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OfflineBleuboxo
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MIMOSA BARK IN MAiLBOX!!!!!
    #321571 - 05/20/01 08:43 PM (21 years, 4 months ago)

good afternoon friends, my mimosa bark is in my hands as we speak!! im so excited to try this stuff out! its already ground up too! so im just waiting for my jars to fully colonize now. i have rye jars, innoculated them with dextrose mycelia, so it should be done in about 4-5 days i guess. ill keep posting.

" Insanity is just a thread of reality...the make-as-you-go part of living, the bare second reflex of dying _Stavros Christou... by_"


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" Insanity is just a thread of reality...the make-as-you-go part of living, the bare second reflex of dying _Stavros Christou... by_"


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OfflineThe Messiah
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Bleuboxo]
    #321656 - 05/20/01 10:18 PM (21 years, 4 months ago)

excellent! *sinister look*

peace, -=To0dLeS=-
TM


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[green]peace[/green], [blue]~(To0dLeS)~[/blue]
TM


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OfflineoOjonahOo
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: The Messiah]
    #321718 - 05/20/01 11:30 PM (21 years, 4 months ago)

How are you going to compare the psilocybin levels? Bioassay is adequate but certainly not definitive. Do a double blind taste test and ask your friends to evaluate.

It would be nice to settle this questions once and for all.

Personally, though, If i was bothering to order Mimosa Hostilis I would just extract the DMT...I can always grow more mushrooms, potentcy is not that important to me...i'd rather try to see god or at least some mischevious little elves.





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Ultimate Mycodirectory v4.0


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Offlinehubertd8
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: oOjonahOo]
    #323641 - 05/23/01 03:39 AM (21 years, 4 months ago)

i agree, DMT seems like such a more powerful trip, why would you waste it on trying to increase psilocybin levels when DMT is worth so much more, interms of $$$ and effects. Growing more mushies or just more potent one is alot easier. Plus what mushie strain is he mixing it with?



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"There is much pleasure to be gained from useless knowledge."

Bertrand Russell


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OfflineDarK_SavioR
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Bleuboxo]
    #323716 - 05/23/01 05:13 AM (21 years, 4 months ago)

awesome bleuboxo... one thing though, didn't someone mention that the dmt needs to be in the cakes or whatever you innoculate in order for the mycelium to take full advantage of it? I thaught someone said something about reading about Tihkal's experiment with using the dmt to increase the potency of the shrooms and said that casing with the dmt isn't nearly as effective as putting the dmt in the cakes or compost or whatever. Either way, good luck with it and I'll be looking for a post with the results ; )



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Vitamin C chase, kill the taste. You can tell its nasty by the look on my face.
Ralphster44 & The FSR!
All thats stated above is for humor and a lie!!


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InvisibleTripsAreForKids
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: DarK_SavioR]
    #324349 - 05/24/01 01:03 AM (21 years, 4 months ago)

yea, i read that too.. the guy said to mix it in with the jars or the mycelia wont convert as much of it as it would if you mixed it in at the beginning....

Keep Trippin' and Blazin'


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Everything posted above was out of fun, none should be taken serious. I am currently under the influence so take that into consideration.


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OfflinePsilocybin
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: oOjonahOo]
    #324562 - 05/24/01 05:01 AM (21 years, 4 months ago)

Converting the DMT to psilocin is more efficient in terms of psychedelic effects. Psilocin is many times more potent than DMT. I would suggest you do an extraction to obtain the DMT then mix in 10 mg to a substrate, just make some grain spawn, pasteurize some dung/compost and right before you mix in your grain spawn add 10mg of DMT to your dung/compost. This is not alot you will be losing to gain a large increase in mushroom potency, as a normal dose of DMT is 40-50 mg smoked and 100-200mg in an oral dose, you will only be losing less than one smoked dose to produce a lot more mushroom fun.

note: How would you go about adding it before you make the substrate? Do you mean you have to start the spores on the DMT, or that it cannot be added through injections or a casing layer?

Edited by Psilocybin on 05/23/01 10:04 PM.



