Rhythmic Diurnal Synthesis And Signaling Of Retinoic Acid In The Rat Pineal Gland And Its Action To

Terma

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@Amazoniac: Follow-up to Anti-Peat - Grant Genereux's Theory Of Vitamin A Toxicity

Rhythmic Diurnal Synthesis and Signaling of Retinoic Acid in the Rat Pineal Gland and Its Action to Rapidly Downregulate ERK Phosphorylation
Vitamin A is important for the circadian timing system; deficiency disrupts daily rhythms in activity and clock gene expression, and reduces the nocturnal peak in melatonin in the pineal gland. However, it is currently unknown how these effects are mediated. Vitamin A primarily acts via the active metabolite, retinoic acid (RA), a transcriptional regulator with emerging non-genomic activities. We investigated whether RA is subject to diurnal variation in synthesis and signaling in the rat pineal gland. Its involvement in two key molecular rhythms in this gland was also examined: kinase activation and induction of Aanat, which encodes the rhythm-generating melatonin synthetic enzyme. We found diurnal changes in expression of several genes required for RA signaling, including a RA receptor and synthetic enzymes. The RA-responsive gene Cyp26a1 was found to change between day and night, suggesting diurnal changes in RA activity. This corresponded to changes in RA synthesis, suggesting rhythmic production of RA. Long-term RA treatment in vitro upregulated Aanat transcription, while short-term treatment had no effect. RA was also found to rapidly downregulate extracellular signal-regulated kinase (ERK) 1/2 phosphorylation, suggesting a rapid non-genomic action which may be involved in driving the molecular rhythm in ERK1/2 activation in this gland. These results demonstrate that there are diurnal changes in RA synthesis and activity in the rat pineal gland which are partially under circadian control. These may be key to the effects of vitamin A on circadian rhythms, therefore providing insight into the molecular link between this nutrient and the circadian system.

So I had to tweak my thoughts on the circadian nature of retinoic acid. Although it is tied to food consumption through the environmental and visual cue of carotenoids, RA synthesis (RALDH) mostly seems to serves as a morning signal and should be interacting with light cues (combined with CLOCK/BMAL1). It is less about RA being high all day or in response to food (which might not happen as dramatically in healthy circumstances) and more about it hitting a peak in the late night/morning. This makes sense after all since in organs like the liver NAD+/NADH ratio required for RALDH is highest in the morning with high levels before waking (as the electrons are expected to be lowered - for one - after having been donated to fuel skeletal growth/restructuring).

This would make sense as a way to counteract the growth-hormone-ablated GNMT expression that serves to increase methyl group availability during the night; something has to counteract this. RA stops methylation, restores the transsulfuration pathway and we begin the day. CYP26 may be especially relevant to help restrain the RA increase, with an input from NADPH availability (which may affect timing of breakdown), but I'm not sure exactly what shape it gives the curve.

More importantly: RA would promote the processes of differentiation at the end of the night, after growth hormone did its work. It completes the nighttime processes. GH has a limited window to work at the start of the night using the nutrients acquired during the day.

This could also explain another reason why RA has the potential to increase cortisol (see various studies on its HPA axis effects - despite the doses): it helps propel the "stop healing, wake up" signal, or the transition from sleep/growth to wake.

At the same time, SIRT1 increases in organs like the liver in the morning due to the higher NAD+/NADH ratio - and it's known that RAR and SIRT1 interact:
Reciprocal roles of SIRT1 and SKIP in the regulation of RAR activity: implication in the retinoic acid-induced neuronal differentiation of P19 cells
SIRT1 seems to antagonize RAR; I imagine SIRT1 and RALDH (retinaldehyde->RA) may compete for NAD+ availability or there may be a transcription effect.

I'm not 100% sure about this and it's not essential to understand, but you can almost piece it together:
http://www.jbc.org/content/287/50/42195.full.pdf
SIRT1 seems to increase when differentiation goes from RAR (early) and progresses to PPARdelta (which increases fatty acid utilization for differentiation). I'm not sure about the causality but it seems like PPARdelta and SIRT1 function to help finish differentiation.

Finally, from the original article (they put all their focus on this in the abstract), after 48 hours RA modulates melatonin release through being lowered at the start of the night and increasing toward the end, seemingly by increasing AANAT mRNA at those times and maintaining rhythmicity (not totally clear).

I'll probably have to tweak this again, but from this view it starts to make a lot more sense as it gives RA at least one clear role and timeframe, though there could be organ-specific differences. Of course carotenoids are an important environmental and visual cue for nutrient abundance for mammals, so they should tie into this as well. At the same time you can imagine all the ways this might go wrong, because cellular differentiation is not a simple process and it needs to be timed, much like everything about sleep.

There could be some misinterpretations in this - because RA circadian studies are some of the most complicated I've ever read, and there are likely to be organ-specific difference, and rats/mice vs humans adds extra layers of confusion/complexity in circadian studies - but I'm getting a little more confident as time goes on that it's something along these lines.
 

