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WASF3 disrupts mitochondrial respiration and may mediate exercise intolerance in myalgic encephalomyelitis/chronic fatigue syndrome

datadragon

Senior Member
Messages
389
Location
East Coast, USA
Add Homocysteine to the list.

Yes, Homocysteine induced ER stress and depleted cellular thiols in a dose-dependent manner. https://www.sciencedirect.com/science/article/abs/pii/S2773044122000699 High homocysteine induces endoplasmic reticulum (ER) stress as mentioned by Mario awhile back and earlier in this thread https://pubmed.ncbi.nlm.nih.gov/15243582/ but it also increases NLRP3 inflammasome (which lowers zinc and butyrate for example that increase ER Stress) and downregulates peroxisome proliferator-activated receptor (PPAR) expression (PPAR-a and PPAR-y) https://pubmed.ncbi.nlm.nih.gov/28394319/ https://link.springer.com/article/10.1186/1475-2891-3-4 A lot of that would be downstream effects during inflammation/infection since methylation gets impaired leading to higher homocysteine levels.


TD is shown to cause oxidative stress and the disruption of intracellular calcium concentration. Therefore, it is likely TD may induce ER stress through these mechanisms.

We demonstrated here that Thiamine deficiency up-regulated several markers of ER stress, such as GRP78, GADD153/Chop, phosphorylation of eIF2α and cleavage of caspase-12 in the cerebellum and the thalamus of mice. Furthermore, ultrastructural analysis by electron microscopic study revealed an abnormality in ER structure. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1819404/ Recent research evidence indicates that TD causes oxidative stress, endoplasmic reticulum (ER) stress and autophagy in the brain, which are known to contribute to the pathogenesis of various neurodegenerative diseases. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5337452/

I think we can add Nitric Oxide to the list of suspected players;
Endoplasmic reticulum (ER) stress inhibitors attenuated the NO-induced repressive effects on Ppara gene expression in 10T1/2 adipocytes.
https://pubmed.ncbi.nlm.nih.gov/37739218/

Later it was found that ER stress pathway is also activated by various cellular stresses to protect cells, but when stresses are severe, apoptosis is induced to remove damaged cells. It is reported that NO and reactive oxygen species disturb ER functions, then ER stress-mediated apoptosis pathway is activated. Excess NO causes apoptosis through the ER stress pathway in some types of cells. It was found that the ER stress pathway is activated by various stresses, and when stresses are severe, apoptosis is induced. The NO-induced ER stress pathway may be involved in pathogenesis of various vascular diseases. https://www.ahajournals.org/doi/10.1161/01.ATV.0000223900.67024.15 and https://pubmed.ncbi.nlm.nih.gov/27030287/

Zinc decreases cytokine-induced iNOS expression in endothelial cells. Zinc inhibits iNOS promoter activity.
NF-kB silencing abolishes cytokine-induced iNOS expression. Zinc inhibits the transactivation activity of NF-κB. https://www.sciencedirect.com/science/article/pii/S2213231714000834

Tetrahydrobiopterin (BH4) is a critical cofactor for the rate limiting enzymes in the synthesis of the monoamine neurotransmitters. BH4 is necessary for the conversion of phenylalanine to tyrosine by PAH, tyrosine to L-DOPA by tyrosine hydroxylase (TH) leading to the production of dopamine and norepinephrine, and tryptophan to 5-HTP leading to the production of serotonin.

The PPAR-α agonist Fenofibrate upregulates Tetrahydrobiopterin (BH4) level through increasing the expression of Guanosine 5′-Triphosphate Cyclohydrolase-I in human Umbilical Vein Endothelial Cells. Guanosine 5′-triphosphate cyclohydrolase-I(GTPCH-I), encoded by the GCH-I gene, is the rate-limiting enzyme in BH4 synthesis. https://www.hindawi.com/journals/ppar/2011/523520/ Our previous study demonstrated that homocysteine impairs coronary artery endothelial function by decreasing the level of BH4 in patients with hyperhomocysteinemia. Our previous study also showed that plasma level of BH4 was significantly increased by PPARα agonist fenofibrate in patients with hypertriglyceridemia https://journals.physiology.org/doi/full/10.1152/ajpendo.00367.2010 All PPAR agonists tested lost their potency to downregulate the TNF-α–induced inflammatory response in zinc-deficient cells. However, if zinc was added back, all PPAR agonists significantly downregulated the TNF-α–mediated induction of inflammatory transcription factors NF-κB and AP-1 and significantly reduced the expression of their target genes, VCAM-1 and IL-6 https://www.sciencedirect.com/science/article/pii/S0022316623029346?via=ihub

