Low Toxin Diet Grant Genereux's Theory Of Vitamin A Toxicity

[Travis]

Very interesting case studies. I'll be taking a closer look at them in time, but for now I'd say it's especially interesting to ponder the ones which show eye problems and night blindness from isotretinoin as those are classic symptoms purported to be caused by Vitamin A deficiency.

Grant also quoted (p 108 of PFP) some other research that confirms these as side effects of isotretinoin:

"Effects on vision and central nervous system. In addition to the xerophthalmia commonly experienced, and meibomian gland atrophy and corneal opacities reported with isotretinoin use, photophobia and decreased dark adaptation/night blindness can also occur. The loss of the dark adaptation maybe permanent."

Source: Retinoids and Carotenoids in Dermatology Anders Vahlquist, Madeleine Duvic
CRC Press, Jun 20, 2007 – Medical
See page 116​

This gets at the paradox that Grant highlights: the illogical explanation that both too little and too much VA could result in the exact same symptoms (e.g. night blindness and xerophthalmia).

Some will say: But there are many cases where too much and too little both cause problems!

Okay, but where does too much and too little cause the exact same problems?

For example, too much water and too little water both cause problems, but not the exact same problems — in fact, they cause the opposite problem — and the treatment will be the opposite — which is would you'd logically expect.

Imagine if eating too much and eating too little both caused people to become emaciated. That would be odd, wouldn't it? What we expect to see is that eating too much makes you obese and starvation makes you emaciated.

We logically expect opposite actions to yield opposite results. So when opposite actions (deficiency and excess) yield the same result — that's illogical, and we should examine that closely because there is likely a major error somewhere.

Grant's simple explanation: it's not a paradox — there was simply a mistake, a blunder, a human error which lead to a false conclusion.

It was never a deficiency in VA that caused these problems, it was always excess retinoic acid — which you can get by direct RA intake or by the natural conversion of VA to RA via chronic excess VA intake.

The documented side effects of isotretinoin and tretinoin establish that RA directly causes these symptoms, so to explain this "paradox" all we need to demonstrate is that in every study where a "VA deficiency" was purported to cause "deficiency" symptoms, there was a plausible source of excess VA or RA in the diet and/or a lack of protection from it.

This does seem interesting. Although he seems wrong about certain things, the book could also of course be full of insightful and novel points. He seems to assume that retinol oxidized in the manner of the classic studies he cites becomes retinoic acid, yet it seems reasonable that retinoic acid would be a minor product of nonenzymatic oxidation. However, many other oxidation products would be similar in form and could have retinoid activity. The symptoms observed in the classic studies—officially assumed 'vitamin A deficiency'—could have been due to an unspecified retinol oxidation product, or a few different ones.

 

[Travis]

Most of the things you mentioned are also related to vitamin D deficiencies.


Do you not think the amounts of vitamin A discussed by Grant etc are ridiculous though? For example I struggle to think that 100 IU vitamin A could cause people massive issues, unless something else is at play, for example severe vitamin D deficiencies. I could see excessive amounts having problems though I get that bit

I think only if a person's liver is saturated or near-saturated because 100 IU is a relatively small dose. Some dietary retinol appears to be incorporated into liposomes that bypass the liver and enter circulation. The liver appears responsible for gauging serum retinol and also storing any excess as triglyceride esters with ostensible intent of keeping serum retinol in a very tight range. A person having a retinol-saturated liver could perhaps expect otherwise-minor amounts to induce a substantial increase over the mean, leading to high peripheral retinol and the classic signs of skin peeling. Autopy data reveals that liver retinol ranges from zero to 800 μg/g. The liver appears to take-up the burden of any excess retinol, and its concentrations vary far more than serum retinol—which ranges only threefold between 95% of subjects. Since the liver retinol concentration has been determined as being literally zero in apparently-normal subjects, it is difficult to gauge where pathology begins and ends. Hypervitaminosis A is defined by serum retinol concentrations, and liver biopsies can be seen as invasive.

I don't think anyone would argue that humans have unnatural eating patterns, and looking at the liver concentrations of our evolutionary near-relatives could be a worthwhile approach. A person who believes 'humans are omnivores' would likely focus on different animals, and might even take the substantial amount found in polar bears as proof of liver retinol safety. Perceptions about retinol could depend somewhat on a person's dietary bias, perhaps why the issue is so contentious.

 

[InChristAlone]

I do want to get through his book at some point but I honestly don't want to go back on another restrictive diet. His diet is extremely restrictive. Extremely. People with a history of eating disorders could relapse with this fear of vitamin A.

His low cholesterol is worrying to me. Hopefully as he gets old it will get higher as he would have a great chance of dieing of an infection with it that low in old age. Maybe he isn't thinking that long term. All he cares about is having good skin. I'd like to see his thyroid numbers and vitamin D. His pulse is low. I honestly just think he was suffering hypothyroidism and low vit D. No wonder he feels so much better, he eliminated probably the foods he was reacting so strongly to and added in some nutrient dense food like meat. But clearly he's not converting to t3 very well or his pulse would be higher.

 

[raypeatclips]

I'd like to see his thyroid numbers and vitamin D. His pulse is low. I honestly just think he was suffering hypothyroidism and low vit D.

I've been wondering and asking this since page 1 of this thread, although a book that simply reads "Oh, I had low vitamin D" Probably isn't radical enough!

 

[InChristAlone]

I've been wondering and asking this since page 1 of this thread, although a book that simply reads "Oh, I had low vitamin D" Probably isn't radical enough!

Yep! Plus if he did figure it out he'd have to come out and say he messed up, but when you are that invested in your theory being right it can be really hard to admit you were wrong. The D would explain why northerners would get bone problems.