LeeLemonoil

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Thanks for sharing these thoughts and work Terma. Comprehending RA is a worthwhile attempt.
 
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Terma

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It's definitely the most complicated "vitamin" (quotes for the benefit of others).

But look, you can easily tie this idea into the other thread:
Sarcosine Is Uniquely Modulated By Aging And Dietary Restriction In Rodents And Humans

RA -> GNMT -> Sarcosine -> Autophagy

No one will debate me if I claim the most important time in the day for autophagy is the end of the night and the morning.

Furthermore, it makes even more sense when tied in with GH: if spermidine - a methylation product - increases autophagy, then when methylation stops, you need something to take its place for the rest of the night: sarcosine.
 
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@Amazoniac @SB4 @Blue Jefe

Perhaps at the end of the human waking day, when niacinamide is higher from use of NAD as a reactant in enzymes during the day (e.g. SIRT1 in liver, which concomitantly falls), around the same time that SAMe availability - the methylation cycle - should logically begin to increase (in response to the largest meal of protein as well and later the induction of growth hormone, as well as the notion that it may be a brain-priorizing mechanism, given the frequent effect on lipophilicity of the methylation of compounds, as Travis once pointed out)... NNMT in specific or various tissues may methylate the pool of niacinamide to make it more lipophilic. This might make it gravitate toward lipid-rich depots and tissues... In other words it might help keep or move the niacinamide into the brain so that it can heal, survive the long night, and begin NAD+ regeneration for the morning.

Pet theory, shamelessly pirated from a man of science: Methylation increases at the beginning of the night through growth hormone release and other mechanisms as well as after the last meal of the day, and it generally produces especially lipophilic compounds: carnitine, CoQ, choline, etc. This helps implement part of our brain prioritization program, which is to send nutrients to the brain or otherwise prioritize the brain for specific or general pools of nutrients. This is very important during nighttime.

In addition to the brain, it probably targets the skeleton and bone marrow, to support and maybe simply implement the effects of growth hormone.

Carnitine - and a good reason lysine supply must be sufficient, or rather, not rate-limiting in the synthesis of its various compounds - is an excellent skeletal growth-promoting nutrient:
Sci-Hub | L-Carnitine and Isovaleryl L-Carnitine Fumarate Positively Affect Human Osteoblast Proliferation and Differentiation In Vitro. Calcified Tissue International, 76(6), 458–465 | 10.1007/s00223-004-0147-4
Sci-Hub | l-Carnitine Fumarate and Isovaleryl-l-Carnitine Fumarate Accelerate the Recovery of Bone Volume/Total Volume Ratio after Experimetally Induced Osteoporosis in Pregnant Mice. Calcified Tissue International, 82(3), 221–228 | 10.1007/s00223-008-9109-6
That carnitine is thought to be able activate the GR receptor is not surprising: cortisol is already low at nighttime - replaced by GH - and the absence of GR stimulation or other non-transcriptional effects might otherwise lead to either homeostatic problems or too much (or even too little of some kinds of) inflammation. Carnitine is critical.

Naturally, the second part of our brain prioritization program is the prioritization of the glucose supply toward the brain. This is roughly what GH does but so do several other things through FFA release and (pseudo-)insulin resistance. Of course carnitine also relates to this.

The entire story is scripted in our sleeps: the growth hormone spike together with the methylation cycle implements the nighttime processes: growth and recovery, prioritization of the brain, replenishment of the nutrient pool... In fact, methylation is likely also reused in stressful or other daily situations, and it's because the body is trying to achieve one of these things, during something such as over-exhaustion; otherwise it gets impeded by several things though it is logically supported by dietary nutrient intake.

That is why when you don't sleep properly, you kill yourself slowly: you do not produce as much CoQ, choline, creatine; you give fewer nutrients to your brain; you regenerate less. This is part of why people sleep better with a minimum of fat, fat-soluble compounds, methionine, lysine, ornithine (for polyamine synthesis) in their last meal of the day. (Of course tryptophan also plays a part but it's kind of tangential)

Of course, you would then expect that later in the night retinoic acid gets synthesized to finish the growth/recovery process... This has to begin after NAD+ resynthesis begins during the next, so a bit later. But you see? It all connects: at this point, it should be able to synthesize new NAD+ thanks to the niacinamide that was methylated earlier by NNMT.

While this happens, tryptophan availability should also be have been higher at this time of day, and serotonin proceeds to conversation to melatonin thanks to - guess what? - good o' SAMe: Acetylserotonin O-methyltransferase - Wikipedia

What else? Histamine N-methyltransferase - Wikipedia

Melatonin synthesis + histamine breakdown. (Note: melatonin on its does own not necessarily succeed in inducing or barely even promoting sleep in some people and configurations; this suggests a partial explanation why some people do not feel sleepy from methylation supplements, while some even get excitatory reactions from changes in catecholamines, serotonin, metabolic changes and other, while yet others such as myself could be put to sleep almost immediately from MB12).

