I downloaded the article and put it on my school fileserver https://homepage.usa...884/imminst.pdf
tlm884, thanks a bunch.
Posted 14 June 2009 - 02:40 AM
I downloaded the article and put it on my school fileserver https://homepage.usa...884/imminst.pdf
Posted 14 June 2009 - 07:10 AM
I downloaded the article and put it on my school fileserver https://homepage.usa...884/imminst.pdf
tlm884, thanks a bunch.
Posted 14 June 2009 - 07:41 AM
We don't really want to maximize serum concentration, since MB only improves mitochondrial function in a specific range.By the looks of things, if you want the most out of methylene blues effects on the brain IV administration would be the best bet. However, this is most likely not the easiest route for people. Oral administration results in high concentrations of methylene blue in the liver and intestines and low serum concentrations. However, the LD50 (1180mg/kg in rats) is high enough that you could probably boost your dose a bit to get higher serum concentrations. Multiple daily dosing is also needed as the half life is ~5hours. 2-3 times a day would be sufficient to keep serum levels from changing too much throughout the day but OD dosing would result in extended periods of low serum concentration.
Posted 14 June 2009 - 07:58 AM
We don't really want to maximize serum concentration, since MB only improves mitochondrial function in a specific range.By the looks of things, if you want the most out of methylene blues effects on the brain IV administration would be the best bet. However, this is most likely not the easiest route for people. Oral administration results in high concentrations of methylene blue in the liver and intestines and low serum concentrations. However, the LD50 (1180mg/kg in rats) is high enough that you could probably boost your dose a bit to get higher serum concentrations. Multiple daily dosing is also needed as the half life is ~5hours. 2-3 times a day would be sufficient to keep serum levels from changing too much throughout the day but OD dosing would result in extended periods of low serum concentration.
It's nice that Oral administration actually yields a more steady serum level, which is what we're looking for here.
Posted 23 June 2009 - 02:26 AM
Effect of Methylene Blue on pyridine nucleotides and insulin secretion of rat pancreatic islets H. P. T. Ammon1, 2 and E. J. Verspohl1, 2
(1) Abteilung für Pharmakologie der Universität Erlangen-Nürnberg, FRG<a name="Aff2">(2) Pharmazeutisches Institut der Universität Tübingen, FRGReceived: 12 September 1978 Revised: 5 February 1979
Summary Methylene Blue, which is known to oxidise NADPH in red blood cells, was used to assess a possible role of NADPH in the glucose-stimulated secretion of insulin. When islets from rats were incubated with 3 mg/ml of glucose, Methylene Blue (0.5, 1.0, 2.0 or 5.0 g/ml) significantly decreased the concentration of NADPH, increased that of NADP+ and decreased the NADPH/NADP+ ratio in a dose-dependent manner. This effect was associated with inhibition of the glucose-induced insulin release. No significant change of NADH, NAD+ and ATP could be observed. It is suggested that the secretory response of the pancreatic islet to glucose stimulation depends on the NADPH/NADP+ ratio.
Posted 23 June 2009 - 04:32 AM
Hmmm, Methylene Blue reduces insulin response to glucose.
I presume this isn't a problem if you're on a low-carb diet, but is this a significant problem ?
The concentration they're using is about 1.5 nM to 10 nM
http://www.springerl...0v305830357303/Effect of Methylene Blue on pyridine nucleotides and insulin secretion of rat pancreatic islets H. P. T. Ammon1, 2 and E. J. Verspohl1, 2
(1) Abteilung für Pharmakologie der Universität Erlangen-Nürnberg, FRG<a name="Aff2">(2) Pharmazeutisches Institut der Universität Tübingen, FRGReceived: 12 September 1978 Revised: 5 February 1979
Summary Methylene Blue, which is known to oxidise NADPH in red blood cells, was used to assess a possible role of NADPH in the glucose-stimulated secretion of insulin. When islets from rats were incubated with 3 mg/ml of glucose, Methylene Blue (0.5, 1.0, 2.0 or 5.0 g/ml) significantly decreased the concentration of NADPH, increased that of NADP+ and decreased the NADPH/NADP+ ratio in a dose-dependent manner. This effect was associated with inhibition of the glucose-induced insulin release. No significant change of NADH, NAD+ and ATP could be observed. It is suggested that the secretory response of the pancreatic islet to glucose stimulation depends on the NADPH/NADP+ ratio.