And I'll say it, how long could it take to shows signs of A deficiency? Could be 10 or more yrs. He's at 4. Not long enough in my opinion. Plus with low t3 he won't be using up stores very fast. So he could just be running on the A he does have.

 

[charlie]

I find it interesting that the Coimbra Protocol has a 95% success rate with auto-immune conditions and they use high doses of vitamin D and no vitamin A at all. @Makafre is on the protocol and is having success with healing his MS.

 

[raypeatclips]

I find it interesting that the Coimbra Protocol has a 95% success rate with auto-immune conditions and they use high doses of vitamin D and no vitamin A at all. @Makafre is on the protocol and is having success with healing his MS.

+1 I believe Vitamin D is the missing link in all of this, and none of the, Vitamin A is the issue crowd, have yet given a solid answer as to why this is wrong IMO.

 

[postman]

I just had some liver and it powerfully decreased my multiple chemical sensitivites. Talking about burning or the throat and nose, brainfog, when being exposed to dust, laundry detergents, perfumes, computer fumes etc.

I had the same experience when I took fermented cod liver oil so I'm pretty sure it's the retinol that is the factor. Why would retinol be helpful for this, what would be the proposed mechanism @Frank Why do some people get strong symptom relief from various things when consuming retinol?

 

[Travis]

Linked below is a short review article, followed by some of its' best quotes:

Penniston, Kristina. "The acute and chronic toxic effects of vitamin A." The American journal of clinical nutrition (2006)

'The acute and chronic effects of vitamin A toxicity are well documented in the literature. Emerging evidence suggests that subtoxicity without clinical signs of toxicity may be a growing concern, because intake from preformed sources of vitamin A often exceeds the recommended dietary allowances (RDA) for adults, especially in developed countries. Osteoporosis and hip fracture are associated with preformed vitamin A intakes that are only twice the current RDA.' ―Penniston

'The cleavage of provitamin A carotenoids to retinal is a highly regulated step, and vitamin A toxicity from provitamin A sources is largely impossible. In contrast, absorption and hepatic storage of preformed vitamin A occur very efficiently until a pathologic condition develops.' ―Penniston

'Retinol is absorbed by intestinal epithelial cells, where it is reesterified to long-chain fatty acids and incorporated into chylomicra, which circulate in the intestinal lymph before moving into the general circulation. As the chylomicra lose their triacylglycerol and other constituents, most of the retinyl esters remain within, and the particles become chylomicron remnants. Chylomicron remnants are cleared mainly by the liver,' ―Penniston

'Retinyl esters in serum are normally below 0.2 μmol/L in the fasting state (8), but they increase significantly after a large influx of vitamin A, such as occurs after a vitamin A–rich meal.' ―Penniston
​

Even though serum retinoic acid concentrations remain in a relatively tight range as compared to most other vitamins—as does mean serum retinol values within populations—there still can be a significant postprandial increases of individual serum retinol concentrations.

'Research on vitamin A toxicity has been carried out primarily in animals, and most studies have been short-term and have focused on acute effects (11–13).' ―Penniston

'In developed nations, the increasing availability of and interest in fortified foods and supplements resulted in a large percentage of the population with preformed vitamin A intakes higher than recommended (16). Indeed, observational studies suggest that more than 75% of people may be routinely ingesting more than the recommended dietary allowance (RDA) for vitamin A, much of it as preformed vitamin A (16). Until recently, little research investigated hypervitaminosis A in these situations' ―Penniston

'Fossilized skeletal remains of early humans suggest that bone abnormalities may have been caused by hypervitaminosis A (17, 18)' ―Penniston

'The range of serum retinol concentrations under normal conditions is 1–3 μmol/L (2), and, because of homeostatic regulation, that range varies little with widely disparate vitamin A intakes (10). Case reports of vitamin A toxicity have shown serum retinol concentrations within normal limits (22–24), which suggests that serum retinol is not a good measure of vitamin A status during toxicity.' ―Penniston

'A postprandial increase in serum retinol concentration may be blunted when vitamin A is ingested with either food or ample dietary fat, whereas a significant amount of free (unesterified) retinol may circulate when vitamin A is consumed without dietary fatty acids, which leads to excessive production of retinoic acid (28).' ―Penniston
​

Consuming retinol without fatty acid trigylcerides results in higher retinoic acid concentration, likely because the retinol then has no glyceride esterify-with the intestines. Ostensibly, free retinol and/or it glucorinide are/is better substrate(s) for retinal oxidase than retinol triglyceride or retinol cholesterol esters. This is actually in concord with data on supplementation and the risk of bone fracture; water-soluble forms of retinol are consistently found greater risk factors.

'An acute elevation of retinoids other than retinyl esters—eg, retinoic acid—occurs after the ingestion of a large amount of vitamin A, possibly because the intestinal absorptive capacity is overwhelmed, which leads to the oxidation of retinol to retinoic acid by the intestinal enterocytes (30) and to the rapid formation of retinoic acid from retinol in certain cells (5). Whereas retinoic acid can be produced from excentric cleavage of -carotene in humans (31), it is generally considered a minor contributor to circulating concentrations, at least in normal, healthy persons.' ―Penniston

'Retinyl ester metabolism differs between species. Studies by Schweigert (46) suggested that strict carnivores and birds have a high percentage of vitamin A circulating as retinyl esters, and this appears to be a major way of transporting vitamin A in some species. In contrast, herbivores and omnivores normally circulate vitamin A as retinol bound to RBP. Elevated amounts of retinyl ester in the fasting state have been used as markers for chronic hypervitaminosis A in humans and monkeys.' ―Penniston

'Hepatic storage of vitamin A will continue until a pathologic liver condition develops (20, 57). The vitamin A storage capacity of other tissues, however, has not been fully investigated.' ―Penniston

