Calcirol - Liquid Vitamin D3

Discussion in 'IdeaLabs' started by haidut, Dec 10, 2017.

  1. Amazoniac

    Amazoniac Member

    Joined:
    Sep 10, 2014
    Messages:
    7,013
    Gender:
    Male
    Location:
    Not Uganda
    You must've witnessed at least one person suffering from seneffiosis. It's a debilitating condition that has been taking over alternative wealth circles, the hosts are exposed to a vector (S. seneff) and pathogenesis is immediate if susceptible. What exactly makes it to the brain is still under investigation, but the hosts become wired to sulfur, they're driven to it even when stuffed because the more they has it, the closer they is to solving an issue.

    One of the claims is that sulfation makes photosynthesized venom D unique. Below, they found out that killcidiol is sulfated when incubated at civilized concentrations in liver cells and gels.

    - 25OHD3-3-O-Sulfate Is the Major Metabolite Formed from 25OHD3 by Human Hepatocytes

    "25OHD3-3-O-sulfate is reported to be a major circulating metabolite of 25OHD3 in humans, with an average circulating concentration comparable to that of 25OHD3 (Axelson, 1985; Shimada et al., 1995; Higashi et al., 2014), and thus, the sulfonation metabolic pathway might contribute importantly to vitamin D homeostasis. Surprisingly little is known about how and where 25OHD3-3-O-sulfate is formed. This information is crucial if the contribution from the sulfonation pathway to vitamin D homeostasis is to be fully evaluated."

    "The formation of 25OHD3-3-O-sulfate from 25OHD3 could be considered a catabolic process, but some investigators have hypothesized that it also represents an alternative 25OHD3 storage form in the body (Higashi et al., 2010). This is quite plausible because other endogenous steroid sulfate conjugates, such as estradiol-sulfate and dehydroepiandrosterone (DHEA)-sulfate, circulate at relatively high levels, are deconjugated in target tissues, and contribute to certain physiologic functions (Axelson, 1987; Banerjee et al., 2013; Sánchez-Guijo et al., 2015). Similarly, 25OHD3-3-O-sulfate might be retained in the circulation and distributed to different tissues of the body where it could be hydrolyzed to 25OHD3, replenishing the 25OHD3 pool, as needed. With this in mind, we also tested the binding affinity of 25OHD3-3-O-sulfate for DBP and its presence in human urine and bile."

    "Previous studies have demonstrated that at nonphysiologic concentrations (1–5 mM), 25OHD3 is metabolized to 1a,25(OH)2D3, 24R,25(OH)2D3, 4a,25(OH)2D3, 4b,25(OH)2D3, 25OHD3-3-O-glucuronide, 25OHD3-25-O-glucuronide, and putative 5,6-trans-25OHD3-25-O-glucuronide when incubated with human hepatocytes (Wang et al., 2014). However, formation of 25OHD3-3-O-sulfate in human hepatocytes was not determined. For this investigation, a more physiologic concentration of 25OHD3 (50 nM) was applied to cultured human hepatocytes to generate a metabolite profile. As shown in Fig. 3A, all major metabolites of 25OHD3, except 1a,25(OH)2D3, were detected in the incubations. 25OHD3-3-O-sulfate was the most abundant product observed, followed by 24R,25(OH)2D3, 4,25(OH)2D3 [4a,25(OH)2D3 and 4b,25(OH)2D3], and 25OHD3-glucuronides. Formation of these 25OHD3 metabolites occurred in a linear, time-dependent manner, except for 4b,25(OH)2D3 and 4a,25(OH)2D3 formation, which underwent extensive sequential glucuronidation, as previously reported (Wang et al., 2013a). By comparison, renal tubule epithelial cells and LS180 intestinal epithelial cells showed no detectable formation of 25OHD3-3-O-sulfate during a 24-hour incubation with 50 nM 25OHD3 (data not shown). These cells have previously been shown to catalyze the 24-hydroxylation of 25OHD3 under similar culture conditions (Zheng et al., 2012; Weber et al., 2016). Based on the assay limit of detection, culture conditions, and the approximate number of cells per well, it was estimated that 25OHD3 sulfonation activity per renal tubule epithelial cell or LS180 cell was ,10% that of cryopreserved human hepatocytes."

