Updated Research and Knowledge - Cutting Edge

[Seuxin]

Hello,

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

[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.

[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

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

[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]

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

[InBeforeTheCure]

hi ur hardwork is appreciated-

Thank you. Lots of work left to do though.

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]

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:



(link)

This is probably thanks to reversing mTOR inhibition of autophagy.



(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.

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]

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 fruitflies1, 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]

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.

[TheKingofFighters]

inbeforethecure, please read the below:

http://www.educatedtherapists.com/fox01-and-mtorc-what-role-do-they-play-in-acne/

[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 !

[Trackster]

Can we say based on this that STAT3 doesn't seem to function a big role in hair follicle biology? I would think so.... One would expect at least some hair alterations right?

Interesting, I am not so knowledgable in this field, but I read alot of these comments with interest. It is funny how different some of the opinions are.

http://www.hairlosscure2020.com/jefferies-2016-healthcare-conference-dr-neal-walker-confirms-that-topical-jak-inhibiters-will-be-tested-on-androgenic-alopecia-patients/#comment-105085

Wow…. look at these articles in relation to all previous posts I and others have posted

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4653025/

There is definitely something to this whole miRNA-22 and JAK/STAT pathway thing going on in hair. In this other cancerous t-cell population it looks like like JAK/STAT pathway is actually supressing miRNA-22. Specifically this article mentions JAK3/STAT3/STAT5…. Notice how Walker talked about developing a JAK3 inhibitor as apposed to the others…

miRNA-22 functions as a “tumor suppressor”. Aka it stunts the growth of cells. The exact thing we are seeing in the actual hair shaft part of the hair follicle in AGA. Difference between AGA and AA? One is an adaptive immune response (AGA) and the other is an innate response (AA). Which explains the different timescales to lose hair, and explains the pattern somewhat. We also know from above posts that STAT3 is a positive regulator of miRNA gene suppression. And it is known that miRNA-22 is androgen induced from previous posts. And you may look this up if you wish, but estrogen/estrogen receptor and androgen/androgen receptor are regulated differently by miRNA-22 in different cancers.

Based off of in human trials and on mice, we know that lowering/blocking of JAK/STAT signaling allows hair to grow. When it is high, it is a stop signal in the hair follicle. That much is obvious. This not only works in rodents but in humans. We also know that miRNA-22 functions in hair to stop it from growing by suppressing 50+ keratin genes (and probably others)…. see above article for that. Causing the follicle to regress according to above posted article… exactly like the parthenogenesis of AGA. Also the miRNA-22 promoter is located on the same DNA region in mice and in humans per the article I posted above.

miRNA22 is also implicated in endothelial cell cellular senescence…. We know this is going on in AGA dp cells, perhaps once again through the JAK/STAT pathway? Relieving this reverses senescence. See the below abstract

http://www.ncbi.nlm.nih.gov/pubmed/25323119

Also miRNA-22 regulates smooth muscle cell differentiation. Smooth muscle cell is what the erector pili muscle is made of, in which contact is lost in AGA. Could stopping miRNA-22 jak/stat signaling allow the erector pilli muscle to make contact again? See the below abstract.

http://www.ncbi.nlm.nih.gov/pubmed/25722434

Here is where this gets interesting to me….

http://www.genecards.org/cgi-bin/carddisp.pl?gene=MIR22

The gene coding for miRNA-22 is on chromosome 17. And it appears that the genes coding for stat3, stat5a, and stat5b are ALSO on chromosome 17.

http://www.genecards.org/cgi-bin/carddisp.pl?gene=STAT3&keywords=stat3

http://www.genecards.org/cgi-bin/carddisp.pl?gene=STAT5A&keywords=stat5

http://www.genecards.org/cgi-bin/carddisp.pl?gene=STAT5B&keywords=stat5

Note that stat1, stat2, stat4, and stat6 are not on the same chromosome…. So perhaps this micro RNA is promoting stat3 and stat5 over-expression in AGA.

miRNA’s main function is to post transcriptionally regulate gene expression.

STAT’s stands for “SIGNAL TRANSDUCER ACTIVATOR OF TRANSCRIPTION.” It would only make sense that something like this is the main problem in AGA. It looks super complicated from the outside because so much is going on. But perhaps it is one or two master switches causing the cascade of crap to happen. This is what I believe at least.

Man, I am more confident then ever that proper JAK/STAT inhibition via a topical is truly it. I could be completely wrong, but it sure looks like jak/stat signaling has a ton to do with all of hair biology (really, all of biology in general) not just AA.

[InBeforeTheCure]

@InBeforeTheCure,

I pm'ed you, let's get you some extra RAM so you can make that analysis. We need those computational methods indeed to make something of it all.

Right now I'm looking at algorithms that use prior knowledge and incorporate post-translational regulation, as I think that would be better suited for our purposes. Maybe miRNAs too, we'll see.

Btw;

http://www.ncbi.nlm.nih.gov/pubmed/24064061 (rat though)

Further interesting read between the interplay (Skip to 5. TGF-B and ROS interplay) ; http://www.hindawi.com/journals/omcl/2015/654594/

A nasty positive feed back loop. ROS -> TGFbeta -> ROS

TGFbeta probably contributes to HF fibrosis too (Foitzik et. al, 2000).

