Dolichol deprivation theory

[191]

How are you getting on?

[tryingnottoworry]

Hey guys, sorry for the late update. I’ve been really busy with work, taxes, and just life in general. I haven’t had time to properly sit down and dedicate some time for research, but I finally made some time and found some really amazing info.

First let’s look at a particular type of antibiotic called “Tunicamycin”. Here’s what Wikipedia has to say about it:

Tunicamycin

Tunicamycin is mixture of homologous nucleoside antibiotics that inhibits the UDP-HexNAc: polyprenol-P HexNAc-1-P family of enzymes. In eukaryotes, this includes the enzyme GlcNAc phosphotransferase (GPT), which catalyzes the transfer of N-actelyglucosamine-1-phosphate from UDP-N-acetylglucosamine to dolichol phosphate in the first step of glycoprotein synthesis. Tunicamycin blocks the synthesis of all N-linked glycoproteins (N-glycans) and causes cell cycle arrest in G1 phase. It is used as an experimental tool in biology, e.g. to induce unfolded protein response[1]. Tunicamycin is produced by several bacteria, including Streptomyces clavuligerus and Streptomyces Iysosuperficus.

Basically tunicamycin initiates a UPR by shutting down GPT, which completely inhibits glycosylation. Finasteride, however, inhibits SRD5A3 which reduces dolichol production and hinders GPT’s ability to function, which can also cause a UPR. Tunicamycin has a unique similarity to the actions of finasteride, and the myriad of research available on the effects of tunicamycin on glycosylation may help us to understand the underlying cause for our syndrome. Keep in mind, however, that tunicamycin isn’t a drug people can take, and is only used in in-vitro medical research as even small doses can be lethal. If you’d like an analogy: finasteride is to breathing thin air as tunicamycin is to getting your lungs shut down.

Genome-wide analysis of the unfolded protein response in fibroblasts from congenital disorders of glycosylation type-I patients

Using global transcriptome analysis, we have determined for the first time the scope and extent of the UPR in cells isolated from CDG-I patients. Our results indicate that endoplasmic stress in CDG-I cells is similar, though moderate, to the response induced by tunicamycin-mediated inhibition of N-glycosylation. The moderate UPR detected in CDG-I cells indicates that these cells do not accumulate unfolded proteins, which may lead to the collapse of the secretory pathway. Consequently, the symptoms associated with CDG do not appear to be caused by general glycoprotein folding and secretion.
Transcriptional activation of amino acid transporters was observed after tunicamycin treatment, after glucose deprivation, and in CDG-I cells, thus defining a new stress response in human fibroblasts representing a possible component of the UPR. The significance of this amino acid response is unclear. The increased amino acid supply may be required to compensate for the production of misfolded proteins in glycosylation-stressed fibroblasts. A key role in the long-term response of CDG cells is likely played by the p58IPK protein. Future work will address the exact role of p58IPK in maintaining protein translation and cell viability and determine whether it represents a target for therapeutical approaches aimed at attenuating the pathological consequences of chronic ER stress.

It’s very interesting that the secretory pathway may be one of the systems that gets damaged through ER stress. It’s also interesting how a CDG cell tries to compensate with additional amino acids. Studying these systems may lead to new understandings and treatments.

Let’s take another look at the Unfolded Protein Response (UPR).

[url=http://biology.ucsd.edu/classes/old.web.classes/bisp192.WI06.548055/Patiletal.pdf]Intracellular signaling from the endoplasmic reticulum to the nucleus: the unfolded protein response in yeast and mammals[/url]

Although the mammalian UPR still awaits comprehensive genomic analysis, recent identification of novel target genes suggests that similarly broad patterns may well exist in metazoans: the UPR positively regulates several secretory genes and processes, for example, the dolichol pathway essential for protein glycosylation [45], the ER calcium pump required for maintenance of calcium homeostasis [46] and SERP1/RAMP4, which stabilizes membrane proteins and facilitates their glycosylation [47]. Interestingly, mammalian cells upregulate asparagine synthetase in response to ER stress [48], suggesting that the mammalian UPR might also be involved in regulation of nitrogen metabolism.

This further elaborates on the effects of the UPR on gene expressions. Again, overexpression of GPT can potentially cause a psuedo-CDG.