    upload_2020-1-27_11-57-21.png

    "Herein, we confirm that 25OHD3-3-O-sulfate is [] a quantitatively important circulating product of 25OHD3 (Axelson, 1985; Shimada et al., 1995) and we report for the first time that it is generated primarily in the liver by the enzyme SULT2A1." "Thus, interindividual differences in sulfonation activity may be a major source of variation in circulating blood 25OHD3 concentrations (Fig 8), assuming that hepatic clearance of 25OHD3 is an important determinant of 25OHD3 accumulation in the body."

    "SULT2A1 is known to metabolize hydroxysteroids, such as estradiol, DHEA and bile acids (Chatterjee et al., 2005). That it also catalyzes 25OHD3 sulfonation is not surprising, considering structural similarities. The regiospecificity of SULT2A1 toward 25OHD3 sulfonation, almost exclusively at the 3-position, is similar to that observed for hydroxysteroids and oxysterols. Interestingly, the related SULT isoform, SULT2B1, showed little activity toward 25OHD3, which is in contrast to its ability to generate sulfate metabolites of hydroxysteroids and oxysterols that are substrates for SULT2A1 (Falany and Rohn-Glowacki, 2013). SULT2B1 is expressed primarily in extrahepatic tissues (Falany and Rohn-Glowacki, 2013), whereas SULT2A1 is highly expressed in the liver and adrenal cortex and less so in the gastrointestinal tract (Chatterjee et al., 2005). This distribution pattern suggests that there will be limited 25OHD3 sulfonation activity outside of the liver. Consistent with these findings, no detectable 25OHD3 sulfonation was observed in either human intestinal LS180 cells or renal epithelial cells incubated with a physiologically relevant concentration of 25OHD3."

    This is related to what it was posted on the ex-nutrient thread:

    - From Performance Management System to Menopause: Female Hormones in Context

    "Radioactive estrogen has been shown to accumulate selectively in (liver) cancer cells, which is remarkable since that behavior is so untypical of liver cells. One of my first research projects had to do with the fact that estrogen promotes the formation of beta-glucuronidase, an enzyme which can reverse the reaction which normally occurs in the liver, detoxifying estrogen by combining it with glucuronic acid . Irritated tissues, and all cancers, contain beta-glucuronidase, with the capacity to 're-toxify" estrogen in the irritated or cancerous site, depositing it locally and negating the liver's protective function. More recently, breast cancer cells have been found to contain sulfatase enzymes, with the same kind of function, since the liver's other main route of estrogen detoxication is by combining it with sulfate. A systematic anti-estrogen program (including adequate protein to sustain liver function) would help to minimize the cancer-promoting action of this locally deposited estrogen. I think of the appearance of these estrogen-releasing enzymes in irritated tissue as part of a system for promoting regeneration. In the uterus, estrogen promotes simple growth, and progesterone promotes differentiation. I think something analogous happens in other tissues, with a variety of substances supporting differentiation."​

    "25OHD3-3-O-sulfate was found to have a high binding affinity for DBP, which explains its relatively high abundance in plasma and absence from urine. The crystal structure of DBP indicates that the vitamin D binding site is a cleft, which can easily accommodate large substituents at the C-3 position of 25OHD3 (e.g., conjugated moieties) (Verboven et al., 2002). High binding affinity to DBP would reduce the renal excretion of 25OHD3-3-O-sulfate into urine. We speculate that the complex of 25OHD3-3-O-sulfate and DBP is filtered and then reabsorbed in renal proximal tubules by megalin/cubilin-mediated endocytosis, as shown for the 25OHD3-DBP complex (Rowling et al., 2006)."