This is the reason why MTHFR and ME/CFS patients had some luck or at least temporary relief with methylation supplements: together with growth hormone it promotes a (rest + recovery) and a brain-prioritization program. And I believe some of them complained about or had success with GH/IGF-1. GH and methylation cycle might each accentuate different parts of that program, but it seems logical for methylation to be primarily a brain/fat prioritization program, and GH to be a "rest and recovery" signal (implemented alongside/using several neurotransmitters such as GABAA/B, glycine and chloride channels, progresterone, allopregnanolone, endocannabinoids, kynurenines, serotonin, melatonin, IGF-1, amino acids themselves, ...), but this is a gross generalization.

In other words, people with MTHFR defects - because they produce less SAMe - may have an increased need for rest and sleep.

What am I saying? I believe Freddd (over at phoenix rising) is credible about how extremely high-dose methylcobalamin (MB12) injections, super high dose methylfolate (up to 30mg/day), L-Carnitine fumarate andacetyl-L-Carninitine (fumarate just happens to be part of the more effective forms of carnitine in the studies I just linked above), and adenosylcobalamin allowed his neurological damage to deal, with an additional battery of methylation-supporting nutrients - not entirely unlike my crazy THC protocol. A lot of people eventually seemed to discard Freddd's ideas, and I wasn't convinced of several things, but the guy was intelligent. Think about it: He had to take absurdly gigantic amounts of MB12, specifically to fill his brain/CNS with it, to allow myelin and neurological recovery. Free warning: Freddd depleted his copper levels.

That's how I did my improvements: I ate high levels of methyl-ladden protein every day (including chicken, whey and lysine) and virtually always suffered when I lowered consumption. Meanwhile, large doses of glycine seemed to do little but cause me early waking (hmmm... but again, it could be many other things like ammonia, immune modulation and chloride channels). I frequently took MB12 and methylfolate and derivatives... I had both temporary benefits and strong backlash from high dose niacinamide, however, which is interesting; I'm tempted to say that it put a stress on my ribose or purine supply, or maybe I couldn't handle the product methylnicotinamide since it has to be broken down with minerals and things. Meanwhile, I had tremendous benefit from caprylic acid/ketones, which is not guaranteed to but likely suggests increased endogenous niacin synthesis in neurons as well as in the other brain cells, notably migroglia and the immune system attempting to clean up the brain at night (phagocytosis, autophagy, etc.).

I tried to describe earlier how tons of retinoic acid and/or very high NADP+/NADPH ratio might give way to a state that looks like an "endless day": this may be because it effectively ends - or keeps from happening - the (rest + recovery) and perhaps - at least part - or one - of a brain prioritization program(s). Retinoic acid is the "finish healing now let's go" signal - which almost naturally sounds like a stressor, which it factually is if only because it consumes NAD+ and requires NAD(PH/H?) for CYP26 to get rid of.

Note however, that the recycling of methyl groups using serine, sarcosine and maybe DMG might also implement and be considered part of or one brain prioritization and related programs, depending on the dynamics and distributions of the enzymes in all the tissues, together with the effects of the circadian rhythm on serine product by the kidneys. We basically already know what happens with sarcosine.

Major methyl group-carrying compounds such as carnitine, choline, creatine, trimethyl*, serine, sarcosine, DMG should all be suspected in these processes by association.

Think of the liver, which packages VLDL, triglyrices, phospholipids, GNMT, growth hormone as a signal, and has the most intensive job of perhaps any organ during the night: a large part of its life is dedicated to supporting the brain by maintaining glucose levels... The liver needs a mechanism to send other types of compounds - including but not limited to the compounds it produces itself through the liver methylation cycle - to the brain, as well as to other lipophilic depots such as various skeletal tissue, bone marrow, and even for collagen synthesis for tendons and joints (which just happens to be just as well one of the major roles GH/IGF-1 - how could you ignore this?). People coincidentally report relief of joint pain from SAMe supplements:
S-Adenosyl methionine (SAMe) versus celecoxib for the treatment of osteoarthritis symptoms: A double-blind cross-over trial. [ISRCTN36233495]

Processes resembling insulin resistance get turned on to conserve glucose for the brain. This is notably implemented thanks to FoxO nuclear receptors such as FoxO1 in the liver that maintains gluconeogenesis (Insulin-regulated hepatic gluconeogenesis through FOXO1–PGC-1α interaction). That's in fact partly how the liver supports the brain: by sending it nutrients, but also promoting fatty acid oxidation in other tissues competing with it for glucose.

Yes that's what I'm saying: eat carbs during the day, support fat oxidation and glycogen levels at nighttime. Carbs are not designed to support skeletal growth; saturated and monounsaturated fats - perhaps together with ketones for energy, though they're obviously relevant to the brain - are much more appropriate for such purpose.