THE MAMMALIAN UNFOLDED PROTEIN RESPONSE

CONSEQUENCES OF eIF2α PHOSPHORYLATION Phosphorylation of eIF2α by PERK shuts off general translation (12). perk −/− cells are sensitive to ER stress and are partially rescued by translation inhibitors, e.g., cycloheximide (119). Short-lived proteins are cleared from the cell during inhibition of translation. An important example is cyclin D1, which controls exit from G1 and entry into the S phase of the cell cycle through phosphorylation of the retinoblastoma tumor suppressor protein. Cyclin D1 activity is controlled through stimulation of its synthesis by mitogens and, thus, by its abundance in the cell. Loss of cyclin D1 during ER stress arrests mammalian cells in G1 (120). This explains the well-documented observation that depletion of dolichol phosphate and inhibition of N-linked glycosylation cause a G1 arrest in mammalian cells

So dolichol deprivation causes cellular G1 arrest. Let’s see what happens if G1 arrest is prolonged.

Prolonged arrest of mammalian cells at the G1/S boundary results in permanent S phase stasis.

S phase stasis may thus be a ‘trap’ in which cells otherwise competent for S phase have lost a key component required for replication and thus can neither go forward nor retreat to G1 status.

Well that sounds bad. This basically means that cells affected by this have lost their ability to divide, and cannot retreat back into G1 stage. Further research on this could help to understand the resultant effects on the human body.

Let’s take another look at 5 alpha reductase type 3 (SRD5A3).

Patients with Mutation in SRD5A3 Show an Early Defect in Lipid-Linked Oligosaccharide Synthesis

The absence of whole glycan chains on proteins indicated that the metabolic block occurred early in the N-glycosylation path-way, altering synthesis or transfer of the glycan part of lipid linked oligosaccharide (LLO), to recipient proteins. Epitope tagged SRD5A3 localized predominantly to the ER (Figure 2B), where LLO synthesis occurrs (Aebi and Hennet, 2001).
An abnormal composition of the glycan precursor impairs its transfer to acceptor proteins. Accordingly, we investigated the size of the LLO glycans using HPLC after [2-3H]-mannose metabolic labeling with fibroblasts from index patients CVH- 385-IV-11 and CVH-385-IV-13. Since no major structural abnormalities in LLO were detected (Figure S2E), we also determined the amount of radiolabeled LLO. We detected a severe reduction in the amount of newly synthesized LLO in four of the five patients tested compared to three control cell lines (Figure 2C), suggesting that the N-glycosylation block occurs prior the glycan transfer step.
The reduced levels of LLO could be explained by a limited availability of Dol-P. To test this hypothesis, we used an in vitro assay to assess the production of Dol-PP-GlcNAc1 and Dol-PP-GlcNAc2, the first two reactions of LLO synthesis. With fibroblast homogenates used as a source of enzyme and UDP- [14C]GlcNAc as glycosyl donor, all SRD5A3 deficient patient samples showed a reduced synthesis of Dol-PP-GlcNAc1/2 without addition of exogenous Dol-P, compared to controls (Figure 2D). However, when exogenous Dol-P was added to the incubation mixture, formation of Dol-PP-GlcNAc1/2 was increased to levels comparable with or even higher than controls. Fibroblasts from patients with other known CDG-I defects (CDG-Ik or CDG-Io) behaved comparable to controls, showing no evidence of Dol-P mediated rescue (Figure 2D). Elongation of Dol-PP-GlcNAc2 to Dol-PP-GlcNAc2-Man5 was unremarkable. Similarly, OST activity was normal (data not shown). Altogether, the rescue of the enzymatic GlcNAc transferases deficiencies by exogenous Dol-P indicates that the amount of Dol-P is limiting in the patients’ fibroblasts and suggests a defect in Dol or Dol-P biosynthesis.

It appears that by adding exogenous dolichol, CDG cells can begin functioning normally again. This is extremely promising! Since we don’t have an actual CDG, but potentially only a pseudo-CDG that is perpetuating itself due to being stuck in a loop of ER stress, supplementing exogenous dolichol may be the key to breaking us out of this cycle. I’ve theorized this before, and it could definitely explain how the spinach diet was helping me. However Ropren may potentially be a more effective treatment.