    "Although 25OHD3-3-O-sulfate is a major circulating form of vitamin D3, whether it possesses biologic activity directly or indirectly is unclear. A number of studies have been conducted to understand the biologic activities of vitamin D3-sulfate, a conjugated metabolite of vitamin D3 (Higaki et al., 1965; Sahashi et al., 1967a,b, 1969). Vitamin D3-sulfate was synthesized previously (Reeve et al., 1981) and its biologic activity was determined in a vitamin D–deficient rat model. Activity was observed, but only at doses higher than what can be elicited by vitamin D3 (Nagubandi et al., 1981). Later studies also showed less biologic activity of vitaminD3-sulfate than free vitaminD3 in vivo (Cancela et al., 1985). However, in each of these studies, vitamin D3-sulfate was administered, rather than having the metabolite generated in situ, with the uncertainties of bioavailability and access to cellular sites that complicate quantitative comparisons. Thus, it is possible that 25OHD3-3-O-sulfate might undergo hydrolysis, catalyzed by ubiquitous sulfatases and regenerate 25OHD3. This type of hormone conjugate cycling is observed for estrogen and DHEA (Mueller et al., 2015), where the sulfoconjugates are the dominant form in blood circulation and are distributed to peripheral tissues where desulfonation can occur. In the case of DHEA-3-O-sulfate, conversion to DHEA is followed by metabolism to androstenedione and downstream androgens and estrogens (Strott, 2002)."

    "Finally, given the detection of 25OHD3-3-O-sulfate in bile, we are intrigued by the possibility that preferential delivery of the hormone conjugate to the duodenum and upper small intestine might explain the preferential expression of vitamin D receptor target genes, such as CYP3A4, transient receptor potential cation channel subfamily V member 6 (TRPV6), and calbindin D9K, in the upper small intestine (Wang et al., 2013b). Results from unpublished studies indicate that 25OHD3-3-O-sulfate is a substrate for the cell uptake transporter, organic anion transporting polypeptide 2B1 (OATP2B1), which is expressed in the intestinal epithelia (Drozdzik et al., 2014). Once absorbed into mucosal epithelial cells, 25OHD3-3-O-sulfate could be hydrolyzed to 25OHD3 and then undergo 1a-hydroxylation to the active hormone and contribute to the regulation of TPRV6, calbindin D9K, and CYP3A4 (Wang et al., 2013b). With regard to the kidney, 25OHD3-3-O-sulfate bound to the DBP in blood could be filtered in the glomerulus and then reabsorbed in the proximal tubular epithelium through the action of megalin/cubilin, similar to what occurs for the 25OHD3-DBP complex (Negri, 2006). Again, intracellular hydrolysis of the conjugate and bioactivation to 1a,25(OH)2D3 could contribute to the known biologic effects of vitamin D in this tissue. Further work is needed to explore these mechanistic hypotheses."​

    It could be argued that it's sulfatization of killciol. It's been mentioned that its presence in milch in this form supports the argument, yet there are publications that haven't detected any and are ignored.

    The next experiment shows that in a subject whose contamination was mild, utilization was good judging from the appearance of killcidiol after killciol, therefore it's being metabolized and conjugation is lost for it to happen yet levels are maintained elevated in spite of this, so it could've undergoned processing in liver.

    - Vitamin-D Synthesis and Metabolism after Ultraviolet Irradiation of Normal and Vitamin-D-Deficient Subjects

    upload_2020-1-27_11-58-9.png

    - Bioavailability of Alendronate and Vitamin D3 in an Alendronate/Vitamin D3 Combination Tablet

    upload_2020-1-27_11-58-40.png

    Oral intoxication differs (it would be better if all metabolites were measured), but what bothers is the selective sulfuroscopic view to form a solid theory, it's highly suspicious and potentially toxic.