'In fact, adipose tissue is an important storage site for retinol (62). In normal chow-fed rats, the amount averaged 20 nmol vitamin A/g adipose tissue as both retinol and retinyl esters (63). Because adipose tissue and liver represent 15% and 4%, respectively, of the total body mass of normal-weight rats, the total amount of vitamin A in the adipose tissue is ~14% of that in the liver, which contributes significantly to total body stores in normal-weight animals.' ―Penniston

'Extrahepatic tissue vitamin A is characterized in humans (56), small mammals (58–61, 63–66), fish (55), and birds (67, 68). Most studies have focused on concentrations in the kidneys and the lungs, and a few have assayed other tissue. Results of these studies suggest wide interspecies variations in vitamin A concentrations in extrahepatic tissues.' ―Penniston

'The vitamin A concentrations in all tissues ranged widely between the subjects in the study by Schmitz et al, and not all persons were healthy at the time of death. Most of the samples were from persons who had heart disease, cancer, or infections, and these conditions have unknown effects on the distribution of vitamin A among tissues. Therefore, true normal values are unknown.' ―Penniston

'Clinical observations, such as hypercalcemia and elevated alkaline phosphatase, in persons with vitamin A toxicity clearly suggested that vitamin A affects bone. Synthetic retinoids in humans are reported to alter bone metabolism and increase turnover (95, 96). Case reports in children with hypervitaminosis A revealed altered skeletal development (22, 43, 97–100).' ―Penniston
​

In my view, retinoic acid appears to increase protein turnover of bone faster than it can be mineralized. Although retinoic acid and vitamin D antagonize each other nuclear receptor level, there may be pharmacokinetic constraints placed on vitamin D that prevent it from attaining effective relative concentrations.

'Binkley and Krueger (92) noted the consistent occurrence of spontaneous bone fractures associated with hypervitaminosis A and also noted that no compound other than vitamin A is known to be associated with such fractures in animals.' ―Penniston

'Four large, prospective, observational studies (83– 86), conducted in Scandinavia and the United States, where the incidence of osteoporosis is high (108), found associations between preformed vitamin A intake and hip fracture or osteoporosis. The studies differed in many design factors (eg, subject sex and age and years of follow-up), which makes direct comparisons difficult (Table 3). Nonetheless, the findings generated much interest, because calcium intake is generally high in both these areas. It is interesting that the amount of dietary vitamin A associated with this effect was relatively low at 1500 RE, which is much lower than the amount traditionally associated with risk of toxicity and lower than the tolerable upper intake level (ie, 3000 RE),' ―Penniston

'These studies suggest that intakes much lower than 10 times the RDA, the amount conventionally thought to lead to toxicity (2, 20), are needed to increase risk for osteoporosis—ie, ~2 × RDA.' ―Penniston

'The deleterious effects of vitamin A deficiency are known, but further research is needed to ascertain whether subclinical toxicity exists and, if so, what are its effects on overall health and well-being.' ―Penniston ​

 

[Terma]

In my view, retinoic acid appears to increase protein turnover of bone faster than it can be mineralized. Although retinoic acid and vitamin D antagonize each other nuclear receptor level, there may be pharmacokinetic constraints placed on vitamin D that prevent it from attaining effective relative concentrations

There's a lot of focus on Vit D for historical reasons, but what other receptor levels do you think are good candidates to compete with RAR? Anything that dimerizes with RXR? As these guys seem to suggest:
Sci-Hub | Vitamin D and adipogenesis: new molecular insights | 10.1111/j.1753-4887.2007.00004.x
Sci-Hub | Counteraction of retinoic acid and 1,25-dihydroxyvitamin D3 on up-regulation of adipocyte differentiation with PPARγ ligand, an antidiabetic thiazolidinedione, in 3T3-L1 cells | 10.1016/s0024-3205(98)00059-9

Thus, these observations are consistent with the hypothesis that the vitamin D receptor inhibits adipogenesis by blocking PPAR g activity, at least in 3T3-L1 cells, by a mechanism apparently due to VDR monopolizing the limiting amounts of shared endogenous RXR.

 

[Travis]

There's a lot of focus on Vit D for historical reasons, but what other receptor levels do you think are good candidates to compete with RAR? Anything that dimerizes with RXR? As these guys seem to suggest:
Sci-Hub | Vitamin D and adipogenesis: new molecular insights | 10.1111/j.1753-4887.2007.00004.x
Sci-Hub | Counteraction of retinoic acid and 1,25-dihydroxyvitamin D3 on up-regulation of adipocyte differentiation with PPARγ ligand, an antidiabetic thiazolidinedione, in 3T3-L1 cells | 10.1016/s0024-3205(98)00059-9

There's of course the classic antagonism of the nuclear receptors when competing for the same dimer partners, yet it's also curious that both vitamin–hormones bind to the same receptor (RORγ). This class of nuclear receptor had been named 'retinoid orphan receptors' before it's activating ligand had been known, ostensibly on account of RAR homology. The transcribing ligand for all three subforms is now known to be melatonin, yet RORγ is antagonized by the vitamin–hormones A & D. The β-subtype is antagonized only by retinoic acid, while the α-subtype is inhibited by mono- and di-hydroxylated forms of vitamin D (noncalcemic). Since all RORs have the same response element and all three subtypes are found in bone marrow: vitamins A & D could then antagonize eachother though antagonizing, in turn, their respective ROR subtypes. If RORβ were reduced in activity by increasing retinoic acid levels, the two other ROR subtypes should increase in activity by the corresponding amount. Even though this is a different mechanism—i.e. a reciprocal antagonism of ROR-subtypes—than classic dimer competition, it can sill be seen as an instance of vitamin A & D antagonism. Since all RORs subtypes compete for the same dNA binding domains they can be seen to antagonize the rest of their class. Because a molecule that antagonizes an antagonist can be considered an agonist, it's double negative, retinoic acid can then perhaps be seen as a potentiator of RORα. And in a likewise fashion, vitamin D could be expected to increase the activity of RORβ. Below is an excerpt of article I am writing:

