Is Supplementing Vit E Actually Bad For You?

Travis

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@Travis is your interest due to smoking? Are tocotrienol's important?
I think tocotrienols ought to have a different cellular disposition, perhaps associating less with the plasma membrane and more with the mitochondria—like the other prenylated molecules (i.e. coenzyme Q₁₀, vitamin K₂). Of course since the 'head' is the peroxynitrite scavenging part, you'd expect tocotrienols to work for doing that just the same. The water-soluble and nearly tailless γ-CEHC complexes peroxynitrite in other cellular spaces, and phylloquinone metabolism shows that the body can convert an alkane side-chain into a prenylated one: vitamin K₂ is normally synthesized from vitamin K₁ through a 'tail switching' procedure, and this occurs through the tailless water-soluble menadione intermediate. So I think the tocotrienols, of all types, would help to reduce the spontaneous superoxide known to be produced near mitochondria. People eating unrefined diets would have to worry less about vitamin E in general; while smokers, people with autoimmunity, people having cancer (reduces prostaglandin E₂ formation), and those eating mostly animal foods should perhaps focus a bit on γ-tocopherol specifically.
 
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Obi-wan

Obi-wan

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I think tocotrienols ought to have a different cellular disposition, perhaps associating less with the plasma membrane and more with the mitochondria—like the other prenylated molecules (i.e. coenzyme Q₁₀, vitamin K₂). Of course since the 'head' is the peroxynitrite scavenging part, you'd expect tocotrienols to work for doing that just the same. The water-soluble and nearly tailless γ-CEHC complexes peroxynitrite in other cellular spaces, and phylloquinone metabolism shows that the body can convert an alkane side-chain into a prenylated one: vitamin K₂ is normally synthesized from vitamin K₁ through a 'tail switching' procedure, and this occurs through the tailless water-soluble menadione intermediate. So I think the tocotrienols, of all types, would help to reduce the spontaneous superoxide known to be produced near mitochondria. People eating unrefined diets would have to worry less about vitamin E in general; while smokers, people with autoimmunity, people having cancer (reduces prostaglandin E₂ formation), and those eating mostly animal foods should perhaps focus a bit on γ-tocopherol specifically.


So tocotrienols can't hurt to have, could help with oxidative transport (more CO2, more energy). Waiting for my order of Suzy Cohens Full Spectrum E. Thanks again @Travis! You da man!! You deserve the MVP award of the forum! Maybe I will start the MVP thread...
 

Travis

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None of the tocopherols are quinones, so I wouldn't expect them to contribute to electron transport. Vitamin K is a quinone, and is known to increase mitochondrial electron transfer in a manner similar to coenzyme Q₁₀. Either a hydride (∶H⁻) or a hydrogen atom (·H) is given to the ring in this process which eventually serves to separate the hydrogen's electron from its nucleus (H⁺), which is simply one just proton in most cases. Protons define acidity and need to be removed; these waste protons are then shuttled out of the mitochondria as ATP which eventually reaches the cell membrane where it's hydrolyzed into ADP, a phosphate, and our waste proton (H⁺). These of course acidify the extracellular fluid and this too needs to be removed, which is deftly accomplished by the kidneys. The protons are formed ostensibly because the mitochondria wants the electrons: the main event, with ATP somewhat more like a sideshow—an ancillary device mostly there to shuttle protons (H⁺) from the mitochondria to the cell membrane (but given extra energy by confused people to power imaginary 'pumps').

ubiquinone_01.jpg


Microtubules are connected to the mitochondria, and could very well be the main destination for the majority of the electrons originally stripped off of glucose. Microtubules form bundles in the center of nerves and are insulated by myelin, a homogenous material consisting mostly of: saturated fatty acid esters, progesterone, pregenenolone, and microtubule-associated proteins. I think the dispersion of coenzyme Q on the mitochondria is usually modeled with its fatty tail penetrating the membrane and its 'heads' existing at the surface water-phase; an analogous orientation is thought to occur with the tocopherols on the cell's outer membrane. Since the same poly-isoprene 'tail' shared by both coenzyme Q₄ and vitamin K₂ (MK-4) is nearly identical to the tocotrienol tail, being merely one isoprene unit longer, I would expect it to also partition on the mitochondrial surface—though not participating in electron transfer, but merely serving to protect the hydrogen splitting operation (∶H⁻ ⟶ H⁺ + 2e⁻) from free radicals such as superoxide. In the mitochondrial location, you might think that α-tocopherol would be a better choice as it captures superoxide better than γ-tocopherol; however, the latter can be logically reasoned to be more suitable for the outer membrane and especially during times of high peroxynitrite formation (i.e. when the cytokine induction of NADH dehydrogenase and iNOS act to create defensive molecules superoxide (Ȯ₂⁻) and nitric oxide (ṄO), respectively, which then combine together to form the more cell-damaging and prostaglandin-producing peroxynitrite (ONOO⁻)). Peroxynitrite can be neatly captured by γ-tocopherol, which has been proven, and in vitro cellular prostaglandin formation is always lowered in its presence (peroxynitrite is cyclooxygenase's other substrate besides arachidonic acid—also proven).
 
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Obi-wan

Obi-wan

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Thanks @Travis, some say that Vit.K MK4 interferes with Vit. E or vice versa. I take about 15mg of Vit. K2 MK4 per day. Haidut says to just take them a few hours apart
 

Travis

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Thanks @Travis, some say that Vit.K MK4 interferes with Vit. E or vice versa. I take about 15mg of Vit. K2 MK4 per day. Haidut says to just take them a few hours apart
I wouldn't imagine, as MK-has a shorter tail. Leaves have a considerable amount of vitamin K₁ and I don't think herbivores are lacking in vitamin E.
 