Srd5a3 Mutation Is Lethal in Mouse and Results in an Activation of the Unfolded Protein Response Pathway

We found that mice homozygous for a LacZ gene trap (Gt) insertion in intron 3 of Srd5a3 were recovered at embryonic stages, up to embryonic day 12.5 (E12.5) but not beyond (Fig- ures 4A and 4B). At E10.5, Srd5a3Gt(betaGeo)703Lex/Gt(betaGeo)703Lex Gt/Gt embryos (abbreviated Srd5a3 undergo axial rotation observed at E8.5 in WT littermates. Analysis with b-gal colorimetric staining in asymptomatic heterozygous carriers showed strong expression in the yolk sac, eyes, heart, and neural tube (Figures S4A–S4D). In keeping with this, homozygous mutants frequently presented dilated hearts (Figure 4A) and open neural tubes, which is consistent with the broad phenotypes observed in patients.
To identify the misregulated pathways underlying these developmental defects, we carried out expression microarray analysis in Srd5a3Gt/Gt embryos versus WT littermates before morphological differences appeared (Figure 4C). Whole-transcriptome analysis revealed that among the 50 most upregulated transcripts, 20% are involved in the regulation of the unfolded protein response (UPR) or are activated in this pathway (Tables S1 and S2). An activation of the UPR pathway in E8.5 Srd5a3Gt/Gt embryos was confirmed by real-time RT-PCR and with an E9.5 mouse embryonic fibroblast cell line treated with tunicamycin used as a positive control (activates the UPR pathway by blocking N-glycosylation; Figure 4D). A marker of this pathway, BiP, is upregulated at the transcript and protein levels in Srd5A3Gt/Gt embryos, with a particularly high expression in neuroepithelial cells (Figure 4E). Srd5a3 expression was not detected in mutant embryos; however, in cells inhibited for N-glycosylation by tunicamycin treatment, its expression increased significantly (Figure 4D). We also confirmed UPR activation by determining enrichment of gene ontology (GO) categories by all the genes significantly misregulated in Srd5A3Gt/Gt embryos compared with littermate controls. UPR was the biological process most significantly enriched for the genes upregulated (Table S3), whereas genes involved in general cellular metabolic processes and specific embryonic developmental program like regionalization were the most significantly downregulated (Table S3). These observations suggest that Srd5a3 is required for ER protein folding, a primary role of N-glycan during development.

This further elaborates on SRD5A3’s importance in glycosylation and its role in protein folding, as well as the UPR affecting SRD5A3’s expression.

Phenotypic Spectrum Resulting from Disruption of Dolichol Metabolism

Tissues affected in patients with SRD5A3 mutations, such as nervous system, ocular structures, skin, or coagulation factors, reflect sensitivity for alteration in N-glycosylation. Such congenital defects and the detection of a restricted expression pattern of Srd5a3 in mouse embryo suggest a spatial-temporal requirement during development. N-glycan number and branching regulate surface glycoprotein levels, affecting cell proliferation and differentiation (Lau et al., 2007). N-glycosylation may help
regulate specific developmental pathways yet to be discovered. A comparable multisystem disorder has been recently mapped to the same locus, suggesting that these patients have the same genetic defect (Kahrizi et al., 2009).
Although we find defects in the N-glycosylation pathway, dolichol is also required for the synthesis of O-mannose-linked glycans, C-mannosylation, and glycophospholipid anchor synthesis, and some of the pathology may derive from these defects, not explored here. Furthermore, little is known about the glycosylation-independent functions of dolichol, considered as a general membrane component in mammalian cells (Rip et al., 1985).
The pathogenesis and phenotypic specificity of CDGs deserves further investigations. However, our results point to an unsuspected role for a steroid reductase-like enzyme in the pathogenesis of one type of CDG, presumably mediated by a requirement for dolichol synthesis.

This elaborates on the relation between CDG, SRD5A3, and dolichol. It also explains how medical science’s current understanding of dolichol is needs to be further explored.