[Image here depicting Western blot of melatonin-induced expression in bone cells.]​

'There is a considerable amount of redundancy between the transcriptional activity of RORα and RORγ, and so much so that you might wonder ‘why there are even two separate receptors at all?’ Although these two receptors have the same core response element (GGTCA) and can transcribe many of the same dNA sequences, they are differentiated by their antagonists: Retinoid orphan receptor alpha (RORα) can be bound and repressed in nanomolar concentrations by calcidiol, calcitriol, and 24-hydroxycholesterol. Retinoid orphan receptor beta (RORβ), on the other hand, is strongly inhibited only by trans-retinoic acid in the picomolar range; this receptor can be found in the retina, pineal gland, suprachiasmic nucleus, pituitary, and in stem cells—yet most often will loses expression upon differentiation. The retinoic orphan receptor gamma (RORγ) is antagonized by both retinoic acid and the calciferols, a peculiarity which...' ―Travis​

 

[Terma]

There's of course the classic antagonism of the nuclear receptors when competing for the same dimer partners, yet it's also curious that both vitamin–hormones bind to the same receptor (RORγ). This class of nuclear receptor had been named 'retinoid orphan receptors' before it's activating ligand had been known, ostensibly on account of RAR homology. The transcribing ligand for all three subforms is now known to be melatonin, yet RORγ is antagonized by the vitamin–hormones A & D. The β-subtype is antagonized only by retinoic acid, while the α-subtype is inhibited by mono- and di-hydroxylated forms of vitamin D (noncalcemic). Since all RORs have the same response element and all three subtypes are found in bone marrow: vitamins A & D could then antagonize eachother though antagonizing, in turn, their respective ROR subtypes. If RORβ were reduced in activity by increasing retinoic acid levels, the two other ROR subtypes should increase in activity by the corresponding amount. Even though this is a different mechanism—i.e. a reciprocal antagonism of ROR-subtypes—than classic dimer competition, it can sill be seen as an instance of vitamin A & D antagonism. Since all RORs subtypes compete for the same dNA binding domains they can be seen to antagonize the rest of their class. Because a molecule that antagonizes an antagonist can be considered an agonist, it's double negative, retinoic acid can then perhaps be seen as a potentiator of RORα. And in a likewise fashion, vitamin D could be expected to increase the activity of RORβ. Below is an excerpt of article I am writing:

[Image here depicting Western blot of melatonin-induced expression in bone cells.]​

'There is a considerable amount of redundancy between the transcriptional activity of RORα and RORγ, and so much so that you might wonder ‘why there are even two separate receptors at all?’ Although these two receptors have the same core response element (GGTCA) and can transcribe many of the same dNA sequences, they are differentiated by their antagonists: Retinoid orphan receptor alpha (RORα) can be bound and repressed in nanomolar concentrations by calcidiol, calcitriol, and 24-hydroxycholesterol. Retinoid orphan receptor beta (RORβ), on the other hand, is strongly inhibited only by trans-retinoic acid in the picomolar range; this receptor can be found in the retina, pineal gland, suprachiasmic nucleus, pituitary, and in stem cells—yet most often will loses expression upon differentiation. The retinoic orphan receptor gamma (RORγ) is antagonized by both retinoic acid and the calciferols, a peculiarity which...' ―Travis​

Hmmm, I didn't know ROR antagonized each other in this way. If that's the case, do you think these various sterols bind strongly enough to be considered antagonists to A & D & Melatonin's actions at these ROR? I'm trying to think of anything else than Vit D since it's already the most famous vitamin in history, despite the supplementation/contentration/conversion problems (I already got a UVB light).

 

[Travis]

Hmmm, I didn't know ROR antagonized each other in this way. If that's the case, do you think these various sterols bind strongly enough to be considered antagonists to A & D & Melatonin's actions at these ROR?

They certainly do bind strongly enough to be considered powerful ligands (IC₅₀≈2·nM), yet the high-affinity binding of sterols is not proportional to reduced activity (EC₅₀≈2·μM). Although many sterols do bind RORα/γ in the low nanomolar range, the very strongest ones—i.e. the 7-hydroxylated sterols—only inhibit activities up to ~40% in concentrations one thousand-fold higher. Yongjun Wang had demonstrated that sterols will bind RORα/γ and decrease their solubility in water, redistributing them towards other cellular compartments. Yet the 7-hydroxylated sterols had not decreased nuclear translocation of RORα at all, even at the concentrations which reduce RORα activity ~40%. Upon further antibody-binding assays on chromosomal fractions, he had concluded that 7-hydroxycholesterol inhibits the dimerization of a transcriptional coactivator. This had been largely independent of the stereochemistry of the 7-carbon; the 7α- and 7β-hydroxylated isomers had been practically equipotent. On account of the foregoing evidence, I think this RORα/γ sterol inhibition would be only physiologically relevant in the liver, the only organ with significant concentrations of 7α-hydroxycholesterol—the precursor of bile salts. I am under the impression that the secosterols—the vitamin D congeners—are more relevant ligands in every location besides.

