentiation of liver cells (link). Notice also that FOXA2 binds to the homeobox HOXA5, which in the oPossum results was one of the most enriched transcription factors in balding DPCs.
> ...


 Great job Inbeforethecure. You bring up an EXCELLENT point with pioneer factors. I have wondered this myself about 20p11. And in my opinion that is the number one enemy overall. It is the the region linked to AGA the strongest if all ethnicities are considered (Ar-Eda2r is monomorphic in Asian AGA). Upon reading on the two associates genes between the potential SNP's at 20p11 it would seem as if FOXA2, not PAX1 is the one that should be implicated. Foxa2 is know to be an androgen metabolic regulator, and another one of its many functions is response to interleukin 6, which has already been shown to be released in response to DHT in AGA DP and cause Catagen of actively growing follicle. So it really confounded me as to why foxa2 isn't expressed in AGA scalp (not in dp or in the rest of the hair follicle itself) but pax1 is. This seems to fit and clear that up. Awesome job. I really enjoy reading your computer data breakdowns as well.

----------


## iaskdumbquestions

so i dont understand a word in this thread, but when u guys guna cure this?

----------


## TheKingofFighters

inbeforethecure:

I do not know yet if Foxa2 is upregulated or downregulated in AGA individuals, not just necessarily in the scalp itself- but possibly in the organs that are vital to the scalp's functioning. (e.g thyroid, parathyroid, thymus- all of which Pax1 itself is heavily expressed in).

But what i do know from studies is that knockout of FOXa2 switches the response to TH2 cytokines- and this is the group of that is responsible for 'AGA itch' and is contained in sebocytes.  So i will assume that FOXA2 is downregulated in AGA individuals. i might be wrong though.

----------


## TheKingofFighters

> Great job Inbeforethecure. You bring up an EXCELLENT point with pioneer factors. I have wondered this myself about 20p11. And in my opinion that is the number one enemy overall. It is the the region linked to AGA the strongest if all ethnicities are considered (Ar-Eda2r is monomorphic in Asian AGA). Upon reading on the two associates genes between the potential SNP's at 20p11 it would seem as if FOXA2, not PAX1 is the one that should be implicated. Foxa2 is know to be an androgen metabolic regulator, and another one of its many functions is response to interleukin 6, which has already been shown to be released in response to DHT in AGA DP and cause Catagen of actively growing follicle. So it really confounded me as to why foxa2 isn't expressed in AGA scalp (not in dp or in the rest of the hair follicle itself) but pax1 is. This seems to fit and clear that up. Awesome job. I really enjoy reading your computer data breakdowns as well.


 Not necessarily so. I use to think that East Asians(the study on 20p11 u read, i presume, was conducted on Han Chinese- East Asians to be exact.) are only affected by 20p11 too. But the 2016 study now reveals the possiblity of other similar AGA-causative genes affecting multi-ethnic groups. For your info, the study also *ruled out EDAR's invovlement* and stated that AR is the *sole causative gene* in *that region*. But what we do know is that the AR gene is not an AGA-causative 1 in East asians.

There is no defnite answer on il-6, especially when it's a https://en.wikipedia.org/wiki/Treg17_cells cytokine utilising the STAT3 pathway- and http://www.pnas.org/content/97/25/13824.full.pdf is critically involved in hair growth. U might be right, though- in a sense that there are 2 possible scenarios regarding IL-6:

1)There is atually an underexpression of STAT3/Th17 responses in the balding scalp , being replaced by TH1/Th2 cytokine responses instead- resulting in the upregulation of  'bad' inflammation genes in the context of hair growth.
2)There really is an overexpression(like what u think) of STAT3/Th17.

----------


## TheKingofFighters

> Not necessarily so. I use to think that East Asians(the study on 20p11 u read, i presume, was conducted on Han Chinese- East Asians to be exact.) are only affected by 20p11 too. But the 2016 study now reveals the possiblity of other similar AGA-causative genes affecting multi-ethnic groups. For your info, the study also *ruled out EDAR's invovlement* and stated that AR is the *sole causative gene* in *that region*. But what we do know is that the AR gene is not an AGA-causative 1 in East asians.
> 
> There is no defnite answer on il-6, especially when it's a https://en.wikipedia.org/wiki/Treg17_cells cytokine utilising the STAT3 pathway- and http://www.pnas.org/content/97/25/13824.full.pdf is critically involved in hair growth. U might be right, though- in a sense that there are 2 possible scenarios regarding IL-6:
> 
> 1)There is atually an underexpression of STAT3/Th17 responses in the balding scalp , being replaced by TH1/Th2 cytokine responses instead- resulting in the upregulation of  'bad' inflammation genes in the context of hair growth.
> 2)There really is an overexpression(like what u think) of STAT3/Th17.


 My take is that the MAPT gene, a gene that causes AGA and Parkinso's- is probably affecting other groups(than Caucasians) as well. i have a family history of neuro diseases- particularly Parkinson's and to my knowledge, all of my relatives who had it were bald.