Mito

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some say that Vit.K MK4 interferes with Vit. E or vice versa.

So, number one, high doses – so vitamin K and vitamin E are metabolized by the same metabolic pathways that are regulated by the same nuclear receptor. Vitamin E stimulates its own destruction at high concentrations as does vitamin K. The way they do that is to activate the same catabolic pathways by the same nuclear receptor, which means that if you’re megadosing vitamin E, you can deplete the body of vitamin K, and if you’re megadosing vitamin K, you can deplete the body of vitamin E. So high doses could deplete vitamin E levels and contribute to oxidative stress.”
https://chrismasterjohnphd.com/2016/12/10/whats-new-with-vitamin-k2/


And there is this....Interactions of vitamin E and K
 
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Travis

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So, number one, high doses – so vitamin K and vitamin E are metabolized by the same metabolic pathways that are regulated by the same nuclear receptor. Vitamin E stimulates its own destruction at high concentrations as does vitamin K. The way they do that is to activate the same catabolic pathways by the same nuclear receptor, which means that if you’re megadosing vitamin E, you can deplete the body of vitamin K, and if you’re megadosing vitamin K, you can deplete the body of vitamin E. So high doses could deplete vitamin E levels and contribute to oxidative stress.”
https://chrismasterjohnphd.com/2016/12/10/whats-new-with-vitamin-k2/


And there is this....Interactions of vitamin E and K
But doesn't the short forms bypass the natural metabolism that the more longer and vitamin E-like phylloquinone experiences? I wouldn't surprise me that that the CEHC conversion and prenylation of vitamin E would be under negative feedback—also the analogous phylloquinone ⟶ mendione convertsion and subsequent prenylation—but I see MK-4 more like the most downstream product that wouldn't need to be sensed by the body (this is somewhat unnatural and herbivores never eaten this is nature and thus could be expected as not having evolved control mechanisms to sense it).
 

Mito

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But doesn't the short forms bypass the natural metabolism that the more longer and vitamin E-like phylloquinone experiences? I wouldn't surprise me that that the CEHC conversion and prenylation of vitamin E would be under negative feedback—also the analogous phylloquinone ⟶ mendione convertsion and subsequent prenylation—but I see MK-4 more like the most downstream product that wouldn't need to be sensed by the body (this is somewhat unnatural and herbivores never eaten this is nature and thus could be expected as not having evolved control mechanisms to sense it).
1D6E5EF1-93C8-4BFC-8A7F-BF5332BC63AB.jpeg

“In summary, vitamins E and K appear to share the same metabolic pathways; the side chain of both are w- hydroxylated, then undergo b-oxidation.8,9 Hypothetically, MK-4 is decreased in response to high hepatic a-tocopherol and, therefore, the most active form of vitamin K, MK-4, is depleted. There are various possible mechanisms that could account for the decreased MK-4 concentrations. Vitamin E may interfere with formation of MK-4 from K1. K1 must be converted to MK-4 by truncation of its side chain to form menadione,11 followed by tail replacement with geranylgeranyl10 to form MK-4. The mechanism for the conversion of K1 to menadione is unknown, despite the recognition that menadione is an intermediate in the con- version of K1 to MK-4.11,43 Vitamin E may increase metabolic pathways and deplete all vitamin K forms. Clearly, more studies are needed.”

http://www.encognitive.com/files/Vitamin E and K interactions – a 50-year-old problem.pdf
 
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Obi-wan

Obi-wan

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I see @haidut changed TocoVit to a MCT base... @Travis I am liking my Full Spectrum E. @bzmazu, you should switch from the Gamma Gems
 
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Braveheart

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I see @haidut changed TocoVit to a MCT base... @Travis I am liking my Full Spectrum E. @bzmazu, you should switch from the Gamma Gems
Thanks Obi...unfortunately just received order...expensive too, as everything double here for me, plus shipping!...and I'm not rich, to put it mildly..but next time, or if some dinero falls from the sky soon...
 

benaoao

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Well the very first sentence of this topic says it all, if one is healthy there’s no need for a vitamin E supplement. I do have 100iu on hand if I eat out, that’s about it
 

Travis

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What immediately stands-out is the author's use of 'VES' to signify α-tocopheryl succinate, a substitution having no place in logical acronym construction (LAC). There is no 'V' anywhere in the word 'α-tocopheryl succinate,' nor is there one in its IUPAC name: (2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-(4,8,12-trimethyltridecyl)]chroman-6-succinate ester. Upon further reflection, the reader finally gets it and becomes amazed that he hadn't thought of 'vitamin E succinate' sooner (yet is still befuddled why Zhang's acronym doesn't match his terminology):

'Here we demonstrate that α-tocopheryl succinate (VES) [?] can suppress the expression of prostate-specific antigen (PSA), a marker for the progression of prostate cancer. VES can also suppress androgen receptor (AR) expression by means of transcriptional and posttranscriptional modulation, but not ligand binding, nuclear translocation, or AR dimerization.' —Zhang

What also stands-out is the authors use of selenomethionine to inhibit cancer proliferation. On the basis of this, you might imagine this to be a somewhat-sophisticated researcher that knows all about polyamines; but if you had thought that you are likely wrong. This researcher had ostensibly used selenomethionine simply on the basis of the a few classic clinical trials, and his continued use of the word 'selenium' to represent 'selenomethionine' demonstrates naiveté.