    --
    - Nongenomic actions of steroid hormones
     
  2. Amazoniac

    Amazoniac Member

    Joined:
    Sep 10, 2014
    Messages:
    7,013
    Gender:
    Male
    Location:
    Not Uganda
    Venom D3 and its IU is toxic nomenclature and unit, should be regulated as drugs. There ainn't a good reason for the '3' and the unit serves for nothing when it could for something, it's just one more unnecessary thing to memorize.

    I was discussing with yerrag how basic concepts are neglected, these are examples.

    A safe margin of intakes exists for all nutrients, no need to be perfect, but if dosing is usually as multiples of thousands of IU, it better be meaningful, otherwise why not something more practical than an irrelevant unit?

    A person may argue that it doesn't matter, it's the biological effect that counts (it's the given amount in whatever unit for a desired outcome). Although this is true, what are the odds that it's ideal to grade it as such? We might have the impression to be working with round figures, but in reality they isn't because the divisions have no significance and it's pointless to keep using them, it's off from relevant parameters (such as multiples required to induce a certain effect in humanoids or derived from our synthesis).

    Another argument to be made is that diet will provide some, which throws off round figures anyway when the total intake is considered. It's also true, but in using an irrelevant unit, it's possible to induce people to use more or less without a decent justification. The more you can divide it, the more control the users will have over the dose even if these are arbitrary (for making it easier to make adjustments). It's why reducing the content per drop to 1000 IU was a step on the right direction. For example, let's say that instead of 1000 IU, 800 IU would be more compatible with our biology because it was graded based on a scale of biological effects. Dosing it twice of each would provide 2000 or 1600 IU, which is quite a significant difference in the long-term. If a reasonable dose is 1600 IU, how can we get them close if the minimum dose is 1000 IU? Now if the minimum is 500 IU or 400 IU, we can obtain 1500 IU or 1600 IU without a problem. It's a matter of deciding how it's best to intoxicate ourselves.

    With the suspicion of its randomness, I camed across the following publication. Low and be hold, we can confirm how arbitrary it is for us because the unit originated from the vitamin D activity of 1 mg of irradiated ergosterol under standardized conditions (I think it was based on an amount that made animals reach a certain bone density). Later, that was defined as 0.025 mcg of venom D2. Then, it was decided that it's best to use venom D3 as the standard. At this point, it's already meaningless for humans. If it was something like the amount produced from irradiated 7-dehydrocholesterol present in skin, a typical area exposed, moderate pigmentation, at noon of the time on equator, and so on, it would be understandable.

    However, given how responses vary between people, metric system would still be preferable in spite of bearing no connection with biology due to the practical aspect of it.

    Toxic independent of the unit.

    - Problems relating to the definition of an international unit for vitamin D and its metabolites

    - Vitamin D

    "Vitamin D1 [520-91-2] is a mixture of vitamin D2 and lumisterol." :thumbsdown:

    "The common name vitamin D is used throughout the pharmaceutical industry for simplicity. The trivial name calciferol has also been used extensively with the prefix ergo- and chole-, which indicate vitamin D2(2)and vitamin D3(4), respectively (see Steroids)."

    "The vitamins D are 9,10-secosteroids, that is, steroid molecules with an opened 9,10 bond of the B-ring."

    "The isolation and structure elucidation of vitamin D are closely related to the efforts to understand and cure rickets and related bone diseases. The advent of the use of soft coal, the migration of people to cities, and the tendency of people and animals to spend less time in sunshine caused a decline in the ability of populations to synthesize sufficient quantities of vitamin D3. This led to the increased incidence of rickets, beginning aroundthe mid-1600s (3)."