----------


## Trouse5858

So Replicel looks like a complete dud. They've stalled with trials to the point that earliest possible release date in Japan is now end of 2019/ 2020 and their stock is in steady decline. In all honesty if Histogen ends up being a disappointment as well, I just see Follica as a legitimate remedy. Other than that, anyone on this forum not maintaining with fin/DUT is a baldy before any new treatments actually come to fruition. Good luck making those gains in the gym fellas cuz we ain't woo'ing the ladies with our hair any time soon

----------


## kuba197

> So Replicel looks like a complete dud. They've stalled with trials to the point that earliest possible release date in Japan is now end of 2019/ 2020 and their stock is in steady decline. In all honesty if Histogen ends up being a disappointment as well, I just see Follica as a legitimate remedy. Other than that, anyone on this forum not maintaining with fin/DUT is a baldy before any new treatments actually come to fruition. Good luck making those gains in the gym fellas cuz we ain't woo'ing the ladies with our hair any time soon


 Hoping for s-equol to bind the dht. It could give us a time. And fingers crossed for PSI being tested on private forum.

----------


## mlamber5

> Not necessarily so. I use to think that East Asians(the study on 20p11 u read, i presume, was conducted on Han Chinese- East Asians to be exact.) are only affected by 20p11 too. But the 2016 study now reveals the possiblity of other similar AGA-causative genes affecting multi-ethnic groups. For your info, the study also *ruled out EDAR's invovlement* and stated that AR is the *sole causative gene* in *that region*. But what we do know is that the AR gene is not an AGA-causative 1 in East asians.
> 
> There is no defnite answer on il-6, especially when it's a https://en.wikipedia.org/wiki/Treg17_cells cytokine utilising the STAT3 pathway- and http://www.pnas.org/content/97/25/13824.full.pdf is critically involved in hair growth. U might be right, though- in a sense that there are 2 possible scenarios regarding IL-6:
> 
> 1)There is atually an underexpression of STAT3/Th17 responses in the balding scalp , being replaced by TH1/Th2 cytokine responses instead- resulting in the upregulation of  'bad' inflammation genes in the context of hair growth.
> 2)There really is an overexpression(like what u think) of STAT3/Th17.


 Ya I think you misinterpreted what I said, I am in agreeance with you. Other regions are involved for Asians besides 20p11, but the region on chromosome x effecting the Ar isn't one of them, and 20p11 is the single strongest at risk region. In fact amongst Europeans, there are 3 different spots between foxa2 and pax1 that are at risk points for AGA. There are only 2 at risk spots between eda2r and Ar (although the single highest at risk SNP is one of these nucleotide variants at one of those spots). Ago
Again the fact that Ar obviously has to do with androgen metabolism and foxa2 does as well fits the phenotype. Some other big spots are HDAC9 and HDAC4 (amongst many others). HDAC4 has been shown in some cell types to aid in Ar gene suppression so perhaps a certain variant in AGA doesn't allow this process to happen as effectively.

----------


## TheKingofFighters

> Ya I think you misinterpreted what I said, I am in agreeance with you. Other regions are involved for Asians besides 20p11, but the region on chromosome x effecting the Ar isn't one of them, and 20p11 is the single strongest at risk region. In fact amongst Europeans, there are 3 different spots between foxa2 and pax1 that are at risk points for AGA. There are only 2 at risk spots between eda2r and Ar (although the single highest at risk SNP is one of these nucleotide variants at one of those spots). Ago
> Again the fact that Ar obviously has to do with androgen metabolism and foxa2 does as well fits the phenotype. Some other big spots are HDAC9 and HDAC4 (amongst many others). HDAC4 has been shown in some cell types to aid in Ar gene suppression so perhaps a certain variant in AGA doesn't allow this process to happen as effectively.


 Ok, so for your info once again, the 2016 study also ruled out both HDAC4 and HDAC9 as AGA-causative genes. Instead, PER2 and TWIST2 in HDAC4's region were the true AGA-causative genes found to be downregulated in balding scalps.

TWIST1 in HDAC9's region was found to be the true AGA-causative gene- upregulated in balding scalps.

----------


## TheKingofFighters

> Ok, so for your info once again, the 2016 study also ruled out both HDAC4 and HDAC9 as AGA-causative genes. Instead, PER2 and TWIST2 in HDAC4's region were the true AGA-causative genes found to be downregulated in balding scalps.
> 
> TWIST1 in HDAC9's region was found to be the true AGA-causative gene- upregulated in balding scalps.


 Unsurprisingly, PER2 and TWIST2 is upregulated by Estradiol and Genistein while TWIST1 is downregulated by estrogenic compounds

----------


## InBeforeTheCure

> atually- the clue is something that u've have already read before: 
> 
> http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4365504/
> 
> SNP
> marker
> Position
> (hg19)
> Alleles IS
> ...


 It's unlikely that both Hnf4a and Foxa2 would be involved in DPC differentiation because expression of those two together turns fibroblasts into hepatocytes. However, even if only one of them is involved, you would likely find that both show up in this sort of analysis because their target genes overlap, and of course we do see that.