Biological Activity of SRD5A3

SRD5A3 sequence predicts a steroid 5a-reductase domain, and some enzymes with this domain are able to reduce a variety of steroid hormones with a delta4,5,3-oxo structure (Russell and Wilson, 1994). Mutation of the SRD5A2 gene in human causes male pseudohermaphroditism as a result of an enzymatic block of testosterone to dihydrotestosterone conversion (Andersson et al., 1991), and mutation of the Srd5a1 gene in mice affects reproduction and parturition, suggesting involvement in androgen metabolism (Mahendroo and Russell, 1999). Interest- ingly, a previous study suggested that cell extract containing overexpressed SRD5A3 was able to reduce testosterone to dihydrotestosterone (Uemura et al., 2008), albeit at a slow rate.
However, both our biochemical and clinical investigations in the patients with SRD5A3 mutations indicate that the nature of the substrate of the SRD5A3 enzyme is not related to the steroids. Our patients displayed no abnormal sexual abnormalities that would have suggested a primary defect of steroid metabolism. Moreover, karyotype analysis excluded the possibility of sex reversal in all (data not shown). These observations lead us to hypothesize that the in vivo substrate of SRD5A3 could be a different lipid. Polyprenols share a common origin with cholesterol because they are also built from isoprene units.
Another enzyme with a predicted steroid 5a-reductase domain, Tsc13/GPSN2, has been shown to be an enoyl reductase involved in the elongation of very long chain fatty acids (Kohlwein et al., 2001). This study also illustrates that the predicted steroid 5a-reductase domain is involved in the reduction of a nonsteroid lipid and suggests that the full spectrum of lipid reduction mediated by steroid 5a-reductase-like enzymes needs further evaluation.

This is super-fascinating! This article discusses the condition of “pseudohermaphroditism”, which is caused by a defective SRD5A2 enzyme, and was the disorder that finasteride was originally modeled after. It explains how a SRD5A3 disorder is unrelated to a SRD5A2 disorder, however in Merck’s “brilliance”, they designed a drug that inhibits both SRD5A2 AND (unwittingly) SRD5A3. THOSE BUTTFACES.

Residual Dolichol in SRD5A3 Mutants

Dolichol was still detected in human, mouse, and yeast SRD5A3/ DFG10 mutants, suggesting the existence of another de novo biosynthetic pathway for dolichol production. The presence of dolichol in these mutants is not explained by dietary contribution, which was reported to be negligible in rat (Keller et al., 1982), and the nature of the mutations in human, mouse, and yeast suggests that these organisms have null mutations for this gene. These observations indicate the existence of an alternative pathway for de novo synthesis of dolichol in eukaryotic cells. Disruption of LLO biosynthesis due to mutation in Dpagt1 in mouse results in embryonic lethality at E5 (Marek et al., 1999), a more severe phenotype than that observed in Srd5a3 mutant mouse embryos, consistent with an alternative pathway. One candidate is the TSC13 gene, the only other gene in S. cerevisiae encoding a steroid 5a-reductase domain (Pfam database). We tested whether the tsc13 mutant had abnormal CPY glycosylation and whether the dfg10/tsc13 double mutant showed further increase of the polyprenol/dolichol ratio, but found no effect of either (Figure S3), suggesting that the alternative pathway for dolichol synthesis is independent of these genes. Interestingly, among the pathways activated in embryonic mouse mutants was the mevalonate pathway, including the isoprenoid biosynthetic enzymes (Tables S1 and S2). This could suggest a positive feedback mechanism, which might help organisms overcome a partial block of these pathways.

Holy crap, this is amazing! This suggests that alternate pathways of dolichol (mevalonate is the polyprenol precursor to dolichol) can allow an organism to overcome a block on these pathways. It’s possible we may lack the alternate pathways which prevent dolichol deprivation from happening. This could explain why some people are symptom free from Finasteride, and why we got the shaft.

It’s been about two and a half weeks since my last progress report, and I’ve been doing really well. I think I was on the verge of an iron overload when I was supplementing 500mg of vitamin C every day with my spinach. I was throwing up, having muscle pain all over, and having a fast & weak heart rate, but these symptoms slowly went away after a week or two of discontinuing vitamin C supplements. I’ve also been slowly weening off of the spinach diet (I decided to ween off extra slow), and I’m down to eating about 1/8th of a bag a day. I’ll probably stop it completely later this week. I’m actually starting to feel amazing again, as my libido and energy are continuing to improve. Erections are frequent, easily maintained, sensitivity is improving, kegel contractions seem to be stronger and in a better rhythm during ejaculation, and I’m just feeling great in general.