Btw, there are people walking around with either loss of function or gain of function mutation of STAT3. No hair alterations are observed in those people. Interestingly tooth and bone development is altered.

Can we say based on this that STAT3 doesn't seem to function a big role in hair follicle biology? I would think so.... One would expect at least some hair alterations right?

I guess the argument then would be that there are multiple STATs at work, and maybe there are some that are more important to AGA than STAT3, or there are redundancies between them that compensate for the knockout of one of them. Either way, according to the Chew data, all STATs are downregulated* at the mRNA level in balding DPCs, but it's still possible that they are nevertheless phosphorylated at a higher rate. I'm far from convinced of the Jak-Stat hypothesis though.

* There was a study from 2004 that contradicts this one. Contrary to Chew et. al, they found STAT1 to be upregulated in balding DPCs.

Take loss of function in the VDR and alopecia very often develops; http://www.nature.com/bonekeyreports...ekey20145.html

Other example is loss of function in HR; http://onlinelibrary.wiley.com/doi/1...09.01042.x/pdf

Then you have APCDD1 etc....

Interesting then that "response to vitamin D" is the 2nd most enriched biological process for genes upregulated in balding DPCs in PANTHER. Chemical had some stuff on VDR earlier in this thread as well.

It has been hypothesized that the role of the VDR in the hair cycle is to repress the expression of a gene(s) in a ligand-independent manner.46, 55, 89, 91, 93 The ligand-independent activity requires that the VDR heterodimerize with RXRα and bind to DNA.46, 90 The corepressor actions of HR may also be required in order for the unliganded VDR to repress gene transcription during the hair cycle. Mutations in the VDR that disrupt the ability of the unliganded VDR to suppress gene transcription are hypothesized to lead to the derepression of a gene(s) whose product, when expressed inappropriately, disrupts the hair cycle that ultimately leads to alopecia.46, 55, 89, 91, 93 Inhibitors of the Wnt signaling pathway are possible candidates.13, 95, 96, 97

This is oPOSSUM for the top 1,000 downregulated genes in balding vs. non-balding DPCs:

TF	Z-score
NFATC2	17.98
ELF5	15.897
RXRA::VDR	15.677
Hand1::Tcfe2a	15.49
HOXA5	14.369
SPIB	13.276
TAL1::TCF3	12.972
FEV	12.58
REL	12.51
Tal1::Gata1	12.244
RELA	11.862
Gfi	11.796
MAX	11.792
RUNX1	10.994
AP1	10.697
Nobox	10.636
IRF1	10.348
SRY	10.206
ARID3A	9.57
RORA_2	9.492
Pdx1	9.492
SPI1	9.255
FOXF2	8.879
STAT1	8.819
Myf	8.689
MZF1_1-4	8.439
Nkx2-5	8.334
CEBPA	8.037
NF-kappaB	7.907
Myb	7.782
FOXO3	7.592
MZF1_5-13	7.444
IRF2	7.302
PBX1	7.297
Sox5	7.25
Prrx2	7.04
NHLH1	6.799
Nr2e3	6.794
NR4A2	6.589
Gata1	6.416
NR1H2::RXRA	6.404
Foxa2	6.043
Sox17	5.929
Stat3	5.891
Ddit3::Cebpa	5.625
FOXA1	5.237
Tcfcp2l1	5.062
Nkx3-2	4.999
Spz1	4.96
Arnt::Ahr	4.668
Sox2	4.658
USF1	4.477
HLF	4.441
NFKB1	4.252
MYC::MAX	4.13
NFIL3	4.088
CTCF	4.065
ZNF354C	4.062
TBP	3.998
SP1	3.955
Lhx3	3.926
INSM1	3.925
RORA_1	3.892
RREB1	3.806
Pou5f1	3.754
HNF4A	3.748
EBF1	3.642
CREB1	3.423
MEF2A	3.228
TEAD1	2.982
SRF	2.92
ELK1	2.828
ESR1	2.805
FOXI1	2.764
T	2.716
E2F1	2.366
HIF1A::ARNT	2.284
Klf4	2.03
SOX9	2.026
NR3C1	1.983
HNF1B	1.958
NR2F1	1.744
NFE2L2	1.424
RXR::RAR_DR5	1.279
TLX1::NFIC	1.11
FOXD1	1.074
Arnt	0.899
Foxq1	0.781
NKX3-1	0.478
Ar	0.308
Mycn	0.298
Foxd3	0.146
YY1	0.103
Esrrb	0.041
NFYA	-0.511
Pax4	-0.593
ESR2	-1.383
Evi1	-1.674
HNF1A	-1.712
PLAG1	-1.754
Myc	-2.201
Pax6	-2.239
EWSR1-FLI1	-2.322
Pax5	-2.345
PPARG	-2.566
TP53	-2.905
Egr1	-3.389
PPARG::RXRA	-3.586
ZEB1	-3.846
MIZF	-4.012
znf143	-4.448
Zfx	-4.475
GABPA	-5.105
ELK4	-5.993
Zfp423	-6.415
REST	-8.525

If RXRA::VDR acts mostly as a repressor, then interestingly enough, it seems like it may be more active in AGA DPCs. Also, the PPARG::RXRA complex is underrepresented in the same set. It makes me wonder if, for some reason, RXRA is associating with VDR at the expense of PPARG. Of course, this is just in DPCs and VDR expression is much higher in ORS cells according to the Hair-Gel site, so things may be different there.