'Moreover, selenomethionine (SM), a prostate cancer treatment adjuvant, shows an inhibitory effect on LNCaP cell growth, yet has no effect on the AR/PSA pathway.' —Zhang

'VES, but Not Selenium, Affects AR and PSA Expression. In previous research, selenium has been combined with vitamin E to study their antitumor activity, especially in epidemiological studies (4, 28). Therefore, we were interested in testing whether selenium could function similarly to vitamin E, which inhibits AR expression in LNCaP cells.' —Zhang

'Vitamin E, Selenium, and Prostate Cancer. In addition to vitamin E, a growing body of evidence suggests that a higher serum level of dietary supplemental selenium substantially reduces the incidence of lung, colon, or prostate cancer (40, 41). Other studies also reported that the selenium compounds inhibit the growth of prostate cancer cells through the induction of apoptosis.' —Zhang

He could not demonstrate a binding site for vitamin E succinate yet had eliminated the androgen receptor as a candidate, leaving the reader only to speculate on how it works. After thinking about this molecule for a few minutes, and the succinate moiety in particular, I had speculated that it could be acting as a calcium ionophore—or that is: The 6-succinate ester of vitamin E would be expected to chelate calcium from the extracellular fluid and carry it into the cytosol.

'Cell Culture and VES Treatment. The LNCaP and COS-1 cells were purchased from the American Type Culture Collection (Manassas, VA). Fibroblast cells were primarily cultured from normal prostate tissue. LNCaP cells were grown in phenol red-free RPMI medium 1640 . . .' ―Zhang

Sigma Aldrich informs me that 'RPMI-1640 contains 0.42 mM calcium' and Yuewen Gong informs me that 'intracellular calcium seems to be a potent regulator of AR gene expression in LNCaP cells.' On the basis of these foregoing facts, perhaps it would be worth reading his entire article:

Gong, Y. "Calcium regulation of androgen receptor expression in the human prostate cancer cell line LNCaP." Endocrinology (1995)

'To determine whether intracellular levels of calcium affected the expression of the AR, the AR-containing cell line LNCaP was treated with the calcium ionophores A23187 (10⁻⁶ M) or ionomycin (10⁻⁶ M), or the intracellular Ca*+- ATPase inhibitor thapsigargin (10⁻⁷ M), for 16 h.' ―Gong
Yuewen Gong had used the same culture media—RPMI 1640—as the Zhang study but had used two different calcium ionophores, albeit at lower concentrations than Zhang's vitamin E acetate (10⁻⁶ M vs 10⁻⁵ M). As he had hinted-about in the absract, intracellular calcium was found capable of reducing the expression of the androgen receptor:

'In the current study two compounds, A23187 and thapsigargin, were used to increase intracellular calcium levels in LNCaP cells. A23187 is a Ca²⁺ ionophore, widely used to analyze the role of Ca²⁺ in the regulation of gene expression (3436).' ―Gong

'The time dependencies of A23187 and thapsigargin downregulation of AR were examined next. Levels of AR transcripts in LNCaP cells began to decrease after 6-8 h of incubation with A23187 (10⁻⁶ M), reached a nadir after 16 h of incubation (29% of control), and increased to 59% of the control level at 24 h. Thapsigargin (10⁻⁷ M) treatment also resulted in a reduction in AR transcripts, decreasing after 6-8 h of incubation, reaching 23% of the control level after 10 h, and increasing to 68% of control by 24 h.' ―Gong

calcium.png


'In conclusion, increases in intracellular calcium concentration by the calcium ionophore or thapsigargin effectively reduce AR expression in LNCaP cells. Calcium inhibition of AR expression seems to precede calcium-induced apoptotic cell death. Furthermore, the mechanism of this reduction seems to be different from a pathway in which the calcium signal is transduced via a phosphorylation mechanism.' ―Gong

Although you'd almost be certain by the molecular structure that α-tocopheryl succinate is a calcium ionophore, we don't have to speculate since this had be proven in 1985.

Fariss, M. "Vitamin E reversal of the effect of extracellular calcium on chemically induced toxicity in hepatocytes." Science (1985)

'After a 1-hour incubation, the a-tocopherol content of control cells incubated with Ca²⁺-free medium and vitamin E succinate was approximately 1.1 nmol per 10⁶ cells or 1200 percent of the initial value (Table 1).' ―Fariss

calcium 2.png


'We therefore examined the effect of exogenous vitamin E on chemically mediated cell death in the presence or absence of extracellular Ca²⁺. The results show that the addition of vitamin E succinate to the incubation medium protected isolated hepatocytes incubated in Ca2²⁺ free medium against chemically induced cell death. Hence chemically induced cell death may be dependent not on the presence of extracellular Ca²⁺ but rather on cellular α-tocopherol (Table 1).' ―Fariss

Marc Fariss proves that vitamin E succinate responds powerfully to calcium levels, yet explains its ability to protect cells against calcium-related cell death in a calcium-independent manner. He seems entirely oblivious to the fact that he's using a calcium chelator and keeps referring to this as simply 'vitamin E,' as if the succinate moiety is irrelevant.