Think about it: why does the liver have the most productive methylation cycle of any organ, all the while its major role as VLDL/lipoprotein exporter is actually primarily limited by phospholipid synthesis (CDP-choline, choline, DAG, uridine, etc.) and choline sufficiency is usually within reach? Because it corresponds to its primordial importance of keeping the brain alive at night and in general as well as providing sufficient pools of nutrients and hormones for other tissues to recover at night.

Guess what? Along with GH/IGF-1 and amino acids, SAMe enhances the mTorC1 pathway for growth and protein synthesis:
SAMTOR is an S-adenosylmethionine sensor for the mTORC1 pathway. - PubMed - NCBI
SAMTOR joins the family

What else? Polyamines.

There is no question: methylation is a recovery/reprioritization program. Freddd proved that it works. Except everyone was trying to do it during the day, probably with numerous cortisol issues, and it's conceivable that too much daytime . So the difference is I think the methylation can be demonstrated to be driven by circadian signals (food and factors affecting GNMT, MAT1A, etc.) and may interface in circadian regulation if misused. I knew something was off when methylcobalamin kept putting me to sleep (melatonin synthesis, it's presumed) while others were saying they were getting adrenaline-type responses (from some SAMe targets and probably glucose regulation failure).

Furthermore, I think Travis had the right idea that maybe NNMT helped drive niacinamide toward lipid-ladden tissues, and this might imply that insufficient evening/nighttime methylation could leave a B3 shortfall in the brain or otherwise in the wrong cells, fluids or tissues... and perhaps impede their ability to recover, or perhaps more appropriately, finish recovering.

Furthermore furthermore, the liver might methylate niacinamide to get it away from itself - the liver is a relatively lean tissue in normal circumstances. Perhaps fatty liver causes issues or interferes with this, and eventually high level of methylnicotinamide in fatty liver depots may become involved in promoting liver cancer.

This would curiously coincide with the success in patients of all diseases of treatments using caprylic acid/ketones (through Trp uptake and perhaps lipoic acid sufficiency), lipoic acid, and other things that increase NAD+ availability in the brain:
α-Lipoic acid regulates lipid metabolism through induction of sirtuin 1 (SIRT1) and activation of AMP-activated protein kinase. - PubMed - NCBI
AIMS/HYPOTHESIS: Sirtuin 1 (SIRT1) is a longevity-associated protein, which regulates energy metabolism and lifespan in response to nutrient deprivation. It has been proposed to be a therapeutic target for obesity and metabolic syndrome. We investigated whether α-lipoic acid (ALA) exerts a lipid-lowering effect through regulation of SIRT1 activation and production in C(2)C(12) myotubes.
METHODS: ALA-stimulated AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), adipose triacylglycerol lipase (ATGL) and fatty acid synthase (FAS) production, as well as intracellular triacylglycerol accumulation and fatty acid β-oxidation were analysed in the absence or presence of a SIRT1 inhibitor (nicotinamide), SIRT1 small interfering (si) RNA and an AMPK inhibitor (compound C) in C(2)C(12) myotubes. Mice with streptozotocin/nicotinamide-induced diabetes and db/db mice fed on a high-fat diet were used to study the ALA-mediated lipid-lowering effects in vivo.
RESULTS: ALA increased the NAD(+)/NADH ratio to enhance SIRT1 activity and production in C(2)C(12) myotubes. ALA subsequently increased AMPK and ACC phosphorylation, leading to increased palmitate β-oxidation and decreased intracellular triacylglycerol accumulation in C(2)C(12) myotubes. In cells treated with nicotinamide or transfected with SIRT1 siRNA, ALA-mediated AMPK/ACC phosphorylation, intracellular triacylglycerol accumulation and palmitate β-oxidation were reduced, suggesting that SIRT1 is an upstream regulator of AMPK. ALA increased ATGL and suppressed FAS protein production in C(2)C(12) myotubes. Oral administration of ALA in diabetic mice fed on a high-fat diet and db/db mice dramatically reduced the body weight and visceral fat content.
CONCLUSIONS/INTERPRETATION: ALA activates both SIRT1 and AMPK, which leads to lipid-lowering effects in vitro and in vivo. These findings suggest that ALA may have beneficial effects in the treatment of dyslipidaemia and obesity.

Cancer is an exploitation or improper activation of the nighttime recovery mechanisms, which further include NADPH production for fatty acid, cholesterol, GSH regeneration, NAD+ synthesis from Trp (KMO), Cytochrome P450 enzymes (which also support retinoic acid catabolism to control its levels); you then combine that with a noticeable retinoic acid shortfall (not the least from increased NADPH levels) leading to loss of differentiation, along with a deficit in overall acetylation, eventually leading to immune tolerance due to loss of membrane immune markers such as Hsp70.