    "The bone disease rickets in children, and a similar condition in adult known as osteomalacia, is characterized by the body’s inability to calcify the collagen matrix of growing bone, resulting in wide epiphysealplates and large areas of uncalcified bone called osteoid. The resultant lack of rigidity of bones leads to theends becoming twisted and bent, particularly in long bones. The ribs develop a bumpy and uneven texture known as rosary ribs and the legs become bowed. Also, the cranium becomes soft and misshapen. In adults, no long-bone growth occurs, but new bone, which is being continually remodeled, activates cells to resorb bone, followed by osteoblast-mediated bone-growth replacement (4, 5)."

    "Although there is evidence that rickets was manifest in humans as early as 800BC, it was not until 1645 that it was first described (6). The progress of finding a cure for rickets was relatively slow until the late 1800s, when a sufficient number of scientific developments began to allow workers to unravel the difficult puzzle of vitamin D metabolism. Tarret in 1889 had isolated ergosterol from ergot of rye and demonstrated that it was different from cholesterol. The similarity of the structures, however, led to difficulty in the structural elucidation of the vitamin D molecule. Mellanby (7) demonstrated the lack of a dietary component could be used to develop rickets and was able to raise D-deficient animals. This allowed research aimed at finding the antirachitic factor to be carried out."

    "The irradiation of calciferol in the presence of iodine leads to the formation of 5,6-trans-vitamin D2 [14449- 19-5] 5 or D3 [22350-41-0] 5 (67, 68). 5,6-trans-Vitamin D as well as vitamin D (2) or (4) can be converted to isovitamin D by treatment with mineral or Lewis acids."

    "Vitamin D and its products are sensitive to uv light, heat, air, and mineral acids. Its sensitivity to these conditions is exaggerated by the presence of heavy-metal ions, eg, iron."

    "Vitamin D is introduced into the bloodstream either from the skin after natural synthesis by the irradiation of 7-dehydrocholesterol stored in the epidermis (172) or by ingestion and absorption of vitamin D2 or vitamin D3 through the gut wall (40). Between 60 and 80% of the vitamin introduced in the blood is taken up by the liver, where cholecalciferol is transferred from chylomicrons to a vitamin D-binding protein (DBP), an α-globulin specific for vitamin D and its metabolites but one which does not bind with previtamin D in the skin (173). Cholecalciferol is hydroxylated in the liver at the C-25 position (51, 141, 174). This hydroxylation occurs in the endoplasmic reticulum and requires NADPH, a flavoprotein, cytochrome P-450, Mg2+, and O2 (175)."
     
  3. Amazoniac

    Amazoniac Member

    Joined:
    Sep 10, 2014
    Messages:
    7,013
    Gender:
    Male
    Location:
    Not Uganda
    They question below the constance of the enzyme(s?) that add a hydroxyl group on carbon 25 (the 25-hydroxylases that convert killciol to killcidiol), because so far it's been assumed that only the next steps of its metabolism are impacted according to the state of the victim. That there are about 50 variations of this pollutant and 'only 1 out of 3-10 molecules' of killciol ends up being converted to killcidiol. Also, (mostly for excretion) it can be sulfated, glucuronidated, taurinated, fatty acidated ('most vitamin D found in fish is in the form of fatty acid esters') and there may be recovery from what's excreted in the intestines (the enterohepatic recirculation). You'll find other topics discussed that make it worth reading:

    - Vitamin D Metabolism Revised: Fall of Dogmas
     
  4. Amazoniac

    Amazoniac Member

    Joined:
    Sep 10, 2014
    Messages:
    7,013
    Gender:
    Male
    Location:
    Not Uganda
    One potential concern in not using absorption enhancers is requiring large doses of venom D that could build up at the upper layers of the skin, being metabolized locally before entering the circulation, leading to aqvcrsc conscdncuccs. However, in the experiment below they've used large amounts and the outer skin layer content was still low relative to the dose. The other layers had it even lower. They have disconsidered formulations without the boosters.

    - Skin layers

    upload_2020-2-9_11-48-50.png
    Source: the internet.