In 1990, the unique effect of vitamin E succinate to reduce cell proliferation had been proven yet again. To account for this effect, Kimberly Kline had rattled-off a litany of mechanisms while not committing to any one in particular. Of these mechanisms, Doctor Kline had toyed with the 'vitamin E succinate as a calcium ionophore' idea for awhile, yet had disregarded it (based on imprecise logic):

Kline, K. "Growth‐inhibitory effects of vitamin E succinate on retrovirus‐transformed tumor cells in vitro." (1990)

'Five antioxidants, including a synthetic analogue of vitamin E, Trolox, as well as the active vitamin form, DL-α-tocopherol, were incapable of inhibiting C4#1 tumor cell growth, indicating that a mechanism of action other than or in addition to functions as an antioxidant may be operating.' ―Kline

'These studies demonstrated that vitamin E, in the form of vitamin E succinate, inhibited the growth of retrovirus-transformed tumor cells in vitro and suggested that the antiproliferative effects of vitamin E succinate did not involve antioxidant properties but rather, as yet, unidentified mechanisms leading to cell cycle blockage.' ―Kline

succinate.png


'The mechanism(s) whereby vitamin E succinate inhibits tumor cell proliferation is unknown. One well-characterized function for vitamin E is its antioxidant effect (27,28). Although vitamin E succinate does not exhibit any antioxidant properties unless the succinate is clipped off (29)...' ―Kline

'Considering that DL-α-tocopherol did not exhibit any antiproliferative activity at a concentration 20-fold greater than vitamin E succinate suggests that the antiproliferative activity exhibited by vitamin E succinate involves a mechanism that does not correlate with the classically defined units of tocopherol biologic activity.' ―Kline

'Regardless of the amount of calcium in the media in which the C4#1 cells were cultured (from 0.4 mM normally contained in serum-free RPMI-1640 to 2.0 mM), no differences were seen in either the ability of the C4#1 tumor cells to proliferate or in the cytostatic properties of various vitamin E succinate treatments. If it is speculated that vitamin E succinate is acting as a calcium ionophore, it would be predicted that vitamin E succinate treatment of C4#1 cells in the presence of increasing amounts of calcium in the media (0.4 mM to 2.0 mM) might result in increased cellular uptake of calcium. This would result in increased cytostatic effects (i.e., increased percent inhibition of tumor cell proliferation) and perhaps even cell kill at the highest calcium concentrations. The absence of any effects suggests that vitamin E succinate is not acting as a calcium ionophore in this situation.' ―Kline
Kimberly Kline fails to note that the extracellular calcium concentration would only matter when it's limiting, or when in the presence of a vitamin E succinate concentration that is greater than the calcium concentration. The highest concentration of vitamin E acetate she'd used (19·μM) had still been ~20× less than her lowest Ca²⁺ concentration (0.4·mM). On account of there being an excess of Ca²⁺ at both concentrations you wouldn't expect to see a difference anyhow, and would only expect to see one when the vitamin E acetate concentration approaches that of the Ca²⁺ concentration.

However, she did cite authors sympathetic to the 'ionophore' idea.

Slack, R. "Studies on the effects of vitamin E on neuroblastoma N1E 115." Nutrition and cancer (1989)

'It is quite possible that the free carboxylic acid group of tocopherol succinate is functional in conveying growth-arresting properties to this vitamin E derivative. Located in the membrane, this derivative could act much like free fatty acids do in biomembranes (i.e., as calcium ionophores) (36). Increases in calcium uptake by the cell as well as changes in levels of this divalent cation in the medium have been shown to greatly influence cell proliferation and differentiation processes (37-39). The functionality of the carboxyl group of tocopherol succinate with respect to calcium fluxes needs to be further investigated.' ―Slack
Finally, there is another person who sees things this way . . . and another:

Gogvadze, V. "Involvement of Ca²⁺ and ROS in α‐tocopheryl succinate‐induced mitochondrial permeabilization." International journal of cancer (2010)

'In particular, in addition to ROS production, α-TOS stimulates rapid Ca²⁺ entry into the cells with subsequent accumulation of Ca²⁺ in mitochondria—a prerequisite step for MPT induction.' ―Gogvadze

'Jurkat T-lymphocytes were cultured in RPMI 1640 complete medium...' ―Gogvadze

'Addition of α-TOS to transfected Tet21N cells caused a time-dependent stimulation of mitochondrial H₂O₂ production, which was blocked by the antioxidant N-acetylcysteine.' ―Gogvadze

'To investigate the ability of α-TOS to stimulate MPT, sequential pulses of Ca²⁺ were added to isolated rat liver mitochondria in the presence of α-TOS. Addition of Ca²⁺ to mitochondrial suspension led to a rapid increase in the level of this cation in the incubation buffer followed by a return to the initial level as mitochondria took up the added Ca²⁺.' ―Gogvadze

Gogvadze had proved vitamin E succinate was a calcium ionophore and had even said as much, yet he'd been focused on using rather high concentrations (50·μM) to draw-in enough Ca²⁺ to increase mitochondrial hydrogen peroxide concentrations—quite a bit different than the lower-dose transcriptional shifts noted by Zhang. Although his mechanisms and descriptions seem solid, these are not the same mechanisms noted by the other scientists and represent an entirely non-selective 'Ca²⁺ overkill' approach to inhibiting cancer cells.