Did I mention autophagy? Several years ago I was perplexed about a few cases of mTor stimulation where both AMPK and mTor were simultaneously upregulated (there was an involved of mTorC2 and of course interaction with FoxO), which is odd because they normally oppose each other. Well: if growth hormone and methylation support spermidine synthesis, they may effectively contribute to autophagy, as well as through other signals most likely including FoxO nuclear programs. However, they simultaneously support mTorC1 synthesis. Moreover, this effect involves mTorC2, which increases glucose uptake in some tissues and curiously happens to be distributed in the body in patterns and activation that suggest it's used in skeletal restructuring:
mTORC2 - Wikipedia
mTORC2 has been shown to function as an important regulator of the cytoskeleton through its stimulation of F-actin stress fibers, paxillin, RhoA, Rac1, Cdc42, and protein kinase C α (PKCα).[2]

mTORC2 also regulates cellular proliferation and metabolism, in part through the regulation of IGF-IR, InsR, Akt/PKB and the serum-and glucocorticoid-induced protein kinase SGK. mTORC2 phosphorylates the serine/threonine protein kinase Akt/PKB at a serine residue S473 as well as serine residue S450. Phosphorylation of the serine stimulates Akt phosphorylation at a threonine T308 residue by PDK1 and leads to full Akt activation.[4][5] Curcumin inhibits both by preventing phosphorylation of the serine.[6] Moreover, mTORC2 activity has been implicated in the regulation of autophagy[7](macroautophagy[8] and chaperone mediated autophagy).[9] In addition, mTORC2 has tyrosine kinase activity and phosphorylates IGF-IR and insulin receptor at the tyrosine residues Y1131/1136 and Y1146/1151, respectively, leading to full activation of IGF-IR and InsR.[10]

Why autophagy, mTorC1 and mTorC2 needed at the same time? Growth/proliferation of course, but specially: cellular remodeling. Nighttime cellular remodeling.

These ideas are largely inspired by Paul Jaminet's insistence of the circadian rhythm as one of the most critical factors in health, as well as his insistence on saturated fat as a large part of the diet. However, he had an additional focus on the gut perspective, which is a whole other topic (it maintains things like butyrate and acetate supplies).

It's all here Amazony, all courtesy of some pirate lady from Kazakhstan... The answer's in the rhythm, of our days, the rhythm of the city - boy once you get it down, you can own this town, you can wear the crown

Edit: You can also see this creates a clear relation between GH and TH (thyroid hormone): GH and TH can both downregulate GNMT, but in another sense, GH provides methylation products for for TH to work with, notably on its regenerative abilities. It's no coincidence that GH was thought to increase T4->T3 conversion. On the other hand, near the end of the night and beginning of the day, TH is present and supported by regenerated NAD+ levels in the liver, while retinoic acid and probably cortisol combat GH and TH's downregulatory effects on GNMT.
 
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Terma

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You can also see this creates a clear relation between GH and TH (thyroid hormone): GH and TH can both downregulate GNMT, but in another sense, GH provides methylation products for TH to work with, notably to help implement its regenerative abilities - many of which work using fat oxidation (T3 is known to be able to induce FoxO in the right circumstances and maybe even in general). It's no coincidence GH was noted to increase T4->T3 conversion. On the other hand, near the end of the night and beginning of the day, TH is present and supported by regenerated NAD+ levels in the liver, while retinoic acid (also supported by NAD+) and probably cortisol combat GH and TH's downregulatory effects on GNMT.

Everything fits like a glove: It is very logical for retinoic acid to consume NAD+ in the morning: this produces NADH, which might contribute to shifting 11b HSD2 in the direction of cortisol activation, besides other things like changes in steroid synthesis.
 

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Thank you for the treatise.

Sometimes there's a publication that you doesn't has access to, only the references, forcing you to picture the development of the text based on their order. It's interesting to reverse this process and try to understand what a poster had in mind. You really liked the 'Nicotinamide: a double-edged sword', didn't you?

These don't seem but are relevant:

- Insights into the impact of N- and O-methylation on aqueous solubility and lipophilicity using matched molecular pair analysis
Can you recover nicotinamide after it has been methylated?

- Biochemical Basis of Medicine (0-7236-0722-2)

upload_2019-8-18_20-55-23.png

- Heterocyclic Chemistry
- Aromaticity - Wikipedia (obvious for niacin since it has its characteristic smell)

- Hypolipidemia, Low Cholesterol, And The Art Of Being Hormoneless "a.m."
What do you mean by people sleeping better with a minimum of fatty compounds? They can be helpful in contributing to the raw materials.

- The Metabolic Capacity For Glycine Biosynthesis Does Not Satisfy The Need For Collagen Synthesis

- Influences of diet and the gut microbiome on epigenetic modulation in cancer and other diseases
You mentioned acetate and butyrate, also isotretinoin harm elsewhere. Have you ever come across a report of direct effect?

- Vitamin D's role in cell proliferation and differentiation

FFA and temporary insulin resistance reminded me of Orion mentioning on Grant's thread that he used to run on adrenaline before eliminating toxins from his diet.


