Oxindole

[SB4]

@Terma I didn't take much histidine at all, just recommended dose. Don't really have allergies MCAS either.

Did you notice problems before with carb tolerance? I have massive problems with it unless I am doing this other stuff. Mainly heart problems, my heart pounding kicks up massively, and I just generally overall feel much worse.

I messed around with TUDCA in the past and I improved in that time frame too but could never say if it was due to that or not.

I have not read the study you posted early. Could you sum up for me the relationship between ketones, TRP, and Oxindole?

 

[Terma]

@LeeLemonoil I definitely avoid all those, yep

@SB4 What were the severities of the patients in that study? They don't even mention the term "ME", sometimes a red flag. Indeed it does look more like just a ratio change just from that graph. This suggests again an NADH/NADPH insufficiency to me, at least for those people. I think they assume it's from too much ROS/peroxidation, I can't read it all. In that case GSH helps you.

Running out of NADPH is sort of like being in an "endless day" or "out of control" oxidation [of course it depends which enzymes are worst affected - my generalization would say that short-term emergencies like ROS get prioritized while the rest suffer, but it's more complicated than that... I think it's studied somewhere, I still have to look more].

I'm not super interested in NADH absorption; you can take all the precursors for NAD synthesis (niacin, niacinamide, ribose) and then add reduction using ascorbic acid, NAC, etc. which all those patients end up taking. They likely need extra ribose regardless of that study. Not unlikely the success of ribose is due to NAD synthesis as well as lowering quinolinic acid (PRPP cofactor).

That's interesting about histidine. I will try to see later if there is some more relation between histidine/histamine and TRP or other pain-sensing channels (not sure which are the others).

Carb intolerance I believe I might have some in my brain, but otherwise my glucose regulation hasn't seemed to be an issue. Tons of magnesium and taurine help prevent those issues. It's slightly easier to prevent than to fix.

It turns out ketones actually make more Trp available to neurons and astrocytes, where it turns into NAD and KYNA and their interactions involve lactic acid. To me this seems an area where carnosine/beta-alanine might be a valuable tool. I need to re-read it again, I don't want to give misinterpretation of the details. Oxindole wasn't even an important element in the study.

Edit: Does Oral Coenzyme Q10 Plus NADH Supplementation Improve Fatigue and Biochemical Parameters in Chronic Fatigue Syndrome?

A total of 113 consecutive patients who had attended to CFS Clinical Unit (Vall d'Hebron Hospital, Barcelona, Spain) from January to December 2013 with a diagnosis of CFS according to the 1994 CDC Fukuda's criteria were initially evaluated and potential enrolled for eligibility in the study.

Yeah I'm not too convinced about the cohort. And I'm not sure CoQ10 helps us interpret these results here... Not the best study. I'd ask the other guys (nanxidon/Hip/jefe) if this was really a good study for ME/CFS, because I can't remember too good.

[Keep in mind also that problems in ME/CFS do seem to have cell-type specific characteristics, and NADPH modulates immune cells, so it could be more at that level than muscle cells or other cells, or vice-versa depending on the effect/pathway that's studied]

[Maybe yet another way to interpret that study - which I kind of doubt is very good for this topic - is that increased reduction and/or citrate synthase might help increase fat storage and prevent lipid oxidation or peroxidation that way? PPARgamma does something similar and is known to help this sort of thing]

 

[SB4]

@Terma Thanks. Yeah, I'm not too interested in NADH as a supp for ME but what I am interested in is the NAD+/NADH ratio going down and correlating with symptom improvement. This is going against my understanding of NAD+/NADH ratios. Now it doesn't seem to say where the ratio went down (cytosol/nucleus/mito) but I suspect it is all 3 totaled.

In the study as far as I can see (although I haven't looked hard) it just says NADH supplement which I assume is just that, NADH. I thought it would be broken down in the gut to NR, then changed in the liver to Nicotinamide which is then distributed to tissues with the rest being methylated. This is what happens with NAD+ I think so can't see why NADH would be any different. Anyway I will stop talking about this now as I have already derailed this thread too much.