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Offlinegray1
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Psilocybin]
    #325086 - 05/24/01 10:07 PM (21 years, 4 months ago)

i would reccomend extracting the dmt and including it/exposing it to the fungi organism as early as possible. in order to be effective in increasing the psilocybin content (which we have shown numerous times as theoretically possible based on the simple cellular metabolic conversions) it must be bioavailable to the organism. i recently read a pare concerning incorporation of substances into the fungi based on radiolabelling, and it appears as if dmt is poorly absorbed/transported into the organism. therefore, make it as easy as possible by providing it in a pure form and exposing the organism to it as long as possible.

i also mention this in the doubters of the dmt method post
"tryptamine, whcih is readily formed from tryptophan by P. cubensis, serves as a better precursor of psilocybin than tryptophan. N-methyltryptamine is a still better progenitor of psilocybin, but n, n dimethyltryptamine is rather poorly incorporated as judged from the dilution factors, however, if the poor absorbtion of this compound by the fungus, less than 5%, is taken into account, the high dilution factor does not make it an unlikely intermediate."

best of luck, please post results when you get them, along with exactly how you did the experiment, and how you tested the potency.

gray1
an exerpt from the journal

c12h16n24ohdmt


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Invisiblepaddoholland
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Bleuboxo]
    #328518 - 05/29/01 01:55 PM (21 years, 4 months ago)

I suppose there will be a rise of levels, but non will be measuered. I doubt if you'd notice it. Extraction takes longer than making some extra shrooms so you can eat more and trip as hard as with super potent ones!

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OfflineBleuboxo
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: paddoholland]
    #329295 - 05/30/01 07:46 AM (21 years, 4 months ago)

damn, long time i havent posted...10 days!!! 1094 new posts!!! whoa!!!! im in florida at the moment and my roommate said that all the jars are completely colonized so i just told him to go ahead and case them. so far they have been cased for two days now, and oh yeah, i didnt know about adding it to the jars first unless i would have waited and done that. but the quantity wasnt rerally that much, so two casings with mimosa are now in progress. hope to see results in about 10 days or so. ill be back home monday night, the 4th of june. time is really flying because i havent been around my mushies almost two weeks it seems like i forgot everything about growing....NOT lol. just seems weird though, just curious to see how everything is going. well, wish me luck and ill post on monday. have a good trip!

" Insanity is just a thread of reality...the make-as-you-go part of living, the bare second reflex of dying _Stavros Christou... by_"


--------------------
" Insanity is just a thread of reality...the make-as-you-go part of living, the bare second reflex of dying _Stavros Christou... by_"


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Invisiblesynaptic
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Bleuboxo]
    #329704 - 05/30/01 10:18 PM (21 years, 4 months ago)

Heh, I just got my pre-ground MHRB in the mail too... 150g total from Brazil (via the netherlands)..

Are you going to extract the elf spice? I've been reading quantum tantra's a/b extraction procedure but 39 days for an extraction is a long time to wait for 30 minutes of space cadeting.. saw something at the hive about having at least some product available in 2 hours (basically, you do your first batch right away then reextract the remaining stuff over the next few weeks)..

Let me know how it goes!

synaptic



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Offlinethe universe
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #330276 - 05/31/01 09:22 AM (21 years, 4 months ago)

So what you're saying (hypothetically) is if I got some L-Tryptophan from the football league, that would work better than DMT? Bada-bing bada-boom bada-potent-ass-shroom? That sure would save a guy a lot of money.

"If a man has character, he has also his typical experience, which always recurs."-Nietzche


--------------------
"If you had a million years to do it in, you couldn't rub out even half the 'Fuck you' signs in the world."- J. D. Salinger


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Invisiblesynaptic
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: the universe]
    #330313 - 05/31/01 10:14 AM (21 years, 4 months ago)

The amino acid tryptophan is the immediate precursor of tryptamine and brown rice contains lots of it. I've still never been able to find out how much tryptophan BRF contains as a % by weight or anything. While I suspect that doping the substrate with additional tryptophan will increase alkaloid levels, I'm betting there are some other amino acids involved too that might also need to be supplemented.

Does anyone have access to journal archives or have the enzymatic biosynthesis of psilocybin/in article laying around? I'd really like to look at this and Elsevier is the grinch when it comes to looking at their stuff online.

Agurrel, S. and J.L.G. Nilsson. "A Biosynthetic Sequence from Tryptophan to Psilocybin." Tetrahedron Letters 9: 1063-1064 (1968).

Of course, subjective bioassay is only interesting to a point. Without some mechanism for quantitatively measuring the different amounts of psilocybin/in in the mushrooms, I think the results would still be rather inconclusive and potentially wrong. Other factors also come into play in the development of the actual mushroom fruit body as alkaloid levels vary from one to another depending on when it was harvested, how it was grown, and so on. On the other hand, I think incubation could be fairly controlled.