The original theory of the spinach diet (based off of the dolichol deprivation theory) was that by adding exogenous dolichol by eating abnormally large amounts of spinach, you could potentially break the cycle of a psuedo-congenital disorder of glycosylation caused by the overexpression of GlcNAc phosphotransferase caused by stress of the endoplasmic reticulum caused by dolichol deprivation caused by the inhibition of SRD5A3 caused by finasteride. The idea was that if you could overcompensate with dolichol, you could potentially convince the ER to reverse the overexpression of GPT. Symptomatic wise, throughout this treatment I went from feeling terrible to feeling great, to starting to feel lousy again, and then slowly back to feeling well again. Could this be the end of this nightmarish rollercoaster for me? I suppose only time can tell.

I’ll keep you guys updated whenever I can, and please feel free to help me further the research of the dolichol deprivation theory (I suppose a more suitable name at this point would be the psuedo-CDG theory, but dolichol deprivation will always have a special place in my heart ) Thank you again for your prayers and support, and happy Easter!

[gefinauser]

Yeah, that sounds like a sound theory. Good stuff there TNNTW! Definitely keep us up to date about your condition. Especially now that the time that i get into next bloodtests gets prolonged, i’m more and more concerned of the effects of the prolonged pfs (or…glycosylation disorder). It’s important to know whether your recovery is sustained.

[191]

I think there is definitely something to explore here. Since stopping spinach i have declined in function even more so. Semen has become watery again and there has been some shrinkage. Spinach was not as effective as doxy for me but this could be a dose related thing. The problem with spinach is that we can’t eat too much of it because of iron problems. I would like to able to try a very large dose but not sure this is possible. Anyone know how much is safe to eat?

Good work tntw!

Just so everyone knows I support any theory with a degree of evidence supporting it. I know I have been a big contributor to the epigenetic thread but I try not to hone in on one theory alone as to do so would be naive. This theory has some weight behind it now. I hope people pay more attention to this.

“These observations suggest that Srd5a3 is required for ER protein folding, a primary role of N-glycan during development.”

Fin inhibits SRD5A3. I have postulated that proteins are where the problem is in another thread and this idea would support that notion.

[Tpenf]

I am trying this, only my 2nd day. I was bored yesterday so started playing around with trying to make this a bit more tasty… I have come up with… Spinach, apple, banana and lemon juice blended with water! Tastes pretty good imo might try adding some fresh mint also. Hope this helps. Cheers

[tryingnottoworry]

@19

The problem with spinach is that we can’t eat too much of it because of iron problems. I would like to able to try a very large dose but not sure this is possible. Anyone know how much is safe to eat?

Possibly. Try eating it alongside iron absorption inhibitors such as milk, calcium, magnesium, and oil of oregano. Avoid eating things with citric acid or vitamin C at the same time. Vitamin C supplements or multivitamins that include Vitamin C can boost iron absorption and prevent iron depletion, so avoid taking those right before, during, or right after eating your spinach. If you need to take them, I’d probably wait at least an hour before/after any meals.

@Tpenf

I am trying this, only my 2nd day. I was bored yesterday so started playing around with trying to make this a bit more tasty… I have come up with… Spinach, apple, banana and lemon juice blended with water! Tastes pretty good imo might try adding some fresh mint also. Hope this helps. Cheers

I definitely think it’s good to mix up your recipes. Eating the same thing day in and day out can get a little tiring, so finding what tastes best to you will definitely make it a less grueling process. Just keep in mind that citric acid and Vitamin C can boost iron absorption, so be sure to pay attention for signs of iron overload.

[191]

Thought you might find this interesting.

mendeley.com/research/effects-anabolic-androgenic-steroids-serum-ubiquinone-dolichol-levels-among-steroid-abusers/

The effects of anabolic androgenic steroids on serum ubiquinone and dolichol levels among steroid abusers.

We measured serum ubiquinone and dolichol concentrations in 13 men while they abused anabolic androgenic steroids (AAS) and during the following withdrawal period. Serum total, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) cholesterol and triglycerides were also determined. AAS administration increased serum ubiquinone by 68% (P < .001) and decreased serum dolichol by 30% (P < .002). Both nonsterol isoprenoid levels in plasma correlated with the AAS dose, ubiquinone positively (P < .001) and dolichol negatively (P < .002). When the subjects were taking steroids, the ubiquinone to LDL ratio was 42% higher than during the withdrawal period. In conclusion, our study suggests that AAS have an influence on the by-products of the mevalonate pathway.