Since I now have support for the calcium ionophore mechanism, I propose the low-dose transcriptional shifts had likely something to do with calmodulin:

Pelley, R. "Calmodulin-androgen receptor (AR) interaction: calcium-dependent, calpain-mediated breakdown of AR in LNCaP prostate cancer cells." Cancer research (2006)

Karacosta, L. "A regulatory feedback loop between Ca²⁺/calmodulin-dependent protein kinase kinase 2 (CaMKK2) and the androgen receptor in prostate cancer progression." Journal of Biological Chemistry (2012)

Cifuentes, E. "Physical and functional interaction of androgen receptor with calmodulin in prostate cancer cells." Proceedings of the National Academy of Sciences (2004)

'We observed tight binding of AR to CaM when LNCaP cell extracts were subjected to CaM-affinity column chromatography. AR binding to CaM was Ca²⁺-dependent and was inhibited by pretreatment of the cell extracts with W-7. Using immunofluorescence staining and confocal microscopy, we demonstrated colocalization of AR and CaM in the nucleus of LNCaP cells.' ―Cifuentes

'LNCaP cells were grown in RPMI medium 1640 containing 10 nM testosterone, whereas PC3 cells were grown in DMEM.' ―Cifuentes

'The binding of AR to CaM suggested that the two proteins may interact in cells. This hypothesis was examined by immunocytochemistry. Confocal analysis of exponentially growing LNCaP cells showed a diffuse distribution of CaM throughout the cell (Fig. 5). CaM was present in the nucleus, cytoplasm, and membranes. In contrast, AR was found predominantly in the nucleus. Analysis with a colocalization software program revealed that CaM in the nucleus is colocalized with AR (Fig. 5).' ―Cifuentes

'As shown in Fig. 3A, the bulk of cellular proteins passed freely through the column without binding to CaM in the presence of Ca²⁺ (fractions 1–8). However, a minor fraction of protein that was retained on the column in the presence of Ca²⁺ was eluted when Ca²⁺ was chelated with EGTA. The small peak of proteins observed after elution with EGTA-containing buffer represents proteins that exhibit Ca²⁺-dependent binding to CaM. Western blot analysis (Fig. 3B) of an equal amount of protein from each fraction eluted from the column (in the case of fractions 11 and 14, maximum volume was used) revealed a significant presence of AR in the peak fraction that eluted with EGTA. This fraction also contained cyclin A but not -actin, an abundant protein in LNCaP cell extracts (Fig. 3B).' ―Cifuentes

The androgen receptor binds to calmodulin only in the presence of calcium, a fact which provides the most straightforward mechanism for this effect. Based on the foregoing evidence, the ability of vitamin E ionophore to inhibit the proliferation of androgen-dependent LNCaP cells first-and-foremost depends on Ca²⁺ ions. These ions then bind to calmodulin, giving it an increased affinity for the androgen receptor which prevents its translocation into the nucleus.
 
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Obi-wan

Obi-wan

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What immediately stands-out is the author's use of 'VES' to signify α-tocopheryl succinate, a substitution having no place in logical acronym construction (LAC). There is no 'V' anywhere in the word 'α-tocopheryl succinate,' nor is there one in its IUPAC name: (2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-(4,8,12-trimethyltridecyl)]chroman-6-succinate ester. Upon further reflection, the reader finally gets it and becomes amazed that he hadn't thought of 'vitamin E succinate' sooner (yet is still befuddled why Zhang's acronym doesn't match his terminology):

'Here we demonstrate that α-tocopheryl succinate (VES) [?] can suppress the expression of prostate-specific antigen (PSA), a marker for the progression of prostate cancer. VES can also suppress androgen receptor (AR) expression by means of transcriptional and posttranscriptional modulation, but not ligand binding, nuclear translocation, or AR dimerization.' —Zhang

What also stands-out is the authors use of selenomethionine to inhibit cancer proliferation. On the basis of this, you might imagine this to be a somewhat-sophisticated researcher that knows all about polyamines; but if you had thought that you are likely wrong. This researcher had ostensibly used selenomethionine simply on the basis of the a few classic clinical trials, and his continued use of the word 'selenium' to represent 'selenomethionine' demonstrates naiveté.

'Moreover, selenomethionine (SM), a prostate cancer treatment adjuvant, shows an inhibitory effect on LNCaP cell growth, yet has no effect on the AR/PSA pathway.' —Zhang

'VES, but Not Selenium, Affects AR and PSA Expression. In previous research, selenium has been combined with vitamin E to study their antitumor activity, especially in epidemiological studies (4, 28). Therefore, we were interested in testing whether selenium could function similarly to vitamin E, which inhibits AR expression in LNCaP cells.' —Zhang

'Vitamin E, Selenium, and Prostate Cancer. In addition to vitamin E, a growing body of evidence suggests that a higher serum level of dietary supplemental selenium substantially reduces the incidence of lung, colon, or prostate cancer (40, 41). Other studies also reported that the selenium compounds inhibit the growth of prostate cancer cells through the induction of apoptosis.' —Zhang

He could not demonstrate a binding site for vitamin E succinate yet had eliminated the androgen receptor as a candidate, leaving the reader only to speculate on how it works. After thinking about this molecule for a few minutes, and the succinate moiety in particular, I had speculated that it could be acting as a calcium ionophore—or that is: The 6-succinate ester of vitamin E would be expected to chelate calcium from the extracellular fluid and carry it into the cytosol.

'Cell Culture and VES Treatment. The LNCaP and COS-1 cells were purchased from the American Type Culture Collection (Manassas, VA). Fibroblast cells were primarily cultured from normal prostate tissue. LNCaP cells were grown in phenol red-free RPMI medium 1640 . . .' ―Zhang

Sigma Aldrich informs me that 'RPMI-1640 contains 0.42 mM calcium' and Yuewen Gong informs me that 'intracellular calcium seems to be a potent regulator of AR gene expression in LNCaP cells.' On the basis of these foregoing facts, perhaps it would be worth reading his entire article:

Gong, Y. "Calcium regulation of androgen receptor expression in the human prostate cancer cell line LNCaP." Endocrinology (1995)