...Continuing along the de novo BH4 synthesis pathway, H2NTP is next converted to 6-pyruvoyl tetrahydropterin by the zinc-dependent enzyme, PTPS. Although GTPCH is rate limiting to BH4 synthesis in most cells, PTPS has been suggested to be rate limiting in some, most notably human hepatocytes. PTPS may become rate limiting in other tissues and cells, after stimulation with cytokines and other immunological stimuli that induce BH4 synthesis by up-regulation of GTPCH expression https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4038990/
 
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Messages
54
Take the following with a grain of salt, as I do not have any background in this stuff.

The WASF3 article says the following:
Increased ER stress would normally be expected to turn down protein translation, but intriguingly, the inhibitory phosphorylation of protein translation factor eIF2α, a target of PERK kinase activity, was unexpectedly lower (i.e.,
activated) in patient S1 cells (Fig. 6A, lane 2 vs. 1). This finding suggested that there was “ER stress response failure” in patient S1 cells as also observed in the ME/CFS muscle biopsy samples (31). We next examined the effect of reducing ER stress in S1 cells by two different inhibitors, tauroursodeoxycholic acid (TUDCA) and salubrinal (36, 37). While the effect of TUDCA was less apparent as expected of a relatively nonspecific drug, the inhibitor of protein phosphatase 1 (PP1) salubrinal, which prevents the dephosphorylation of eIF2α thereby inhibiting protein translation, was more effective in decreasing the levels of both PERK and WASF3 in S1 cells (Fig. 6A, lanes 3-6 vs. 2). The salubrinal treatment restored the decreased levels of SC III2+IV in patient S1 cells, while their complex V level remained unchanged, demonstrating the targeted and specific nature of the signaling pathway (Fig. 6B).
Salubrinal has not been used in any human trials I could find, so I searched for another drug with the same effect (inhibition of eIF2α dephosphorylation). I found the following:
Mice treated with Nelfinavir display hallmarks of this stress response in the liver, including α subunit of translation initiation factor 2 (eIF2α) phosphorylation, activating transcription factor-4 (ATF4) induction, and increased expression of known downstream targets. Mechanistically, Nelfinavir-mediated ISR bypassed direct activation of the eIF2α stress kinases and instead relied on the inhibition of the constitutive eIF2α dephosphorylation and down-regulation of the phophatase cofactor CReP
https://www.pnas.org/doi/epdf/10.1073/pnas.1514076113

Now, the confusing part is that antiretrovirals like Nelfinavir induce ER stress, and are generally unhealthy:
Increased eIF2α phosphorylation is associated with several pathophysiological conditions including neurodegeneration, cancer, diabetes, and obesity (44–46). Long-term treatment with the HIV-PIs is associated with adverse effects such as hyperlipidemia or hypolipidemia, body fat redistribution, osteopenia and osteoporosis (47), as well as insulin resistance and susceptibility totype II diabetes (48–51).
And yet, I struggle to find reports of Salubrinal causing the same issues, even though it appears to have the same effect (inhibiting eIF2α dephosphorylation). Perhaps it is because Salubrinal's effect inhibit a protein upstream of eIF2α (instead of Nelfinavir, which seems like its effect may be more direct)?:
Salubrinal has been shown to prevent eIF2α dephosphorylation by inhibiting the protein complex GADD34/protein phosphatase 1 (PP1), which consists of the general cellular serine/threonine phosphatase PP1 and the non-enzymatic cofactor GADD34
Or, perhaps it is just because Salubrinal has not been tried in any in vivo trials. I don't know, this is pure guesswork.

The next question is, what is more helpful/achievable in CFS: reducing the ER stress, or buffing the EC stress response? The WASF3 paper makes it sound like the stress response is more uniquely dysfunctional in ME/CFS than the presence of ER stress itself. Could Nelfinavir's activation of the ER stress response (which appears to be insufficient in ME/CFS according to the WASF3 paper) outweigh the downside of increasing the actual ER stress?