@niner: I've seen analytical reports on MB purity for the kind sold to consumers, and all the heavy metal levels where well below the strictest limits. Even if some metals like mercury or lead, for the sake of argument, were in the low ppb range, when you are taking just 100 microliters of that solution, you'll only be getting the equivalent of 100 parts per quadrillion amounts; well below detection, and you probably get an order of magnitude or two more than that in your drinking water. Furthermore, MB is used to treat fish of fungal diseases and other pests, and fish are more sensitive to heavy metals than us, from what I know.
My area of research is directly on a mouse/human metal sensor protein (for zinc), which also responds to heavy metals. Our bodies have a lot of heavy metal defenses. Metals just aren't quite as dangerous (nor is radiation) as we treat them, but better safe than sorry!
Edited by rwac, 23 June 2009 - 04:34 AM.
Posted 28 June 2009 - 02:43 PM
Here's an interesting article on MB and red light to treat cancer. It seems like this idea goes back to 1989.
http://cancerres.aac...ull/63/4/776#B8
"Treatment of cancer by PDT (photodynamic light therapy) usually involves the generation of reactive, excited oxygen molecules (1) A photosensitizing dye absorbs light, exciting it to an energetic triplet state capable of transferring its energy efficiently to ground state, triplet molecular oxygen to form excited, singlet oxygen. .... Because red light penetrates tissue much deeper than that of shorter wavelength, photosensitizers absorbing in the red are preferred. Phenothiazinium dyes such as MB+ or TB+ absorb strongly at approximately 600–670 nm, and have been identified as having potential for cancer diagnosis or treatment. ..... MB+ has been extensively investigated as a photocatalyst of oxidative processes, in experimental PDT in vitro and in vivo, and in limited clinical studies involving PDT (8,9) . We hypothesized that these dyes could be photocatalysts of oxidation of IAA, the commonest plant auxin (growth hormone), the dye triplet state oxidizing IAA to produce a radical-cation (Fig. 1) ....We have shown that products of IAA oxidation are cytotoxic to mammalian cells, including human tumor cells. Thus, the combination of IAA or suitable analogues with these photosensitizers could provide an alternative route to photosensitized cell killing not involving singlet oxygen. ... We show here that IAA, a common, nontoxic plant chemical, is transformed into potent cytotoxins using red light and these phenothiazinium dyes that are already in clinical use. Activating IAA rather than oxygen to form cytotoxins offers the potential to treat hypoxic tumors effectively.
In conclusion, the present results show that the phototoxicity of MB+ and TB+ is enhanced dramatically by the presence of IAA at concentrations likely to be achievable in humans. The use of IAA, or analogues with modified redox properties, with these or other suitable photosensitizers is attractive because of the opportunity for rational selection of both photosensitizer and IAA analogue, based on thermodynamic (redox) and kinetic arguments. In principle, a requirement is that the reduction potential of the photosensitizer triplet state [e.g., of the couple MB+/MB·, where MB· is the radical (semiquinone)] is higher than that of the indolyl radical cation (IAA·+/IAA); ideally, the reduced dye should also be efficiently "redox cycled" to the parent dye by oxygen. These requirements appear to be fulfilled for MB+ and IAA."
I imagine light sticks being for throat and prostate cancer.
and it seems like if you take MB and get a lot of sun light it could kill viruses in the blood: "An increasingly important medical use of MB+ exploiting its photodynamic activity is to reduce viral contamination of blood plasma by MB+ and light."