Wang, Yongjun. "Modulation of RORα and RORγ activity by 7-oxygenated sterol ligands." Journal of Biological Chemistry (2009)

'Recently, we demonstrated that synthetic ligands can bind with high affinity and repress the transcriptional output of both RORα and RORγ, thus functioning as inverse agonists (26). This finding suggests that there might also be natural product ligands that also behave as inverse agonists.' ―Wang

'When 7α-OHC is included, a dose-dependent inhibition of the transcriptional activity of RORα was observed with the maximal efficacy reaching approximately a 40% reduction in RORα-stimulated transcription with an IC₅₀ of 1.3 µM.' ―Wang

'Dose-dependent inhibition of RORα transactivation activity was observed for both 7β-OHC and 7-KC with maximal efficacy of approximately 40% inhibition and IC₅₀s in the range of 0.7 to 1.4 µM' ―Wang

'Similar to RORα, the 7-oxygenated sterols suppressed the constitutive activity of GAL4-RORγ LBD while other oxysterols did not. All three 7-oxgenated sterols (7α-OHC, 7β-OHC, and 7-KC) dose-dependently repressed the transactivation activity of GAL4-RORγ with IC₅₀s in the range of 0.62 to 2.2 µM.' ―Wang

'Fig. 5C shows that the 7-substituted oxysterols display similar affinities for RORα (Kᵢ values ranged from 12 to 18 nM) while cholesterol exhibited only very limited ability to displace the radiolabeled oxysterol. Cholesterol sulfate bound to RORα, but the degree of displacement of [³H]-25-OHC was less than for the 7-oxygenated sterols and as was the case for 25-OHC, cholesterol sulfate was ineffective in modulating the transcriptional activity of RORα.' ―Wang

'The apparent difference between the binding affinity (biochemical assay on purified LBD) and potency in cell-based assays (cells treated with ligand) is not unexpected since it is also seen with the LXR class of oxysterols receptors. For example 24,25-dihydroxycholesterol binds to LXRα with a Kᵢ of 200 nM but displays a 4 µM EC₅₀ in a cotransfection assay (46).' ―Wang

'Treatment of the cells with 7- oxygenated sterols resulted in a suppression of the activity of RORα and RORγ (30-40%) consistent with the results from the GAL4- ROR LBD studies.' ―Wang

'As shown in Fig. 6G, a 40 to 48% decrease in SRC-2 occupancy was detected upon treatment of cells with the 7-oxygenated sterols, which is consistent with the magnitude of decrease in G6Pase expression caused by these ligands.' ―Wang

'Treatment with 7αOHC did not affect the level of RORα occupancy of the G6Pase promoter; however, in the reChIP experiment using the SRC-2 antibody a 36% decrease in the amount of SRC-2 occupancy was noted in the presence of the ROR ligand demonstrating that 7α-OHC decreased the ability of RORα to recruit this coactivator to the G6Pase promoter and thus decreasing the expression of the gene.' ―Wang

'It is interesting to note that 7αOHC caused a nearly identical decrease in SRC-2 recruitment to RORα at the G6Pase promoter (36%) compared to the decrease in G6Pase mRNA expression (33%).' ―Wang

'7α-Hydroxycholesterol Regulates Hepatocyte Glucose Output Acting as a RORα/γ Inverse Agonist – Based on the significant role RORα and RORγ play in regulation of glucose metabolism, as well as our data illustrating 7-oxygenated sterol regulation of G6Pase expression, we assessed the physiological relevance of 7α-OHC as a RORα/γ ligand in primary mouse hepatocytes.' ―Wang

'In cell-based assays, in contrast to cholesterol and cholesterol sulfate that did not modulate RORα or RORγ activity, the 7-oxygenated sterols acted as inverse agonists decreasing the transcriptional activity of these two nuclear hormone receptors.' ―Wang

'Although the 7-oxygenated sterols only repress ROR transcription by 30–40% in the Gal4 cotransfection assay this level of efficacy is consistent in all of the assays and most importantly, underlies the ability of 7αOHC to suppress hepatocyte glucose output by 30%, a physiologically significant decrease.' ―Wang

'...physiologically relevant level of 7α-OHC in a ROR target tissue such as the liver.' ―Wang

'Interestingly, administration of glucose to fasting rats results in a significant increase in the production of 7α-OHC in the liver and it is possible that production of this ROR ligand may play a role in regulation of glucose metabolism in response to feeding. This is supported by data from both animal models and in humans.' ―Wang

'Interestingly, the rate limiting enzymes for biosynthesis of both heme and 7α-OHC are regulated in a circadian fashion.' ―Wang

'Since serum 7α-OHC levels correlate with level of bile acid synthesis it is possible that this RORα ligand may modulate the mammalian clock in many tissues.' ―Wang​

 

[Terma]

They certainly do bind strongly enough to be considered powerful ligands (IC₅₀≈2·nM), yet the high-affinity binding of sterols is not proportional to reduced activity (EC₅₀≈2·μM). Although many sterols do bind RORα/γ in the low nanomolar range, the very strongest ones—i.e. the 7-hydroxylated sterols—only inhibit activities up to ~40% in concentrations one thousand-fold higher. Yongjun Wang had demonstrated that sterols will bind RORα/γ and decrease their solubility in water, redistributing them towards other cellular compartments. Yet the 7-hydroxylated sterols had not decreased nuclear translocation of RORα at all, even at the concentrations which reduce RORα activity ~40%. Upon further antibody-binding assays on chromosomal fractions, he had concluded that 7-hydroxycholesterol inhibits the dimerization of a transcriptional coactivator. This had been largely independent of the stereochemistry of the 7-carbon; the 7α- and 7β-hydroxylated isomers had been practically equipotent. On account of the foregoing evidence, I think this RORα/γ sterol inhibition would be only physiologically relevant in the liver, the only organ with significant concentrations of 7α-hydroxycholesterol—the precursor of bile salts. I am under the impression that the secosterols—the vitamin D congeners—are more relevant ligands in every location besides.