I have no idea, and certainly do not recommend trying Nelfinavir based on this. Just a thought experiment.
 

Dakota15

Senior Member
Messages
293
Location
Midwest, USA
Sharing with permission, a message by Dr. Hwang. I thought it may be beneficial for visibility for the community to view:

"I have received a large volume of emails since the publication of the Washington Post article by Brian Vastag about my research. With a desire to be responsive, I have listed below information that addresses common themes within the emails I am receiving.

1) While our research study is not currently recruiting clinical participants, our work continues in the lab to better to understand the “biology” of the protein called WASF3, which may give us some ideas about the mechanism and/or etiology of myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS).

2) To be clear we do not believe WASF3 is the cause of ME/CFS but is one of the factors mediating the energy deficiency in muscle. Understanding its biology and regulation, for example, may help us understand the causes of ME/CFS.

3) We observed increased levels of WASF3 in a subset of the 14 ME/CFS patients. Please understand this is a small sample size. So WASF3 levels in muscle may explain fatigue in only a subset of patients with ME/CFS diagnosis. There could be other factors mediating fatigue in people with ME/CFS.

4) There are no clinical or blood tests available to check for WASF3 levels, and our experiments have not observed WASF3 in blood. In the publication reported publicly through the Washington Post, we used skin biopsy cells grown in tissue culture and muscle tissue samples obtained by needle biopsy to measure WASF3 levels. These tests cannot be done in the clinics by your physicians. But even if they could, more studies are needed in order to understand the meaning of these results.

5) We studied WASF3 levels only in patients diagnosed with ME/CFS. We have not examined patients with Long COVID, so it remains unknown whether the protein WASF3 is involved in Long COVID. Future studies are needed to address this question.

6) As you may know, there are many different medical conditions associated with fatigue. My research examines only one, ME/CFS. I cannot speak to the association of the WASF3 protein to other conditions. My research will continue to focus on the ME/CFS condition through other studies – including exploratory studies.

7) As far as next steps, it is difficult to anticipate all the different issues that will need addressing with getting a clinical trial approved. So, I hesitate to predict timing.

8) In an exploratory treatment study, we would plan to focus on the ME/CFS condition. Therefore, I would expect that an inclusion criterion to participate might be having had a formal diagnosis of ME/CFS after evaluation by a physician at an outside medical center, but again it is difficult to anticipate the issues that might arise in the design and approval processes of a clinical study.

Patient input is critical to better understanding ME/CFS. The published research article provides important insights that will serve to encourage more research into ME/CFS and advances our understanding of this debilitating condition.

Sincerely,

Paul Hwang, MD, PhD"
 

SlamDancin

Senior Member
Messages
511
I don’t want to derail the thread too much but I am curious if there is a connection here to the 2022 study by Peterson et al that showed increased ATG13, mTORc1 activation and impaired autophagy in me/CFS serum causing increased release of ROS and NO from microglia. ROS is crucial mechanism for NLRP3 activation and NO is an NLRP3 activator. It’s especially interesting because autophagy, as far as I understand it, is an ER heavy process, and ER stress is often caused by impaired autophagy and Vice versa in a vicious cycle. I’m going to do some reading in this direction, @datadragon may want to look into these autophagy targets.
RAGE and NLRP3 are on the same signaling axis. So activation of RAGE would ostensibly activate NLRP3 as well.

Elevated ATG13 in serum of patients with ME/CFS stimulates oxidative stress response in microglial cells via activation of receptor for advanced glycation end products (RAGE)
 
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Dakota15

Senior Member
Messages
293
Location
Midwest, USA
Dr. Hwang’s reply to me today.

I asked, “Do you think that Dr. Gibbons and NHLBI will support your pursuit for an exploratory treatment trial in this research?“

“Yes, I am sure that our clinical protocol once it is developed will be supported. I believe the NIH realizes the importance of considering and following up on every treatment lead for ME/CFS, including our specific finding.”
 
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datadragon

Senior Member
Messages
389
Location
East Coast, USA
To be clear we do not believe WASF3 is the cause of ME/CFS but is one of the factors mediating the energy deficiency in muscle. Understanding its biology and regulation, for example, may help us understand the causes of ME/CFS.

On that note...