"The prevalence of West Nile virus (WNV) infections and associated morbidity has accelerated in recent years. Of particular concern is the recent demonstration that this virus can be transmitted by blood products and can cause severe illness and mortality in transfusion recipients. We have evaluated methylene blue (MB) + light as a safe and cost-effective means to inactivate WNV in vitro. This regimen inactivated WNV with an IC50 of 0.10 μM. Up to 107 pfu/ml of WNV could be inactivated by MB + light with no residual infectivity. MB + light inactivated three primary WNV isolates from the years 1999, 2002 and 2003 and prevented mortality in a murine model for WNV infection. Since MB is already approved for human use at a dose of 100 mg/kg/day, we conjecture that MB + light treatment of blood products for high-risk patients will be efficacious and suitable for use in resource-limited settings."
Posted 23 July 2009 - 05:45 PM
Perhaps good if collagen production is not desirable in response to lipid peroxidation?Ascorbic acid (0.2 mmol/L) induced lipid peroxidation and stimulated collagen alpha 1(I) gene transcription in cultured human fibroblasts. Inhibition of the ascorbic acid-induced lipid peroxidation in cultured human fibroblasts with alpha-tocopherol (50 mumol/L) or methylene blue (10 mumol/L) prevented the stimulation of collagen gene expression.
Edited by stephen_b, 23 July 2009 - 05:45 PM.
Posted 23 July 2009 - 10:48 PM
Methylene blue prevents the stimulation of collagen gene expression (PMID 1962560).
Perhaps good if collagen production is not desirable in response to lipid peroxidation?
Posted 24 July 2009 - 02:40 PM
Posted 24 July 2009 - 03:50 PM
We're all waiting for somebody to test whether mice live longer.?!? with this kind of view, pretty much any effects from anything is good, as long as you find the rare specific disease that it possibly alleviates
personally i'm many more times interested in knowing whether mice live longer under methylene blue
Posted 25 July 2009 - 12:41 PM
me neitherI don't see the problem with discussing other effects
i'm not sure preventing collagen production is good in general. Plus this is in vitro results under very specific tests. concluding that MB is beneficial in general, based on such minor observation is IMO very biased. this is what i meantand MB has many many beneficial effects
We're all waiting for somebody to test whether mice live longer.
haven't you seen the MPrize @ home threads?btw, what are you upto with your mice ?
Posted 04 October 2009 - 02:42 PM
Posted 06 October 2009 - 05:40 AM
The MB-dependent increase in oxidant production at the high concentration of MB could be due to MBH2 auto-oxidization by O2 to form a superoxide radical. Because it is known that MBH2 preferentially reduces cytochrome c over O2 (40), we suspected that the levels of cytochrome c in the mitochondrial lysate become limiting in the presence of 1 or 10 µM MB. Therefore, we tested whether cytochrome c added in excess to the mitochondrial lysate could block the production of oxidants by 10 µM MB. We found that adding cytochrome c to the mitochondrial lysate did, in fact, eliminate the MB-dependent increase in the oxidation of DCFH
One of the earliest examples of pharmacological preconditioning was the mild inhibition of the respiratory chain with nitropropionic acid or acetylsalicylic acid,http://www.pubmedcen...tid=2668955#R38 which induced cellular changes such as those seen with hypoxia. Another relevant example is the use of the iron chelator desferrioxamine, which induces nuclear translocation of the transcription factor hypoxia-inducible factor 1, with consequent expression of a plethora of hypoxia-inducible genes, including erythropoietin, vascular endothelial growth factor, and hexokinase
Examples include inhibitors of mitochondrial respiration, stabilisers of hypoxia-inducible factor 1 (signal: hypoxia), or lipopolysaccharide (signal: infection)
Posted 19 October 2009 - 12:42 PM
We're all waiting for somebody to test whether mice live longer.
I don't see the problem with discussing other effects until then, and MB has many many beneficial effects.