I am interested in the NADPH deficiency idea. I experience lactic acid buildup from doing next to nothing. I am unsure whether this is simply poor blood flow as my main symptoms are all heart based or if it is a metabolic shift to a more Warburg type metabolism. If it is the Warburg type then this leads to increased NADPH to deal with excess ROS. Perhaps we cannot produce enough NADPH as the ROS is too high for whatever reason (chronic inflammation due to chronic enterovirus?, gut flora source?). This is inline with the Peroxynitrite ME theory.

It turns out ketones actually make more Trp available to neurons and astrocytes, where it turns into NAD and KYNA and their interactions involve lactic acid. To me this seems an area where carnosine/beta-alanine might be a valuable tool. I need to re-read it again, I don't want to give misinterpretation of the details. Oxindole wasn't even an important element in the study.

Yeah I thought it said that but that should make us feel worse when ketosis tends to make ME better. Interesting. I do have some unused beta-alanine, I was going to take it in conjunction with histidine but I stopped it after day 1.

 

[Terma]

Right, well I just assumed it breaks down to niacinamide and ribose (or NR). NAD can also be given intravenously.

Quick disclaimer: there are times I might use NADH and NADPH interchangeably by mistake/forgetfulness - this is not necessarily correct at all - they do interconvert by mitochondrial enzymes (both ways, circumstantially) but of course they decouple by the P and I imagine the enzymes might even get faulty (or low ATP/ions). For example to correct the above, it might be more accurate to call NADH one "end of day" signal, while NADPH might be better seen as "recovery" given its anabolism and link to feeding (except then there's a reason dinner is usually the largest meal... feeding and circadian rhythm are friends) - although in some pathways it might express more a resource "sufficiency". That said these are fundamentally generic molecules (and interpretations likely depend on cell type).

And if anyone finds clear mistakes in this please point them out. You can't do this without being wrong sometimes. Just can't promise I'll be around at the time.

Anyway the study I really wanted shine a light on was the first one in this post, because I haven't read it myself yet: Oxindole

 

[Terma]

Quick remark (I don't have the time to vet this), for diagnostic purposes (self-experimentation or with a specialist), if you wanted to bypass IDO/TDO, you could use a strategy like this - which is administering pre-formed kynurenine together with body KMO/KAT inhibitors (I'm assuming these don't cross the BBB) and for some reason probenecid is also needed - to get it into the brain, which apparently has already been tried on mice with migraines, though I've never heard of these:

Endogenous neuroprotection in chronic neurodegenerative disorders: with particular regard to the kynurenines

From a therapeutic aspect, enhancement of the effects of KYNA would be a successful therapeutic strategy providing protection against the effects of neurotoxic 3-OH-L-KYN and QUIN. This may be achieved by administration of the KYNA precursor L-KYN. Thus, a combination of the blood–brain barrier-penetrable L-KYN with nicotinylalanine, an agent that inhibits both kynurenine 3-hydroxylase and kynureninase activity, and with probenecid, an inhibitor of organic acid transport for enhancement of the brain KYNA concentration, exerted protective effects in the SNpc against both NMDA and QUIN-induced excitotoxicity [236], and in the striatum against QUIN-mediated neurotoxicity [237], possibly via the elevated level of KYNA. However, the co-administration of probenecid seems necessary to achieve a considerably elevated KYNA level in the field of interest. Although L-KYN alone was able to induce amelioration in animal models of cerebral ischemia [238, 239], it is mainly used in combination with probenecid to achieve neuroprotection (e.g. in a migraine model: [240]). However, it has recently been shown that probenecid alone can mitigate the alterations in that migraine model [241]. Further, probenecid is protective in a transgenic mouse model of HD [242], whereas L-KYN alone is ineffective (our unpublished data). In an MPTP model of PD, we did not find any protection with pure L-KYN either. Nevertheless, the available data suggest that L-KYN in combination with probenecid is perhaps capable of enhancing neuroprotection in chronic neurodegenerative disorders.

(There are various other interesting things in that article)

But the only person in the world who'd try that on his own is Hip.