During ER stress, calcium stores are depleted due to release from the ER. Increased level of mitochondrial calcium alter metabolism and eventually ROS production. This increase in oxidative phosphorylation and ROS generation induces the mitochondria to work faster and consume more oxygen. In addition, under high mitochondrial ROS generation, as oxygen consumption increases, nitric oxide synthase is stimulated by calcium and inhibits the activity of complex IV, which further increases ROS production. This signaling axis usually operates at physiological concentrations of nitric oxide (NO). At the same time, NO and high-calcium mitochondria can inhibit complex I, open MPTP to release cytochrome c to block the respiratory chain at complex III, and thus initiate ROS generation. Calcium also modifies the redox environment by disrupting GSH (Glutathione) inhibition.

https://www.mdpi.com/1422-0067/17/3/327 Studies are linked to in this study link from that text paragraph above for the researchers. The previously mentioned zinc, vitamin A, copper and their effects on mitochondrial function also should be looked at further to add to what I posted.
 
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I think we need to understand exactly what's going on in the WASF3 paper, before we can propose what the goal is. A lot of drugs/supplements ME/CFS patients have tried can "reduce ER stress" by general (e.g. upstream, high-level) mechanisms, but these don't yield impressive results over the long-term.

Normal Unfolded Protein Response (UPR)
Overview from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8146082/
The unfolded protein response (UPR) is the cells’ way of maintaining the balance of protein folding in the endoplasmic reticulum, which is the section of the cell designated for folding proteins with specific destinations such as other organelles or to be secreted by the cell. The UPR is activated when unfolded proteins accumulate in the endoplasmic reticulum. This accumulation puts a greater load on the molecules in charge of folding the proteins, and therefore the UPR works to balance this by lowering the number of unfolded proteins present in the cell. This is done in multiple ways, such as lowering the number of proteins that need to be folded; increasing the folding ability of the endoplasmic reticulum and by removing some of the unfolded proteins which take longer to fold. If the UPR is successful at reducing the number of unfolded proteins, the UPR is inactivated and the cells protein folding balance is returned to normal. However, if the UPR is unsuccessful, then this can lead to cell death.
Now, what does that process look like in terms of the protein kinases? I found a good excerpt from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3578356/:
PERK’s lumenal stress-sensing domain is structurally and functionally related to Ire1’s, though the sequence identity is low, and PERK also contains a cytosolic kinase domain that undergoes trans-autophosphorylation in response to ER stress. Unlike Ire1, however, PERK also phosphorylates the eukaryotic translation initiation factor eIF2α. Phosphorylation of eIF2α results in a reduction of general protein synthesis and thus a decrease in the load of proteins entering the ER (Harding et al. 1999). Under such conditions, mRNAs containing inhibitory upstream open reading frames in their 5′-untranslated region are preferentially translated (Jackson et al. 2010). One such mRNA encodes the transcription factor ATF4 that activates downstream UPR target genes, including GADD34 (growth arrest and DNA damage-inducible 34) and CHOP (transcription factor C/EBP homologous protein) (Harding et al. 2000; Scheuner et al. 2001). GADD34 encodes the regulatory subunit of the protein phosphatase PP1C complex that dephosphorylates eIF2α (Novoa et al. 2001), comprising a negative feedback loop to reverse the translational attenuation mediated by PERK. The downstream transcription factor CHOP activates genes involved in apoptosis (Wang et al. 1998; Zinszner et al. 1998). Thus, the PERK branch first mediates a prosurvival response, which switches into a proapoptotic response on prolonged ER stress (Walter and Ron 2011).
Takeaways of the stress response:
  • PERK is a sensor of ER stress (e.g. of unfolded protein accumulation in the ER)
  • When activated by stress, PERK's job is to deactivate (by phosphorylation) eIF2α
  • eIF2α positively regulates protein synthesis
  • Decreasing the total number of proteins that need to be folded is one method to reduce accumulation of unfolded proteins in the ER, which are the cause of ER stress
Now, what happens when there is no more ER stress? PERK should automatically become disabled, because it is a direct sensor of ER stress (or almost direct). So, PERK will stop phosphorylating eIF2α, but something still needs to dephosphorylate eIF2α to bring protein synthesis back to normal. That 'something' is PP1 and GADD34: (https://www.pnas.org/doi/10.1073/pnas.1501557112)
Following relief of the stress, the growth arrest and DNA damage-inducible protein GADD34 associates with the broadly acting serine/threonine protein phosphatase 1 (PP1) to dephosphorylate eIF2α.