Methylene blue trihydrate has a variety of biomedical and biologically therapeutic applications. Groups of 50 male and 50 female rats and mice were administered methylene blue trihydrate in 0.5% aqueous methylcellulose solution by gavage at doses of 0, 5, 25, or 50 mg/kg bw/day (rats) or 0, 2.5, 12.5, and 25 mg/kg bw/day (mice), 5 days per week for 2 years. In rats survival of all dosed groups was similar to that of the vehicle controls, whereas mice exhibited a dose-dependent increase in survival. Rats receiving 25 and 50 mg/kg bw/day and mice receiving 25 mg/kg bw/day developed mild anemia. The incidences of pancreatic islet cell adenoma and adenoma or carcinoma (combined) were increased in all dosed groups of male rats, but increases were statistically significant in 25 mg/kg bw/day males only and the dose-response was non-linear. There was a corresponding increase in the incidence of pancreatic islet cell hyperplasia but statistically significant only in the 50 mg/kg bw/day male rats. There were no significant increases in neoplastic transformation observed in the mice; however, positive trends were noted for adenoma or carcinoma (combined) of the small intestine and malignant lymphoma.
Posted 19 October 2009 - 04:26 PM
Not the same as living longer but it points in the right direction. Are the dosages used too large to cause concern about the noted carcinomas?
Edited by rwac, 19 October 2009 - 04:27 PM.
Posted 19 October 2009 - 05:13 PM
Posted 20 October 2009 - 02:53 AM
Maybe I'm overly cautious, but I'd consider that mouse study a negative. Without human studies, isn't there some concern over long term use? The carcinoma thing would worry me... actually, taking fish medicine would worry me too...
Perhaps it is simply the MAO Inhibition affect causing those taking MB to perceive a benefit? A placebo test would also be interesting, although that could be difficult to run -- guess some blue dye drops might work.
Edited by abelard lindsay, 20 October 2009 - 02:58 AM.
Posted 20 October 2009 - 03:27 AM
Since the doses are, what, 5-6 orders of magnitude higher than we're using, I'm not terrifically worried about the carcinomas. The dose-dependent increase in survival is interesting, but given the dose, it makes me think that there is something else going on in the high-dose regime compared to the low-dose regime.Yeah, the doses are much higher than the ones we're talking about.Not the same as living longer but it points in the right direction. Are the dosages used too large to cause concern about the noted carcinomas?
However, a dose-dependent increase in survival would mean that we need *much* larger doses for life extension.
Posted 20 October 2009 - 03:35 AM
True. And I didn't mean to imply the benefits were solely in a person's head (placebo or depression related). I just know some of the reports like tiny tiny doses having a greater effect than normal dosing, or CoQ10 possibly interfering with MB (although apparently not if dosed at a different time, even though half-life of CoQ10 is rather long)... sounds strange.Maybe I'm overly cautious, but I'd consider that mouse study a negative. Without human studies, isn't there some concern over long term use? The carcinoma thing would worry me... actually, taking fish medicine would worry me too...
Perhaps it is simply the MAO Inhibition affect causing those taking MB to perceive a benefit? A placebo test would also be interesting, although that could be difficult to run -- guess some blue dye drops might work.
Its positive effects may also be due to its strong anti-microbial and anti-fungal properties. That's what it's used for after all, to remove fungal, bacterial and parasitical infections from fish tanks.
Posted 20 October 2009 - 03:49 AM
Since the doses are, what, 5-6 orders of magnitude higher than we're using, I'm not terrifically worried about the carcinomas. The dose-dependent increase in survival is interesting, but given the dose, it makes me think that there is something else going on in the high-dose regime compared to the low-dose regime.
Edited by rwac, 20 October 2009 - 03:52 AM.
Posted 01 November 2009 - 08:15 PM
Effect of methylene blue, indigo carmine, and Renografin on human sperm motility.
Sheynkin YR, Starr C, Li PS, Goldstein M.
Department of Urology, Center for Male Reproductive Medicine and Microsurgery, New York Hospital-Cornell Medical Center, New York, New York 10021, USA.