Also, in case this wasn't clear: I dedicated many posts to NADPH but I would think the main characteristic of the disease would rest at IDO or even TDO inhibition, and then NADPH deficiency would give the disease some more autoimmune-like aspects and severity differences (because between IDO and KMO, that's 2 enzymes controlling flux, meaning a few possible layers of severity and symptoms). But I also think because of this, if you tried kyn you would probably want to supplement an NADPH-boosting supplement (assuming the inhibitors don't work by inhibiting NADPH! Then you'd have a problem - check that first). (NAC spares NADPH as per the Lupus study, but it will confound your experiment so you'd have to try it alone first)

 

[Terma]

I'll leave these here from the ketone study, this'll save me some words and it's not really that off-topic (it's missing stuff from the HPA axis and endocannabinoids i.e. bigger picture):

Endogenous neuroprotection in chronic neurodegenerative disorders: with particular regard to the kynurenines

https://sci-hub.tw/10.1016/j.neuroscience.2015.11.029

 

[Terma]

Again I'm not sure if this would be involved in ME/CFS, but it looks serious enough to [manifest as] a disease - these are real patients. Maybe the simplest course is to find what bacteria does this the worst and kill it forever. I still haven't read some of these studies in full, so feel free, mateys (that was my sci-hub captcha).

Peripheral and splanchnic indole and oxindole levels in cirrhotic patients: a study on the pathophysiology of hepatic encephalopathy. - PubMed - NCBI

OBJECTIVES:
Intestinal bacteria metabolize tryptophan into indole, which is then further metabolized into oxindole, a sedative compound putatively involved in the pathophysiology of hepatic encephalopathy (HE). The aim of this study was to measure indole and oxindole levels in patients with cirrhosis with or without HE and to establish whether an intestinal production and a hepatic metabolism of these substances exist.

METHODS:
We studied 10 healthy subjects (controls) and 51 cirrhotic patients: 17 without HE, 14 with a minimal HE, 8 with overt HE, and 12 who had undergone a transjugular intrahepatic portosystemic shunt (TIPS) procedure. In the last group, blood was collected from the artery, and the portal and hepatic veins during TIPS construction and from the peripheral veins before, immediately after, and at weekly intervals during the first month after TIPS.

RESULTS:
Plasma indole levels were significantly higher in patients with overt HE. Oxindole levels were higher in cirrhotics than in controls. Indole and ammonia were significantly correlated (r=0.66). Peripheral and splanchnic determinations showed that indole was produced in the intestine and cleared by the liver, similar to ammonia. TIPS implantation increased both indole and ammonia levels. After TIPS, the psychometric performance worsened in 4 of the 12 patients. The increase in indole plasma concentrations in these four patients was higher than in those who remained stable after undergoing TIPS.

CONCLUSIONS:
Indole correlates with HE and has a significant intestinal production and hepatic extraction; its level increases after TIPS and is related to psychometric performance. These data suggest that indole may be involved in the pathophysiology of HE.

As mentioned previously, it is reasonable to assume that indole is metabolized into oxindole, the neuroactive metabolite. Oxindole formation (see Figure 1 ) requires the action of oxidative enzymes that are expressed in most mammalian cells and are present in the gut mucosa, the liver, the kidneys, the leukocytes, and even in the brain tissue ( 22 ). This may explain the rather high variability of plasma oxindole levels in cirrhotic patients in whom the liver (and kidney) function is significantly impaired.

On the basis of the above findings, we hypothesize that indole produced in the intestine reaches the systemic circulation when the liver fails and / or large shunts are present and may be transformed into oxindole in the brain, thereby fitting all the features in order to be considered a substance with potential involvement in the pathogenesis of HE. In fact, oxindole, the main indole metabolite, interacts with neuronal voltage-operated sodium channels, thus reducing cell excitability ( 21 ) and contributing to the changes in brain function found in HE. As ammonia has been shown to interact with different neuronal and glial targets (for instance, with molecular mechanisms controlling glutamate synthesis and release ( 12 )), it is possible that the basic actions of oxindole and ammonia additively contribute to the development of the signs and symptoms found in HE.

They even advise a diet low in tryptophan:

Considering the gut origin of indole, it is possible that, similar to what occurs in the case of ammonia, plasma indole levels would also be reduced by most of the treatments traditionally used in patients with overt HE, such as gut cleansing ( 34 ), non-absorbable disaccharides, or low-absorbable antibiotics ( 29 ). Moreover, as tryptophan is an indole precursor, a vegetable diet low in tryptophan content should decrease indole formation. Finally, an attempt should be carried out to modify the gut flora by administering probiotics without tryptophanases (unable to metabolize tryptophan into indole, see Figure 1 ).