'To determine whether intracellular levels of calcium affected the expression of the AR, the AR-containing cell line LNCaP was treated with the calcium ionophores A23187 (10⁻⁶ M) or ionomycin (10⁻⁶ M), or the intracellular Ca*+- ATPase inhibitor thapsigargin (10⁻⁷ M), for 16 h.' ―Gong
Yuewen Gong had used the same culture media—RPMI 1640—as the Zhang study but had used two different calcium ionophores, albeit at lower concentrations than Zhang's vitamin E acetate (10⁻⁶ M vs 10⁻⁵ M). As he had hinted-about in the absract, intracellular calcium was found capable of reducing the expression of the androgen receptor:

'In the current study two compounds, A23187 and thapsigargin, were used to increase intracellular calcium levels in LNCaP cells. A23187 is a Ca²⁺ ionophore, widely used to analyze the role of Ca²⁺ in the regulation of gene expression (3436).' ―Gong

'The time dependencies of A23187 and thapsigargin downregulation of AR were examined next. Levels of AR transcripts in LNCaP cells began to decrease after 6-8 h of incubation with A23187 (10⁻⁶ M), reached a nadir after 16 h of incubation (29% of control), and increased to 59% of the control level at 24 h. Thapsigargin (10⁻⁷ M) treatment also resulted in a reduction in AR transcripts, decreasing after 6-8 h of incubation, reaching 23% of the control level after 10 h, and increasing to 68% of control by 24 h.' ―Gong

View attachment 9399

'In conclusion, increases in intracellular calcium concentration by the calcium ionophore or thapsigargin effectively reduce AR expression in LNCaP cells. Calcium inhibition of AR expression seems to precede calcium-induced apoptotic cell death. Furthermore, the mechanism of this reduction seems to be different from a pathway in which the calcium signal is transduced via a phosphorylation mechanism.' ―Gong

Although you'd almost be certain by the molecular structure that α-tocopheryl succinate is a calcium ionophore, we don't have to speculate since this had be proven in 1985.

Fariss, M. "Vitamin E reversal of the effect of extracellular calcium on chemically induced toxicity in hepatocytes." Science (1985)

'After a 1-hour incubation, the a-tocopherol content of control cells incubated with Ca²⁺-free medium and vitamin E succinate was approximately 1.1 nmol per 10⁶ cells or 1200 percent of the initial value (Table 1).' ―Fariss

View attachment 9400

'We therefore examined the effect of exogenous vitamin E on chemically mediated cell death in the presence or absence of extracellular Ca²⁺. The results show that the addition of vitamin E succinate to the incubation medium protected isolated hepatocytes incubated in Ca2²⁺ free medium against chemically induced cell death. Hence chemically induced cell death may be dependent not on the presence of extracellular Ca²⁺ but rather on cellular α-tocopherol (Table 1).' ―Fariss

Marc Fariss proves that vitamin E succinate responds powerfully to calcium levels, yet explains its ability to protect cells against calcium-related cell death in a calcium-independent manner. He seems entirely oblivious to the fact that he's using a calcium chelator and keeps referring to this as simply 'vitamin E,' as if the succinate moiety is irrelevant.

In 1990, the unique effect of vitamin E succinate to reduce cell proliferation had been proven yet again. To account for this effect, Kimberly Kline had rattled-off a litany of mechanisms while not committing to any one in particular. Of these mechanisms, Doctor Kline had toyed with the 'vitamin E succinate as a calcium ionophore' idea for awhile, yet had disregarded it (based on imprecise logic):

Kline, K. "Growth‐inhibitory effects of vitamin E succinate on retrovirus‐transformed tumor cells in vitro." (1990)

'Five antioxidants, including a synthetic analogue of vitamin E, Trolox, as well as the active vitamin form, DL-α-tocopherol, were incapable of inhibiting C4#1 tumor cell growth, indicating that a mechanism of action other than or in addition to functions as an antioxidant may be operating.' ―Kline

'These studies demonstrated that vitamin E, in the form of vitamin E succinate, inhibited the growth of retrovirus-transformed tumor cells in vitro and suggested that the antiproliferative effects of vitamin E succinate did not involve antioxidant properties but rather, as yet, unidentified mechanisms leading to cell cycle blockage.' ―Kline

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'The mechanism(s) whereby vitamin E succinate inhibits tumor cell proliferation is unknown. One well-characterized function for vitamin E is its antioxidant effect (27,28). Although vitamin E succinate does not exhibit any antioxidant properties unless the succinate is clipped off (29)...' ―Kline

'Considering that DL-α-tocopherol did not exhibit any antiproliferative activity at a concentration 20-fold greater than vitamin E succinate suggests that the antiproliferative activity exhibited by vitamin E succinate involves a mechanism that does not correlate with the classically defined units of tocopherol biologic activity.' ―Kline

'Regardless of the amount of calcium in the media in which the C4#1 cells were cultured (from 0.4 mM normally contained in serum-free RPMI-1640 to 2.0 mM), no differences were seen in either the ability of the C4#1 tumor cells to proliferate or in the cytostatic properties of various vitamin E succinate treatments. If it is speculated that vitamin E succinate is acting as a calcium ionophore, it would be predicted that vitamin E succinate treatment of C4#1 cells in the presence of increasing amounts of calcium in the media (0.4 mM to 2.0 mM) might result in increased cellular uptake of calcium. This would result in increased cytostatic effects (i.e., increased percent inhibition of tumor cell proliferation) and perhaps even cell kill at the highest calcium concentrations. The absence of any effects suggests that vitamin E succinate is not acting as a calcium ionophore in this situation.' ―Kline
Kimberly Kline fails to note that the extracellular calcium concentration would only matter when it's limiting, or when in the presence of a vitamin E succinate concentration that is greater than the calcium concentration. The highest concentration of vitamin E acetate she'd used (19·μM) had still been ~20× less than her lowest Ca²⁺ concentration (0.4·mM). On account of there being an excess of Ca²⁺ at both concentrations you wouldn't expect to see a difference anyhow, and would only expect to see one when the vitamin E acetate concentration approaches that of the Ca²⁺ concentration.