[troubledfinuser2]

[qoute]
5 alpha reductase type 2 has been, for the longest time, the focal point of our research and understanding of PFS, but it isn’t until now that we can see the effects of inhibiting 5 alpha reductase type 3. Unlike what most other theories suggest, a glycosylation disorder would be our “root” cause, as it is upstream of basically every higher function in the body, including cortisol production, thyroid function, liver function, hormone function, and nearly everything else. Every other theory may address the cause of our symptoms, but what’s causing the cause? For example, we almost certainly get many of our symptoms due to a dysfunctional endocrine system, but what is causing our endocrine systems to be (and remain) dysfunctional? Why do treatments not correct the problem like they should? Why does TRT not solve our problem? Why do we have such a wide range of symptoms? Why can anti-biotics cause a positive change? Why can tyrosine cause a positive change? Why do we share many of the same symptoms as statin users? Why does caffeine and alcohol affect us? A glycosylation disorder, brought on by an altered gene expression, brought on by dolichol deprivation, caused by finasteride inhibiting SRD5A3 explains everything, and has a direct link through and through.
[/qoute]

Lots of good questions that likely will never be awnsered satisfactory. Perhaps some get better from tyrosine (ive never tried it) because it potentially changes 3b hsd activity and thus dht metabolism?

ncbi.nlm.nih.gov/pubmed/21166213

[tryingnottoworry]

I was asking those questions rhetorically. If you read further back in this thread you’ll see I already addressed them.

To answer the red highlighted question, see page 2.

[tryingnottoworry]

@19

Thought you might find this interesting.

mendeley.com/research/effects-anabolic-androgenic-steroids-serum-ubiquinone-dolichol-levels-among-steroid-abusers/

The effects of anabolic androgenic steroids on serum ubiquinone and dolichol levels among steroid abusers.

That’s a pretty interesting connection. It’s well known that steroid abuse can screw you up pretty badly, though finasteride is probably a more potent dolichol depriver since it directly inhibits the enzyme that produces it. It certainly makes you wonder why steroid withdrawal can deplete dolichol, and how this aspect affects the post steroid syndrome as a whole. That article was written 15 years ago, I wonder if they’ve found any connections to SRD5A3 since then, or if it just disrupts the mevalonate pathway like statins do.

[191]

Its so hard to pinpoint where tyrosine helps as it affects so much. Its a precursor to many things so I wouldn’t rest theories on the basis of tyrosine.

I think its interesting to see why taking hormones can be counterproductive.

[tryingnottoworry]

Tyrosine is involved with O-Glycosylation, which is further down the pathway from N-Glycosylation. If N-Glycosylation is disrupted by a psuedo-CDG, then O-Glycosylation will be disrupted as well. Supplementing tyrosine may be temporarily patching the deficiency left by the disruption of N-Glycosylation.

However there probably exists complexities and interactions none of us can explain with certainty, as we’d need proper research studies to properly understand the scope of our condition. We’ve only recently discovered the glycosylation link to finasteride (less than 2 months ago), so there’s probably still a mountain of complexities still yet to be uncovered. However I feel like we’re finally looking in the right place, and hopefully we can solve this syndrome once and for all.

[191]

Yes it is but it also is involved in lots of other things. For example: Tyrosine kinases are involved in neurotrophic factor-mediated AR activation.

viewtopic.php?f=5&t=3901&start=280

My point is it may fit in with a theory but it doesn’t make the theory true. Thats all.

[tryingnottoworry]

Gah, you ninja’d a reply before I could finish editing my post

Indeed. We can find connections to why it can help, but there’s almost certainly far more things going on that we don’t know about. From what we’ve gathered from our own anecdotes, tyrosine has given people complete recoveries, but they’re always temporary. It leads me to believe that the underlying problem would be deeper than what tyrosine is involved with, yet still somehow connected to the root problem (such as N-Glycosylation). Also keep in mind that if Tyrosine kinase was being disrupted by a pseudo-CDG, then it would also disrupt its functionality of neurotrophic factor-mediated AR activation. These theories certainly aren’t mutually exclusive. But that’s about the extent of my understanding. My iMac can do wonders, but it can’t test cellular functions