So you probably want to restrict tryptophan during liver failure.

This could even add a twist to the function of the Kynurenine pathway: theoretically if indoles were an issue in the past, the immune system could even have evolved to try to catabolize Trp in the presence of bacteria to prevent indole formation. This is going out on a limb though and obviously it evolved for other reasons as well.

More:
Microbial Degradation of Indole and Its Derivatives

This does begin to make some sense: someone could conceivably end up with a real Trp deficiency from this if their gut were terrible - or at least not enough Trp to fuel NAD in neurons and KYNA in astrocytes (hence the ketones). But more importantly, if that person consumed Trp as a pure amino or even whey, they could get a benefit because these may absorb much faster than food, before gut bacteria can metabolize them to indoles as much. However, their liver might also be compromised from years of LPS and other issues, leading to the above... at that point, perhaps the issue at hand - indoles and oxindole in particular - moves from the liver to the brain?

 

[Terma]

@SB4 This is highly anectodal, but: I did know someone with MCAS/allergies and osteoporosis and my guess is they ended up copper-deficient in the long run (or even inorganic copper problems due to copper in water pipes), and it results in severe bone pain. So if histidine hurts you that much, I'd watch out for functional copper utilization issues (not necessarily "deficiency") and osteoporosis. Might not be that, I didn't look deeper yet, stuff is piling up (I need like 4 quantum engines).

 

[Terma]

@SB4 It would even seem plausible to me for it to be copper overload, but again it comes back to utilization/buffering. You can get some hints from papers like this (histidine can move copper, you need GSH - or some unknown alternative, not sure how likely that is - to handle it - they don't even know all the carriers that exist):
Liver as a key organ in the supply, storage, and excretion of copper

 

[SB4]

@Terma thanks although not sure how accurate these tests are, my hair mineral analysis came back with copper on the low end. 1.2 (0.9 - 3.9). Then again on that test most of my minerals came back low.

 

[Terma]

I think this is important to point out again:
Cortisol - Wikipedia

11-beta HSD1 uses the cofactor NADPH to convert biologically inert cortisone to biologically active cortisol
11-beta HSD2 uses the cofactor NAD+ to convert cortisol to cortisone

1. NAD+ sustains cortisol deactivation in 11-beta HSD2-expressing cells. This means that an increase in the amount of Trp dedicated toward the kynurenine pathway of those cells - with the requirement that KMO (NADPH) is functional and not saturated, in the same cells - may help control cortisol levels.
2. The NADPH-consuming enzyme KMO that dedicates kynurenine toward NAD+ production is theoretically at odds with cortisol activation by NADPH-consuming 11-beta HSD1 (it depends on the inter- and intra-cellular distribution of these enzymes and the NADPH pool).

This could help explain occasional positive effects of Trp administration (not 5-HTP) in cortisol levels in experiments as well as subjective reports from people. It suggests a kind of feedback mechanism for free fatty acids increasing brain Trp update, further explains the subjective effects of ketosis, and other things. It's not really new or complete (due to cell type specificity) but it bears repeating.

***

If you factor in indoles/oxindole, you have that the kynurenine pathway in neurons may be unable to produce enough NAD+ (or NADPH to recover the NADPH lost at the KMO enzyme), and of course KYNA by astrocytes, due to competition, not impossibly even at TDO (not even IDO).

A high level of stress in the organism along with feedback GR insensitivity and attempt to manufacture hormones such as cortisol, compensatory GSH, detoxification and other products (even H2O, produced by monooxygenases) puts an even greater strain on the NADPH pool of cells, worsening again KMO performance and NAD+ production. At some point, the Trp that does get committed by KMO toward niacin synthesis will get stuck as quinolinic acid from lack of PRPP due to ribose deficits. This could be before or after NAD+ synthesis from niacin crashes, because that one is dependent on both ribose and purines. There might or might not be picolinic acid over- or under-production, I'm not sure. You would probably try to throw a bunch of vitamin B3 at it, but it will not work for long or if you start too late because ribose gets depleted eventually, and then B3 could go so far as to hurt your state.