Known Pathological UPR
Now the dual nature of eIF2α becomes more apparent: turning down total protein synthesis, while helpful during ER stress, is probably not ideal long-term: (https://www.cell.com/trends/cell-biology/fulltext/S0962-8924(20)30101):
Although activation of UPR aims to restore cellular function, prolonged ER stress response can activate apoptotic signals, leading to the robust expression of downstream signaling, which damage the target cells (Figure 1A ). Indeed, increasing evidence demonstrates that metabolic disorders, such as obesity, type 2 diabetes, and age-associated pathogenesis, are associated with chronic ER stress.
But, the most important point is that the cell's high-level goal of activating the ER stress response was not necessarily the problem:
However, the ER stress response or sensing failure also contributes to the progression of metabolic diseases where the downstream molecules involved in ER stress responses fail to be fully activated despite the activation of the upstream ER stress sensors
This last bolded sentence is the crux of the WASF3 paper's hypothesis. But, it has also been observed in other diseases, such as in diabetic/obese mice:
Similarly, the tendency of the phosphorylated ER stress sensor PERK (protein kinase R-like ER kinase) is high, while the downstream target phosphorylation of eIF2α (eukaryotic initiation factor 2-α-subunit) and DDIT3 (DNA damage-inducible transcript 3)/GADD153 (growth arrest- and DNA damage-inducible protein 153), or CHOP [CCAAT/enhancer-binding protein (C/EBP) homologous protein], are decreased
It's like the thermostat (PERK) is set to cool things down, but the furnace (eIF2α) never turns off fully. So, the thermostat stays pathologically activated, forever.

Pathological UPR in ME/CFS
Now the significance of the WASF3 paper are clear: it's essentially the same state as the diabetic/obese mouse model:
Increased ER stress would normally be expected to turn down protein translation, but intriguingly, the inhibitory phosphorylation of protein translation factor eIF2α, a target of PERK kinase activity, was unexpectedly lower (i.e.,
activated) in patient S1 cells (Fig. 6A, lane 2 vs. 1).
And they found that inhibiting PP1, which I explain in the beginning of this post, was effective by directly intervening with the downstream molecules (namely eIF2α):
While the effect of TUDCA was less apparent as expected of a relatively nonspecific drug, the inhibitor of protein phosphatase 1 (PP1) salubrinal, which prevents the dephosphorylation of eIF2α thereby inhibiting protein translation, was more effective in decreasing the levels of both PERK and WASF3 in S1 cells (Fig. 6A, lanes 3-6 vs. 2).

Relation to ATG 13 paper (@SlamDancin 's question)
I don't pretend to understand the depths of this stuff well enough to give a confident answer here. But, here's what I found after googling some stuff.
(https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4849280/):
Inhibition of mTORC1 by rapamycin actually induces stress responses, including reduction in protein synthesis and induction of autophagy, which are protective mechanisms for the cells to survive under stress conditions
(https://academic.oup.com/nar/article/41/16/7683/2411270?login=false)
[...] recent data suggest that, analogous to yeast, the suppression of mTORC1 activity permits ULK1 to form an autophagy initiation complex with ATG13 (2). The phosphorylation of eIF2α, which also occurs with amino acid deprivation as well as with endoplasmic reticulum (ER) and other cellular stresses, similarly attenuates global protein synthesis (reviewed in (3)). However, eIF2α phosphorylation also paradoxically increases the translation of select mRNAs, including the transcription factor ATF-4 which transactivates the autophagy genes LC3B and ATG5
If I were to force a conclusion, maybe the upregulated ATG13 found in the ME/CFS paper (https://www.sciencedirect.com/science/article/abs/pii/S1044743122000379?via=ihub) could represent a parallel effort to attenuate protein synthesis. And, it would stay pathologically activated for the same reason that PERK remains pathologically activated: the downstream molecules aren't cooperating to get the job done.

Next Steps in Understanding

There are a lot of other proteins and "chaperones" mentioned in the WASF3 paper, like BiP, that I don't fully understand yet. I think some of these chaperones are intended to help with protein folding, which sort of addresses the "root" of the problem more than reducing global protein synthesis, which might be seen as more of a "workaround". But I'm not sure if this interpretation is quite right.
 