    - Investigating Transdermal Delivery of Vitamin D3

    upload_2020-2-9_11-49-13.png


    upload_2020-2-9_11-49-19.png

    "Compared to F1, formulation F2 delivered four times more compound into the epidermis and two times more into the stratum corneum, while F3 delivered eight times more into the epidermis and ten times more into the stratum corneum."

    Enhancers:
    • Nothing
    Outer layer: 1x
    Upper layers: 1x
    Ratio: 1​
    • Oleic acid
    Outer layer: 2x
    Upper layers: 4x
    Ratio: 0.5​
    • Dodecylamine
    Outer layer: 10x
    Upper layers: 8x
    Ratio: 1.25​

    However, it didn't correlate with serum intoxication:


    "The cumulative amounts of vitamin D3 penetrated through the skin into the receptor solution (pig serum) over 24 h from F1, F2, and F3 were found to be 170 ± 48, 0, and 360 ± 260 ng/cm2, respectively. For F2, no detectable increase in the concentration of vitamin D3 in the porcine serum was observed."​

    From the artificial part of the experiment:

    "[..]oleic acid—the penetration enhancer in the F2 ointment—decreased the release of vitamin D3 compared to the control. This is most likely due to the lipophilic complex formed between oleic acid and vitamin D3 (48,49)."​

    I should have included this on the other thread:

    "In patients with celiac disease, biliary obstruction and chronic pancreatitis, the absorption of tritium-labeled (3H)-vitamin D3 fell to 50%, <28%, and <18%, respectively. This is significantly lower than normal subject absorption, which ranges from 62 to 91% (4,5). In another publication, it has been reported that only 50% of the vitamin D3 dose is actually absorbed in cystic fibrosis patients (6). Furthermore, long-term use of bile acid binding medications such as colestipol and cholestyramine will decrease vitamin D3 absorption (5)."​

    Results above are odd considering the following..

    - Be Wary Of Vitamin D Supplementation
     
  5. Amazoniac

    Amazoniac Member

    Joined:
    Sep 10, 2014
    Messages:
    7,013
    Gender:
    Male
    Location:
    Not Uganda
    Questioning the seneffiotic argument:

    - Water-Soluble Vitamin D in Human Milk: A Myth

    "The earlier reports of large amounts (400 to 950 IU/liter) of vitamin D sulfate in human milk were based on a precipitation of vitamin D sulfate with barium hydroxide, and ultimately a calorimetric reaction with antimony trichboride which is very sensitive to, but not specific for, vitamin D. In these early reports it is unclear exactly what compound(s) was precipitated with barium hydroxide. More precise techniques for separation of vitamin D metabolites (including high-pressure liquid chromatography) have now shown negligible amounts of vitamin D sulfate in the whey fraction of human milk.[9,11]"

    "When [] purified vitamin D sulfate was given either orally or intraperitoneally to vitamin D-deficient rats,[8,10] it had less than 5% of the biologic activity of vitamin D3 in terms of mobilizing calcium from bone and less than 1% of the activity in supporting bone calcification[10] or stimulating the intestinal transport of calcium.[8,10]"​

    - Vitamin D of human milk: identification of biologically active forms

    Both by Heitor and friends.

    The strange part is pushing venom D on fetuses. Dangerous, we know what should be done.
     
  6. Peatogenic

    Peatogenic Member

    Joined:
    Sep 11, 2017
    Messages:
    476
    Gender:
    Male
    Your communication style is very cryptic. I can't tell if you prefer it that way.
     
  7. Amazoniac

    Amazoniac Member

    Joined:
    Sep 10, 2014
    Messages:
    7,013
    Gender:
    Male
    Location:
    Not Uganda
    - Sulfur Deficiency | The Weston A. Price Foundation

    "The form of vitamin D that is present in both human milk[19] and raw cow’s milk[2] is vitamin D3 sulfate (pasteurization destroys it in cow’s milk)."​
     
Loading...