If during that process you developed a P5P deficiency (due to broken conversion from B6 or other), xanthurenic acid would presumably increase and basically start shutting down the system although you might feel some form of relief or better. This is a dramatic change: you go from excitatory quinolinic acid to a parking brake.

Finally, metabolism including thyroid must be supported to maintain FAD production from B2 (necessary for KMO), and at any point that can crash, independently or dependently.

At a certain point you don't make as much ATP anymore. Then or even before, cells lose magnesium, which will cripple a few enzymes, notably possibly the quin->niacin conversion.

Theoretically, either indoles/oxindole or extreme stress could cripple metabolism this way, by impeding the best route of tryptophan usage, and if it did it long enough for ribose production to collapse you'd be in a trap. Meanwhile you get tons of new issues due to Trp repartitioning: KYNA, serotonin, tryptamine, IDO/immunity, possibly infection... At some point you can't even make cortisol (or it inverts, since some of these disorders may involve or even fully be due to circadian disorders).

Note: For the sake of "brevity", I've omitted considerations about inter-cellular and inter-compartmental differences; they may pose some obstacles or minimum concentrations, but to my knowledge it is possible for much of this to happen in the same cell.

***

From a different perspective:

If you supplement niacin, you spare NADPH and PRPP because you're skipping KMO and quinolinic acid. However, you increase the Trp burden on all the other pathways and possibly worsen problems with indoles; this amplifies the more you deplete PRPP or purines (adenine). Meaning: B3 supplementation requirements should be highly disease- and state-dependent. It is a good idea to take it primarily if the kynurenine pathway is broken but PRPP and purines are not.

***

These explanations validate the THC protocol I am using further: DHEA + progesterone + magnesium + others help control cortisol and affect steroid conversions, but on top of that, the increase in NAD+ production from Trp by the ketones will further lower cortisol by/in 11b HSD2 cells; since NAD+ is obviously a precursor for NADPH, it helps ensure that supply for 11b HSD1. Ribose, assuming it reaches the cells - or delivered/spared/synthesized in any way - gains further importance in the synthesis of PRPP (and thus NAD+/NADPH) to support this NAD+ synthesis; furthermore I would expect ribose to help normalize cortisol levels even greater than the effect of NAD+ production due to the effects of lowering quinolinic acid and other.

This metabolic usage of Trp is likely to steal the pool of it away from other Trp-consuming pathways in the body, since classically TDO is one of the biggest consumers of Trp (although iirc it is biggest in the liver?) - it can conceivably compete with IDO at least indirectly (you might also suspect that IDO activity in immune cells might have been starving TDO of Trp in neurons/astrocytes and IDO in astrocytes iirc). THC helps prevent the Trp influx from causing serotonin signaling, while anti-inflammatory effects of the various substances may lower IDO including some Trp metabolites themselves. In other words, this is the mechanism that may help counter the pro-cortisol effects of THC. It may well balance out. Also I think DHEA(S) is doing something special beyond what I currently know, and there's something about pregnenolone vs progesterone I can't quite put my finger on...

On the other hand, maybe I just activated a magical Trp->tryptamine conversion pathway and I'm tripping balls. **** maybe this is the formula for endogenous dimethyltryptamine synthesis? I do get more than enough methyl as it is. Wouldn't that be funny? We can transcend now guys.

 

[Terma]

Note: I definitely believe you could have detrimental Trp deficiencies localized to certain brain areas and circuits. It's frequently postulated that IDO activity in immune cells could rob TDO in neural cells of Trp. This is my reason for well-receiving the news that ketones can drive Trp into the brain, instead of freaking out automatically. It also makes BCAAs potentially counterproductive, but more or less exonerates carbs.

But you could even have an area so commonly depleted of bioavailable Trp that some post-synaptic 5-HT receptors become oversensitive there. In this situation, you'd have a lower "baseline" of serotonin, but then if you get an injection of Trp due to any kind of stressor (like tissue damage), TPH2 would be more than happy to fuel big spikes of serotonin. Then you'll get anxiety, OCD, social longing and clinginess, or any other behavior/symptom encouraged by post-synaptic 5-HT1a, 5-HT2a/c, etc. depending on the brain area and circuit.