However, she did cite authors sympathetic to the 'ionophore' idea.

Slack, R. "Studies on the effects of vitamin E on neuroblastoma N1E 115." Nutrition and cancer (1989)

'It is quite possible that the free carboxylic acid group of tocopherol succinate is functional in conveying growth-arresting properties to this vitamin E derivative. Located in the membrane, this derivative could act much like free fatty acids do in biomembranes (i.e., as calcium ionophores) (36). Increases in calcium uptake by the cell as well as changes in levels of this divalent cation in the medium have been shown to greatly influence cell proliferation and differentiation processes (37-39). The functionality of the carboxyl group of tocopherol succinate with respect to calcium fluxes needs to be further investigated.' ―Slack
Finally, there is another person who sees things this way . . . and another:

Gogvadze, V. "Involvement of Ca²⁺ and ROS in α‐tocopheryl succinate‐induced mitochondrial permeabilization." International journal of cancer (2010)

'In particular, in addition to ROS production, α-TOS stimulates rapid Ca²⁺ entry into the cells with subsequent accumulation of Ca²⁺ in mitochondria—a prerequisite step for MPT induction.' ―Gogvadze

'Jurkat T-lymphocytes were cultured in RPMI 1640 complete medium...' ―Gogvadze

'Addition of α-TOS to transfected Tet21N cells caused a time-dependent stimulation of mitochondrial H₂O₂ production, which was blocked by the antioxidant N-acetylcysteine.' ―Gogvadze

'To investigate the ability of α-TOS to stimulate MPT, sequential pulses of Ca²⁺ were added to isolated rat liver mitochondria in the presence of α-TOS. Addition of Ca²⁺ to mitochondrial suspension led to a rapid increase in the level of this cation in the incubation buffer followed by a return to the initial level as mitochondria took up the added Ca²⁺.' ―Gogvadze

Gogvadze had proved vitamin E succinate was a calcium ionophore and had even said as much, yet he'd been focused on using rather high concentrations (50·μM) to draw-in enough Ca²⁺ to increase mitochondrial hydrogen peroxide concentrations—quite a bit different than the lower-dose transcriptional shifts noted by Zhang. Although his mechanisms and descriptions seem solid, these are not the same mechanisms noted by the other scientists and represent an entirely non-selective 'Ca²⁺ overkill' approach to inhibiting cancer cells.

Since I now have support for the calcium ionophore mechanism, I propose the low-dose transcriptional shifts had likely something to do with calmodulin:

Pelley, R. "Calmodulin-androgen receptor (AR) interaction: calcium-dependent, calpain-mediated breakdown of AR in LNCaP prostate cancer cells." Cancer research (2006)

Karacosta, L. "A regulatory feedback loop between Ca²⁺/calmodulin-dependent protein kinase kinase 2 (CaMKK2) and the androgen receptor in prostate cancer progression." Journal of Biological Chemistry (2012)

Cifuentes, E. "Physical and functional interaction of androgen receptor with calmodulin in prostate cancer cells." Proceedings of the National Academy of Sciences (2004)

'We observed tight binding of AR to CaM when LNCaP cell extracts were subjected to CaM-affinity column chromatography. AR binding to CaM was Ca²⁺-dependent and was inhibited by pretreatment of the cell extracts with W-7. Using immunofluorescence staining and confocal microscopy, we demonstrated colocalization of AR and CaM in the nucleus of LNCaP cells.' ―Cifuentes

'LNCaP cells were grown in RPMI medium 1640 containing 10 nM testosterone, whereas PC3 cells were grown in DMEM.' ―Cifuentes

'The binding of AR to CaM suggested that the two proteins may interact in cells. This hypothesis was examined by immunocytochemistry. Confocal analysis of exponentially growing LNCaP cells showed a diffuse distribution of CaM throughout the cell (Fig. 5). CaM was present in the nucleus, cytoplasm, and membranes. In contrast, AR was found predominantly in the nucleus. Analysis with a colocalization software program revealed that CaM in the nucleus is colocalized with AR (Fig. 5).' ―Cifuentes

'As shown in Fig. 3A, the bulk of cellular proteins passed freely through the column without binding to CaM in the presence of Ca²⁺ (fractions 1–8). However, a minor fraction of protein that was retained on the column in the presence of Ca²⁺ was eluted when Ca²⁺ was chelated with EGTA. The small peak of proteins observed after elution with EGTA-containing buffer represents proteins that exhibit Ca²⁺-dependent binding to CaM. Western blot analysis (Fig. 3B) of an equal amount of protein from each fraction eluted from the column (in the case of fractions 11 and 14, maximum volume was used) revealed a significant presence of AR in the peak fraction that eluted with EGTA. This fraction also contained cyclin A but not -actin, an abundant protein in LNCaP cell extracts (Fig. 3B).' ―Cifuentes

The androgen receptor binds to calmodulin only in the presence of calcium, a fact which provides the most straightforward mechanism for this effect. Based on the foregoing evidence, the ability of vitamin E ionophore to inhibit the proliferation of androgen-dependent LNCaP cells first-and-foremost depends on Ca²⁺ ions. These ions then bind to calmodulin, giving it an increased affinity for the androgen receptor which prevents its translocation into the nucleus.