Why is the body having trouble storing those for later (nighttime) use?
Choline fulfills everything that you have in mind (slow metabolism, GH, etc), when did you realize it was no longer beneficial in your case?
 
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One more - I think important - piece of the puzzle:

Histamine H3 receptor - Wikipedia
Histamine H3 receptors are expressed in the central nervous system and to a lesser extent the peripheral nervous system, where they act as autoreceptors in presynaptic histaminergic neurons, and also control histamine turnover by feedback inhibition of histamine synthesis and release.[5] The H3 receptor has also been shown to presynaptically inhibit the release of a number of other neurotransmitters (i.e. it acts as an inhibitory heteroreceptor) including, but probably not limited to dopamine, GABA, acetylcholine, noradrenaline, histamine and serotonin.

Meanwhile:
Production by R-alpha-methylhistamine of a histamine H3 receptor-mediated decrease in basal vascular resistance in guinea-pigs.

This is the reason that one of the older studies found a paraxodical effect whereby administration of histidine together with methionine somehow counteracted the effects of histamine: methylhistamine formed by HNMT is selective for the H3 feedback receptor, which counteracts histamine release - for the nighttime.

This is a double-whammy: normally we think of histamine as 'deactivated' by HNMT, but in fact methylhistamine may be a significant halter/modulator of brain metabolic activity on its own.

There is also 4-methylhistamine that activates H4 instead, it looks as if part is also an immune program, which happens to make some sense as the immune system is important at night:
Evaluation of Histamine H1-, H2-, and H3-Receptor Ligands at the Human Histamine H4 Receptor: Identification of 4-Methylhistamine as the First Potent and Selective H4 Receptor Agonist

In other words, histidine + SAMe = sleep mode, by 2 possible effects.

Lysine also critically supports nighttime growth through carnitine synthesis but also many other things, and generally supports sleep. Glycine is also a supporting factor for creatine synthesis and assuring GNMT is functional in the late night or on demand. It is heavily about amino acids, and their use is coordinated by growth hormone.

Notice how methylhistamine ended up in bone marrow:
Urinary N-methylhistamine as an indicator of bone marrow involvement in mastocytosis. - PubMed - NCBI

Histidine and histamine are more fundamental molecules to life than I realized, but I can't go deeper into it.
 
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I'm not sure how, but many methylations are reversible one way or another.

What do you mean by people sleeping better with a minimum of fatty compounds? They can be helpful in contributing to the raw materials.
Saturated and some monounsaturated fatty acids like oleic, palmitic and stearic are the energy substrates of skeletal growth and restructuring. Even for collagen: chondrocytes and processes may form a protein and carb-derived material, but the chondrocytes and other cells need raw ATP and energy to do work. That energy comes from saturated fats during the night.

They need to be available and supported by all the cofactors for fatty oxidation such as carnitine, PPARa/d/g modulators, etc. These cofactors typically localize in the fatty portions of foods.

Yes very familiar with that. Unfortunately 10g/day distorts your perspective on glycine relative some other amino acids and their contributions on the same processes such as coincidentally collagen synthesis and usage. There are other aminos and aspects to this balance, including the cost of metabolizing glycine itself, which falls on the kidneys (requiring lipoic acid + more, producing ammonia).

- Influences of diet and the gut microbiome on epigenetic modulation in cancer and other diseases
You mentioned acetate and butyrate, also isotretinoin harm elsewhere. Have you ever come across a report of direct effect?
Yes but it's several years ago, no references on hand.

Choline fulfills everything that you have in mind (slow metabolism, GH, etc), when did you realize it was no longer beneficial in your case?
Choline cannot be taken in any significant dose in the evening given acetylcholine synthesis. Sure it would increase GH itself, but you simply do not sleep.

For choline I delegate to the traditional practice of bacon and eggs in the morning. I never take alpha-GPC past the afternoon. Every part of this must reinforce the circadian rhythm. It would not be illogical for the CDP-choline pathway to be more heavily relied on during the day, if methylation is relatively slower.

Rhetorically, if choline is not expected to contribute largely to the methyl pool in humans, much less the brain, then how would it deactivate histamine (HNMT), or synthesize melatonin? You can try but you'll get a bunch of acetylcholine at the same time.

(Anything skipped is just lack of time, sorry)
 
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Remember: phosphatidylcholine is important in bile for digestion. So you may have burden on PC production by CDP-choline pathway during the daytime. It involves significant amounts and letting PEMT fill in for it is a road to disease. This is a good argument for the circadian approach: consume all your methyl groups and most of your protein at dinner time; avoid earlier.

Contrarily, at nighttime, the gut should be calming down to minimize sleep-disruptive LPS production and to allow gut repair, so there should not be much bile release.

At nighttime, I am sure the CDP-choline pathway continues to be active, so you still should support it, but additional methyl groups are likely needed - you get them from high protein eating no matter what - whether or not PEMT gets involved. It is probably good to try to avoid PEMT regardless of any of this, but it will become impossible to completely prevent likely due to the differences in the species they produce.