Wang, Yongjun. "Modulation of RORα and RORγ activity by 7-oxygenated sterol ligands." Journal of Biological Chemistry (2009)

'Recently, we demonstrated that synthetic ligands can bind with high affinity and repress the transcriptional output of both RORα and RORγ, thus functioning as inverse agonists (26). This finding suggests that there might also be natural product ligands that also behave as inverse agonists.' ―Wang

'When 7α-OHC is included, a dose-dependent inhibition of the transcriptional activity of RORα was observed with the maximal efficacy reaching approximately a 40% reduction in RORα-stimulated transcription with an IC₅₀ of 1.3 µM.' ―Wang

'Dose-dependent inhibition of RORα transactivation activity was observed for both 7β-OHC and 7-KC with maximal efficacy of approximately 40% inhibition and IC₅₀s in the range of 0.7 to 1.4 µM' ―Wang

'Similar to RORα, the 7-oxygenated sterols suppressed the constitutive activity of GAL4-RORγ LBD while other oxysterols did not. All three 7-oxgenated sterols (7α-OHC, 7β-OHC, and 7-KC) dose-dependently repressed the transactivation activity of GAL4-RORγ with IC₅₀s in the range of 0.62 to 2.2 µM.' ―Wang

'Fig. 5C shows that the 7-substituted oxysterols display similar affinities for RORα (Kᵢ values ranged from 12 to 18 nM) while cholesterol exhibited only very limited ability to displace the radiolabeled oxysterol. Cholesterol sulfate bound to RORα, but the degree of displacement of [³H]-25-OHC was less than for the 7-oxygenated sterols and as was the case for 25-OHC, cholesterol sulfate was ineffective in modulating the transcriptional activity of RORα.' ―Wang

'The apparent difference between the binding affinity (biochemical assay on purified LBD) and potency in cell-based assays (cells treated with ligand) is not unexpected since it is also seen with the LXR class of oxysterols receptors. For example 24,25-dihydroxycholesterol binds to LXRα with a Kᵢ of 200 nM but displays a 4 µM EC₅₀ in a cotransfection assay (46).' ―Wang

'Treatment of the cells with 7- oxygenated sterols resulted in a suppression of the activity of RORα and RORγ (30-40%) consistent with the results from the GAL4- ROR LBD studies.' ―Wang

'As shown in Fig. 6G, a 40 to 48% decrease in SRC-2 occupancy was detected upon treatment of cells with the 7-oxygenated sterols, which is consistent with the magnitude of decrease in G6Pase expression caused by these ligands.' ―Wang

'Treatment with 7αOHC did not affect the level of RORα occupancy of the G6Pase promoter; however, in the reChIP experiment using the SRC-2 antibody a 36% decrease in the amount of SRC-2 occupancy was noted in the presence of the ROR ligand demonstrating that 7α-OHC decreased the ability of RORα to recruit this coactivator to the G6Pase promoter and thus decreasing the expression of the gene.' ―Wang

'It is interesting to note that 7αOHC caused a nearly identical decrease in SRC-2 recruitment to RORα at the G6Pase promoter (36%) compared to the decrease in G6Pase mRNA expression (33%).' ―Wang

'7α-Hydroxycholesterol Regulates Hepatocyte Glucose Output Acting as a RORα/γ Inverse Agonist – Based on the significant role RORα and RORγ play in regulation of glucose metabolism, as well as our data illustrating 7-oxygenated sterol regulation of G6Pase expression, we assessed the physiological relevance of 7α-OHC as a RORα/γ ligand in primary mouse hepatocytes.' ―Wang

'In cell-based assays, in contrast to cholesterol and cholesterol sulfate that did not modulate RORα or RORγ activity, the 7-oxygenated sterols acted as inverse agonists decreasing the transcriptional activity of these two nuclear hormone receptors.' ―Wang

'Although the 7-oxygenated sterols only repress ROR transcription by 30–40% in the Gal4 cotransfection assay this level of efficacy is consistent in all of the assays and most importantly, underlies the ability of 7αOHC to suppress hepatocyte glucose output by 30%, a physiologically significant decrease.' ―Wang

'...physiologically relevant level of 7α-OHC in a ROR target tissue such as the liver.' ―Wang

'Interestingly, administration of glucose to fasting rats results in a significant increase in the production of 7α-OHC in the liver and it is possible that production of this ROR ligand may play a role in regulation of glucose metabolism in response to feeding. This is supported by data from both animal models and in humans.' ―Wang

'Interestingly, the rate limiting enzymes for biosynthesis of both heme and 7α-OHC are regulated in a circadian fashion.' ―Wang

'Since serum 7α-OHC levels correlate with level of bile acid synthesis it is possible that this RORα ligand may modulate the mammalian clock in many tissues.' ―Wang​

See this is why I asked you: I would've wasted 1 month on/off trying to figure that out. Thanks. I'd assumed sterols were mainly associated with LXR which they are, but that there could be some kind of ROR-based circadian feedback between A&D and sterols since the former affect production of the latter... probably in circadian manner.

That nuclear translocation bit was particularly interesting for a half a second until it suddenly didn't matter for whatever reason I didn't catch.

Particularly that liver-specific bit of deduction is of value. Good point. Yet the fact it's liver-specific may not diminish it that much (bile problems are ubiquitous, accutane and otherwise; btw the accutane victims have all tried Vit D a thousand times over, so any issue there rests on skin/liver/kidney conversion issues, which they've already suspected but no one can show).

I'll probably have to read up on that more in a few weeks.

(Btw I appreciate the circadian angle, just wasn't what I was focused on at the minute)

 

[Travis]

See this is why I asked you: I would've wasted 1 month on/off trying to figure that out. Thanks. I'd assumed sterols were mainly associated with LXR which they are, but that there could be some kind of ROR-based circadian feedback between A&D and sterols since the former affect production of the latter... probably in circadian manner.