Also a side point...In PANTHER, the top 3 categories in the downregulated genes are

1. response to interferon-alpha
2. response to interferon-beta
3. type I interferon signaling pathway



Perhaps this is something we could investigate. In fact, the first paper I've landed on (this one) says the following:



I know you're aware of this paper:

We detected the significant upregulation of miR-221, miR-125b, miR-106a and miR-410 in balding papilla cells.

The paper mentions the three bolded ones.

"miR221/222 can target STAT1 and STAT2"...Reference chasing takes us here:

Unlike miR-145 which is commonly down-regulated in tumors, miR-221/222 are often up-regulated in cancers [23, 27, 28]. Among the genes whose expression levels are altered by antisense-mediated knockdown of miRs-221/222 in U251 glioma cells, ones in the IFN-α signaling pathway are the most significantly modulated, and this observation is dependent on increased expression of STAT1 and STAT2 [23]. Consistently, overexpression of miR-221/222 in U251 glioma cells interferes with IFN signaling by down-regulating STAT1 and STAT2 [23].

Our very own STAT1/STAT2 inhibitor. That's nice.

"and there are numerous miRNA regulators of STAT3, including...miR106a...and miR125b"

More reference chasing:

In addition, we identified 2 potential miR-125b-binding sites within the 3′UTR of Stat3 (Figure 4B), a transcription factor strongly involved in granulocytic differentiation.35,36 To demonstrate direct regulation by miR-125b, luciferase reporter containing wild-type Stat3 3′UTR sequences or their mutant-derivates with deletion of the putative miR-125b-binding sites were transfected into NIH3T3 cells stably over-expressing miR-125b (NIH3T3/miR-125b; Figure 4B and supplemental Figure 3A). As shown in Figure 4C, miR-125b represses luciferase activity by approximately 40% depending on the presence of miR-125b binding in the Stat3 3′UTR. Finally, Western blotting revealed an approximately 30%-40% reduction of STAT3 protein expression in 32D/miR-125b compared with 32D/miR-ctrl cells (Figure 4D).

(link)

Quantitative real-time PCR and Western blotting demonstrated that miR-106a was upregulated, and STAT3 and phospho-STAT3 were downregulated in the hippocampus at 12 weeks post-OVX, compared with age matched controls and the 6 and 8 weeks post-OVX groups. Transfection of human neuroblastoma SH-SY5Y cells with a miR-106a mimic reduced the expression of STAT3 mRNA, compared to control cells transfected with a scrambled mimic. STAT3 and phospho-STAT3 protein expression was upregulated or downregulated by a miR-106a inhibitor or miR-106a mimic, respectively, indicating that miR-106a negatively regulates STAT3. Luciferase reporter gene assays confirmed that miR-106a directly targets the 3' untranslated region (UTR) of STAT3.

(link)

We have our own STAT3 inhibitors too. Fascinating.

So does this mean the interferon/JAK-STAT pathway is suppressed, or is it actually overactive and we're seeing evidence of negative feedback? I guess we'll have to investigate further.

Lastly, is it fair to say in terms of damage control a "primary" anti-oxidant might help? Something like orgotein (SOD)?

Are there complications to using standard ROS scavengers for this? Something like N-acetylcysteine? I imagine people have tried them before, and if they don't work, either they don't combat oxidative stress in hair follicles effectively or oxidative stress is not necessary for AGA to develop.

[Swooping]

@InBeforeTheCure,

I pm'ed you, let's get you some extra RAM so you can make that analysis. We need those computational methods indeed to make something of it all.

Btw;

http://www.ncbi.nlm.nih.gov/pubmed/24064061 (rat though)

Further interesting read between the interplay (Skip to 5. TGF-B and ROS interplay) ; http://www.hindawi.com/journals/omcl/2015/654594/

Btw, there are people walking around with either loss of function or gain of function mutation of STAT3. No hair alterations are observed in those people. Interestingly tooth and bone development is altered.

Can we say based on this that STAT3 doesn't seem to function a big role in hair follicle biology? I would think so.... One would expect at least some hair alterations right?

Take loss of function in the VDR and alopecia very often develops; http://www.nature.com/bonekeyreports...ekey20145.html

Other example is loss of function in HR; http://onlinelibrary.wiley.com/doi/1...09.01042.x/pdf

Then you have APCDD1 etc....


Lastly, is it fair to say in terms of damage control a "primary" anti-oxidant might help? Something like orgotein (SOD)? Did you ever try it TheKingOfFighters? Seems you have a whole lab at home lol. Might be troublesome with a 30kDa protein though.. Not really practical to regularly inject into the scalp.

[kuba197]

What do you think guys about proteasome inhibitors (PSI)?