For the sake of completeness:
Complex interaction between circadian rhythm and diet on bile acid homeostasis in male rats. - PubMed - NCBI
Desynchronization between the master clock in the brain, which is entrained by (day) light, and peripheral organ clocks, which are mainly entrained by food intake, may have negative effects on energy metabolism. Bile acid metabolism follows a clear day/night rhythm. We investigated whether in rats on a normal chow diet the daily rhythm of plasma bile acids and hepatic expression of bile acid metabolic genes is controlled by the light/dark cycle or the feeding/fasting rhythm. In addition, we investigated the effects of high caloric diets and time-restricted feeding on daily rhythms of plasma bile acids and hepatic genes involved in bile acid synthesis. In experiment 1 male Wistar rats were fed according to three different feeding paradigms: food was available ad libitum for 24 h (ad lib) or time-restricted for 10 h during the dark period (dark fed) or 10 h during the light period (light fed). To allow further metabolic phenotyping, we manipulated dietary macronutrient intake by providing rats with a chow diet, a free choice high-fat-high-sugar diet or a free choice high-fat (HF) diet. In experiment 2 rats were fed a normal chow diet, but food was either available in a 6-meals-a-day (6M) scheme or ad lib. During both experiments, we measured plasma bile acid levels and hepatic mRNA expression of genes involved in bile acid metabolism at eight different time points during 24 h. Time-restricted feeding enhanced the daily rhythm in plasma bile acid concentrations. Plasma bile acid concentrations are highest during fasting and dropped during the period of food intake with all diets. An HF-containing diet changed bile acid pool composition, but not the daily rhythmicity of plasma bile acid levels. Daily rhythms of hepatic Cyp7a1 and Cyp8b1 mRNA expression followed the hepatic molecular clock, whereas for Shp expression food intake was leading. Combining an HF diet with feeding in the light/inactive period annulled CYp7a1 and Cyp8b1 gene expression rhythms, whilst keeping that of Shp intact. In conclusion, plasma bile acids and key genes in bile acid biosynthesis are entrained by food intake as well as the hepatic molecular clock. Eating during the inactivity period induced changes in the plasma bile acid pool composition similar to those induced by HF feeding.
 
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Goobz

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Very interesting stuff.

What are you thoughts about elevated retinol binding protein 4? It seems its elevated in conditions of insulin resistance, diabetes, obesity, sleep apnea, but also very high neuro conditions like lewy body disease.

Ive read that its usually lowered by the actions of things like estradiol, a low or abnormal level of which is associated with all of those pathological conditions.

I also cant work out if supplementing Vitamin A would make elevated RBP4 worse or better. Ive read low RBP is a sign of vit a deficiency, but also that all trans retinoic acid supplemenation lowers it.

Vit A seems to have a relationship with insulin resistance, but perhaps a mixed one. Or maybe I havent read enough.
 
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The binding proteins sometimes but not always follow a circadian clock or pattern. If the body is overproducing it then the clock is one possible cause. I don't remember specifics about RBP4, but there is something similar for vitamin D binding protein. But for vitamin D the body is really just trying to maintain a 24-hour supply a calcium for the body (which is why calcitriol is short-lived but its precursors can be very long-lived). I would expect RBP to increase strategically following the clock, but sometimes it is hard to say between the mRNA levels, protein expression and species differences. On the other hand, binding proteins can sometimes have an inhibitory effect on signaling by creating a bigger sink for retinol/D, almost like a "pulling power" away from the cells (IGF BP works this way a little more clearly).

One remark: retinoic acid is known to compete with endocannabinoids for intracellular binding proteins (FABP). Now I wonder if this does not help it implement waking (in addition to affecting RAR signaling by detecting presence of fatty acids). I suspect since some time that RA may interfere or re-regulate cannabinoid signaling, since it's been known to affect CB1 receptor levels in tissues like liver.
 
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Regarding NNMT: it's also conceivable that - rather than in the evening - during the course of the late night or morning - the usage of NAD+ by enzymes like SIRT1 increases niacinamide enough that it then doubles as a methyl sink toward the morning. I don't know if this is significant enough amounts to compare to GNMT/RA.

The reason this makes sense is that GNMT is mostly in the liver, so the brain could need extra enzymes. Of course other enzymes such as HNMT also soak up methyl in the brain, but if methylhistamine stimulates H3, it might not be related to waking - or alternatively, the breakdown of methylhistamine stimulates waking. There are several possibilities about how brain methyl is handled near the morning. But it all fits in a general way.
 
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I think I get it: people doing methylation had to take high doses of potassium per their own accounts, supposedly to heal. That - along with some other supplements - is a trigger for cortisol release.

During the night normally GH provides or contributes to gluconeogenesis (for brain), FFA release, and other substances modulate immunity.