SlamDancin

Senior Member
Messages
511
I don't know if this question makes sense or not, but how could this topic, WASF3 disrupting mitochondrial respiration, pertain to people who can't even get out of bed?
I mean it’s a really basic illustration but yes, i think it would feel as if your muscles couldn’t breathe. As the WASF3 finding was in me/CFS muscle cells, I would imagine that muscles unable to use oxidative phosphorylation would have to rely on inefficient, and often undesirable, glycolysis for energy. Lactic acid is the byproduct of glycolysis, and when I was at my worst, I would have that lactic acid burning feeling every step up the stairs to my apartment on the rare case I had to go out, for example. From my very basic understanding of the science it seems to make sense with both my experience and others that are even more bedbound. Hope that helps.

There are those here though that report no muscle weakness or pain and no change in function, but still have extreme mental fatigue and PEM
 

Rufous McKinney

Senior Member
Messages
13,189
I would have that lactic acid burning feeling every step up the stairs
it feels like this nearly constantly, in certain muscle groups. They are just not getting oxygenated...somehow.

So I can't walk more than a block, and that has NOTHING to do with the concept of "FATIGUE "that so many are so obsessed with. As if thats the ONLY symptom?
 

SlamDancin

Senior Member
Messages
511
@Rufous McKinney I know exactly what you mean Rufous. My muscles were warped around all my bad connective tissue and as the mechanical stretch got worse, either the inability to get oxygen to the cells kicked off a hypoxemic-ER stress-WASF3 inhibition of mitochondrial respiration cycle or the mechanical stretch itself kicked off the ER stress response as I’ve found it does in several cell types. Either way, as I have over three years done incredibly slowly progressing physical therapy to unwarp my body, the ability to use the muscles has returned. I used to constantly feel like I was not getting enough oxygen systemically but also in the muscles and that has largely been fixed by the PT as well. I should mention I may have Marfan syndrome so it was obvious that my body wasn’t right even before the fatigue made it even more obvious
 

Dakota15

Senior Member
Messages
293
Location
Midwest, USA
Last note from Dr. Hwang

"My lab is focused on continuing our experiments to get additional insights into WASF3 while also trying to translate concurrently these insights into the clinics. I believe it is important to proceed carefully and do the best job possible at each stage, especially because I know how important the stakes are here. We have what we need to do our work at this time, but if and when additional needs arise, you can be sure that I will make requests to the appropriate sources."
 

godlovesatrier

Senior Member
Messages
2,502
Location
United Kingdom
RE talks on butyrate, when my butyrate was high in Febuary on my biomesight testing and my firmicutes were high and my bacteroides were low. Prausnitzii was also healthy for the first time ever I think. I improved to 90% for the first time in 7 years - I was very strong and able to withstand not only exercise without crashing quickly but also tolerated taking new supps and rebounded from high levels of exertion quickly. Here's a graph to show some of the data points:

Febuary: Butyrate 60% (prev 20%), Prausniztii 20% (prev 3%), firmicutes 68% (prev 20 to 30%), bacteroidetes 27% (prev 72%).

1696072683069.png


1696072628178.png

My markers are now nowhere near where they were in Feb everthings flipped again. I've tried everything to get back to where I Was without much luck. TUDCA might be a game changer, but it depends what effect it has on firmicute bacteria, it might kill them, if it does that won't help.

Prior to my 90% state I was eating legumes every single day and prior to that I was taking sodium benzoate 500mg with 500mg of glycine every few hours, which oddly seemed to improve my symptoms a lot. Sodium benzoate kills all kinds of microbiome bacteria, glycine has effects too although not sure what on. I plan to take TUDCA and do an additional microbiome test, might do the same with with benzoate and glycine.

looks like Ken L has one reference:

"bile acid therapy[tauroursodeoxycholic acid (TUDCA), or glycoursodeoxycholic acid (GUDCA)] normalized the colitis-associated increased ratio of Firmicutes to Bacteroidetes Interestingly, administration of bile acids prevented the loss of Clostridium cluster XIVa and increased the abundance of Akkermansia muciniphila, bacterial species known to be particularly decreased in IBD patients.”" https://pubmed.ncbi.nlm.nih.gov/28115375/

Just wanted to add my stats as several of you have discussed the same data points I've also found. I am convinced now that if I can get back to how I was in Feb I'd see 90% again.