You're right, Doctor Wang does talk about this:

Wang, Yongjun. "Modulation of RORα and RORγ activity by 7-oxygenated sterol ligands." Journal of Biological Chemistry(2009)

'Interestingly, the rate limiting enzymes for biosynthesis of both heme (60) and 7α-OHC are regulated in a circadian fashion.' ―Wang

'CYP7A1 expression is also circadian (34) indicating that its product, 7α-OHC, may also display a circadian pattern of production.' ―Wang

'Thus, we propose that the 7-oxysterols may regulate the transcriptional output of the constitutively active RORs on both hepatic glucose production and circadian rhythms.' ―Wang​

And curiously, a small handful of studies that has been published explicitly on the circadian rhythm of bile excretion:

[1] Ho, K. J. "Circadian rhythm of biliary excretion and its control mechanisms in rats with chronic biliary drainage." American Journal of Physiology-Legacy Content (1975)
[2] Mizuta, Koichi. "Survival of rats undergoing continuous bile drainage depends on maintenance of circadian rhythm of bile secretion." Chronobiology international (1999)
[3] Shiesh, Shu‐Chu. "Melatonin prevents pigment gallstone formation induced by bile duct ligation in guinea pigs." Hepatology (2000)​

 

[franko]

'A postprandial increase in serum retinol concentration may be blunted when vitamin A is ingested with either food or ample dietary fat, whereas a significant amount of free (unesterified) retinol may circulate when vitamin A is consumed without dietary fatty acids, which leads to excessive production of retinoic acid (28).' ―Penniston
​

Consuming retinol without fatty acid trigylcerides results in higher retinoic acid concentration, likely because the retinol then has no glyceride esterify-with the intestines. Ostensibly, free retinol and/or it glucorinide are/is better substrate(s) for retinal oxidase than retinol triglyceride or retinol cholesterol esters. This is actually in concord with data on supplementation and the risk of bone fracture; water-soluble forms of retinol are consistently found greater risk factors.

The protective role of fats against retinol and retinoic acid is a key point to consider and Grant references that Penniston paper multiple times.

Just to re-emphasize that quote from Penniston:

"A significant amount of free (unesterified) retinol may circulate when vitamin A is consumed without dietary fatty acids, which leads to excessive production of retinoic acid."
​

There are many interesting implications of this fact. But I will elaborate on those later.

Here is one snippet from Grant on the protective role of fats as seen in Wolbach and Howe, 1925:

"I have no doubt that the higher density fats and cholesterol of the butter helped significantly by emulsifying the retinoic acid hidden within the casein. The beneficial, and protective, aspects of these fats would have far outweighed any additional risk it’s own vitamin-A content may have presented."
- p. 106, PFP​

And another relevant snippet from Grant:

"The next suspicious aspect in the evolution of the deficiency theory was that there were other early researchers who were showing that just having adequate amounts of dietary fats, and especially that of butter fat, could prevent and reverse the same “xerophthalmia” disease conditions of the eye.

Probably the most noted was the work of Dr. Masamichi Mori in Japan (not to be confused with S. Mori of Wisconsin who is another prominent early vitamin-A researcher). In 1904, Dr. Masamichi Mori provided a comprehensive account of the pathogenesis of the Hikan condition he observed in children in his clinic. The term Hikan is the Chinese name for these same diseases of the eye. [...]

The next and the critically important observation made by Dr. M. Mori was that this disease was not at all limited to just the eyes.

By Mori’s observation, and definition of the disease, it included five distinct symptoms (comorbidities if you prefer that term). These were: diarrhea, abdominal expansion, excessive appetite, night blindness, xerosis of the conjunctiva, and thinning of the hair.

In addition to these five specific symptoms, he also documented the children commonly having dry skin, and having significant sensitivity to sunlight too. One other very important little detail he documented was that the condition was more prevalent in the summer months. Therefore, clearly, the Hikan disease condition was much more than just an eye disease. Nonetheless, Mori very successfully treated the disease by just adding more fats to the diets of the children. He was quite certain that it was just the additional fats and not something special hidden within the fats, that was responsible for the recovery from the disease. But, to Mori’s American contemporaries, and modern day historians, their conclusion was that Mori had missed the mark with his hypothesis and that it was not just the additional fats that had ameliorated the disease.

However, what I hope to show you later, is that Mori was indeed correct. It was not some deficiency condition ailing these children at all. Rather it was a poisoning, and the additional butter fat in their diet was emulsifying and therefore protecting them from it."
- p. 15, PFP​

Thus we can form a hypothesis that:

Hypothesis: The explanation for the results of some of the "deficiency" experiments is not that added Vitamin A corrected a deficiency, but that some added substance was protective, to some degree, against retinol and/or retinoic acid poisoning.

But when looking at the studies, there are a lot of different diets and different preventative/curative substances to consider.

And then when a substance was shown to be preventative/curative we don't necessarily know which component of that substance was curative/preventative (or if it wasn't the whole or some combination).

There is a lot more that can be said about this, but I'll leave it at that for now.

 

[franko]

I've been wondering and asking this since page 1 of this thread, although a book that simply reads "Oh, I had low vitamin D" Probably isn't radical enough!

+1 I believe Vitamin D is the missing link in all of this, and none of the, Vitamin A is the issue crowd, have yet given a solid answer as to why this is wrong IMO.

Yep! Plus if he did figure it out he'd have to come out and say he messed up, but when you are that invested in your theory being right it can be really hard to admit you were wrong. The D would explain why northerners would get bone problems.

Ok then please explain this for us by answering one simple question:

By what mechanism does low Vitamin D cause inflammation?

 

[raypeatclips]

Ok then please explain this for us by answering one simple question:

By what mechanism does low Vitamin D cause inflammation?