OBJECTIVES: To evaluate the effect of commonly used intraoperative vasography and tissue staining agents, indigo carmine, methylene blue, and Renografin, on sperm motility. METHODS: Semen from 20 healthy men was obtained after 2 to 4 days of abstinence. Sperm motility was initially evaluated in each specimen. Standard solutions of indigo carmine, methylene blue, and Renografin-60 were diluted 2x and 4x with lactated Ringer's solution. Equal aliquots of sperm were mixed with undiluted and diluted drugs, and sperm motility was assessed. RESULTS: Initial mean sperm motility was 70.3%+/-3.0%. Undiluted methylene blue and Renografin severely depressed sperm motility to 1.1%+/-0.5% and 2.3%+/-0.7%, respectively (P <0.05). Diluted methylene blue depressed motility to 4.9%+/-1.8% and 11.2%+/-3.0% (P < 0.05). Diluted Renografin depressed motility to 25.1%+/-4.1% and 55.3%+/-3.3% (P < 0.05). Although undiluted and 2x-diluted indigo carmine moderately decreased sperm motility (48.9%+/-3.2% and 61.7%+/-3.0%, P < 0.05), 4x-diluted indigo carmine had minimal effect on sperm motility (67.3%+/-2.8%, P > 0.05). Lactated Ringer's solution had no effect on sperm motility. CONCLUSIONS: We found a severe, immediate reduction in sperm motility after exposure to undiluted standard solutions of methylene blue and Renografin. Dilution of Renografin significantly decreased its negative impact on the sperm motility, whereas the adverse effect of methylene blue remained fairly constant even with increasing dilution. Sperm motility should be assessed prior to application of these agents. Sperm should be aspirated for immediate use and/or cryopreservation prior to the use of these agents. Indigo carmine may be safely used as a tissue stain or vasography agent with a minimal effect on sperm motility in dilutions of 4x and higher.
PMID: 9886615 [PubMed - indexed for MEDLINE]
Edited by abelard lindsay, 01 November 2009 - 08:16 PM.
Posted 01 November 2009 - 11:36 PM
Effect of methylene blue, indigo carmine, and Renografin on human sperm motility.
...
CONCLUSIONS: We found a severe, immediate reduction in sperm motility after exposure to undiluted standard solutions of methylene blue and Renografin. Dilution of Renografin significantly decreased its negative impact on the sperm motility, whereas the adverse effect of methylene blue remained fairly constant even with increasing dilution. Sperm motility should be assessed prior to application of these agents. Sperm should be aspirated for immediate use and/or cryopreservation prior to the use of these agents. Indigo carmine may be safely used as a tissue stain or vasography agent with a minimal effect on sperm motility in dilutions of 4x and higher.
PMID: 9886615 [PubMed - indexed for MEDLINE]
Edited by rwac, 01 November 2009 - 11:36 PM.
Posted 02 November 2009 - 01:10 AM
Posted 02 November 2009 - 01:56 AM
If it's being used as a tissue staining agent, then it's probably pretty concentrated. I suspect that at the concentration that we'd be using, the effects on sperm motility would be gone.
Edited by rwac, 02 November 2009 - 01:56 AM.
Posted 02 November 2009 - 04:57 PM
Posted 02 November 2009 - 06:08 PM
It is interesting, and I won't deny the possibility of real benefits, but what I think scares most people off (scares me off, anyway), is lack of long-term safety data, and the fact the only real way to get it is by using fish medicine. I'm somewhat conservative regarding sourcing safe supplements as it is, meant for human use. Going to the pet shop to get MB feels like too big a risk to me.As far using CoQ10 and MB blue together, it really didn't make a difference what time I placed these two apart. For me, they did not work together. So I have to choose to either take MB and CoQ10. But trust me, there is NO placebo effect with MB. This stuff really makes you sharp.
I'm really surprised more people aren't jumping on this, especially since a drug was developed because this worked so well.
Posted 02 November 2009 - 06:21 PM
Going to the pet shop to get MB feels like too big a risk to me.
Posted 04 November 2009 - 06:09 AM
Posted 07 November 2009 - 07:42 PM
Not the same as living longer but it points in the right direction. Are the dosages used too large to cause concern about the noted carcinomas?
Yeah, the doses are much higher than the ones we're talking about.
However, a dose-dependent increase in survival would mean that we need *much* larger doses for life extension.
It's also nice to find the potential trouble spots. Pancreas because MB interferes with ROS signaling, and intestine because it is exposed to large doses of MB which is pro-oxidant.
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