[191]

Yes i agree what we really need is to look into these in a lab. However, us doing the lit review for them would help a bit. Also it gives us a place to look. I am confident that if a lab dedicated time to our cause we could have an answer relatively soon. Your improvement says something for this idea. I am not a fan of hormone driven treatments as I think most of us have normal hormones with the occasional thing out of balance. Also in my own experience of messing with hormones (tribulus - increased sexual function but felt AWFUL, Chrysin - did nothing, tamoxifen - did nothing) it never has helped - if anything made things worse. The problem lies at the epigenetic or protein level. Also my improvement on doxycycline and brief improvement on spinach points away from hormones themselves.

I’m not doing so well at the moment. Everything has got worse - shrunken up, dry skin, poor sleep, very tired. When I cam off doxy I managed to preserve some function somehow and then the spinach helped a little and then I stopped taking it consistently and continued to workout and things got worse. I don’t think I can increase the dose enough of dolichol to make a substantial difference (if this theory holds true). You proved to be quite sensitive to it. I think some of us especially the more seriously affected need a dose a lot higher. Ropren is too expensive at the moment and I’m not sure I could cope with two - three bags of spinach. The iron and vitamin A would be too much!

[gefinauser]

Yeah, i too know a person who have recovered pretty well and he says that tyrosine is one of the few things that actually help him out when taken. Mike Tyson or… My Tyrosine? HMMM.
Though this troubles me:

Lots of good questions that likely will never be awnsered satisfactory. Perhaps some get better from tyrosine (ive never tried it) because it potentially changes 3b hsd activity and thus dht metabolism?

ncbi.nlm.nih.gov/pubmed/21166213

[troubledfinuser2]

@gefinauser

Yeah, i too know a person who have recovered pretty well and he says that tyrosine is one of the few things that actually help him out when taken. Mike Tyson or… My Tyrosine? HMMM.
Though this troubles me:

Lots of good questions that likely will never be awnsered satisfactory. Perhaps some get better from tyrosine (ive never tried it) because it potentially changes 3b hsd activity and thus dht metabolism?

ncbi.nlm.nih.gov/pubmed/21166213

Well that and the fact that many here have hormone abnormalities suggesting increased 3b hsd activity does make one think that the benefits from tyrosine could stem from just that.

Correct me if im wrong but low 3adiol g could be from low 3b hsd activty or high 3b hsd activity since it actually converts dht to 3adiol g and then back aswell (remember reading it could go both way?)

Increased progesterone, 17Oh progesterone and increased androstendione (seen in our bloodtests) could stem from 3 things the way i c it.

1- Increased acth stimulation. This would however show an elevation of all adrenal hormones and especially the end product cortisol wich most of us do not have. Thus i consider this very unlikely.

2- Decreased enzyme activity after progesterone, 17 oh progesterone and androstendione in the cortisol pathway. This would be a form of CAH or in our case since its most lilkely not a birth defect just Adrenal hypertrophy. You can test for this btw so it can be excluded.

3-Increased 3b hsd activty. Thus elevated conversion of Pregnenlone–> Progesterone, 17oh pregnenolone–> 17oh Progesterone and elevated conversion of DHEA—> Androstendione. This im not so sure you could test for but what other reasons could there be for elevated androstendione, prog and 17oh prog othor than CAH or elevated 3b hsd activity? Well tumors could cause increased hormone production but this seems highly unlikely considering so many of us have this and that the elevations seem to be pretty much only 3b hsd reduced hormones.

[191]

How much are you eating? Be wary of vitamin A toxicity.

[191]

Has anyone else tried spinach for a period of time? It has not been helping me recently at all. I am in bad shape.

[gefinauser]

I’m on it currently, second day in. Are you in bad shape because of the spinach or is it PFS? also uh didn’t spinach treatment actually help you out when you tried it? I thought i saw you posting about the mental sides getting easier on it.

EDIT a reminder for myself:

“Alpha Lipoidic Acid (ALA) can be found in foods such as meats and vegetables, especially spinach. It is easily absorbed into the bloodstream, and it can also cross the blood brain barrier. Two of the best natural sources of lipoic acid are yeast and liver, but the body can synthesize it when necessary and it is available as a supplement (Bodybuilding.com).”