Vitamin D is hugely important for regulating many systems within the body, something which is discussed a huge amount on the internet, I am certain you will have seen this talked about before. For example below, vitamin D deficient cells react hugely to endotoxin, while the highest levels of inflammatory inhibition is interestingly found at 50 ng/ml the level Peat mentions people aim for. Many people with low vitamin D levels react to things poorly, such as certain foods, and when they raise their levels, they no longer react to them.

Is it really so far fetched for you to believe that one of the other thing, amongst the countless things that are effected by vitamin D, happens to be vitamin A?

"In the current study researchers examined the specific mechanisms by which vitamin D might act on immune and inflammatory pathways. They incubated human white blood cells with varying levels of vitamin D, then exposed them to lipopolysaccharide (LPS), a molecule associated with bacterial cell walls that is known to promote intense inflammatory responses.

Cells incubated with no vitamin D and in solution containing 15 ng/ml of vitamin D produced high levels of cytokines IL-6 and TNF-alpha, major actors in the inflammatory response. Cells incubated in 30 ng/ml vitamin D and above showed significantly reduced response to the LPS. The highest levels of inflammatory inhibition occurred at 50 ng/ml.

Through a complex series of experiments, the researchers identified a new location where the vitamin-D receptor appears to bind directly to DNA and activate a gene known as MKP-1. MKP-1 interferes with the inflammatory cascade triggered by LPS, which includes a molecule known as p38, and results in higher levels of IL-6 and TNF-alpha.

"This newly identified DNA-binding site for the vitamin-D receptor, and the specific pathways inhibited by higher levels of vitamin D provide a plausible mechanism for many of the benefits that have been associated with vitamin D," said Dr. Goleva. 'The fact that we showed a dose-dependent and varying response to levels commonly found in humans also adds weight to the argument for vitamin D's role in immune and inflammatory conditions.""

https://www.sciencedaily.com/releases/2012/02/120223103920.htm



https://www.hindawi.com/journals/iji/2017/8608716/

"The Vit D pathway is now strongly implicated to be involved in the pathogenesis of inflammatory diseases such as inflammatory arthritis, lupus, and BD."

 

[raypeatclips]

By the way everybody. I asked Grant back in April about the vitamin D connection. He claims it has "nothing to do with vitamin D" Despite telling me he never supplemented D, lives in a country that has "dingy amount of sunlight for 9 months a year." I've now asked him if he's ever had his D levels tested, or planning on doing, and awaiting for his reply now.

Surely as a "scientist" he would want blood tests and measurements to back up his theory, and have as much information at his disposable as possible? In my opinion, he has had far, far too low amounts of vitamin D to combat a deficiency. Drinking some vit D fortified whole milk while you live in a dark country isn't going to do anything. Come on. I cannot believe people take this guy seriously. I'm not convinced by his answer and think he dodges giving a proper answer.


He told me this:

"Thanks for the question. No, my descent into disease hell and my recovery from it had absolutely nothing to do with vitamin D. As I’ve documented in my two e-books, leading up to my descent into disease I was consuming large amounts of whole milk. The milk’s fortified with vitamin D. I was consuming this extra milk trying to offset an excess measure of gum recession my dentist had alerted me to.

So I actually had lots of vitamin D in my diet leading up to my serious disease condition. As a matter fact, the extra milk consumption, with the supplemented vitamin D, vastly contributed to it.

I’ve now recovered from all of this disease condition, and my gums have gone back to being normal, and my teeth are now once again feeling quite strong. I did not supplement with vitamin D at all over this time frame. I did not consume any foods that contained vitamin D. I also live in Canada where we have a very dingy amount of sunlight for over nine months of the year. So, by not consuming any vitamin D, and only by eliminating all vitamin A, was I able to make this recovery.

What’s astonishing to me is that in the face of the massive and overwhelming rate of disease, and the staggering rate increases of diseases in North America people still naïvely believe that all of this is somehow attributable to a deficiency. That “deficiency“ excuse is just a bizarre thought to me. For example, in 1970 there were about a total of fewer than 1000 people with kidney disease in all of the USA. Now there are about 660,000 people with full-blown kidney failure and about 30 million people with emerging kidney disease. It is simply mathematically impossible for that rate increase to have anything to do with a deficiency. People need to wake up and face the obvious here and see this for what it really is. It is simply a ******* poisoning. Then when you consider that many of the early vitamin A toxicity experiments conclusively proved that vitamin A would directly induce this same kidney destruction, and also just about every other named chronic disease, in experimental animals, I don’t think it’s a big challenge to connect the dots. We need to face the obvious; it’s nation wide vitamin A poisoning. I mean seriously, after seeing that over the last 100 years that researchers have repeatedly and systematically quickly poisoned tens of thousands of animals into massive disease and early deaths using a so-called vitamin how can any reasonable, rational thinking person continue to believe the phony theory of it being a vitamin?

We need to stop with the BS excuses and end the pointless debate by more people taking on this diet experiment. The chronic diseases have absolutely nothing to do with a deficiency in vitamin D, or anything else. Vitamin D is just a by-product of the solar breakdown of so-called vitamin-A, and it binds to the same cellular receptors. Therefore, vitamin D only appears to be a vitamin because it is far less toxic than vitamin A. And vitamin D blunts the toxicity of vitamin A by blocking up the same receptors, and thereby rate-limits the conversion of vitamin A into the highly toxic retinoic acid form of the retinoid molecule."

 

[dbh25]

Surely as a "scientist" he would want blood tests and measurements to back up his theory

+1

Grant has just posted a 4 year update on his blog to report on his progress after 4 years on his Vitamin A elimination diet.

LDL=56.4?
Is this from his autopsy?