But these people were using methylation during the day, while likely none of them produce much GH anymore, and they have ablated or inverted cortisol. So they have to stimulate cortisol to achieve immune suppression and resource availability to support the growth and healing afforded by methylation (and presumably impeded by stimulants such as histamine, though that one is a special molecule for sure).

Furthermore cortisol itself influences the direction of the liver methylation cycle.

There are several accounts of people achieving full temporary relief from ME/CFS-like symptoms using glucocorticoids. So I would in fact call it a #1 suspect.

This is why healing is so slow: you have a limited nighttime window to express the body's full regenerative potential, and we all lose it as we age.
 

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Watching interesting thread.
Since I supplement with glycine instead of gelatin, do you think I should also use lysine (I have some laying around that I don't use)?
I plan to use gelatin in the near future, so the isolated aminos are temporary.
 
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Yes and no, lysine is important but more glycine could lead to things such as arginine usage through things like creatine synthesis if it's high (or if you want to support it). I found it was worth trying individual aminos anyway to gauge their effects, lysine especially, since they absorb faster than full proteins, but in the end you'll need a little of all of them. If arginine gets too compromised then things like the urea cycle may be affected, and it's the precursor for ornithine in polyamine synthesis which is crucial for healing (you could get ornithine instead I guess). You probably need a little nitric oxide anyway. So basically yes but you can't negate the others for too long. I make up for this over time by cycling nutrients.

Curiously it was suggested in literature that anti-anxiety effects of lysine for nighttime use had nothing to do with arginine. But they gave arginine anyway. But I would not take an isolated lysine+arginine therapy forever either.

The way I saw before was: it's good to supplement a little lysine on top of gelatin, since it contains more arginine.

There are probably other aspects.

Lysine might be particularly valuable as a supplement if you have trouble absorbing it from food proteins. I haven't done the specific research for that but it's plausible absorption issues develop breaking down and absorbing various different amino acids, with possible interference from or missing gut flora.
 

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Alright. Thanks.
I may try small dose of lysine so it doesn't get wasted. Would I be able to feel any short term side effects?
 
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If you overdo lysine you'll feel off, I forgot all the reasons. If you take tons of the chloride form (HCl) you might have that to deal with. You can use it either high dose for pharmacological effects or just for maintenance, but less than 1g was never enough to do anything for me personally.
 

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How much poisonoic acid is required to have a significant effect in drawing methyl groups through glycine? It's difficult to judge by those experiments because the lowest dose used was the humanoid equivalent to 3.4 mg/d of poisonoic acid (purified in oil) + diet: sugarbaby high.
Yes very familiar with that. Unfortunately 10g/day distorts your perspective on glycine relative some other amino acids and their contributions on the same processes such as coincidentally collagen synthesis and usage. There are other aminos and aspects to this balance, including the cost of metabolizing glycine itself, which falls on the kidneys (requiring lipoic acid + more, producing ammonia).
No, I mean, pounding niacin could be a way to allow greater synthesis of glycine assuming that muscle protein consumption is high as usual.
Choline cannot be taken in any significant dose in the evening given acetylcholine synthesis. Sure it would increase GH itself, but you simply do not sleep.

For choline I delegate to the traditional practice of bacon and eggs in the morning. I never take alpha-GPC past the afternoon. Every part of this must reinforce the circadian rhythm. It would not be illogical for the CDP-choline pathway to be more heavily relied on during the day, if methylation is relatively slower.

Rhetorically, if choline is not expected to contribute largely to the methyl pool in humans, much less the brain, then how would it deactivate histamine (HNMT), or synthesize melatonin? You can try but you'll get a bunch of acetylcholine at the same time.
Have you ever compared the effects of phosphatidyl to glycerophosphocholine?

Do you know anything about digestive rate of each protein and how it affects day cycles? Casein is an example that could perhaps be less disruptive.
 
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Goobz

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No, I mean, pounding niacin could be a way to allow greater synthesis of glycine assuming that muscle protein consumption is high as usual.

I’ve heard the opposite (just from a chris masterjohn podcast) - that nicotinic acid needs to be glycinated to be converted to nicotinamide, which then needs to be processed using methyl donors such as choline or TMG.

He was saying that taking too much niacin can drain ones glycine, and this perhaps lower glutathione levels.
 

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I’ve heard the opposite (just from a chris masterjohn podcast) - that nicotinic acid needs to be glycinated to be converted to nicotinamide, which then needs to be processed using methyl donors such as choline or TMG.

He was saying that taking too much niacin can drain ones glycine, and this perhaps lower glutathione levels.
Indeed the excess is excreted as nicotinuric acid and wastes glycine. However what's curious is that in this experiment, excretory methylated metabolites increased markedly with nicotinic acid treatment as well, just as high as with nicotinamide; it was a single dose. And if you note, apparently there wased far more methyl molecules being consumed in the process than glycine. Not sure what to make out of it.
 

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