> BTW, on the BMPs part, not all BMPs are downregulated in balding DPCs. accoridng to a diagram in the study, BMP4's expression is only present in non-balding scalp DPCs where else BMP2's expression is only present in balding scalp DPCs.
> 
> SOX2 is mainly invovled with the shape of the individual hair follicle:


 I haven't yet looked into Sox2 that carefully, but it's also possible that it's not upregulated or downregulated, but could instead be regulated "differently" due to binding by co-factors.




> How about u list down each of the 10 most differentially-regulated genes for proteins, kinases and transcription factors


 Kinases: https://www.dropbox.com/s/ldnww188yt...inase.csv?dl=0

Transcription Factors: https://www.dropbox.com/s/1yh34r1dir...ts_tf.csv?dl=0

Protein Network: https://www.dropbox.com/s/gtyrkljgwu...twork.sig?dl=0




> Great job Inbeforethecure. You bring up an EXCELLENT point with pioneer factors. I have wondered this myself about 20p11. And in my opinion that is the number one enemy overall. It is the the region linked to AGA the strongest if all ethnicities are considered (Ar-Eda2r is monomorphic in Asian AGA). Upon reading on the two associates genes between the potential SNP's at 20p11 it would seem as if FOXA2, not PAX1 is the one that should be implicated. Foxa2 is know to be an androgen metabolic regulator, and another one of its many functions is response to interleukin 6, which has already been shown to be released in response to DHT in AGA DP and cause Catagen of actively growing follicle. So it really confounded me as to why foxa2 isn't expressed in AGA scalp (not in dp or in the rest of the hair follicle itself) but pax1 is. This seems to fit and clear that up. Awesome job. I really enjoy reading your computer data breakdowns as well.


 Thank you. Actually though, I don't think it's cleared up at all.  :Wink:  Pioneer factors are usually actively expressed in the cells they regulate AFAIK, so this would have to be some form of epigenetic memory if anything. As for IL6 and other inflammatory cytokines, I think it would be most natural to blame stress-induced p38 MAPK activity.




> *p38 and inflammation*
> 
> *A strong link has been established between the p38 pathway and inflammation.* Rheumatoid arthritis, Alzheimer's disease and inflammatory bowel disease are all postulated to be regulated in part by the p38 pathway 87, 88, 89. *The activation of the p38 pathway plays essential roles in the production of proinflammatory cytokines (IL-1beta, TNF-alpha and IL-6)* 90; induction of enzymes such as COX-2 which controls connective tissue remodeling in pathological conditions 91; expression of intracellular enzymes such as iNOS, a regulator of oxidation 92, 93; induction of VCAM-1 and other adherent proteins along with other inflammatory related molecules 18. In addition, a regulatory role for p38 in the proliferation and differentiation of immune system cells such as GM-CSF, EPO, CSF and CD-40 has been established 16, 94.


 (link)

MAPK14 in the network diagram = p38-alpha




> so i dont understand a word in this thread, but when u guys guna cure this?


 Chemical found the cure, but unfortunately the international conspiracy of hair transplant surgeons and minox vendors got to him. If he talks, he's a dead man. That's why he's disappeared.




> inbeforethecure:
> 
> I do not know yet if Foxa2 is upregulated or downregulated in AGA individuals, not just necessarily in the scalp itself- but possibly in the organs that are vital to the scalp's functioning. (e.g thyroid, parathyroid, thymus- all of which Pax1 itself is heavily expressed in).
> 
> But what i do know from studies is that knockout of FOXa2 switches the response to TH2 cytokines- and this is the group of that is responsible for 'AGA itch' and is contained in sebocytes.  So i will assume that FOXA2 is downregulated in AGA individuals. i might be wrong though.


 Foxa2, if it's involved, would most likely confer risk to AGA by acting as a pioneer factor at AR target genes IMO. That's something it's known for.




> In fact amongst Europeans, there are 3 different spots between foxa2 and pax1 that are at risk points for AGA.


 Those SNPs are in high linkage disequilibrium with each other, so you would need a _massive_ sample size to discern multiple risk points.




> Ok, so for your info once again, the 2016 study also ruled out both HDAC4 and HDAC9 as AGA-causative genes. Instead, PER2 and TWIST2 in HDAC4's region were the true AGA-causative genes found to be downregulated in balding scalps.
> 
> TWIST1 in HDAC9's region was found to be the true AGA-causative gene- upregulated in balding scalps.


 Focusing on the 5 SNP model from this study:



Best guesses for the functional variants at these sites: AR, PAX1 or FOXA2, RNF145, MTOR, TWIST1?

----------


## Forestry

> Chemical, why did you stop using oleuropein?


 this 5ar research on ganoderma lucidum  from 2007 is interesting, for guys in particular.  reishi is easy to find / inexpensive:

The anti-androgen effect of ganoderol B isolated from the fruiting body of Ganoderma lucidum

I was able to search on the paper title and found a free pdf eventually, can't find the link now...

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## AHMAD

hy guys new to forum. are you still using oleuropien.

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