So my question @Travis is Vit E succinate better than gamma VitE? Or are both beneficial? Just ordered Now Vit. E dry succinate.
 
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Travis

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So my question @Travis is Vit E succinate better than gamma VitE? Or are both beneficial? Just ordered Now Vit. E dry succinate.

I am skeptical that it would reach internal body sites intact at concentrations needed due to its labile ester group and substantial interaction with calcium, but of course I could be wrong so I'll check to see if there are any animal or human studies on this:

Prasad, K. "α-tocopheryl succinate, the most effective form of vitamin E for adjuvant cancer treatment: a review." Journal of the American College of Nutrition (2003)

'...several publications have shown that α-TS produces similar effects on a variety of human and rodent tumor cell lines without affecting the proliferation of most normal cells in vitro [8–37] and in vivo [10,19,20,38–41].' ―Prasad

[10] Helson L. "Vitamin E and human neuroblastoma." Karger (1983)
[19] Neuzil J. "Induction of cancer cell apoptosis by alpha-tocopheryl succinate: molecular pathways and structural requirements." FASEB (2001)
[20] Weber T. "Vitamin E succinate is a potent novel antineoplastic agent with high selectivity and cooperativity with Apo2 ligand in vivo." Clin Cancer Res (2002)
[38] Shklar G. "Regression by vitamin E of experimental oral cancer." J Natl Cancer Inst (1987)
[39] Malafa M. "Vitamin E succinate promotes breast cancer tumor dormancy." J Surg Res (2000)
[40] Malafa M. "Vitamin E inhibits melanoma growth in mice." Surgery (2002)
[41] Barnett K. "Vitamin E succinate inhibits colon cancer liver metastases." J Surg Res (2002)

One study cited by Prasad (#10) shows efficacy in rats having mammary tumors after i.p. injection . . . yet not after s.c. injection. This study had unfortunately not tried the oral route, but the results after i.p. injection had been notable (Fig. B). This had been the first study on vitamin E succinate in vivo but more were to follow. In 2002, mice were injected with melanoma and had received injections of vitamin E succinate (#40). The results of this had been even more profound than Helson's 1983 study (the other Fig. B), although this too is inconclusive as he too had used the i.p. route. Unless you have a large needle and are willing to self-reënact a Pulp Fiction scene, this route of administration is not an option.

figure B.png
Figure B2.png


This seems to work on all cancers and not just androgen-dependent prostate cancer, making a person wonder whether calmodulin interacts with estradiol receptors as well. This does appear to be the case; the estradiol receptor exhibits the same type of Ca²⁺-dependent binding to calmodulin as does the androgen receptor, while the glucocorticoid receptor is without interaction. This calcium regulation must be a general property of sex steroid receptors, and makes a person wonder if this is how prolactin exerts its effect (prolactin powerfully increases intracellular Ca²⁺).

Castoria, G. "In vitro interaction of estradiol receptor with Ca²⁺-calmodulin." Molecular Endocrinology (1988)

'In the initial experiments, crude as well as highly purified [³H]ER complexes in the presence of 10 nM [³H]estradiol were submitted to CaM-Sepharose chromatography in the presence of either Ca²⁺ or EGTA (Table 1). In the presence of Ca²⁺, about half of crude or purified receptor disappears from the receptor preparations because it binds to the columns, whereas in the presence of EGTA no binding occurs.' ―Castoria

'In contrast with the uterus ER the rat liver glucocorticoid receptor does not interact with CaM-Sepharose.' ―Castoria

'Ca²⁺ mediates the interaction of the receptor with CaM under every experimental condition used in this report. Half-maximal binding of purified ER to CaM-Sepharose is observed at about 0.3 μM Ca²⁺. The ability of ER to form a complex with CaM could explain the association of estradiol and Ca²⁺ activities.' ―Castoria
 
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Travis

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So my question @Travis is Vit E succinate better than gamma VitE? Or are both beneficial? Just ordered Now Vit. E dry succinate.

I think γ-tocopherol has more evidence behind it, but α-tocopherol succinate should work to a degree. I am not even sure they'd compete because the succinate form is more water soluble and could be carrier-independent, perhaps chelating Ca²⁺ in the intestines and going straight into chylomicrons? Either way, this certainly appears safe and it works through a completely different mechanism than γ-tocopherol (NO₂ sponge) and selenomethionine (polyamines)—making the effects synergistic, or noncompetive. I think α-tocopherol succinate could also help to absorb calcium, lowering the requirement for vitamin D.
 

Mito

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IdeaLabs Mitolipin has 30 IU of vitamin E per drop and it is high gamma Vitamin E.
 
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Obi-wan

Obi-wan

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We now have 2 worthy Vit E supplements. High Gamma in MCT oil and dry Vit E succinate and both can be taken on the same day. So my original question "Is Supplementing Vitamin E actually bad for you" is NOT if you are taking the right kind. Choice your Vit E carefully...not synthetic and not in PUFA oil and not alpha only. This thread has been a great education on Vit E.
 
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Obi-wan

Obi-wan

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IdeaLabs Mitolipin has 30 IU of vitamin E per drop and it is high gamma Vitamin E.

"We don't have the complete breakdown of all constituents yet, but we did our own analysis through a lab and alpha tocopherol is ~63%, gamma is ~19%, beta is ~%15 and delta is ~3%."-TocoVit.

"The one used for all other products is a high-gamma version but it is still 95% pure and has no residual PUFA. The TocoVit is high-alpha, since it is derived from wheat germ oil."

You were right. Also both are in MCT oil
 
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