This could potentially reflect an area with hyperactive microglia or other immune cells whose IDO enzymes - assuming not saturated with other indoles - consume most of the available Trp, depending on where the immune cells congregate. This is a realistic scenario: regular Trp deficiency in serotonergic circuits due to immune activity and other Trp sinks - which perhaps combined with overactive TPH or other serotonin synthesis enzymes - perhaps due to compensatory upregulation or other stressors - would cause disproportionately stressful neurotransmission in response to Trp surges.

Of course, any of the enzymes or cofactors in serotonin synthesis could themselves be compromised, also leading to an area of low serotonin synthesis, but in that case surges of Trp availability would not cause serotonin spikes. I'm not sure how far depleted you have to be for this, but BH4, B6, B9 or Fe deficiency can do it. I get the impression that in some strange cases, deficiency of vital cofactors can sometimes provide relief of specific symptoms, and represent a flip in or an alternative disease state.

Here's an interesting case: what if a brain circuit is chronically full of Trp, but one of the enzymes or cofactors in serotonin synthesis is compromised? Then you would have a case where the addition of any of BH4, B6, B9 or Fe to the diet will automatically cause increases in serotonin synthesis - which are reaching post-synaptic 5-HT receptors that have been sensitized due to chronic (overall over the course of a day or week) serotonin depletion. This can explain people who have neurological reactions to food.

I like that last one, because I personally have had issues supplementing B6 and P5P and other things, so it would even fit with my spectacular OCD.

 

[Terma]

Gonna leave this here, don't have time to comment, if you read this in full you'll see this is potentially highly critical, because it could theoretically form a feedback loop (ONOO- from glutamate/NMDA -> soaked up by (L-)kynurenine toward KYNA instead of NAD synthesis -> KYNA antagonizes NMDA) though there are unanswered questions about D- vs L- and localization so I'm not making such claims yet:
Alternative kynurenic acid synthesis routes studied in the rat cerebellum

Kynurenic acid (KYNA), an astrocyte-derived, endogenous antagonist of α7 nicotinic acetylcholine and excitatory amino acid receptors, regulates glutamatergic, GABAergic, cholinergic and dopaminergic neurotransmission in several regions of the rodent brain. Synthesis of KYNA in the brain and elsewhere is generally attributed to the enzymatic conversion of L-kynurenine (L-KYN) by kynurenine aminotransferases (KATs). However, alternative routes, including KYNA formation from D-kynurenine (D-KYN) by D-amino acid oxidase (DAAO) and the direct transformation of kynurenine to KYNA by reactive oxygen species (ROS), have been demonstrated in the rat brain. Using the rat cerebellum, a region of low KAT activity and high DAAO activity, the present experiments were designed to examine KYNA production from L-KYN or D-KYN by KAT and DAAO, respectively, and to investigate the effect of ROS on KYNA synthesis. In chemical combinatorial systems, both L-KYN and D-KYN interacted directly with peroxynitrite (ONOO−) and hydroxyl radicals (OH•), resulting in the formation of KYNA. In tissue homogenates, the non-specific KAT inhibitor aminooxyacetic acid (AOAA; 1 mM) reduced KYNA production from L-KYN and D-KYN by 85.1 ± 1.7% and 27.1 ± 4.5%, respectively. Addition of DAAO inhibitors (benzoic acid, kojic acid or 3-methylpyrazole-5-carboxylic acid; 5 μM each) attenuated KYNA formation from L-KYN and D-KYN by ~35% and ~66%, respectively. ONOO− (25 μM) potentiated KYNA production from both L-KYN and D-KYN, and these effects were reduced by DAAO inhibition. AOAA attenuated KYNA production from L-KYN + ONOO− but not from D-KYN + ONOO−. In vivo, extracellular KYNA levels increased rapidly after perfusion of ONOO− and, more prominently, after subsequent perfusion with L-KYN or D-KYN (100 μM). Taken together, these results suggest that different mechanisms are involved in KYNA production in the rat cerebellum, and that, specifically, DAAO and ROS can function as alternative routes for KYNA production.