So how would one go about measuring the actual levels of tryptamine alkaloids in these samples? You could do extractions but those seem fraught with error. I don't know much about chromatography at all (read a tutorial or two on the web) but wouldn't TLC just give you a boolean answer as to whether a chemical is present or can you actually derive amounts?

Edited by synaptic on 05/31/01 03:16 AM.



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Offlinegray1
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: synaptic]
    #330462 - 05/31/01 03:50 PM (21 years, 4 months ago)

synaptic
the only way that other amino acids would be involved in this process is if they were first converted to tryptophan.
tryptophan is the archetypical building block for psilocybin and psilocin.



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Invisiblesynaptic
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #330552 - 05/31/01 07:11 PM (21 years, 4 months ago)

> the only way that other amino acids would be involved in this process is if they were first converted to tryptophan.
> tryptophan is the archetypical building block for psilocybin and psilocin.

Uhm, I don't think this is the case at all.

There are 20 amino acids that form peptide bonds to create polypeptides. Thousands of these amino acids bond together and create proteins. Most enzymes are proteins. You know how complex DNA is with only four base pairs. Proteins have twenty! These are the precursor chemicals to the enzymes that catalyze the psilocybin and psilocin production from tryptophan.

There has to be some upper limit of tryptophan doping where you're getting tryptophan or something else out because there aren't enough enzymes to catalyze the reaction into our desired psilocybin/in.

So we have to choose from:

alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine.

I think it's naive to think only tryptophan will increase yields. Now to just figure out what the fungi needs to create the enzymes of interest.

synaptic



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Offlinegray1
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: synaptic]
    #330754 - 05/31/01 11:20 PM (21 years, 4 months ago)

please recognize this as an attempt to provide constructive criticism and provoke further thought on this subject.

what is naive is for you to think that supplementing with any other amino acid would have more than minute and certainly un-detectable, distant effect on biosynthesis of psilocybin and/or psilocin.

granted, there are several enzymes involved in this process and enzymes are constructed from amino acids, and it may in fact be benefitial towards the goal of increased production of indole alkaloids to have higher than normal physiological levels of the involved enzymes,
this is what i understand you to be suggesting:
supplement with random amino acid---->increased ammount of involved enzyme---->increased amount of psilocybin/psilocin production.

it is simply unreasonable to suggest that supplementing with an amino acid will specifically increase the concentration of a certain enzyme.
it is reasonable to assume that supplementing with amino acids does increase protein/enzyme production in general (think body builders and protein supplements) however, this is unspecific. supplementing with amino acids/protein simply will supply the organism with the proper building blocks, it will not directly stimulate the biosynthetic pathways (shikmic acid pathway) and the actual effect on psilocybin production would be miniscule and extremely watered down.
in general, supplementing with a broad range of amino acids could be benefitial to overall growth and therefore to psilocybin production, but this is as important as proper humidity, light, temperature, nutrients in substrate...
obviously what is desired is to provide the optimal growing conditions and thereby maximize the organism's inherrent, natural ability to produce all of the secondary metabolites that it does, including psilocybin and psilocin,
however, supplementing with a random amino acid with the intent to increase psilocybin content through the reasoning that enzymes are made of amino acids is foolish.

on the other hand, the aromatic amino acid tryptophan is THE most basic precursor for psilocybin and psilocin, it is a direct building block of the final molecule which is modified several times by enzymatic actions, the intermediates being dmt etc...
and it is quite possible that while tryptophan can be incorporated into many enzymes/proteins or other biosynthetic pathways, direct supplementation has the potential to not only provide for the need of starting material, but may also actually stimulate the process through a feedback process.

i hope that this will clear up your mis-conception.
gray1



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Invisiblesynaptic
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #330928 - 06/01/01 03:02 AM (21 years, 4 months ago)

Good points. I get the distinct feeling using the word "naive" kind of irked you. I didn't mean anything by it..

Anyway, I see what you're saying.. yeah, supplementing the other amino acids may be good for the overall nutrition of the fungi but adding this or that amino acid won't increase the amount of enzymes that produce psilocybin/in. That makes sense.. as long as the fungi was given it's recommended daily allowance :) of whatever, it'll produce what it needs and the rest will be wasted. That there's some switch that says "hey, make this much of this enzyme" in it's DNA or something?

While on the other hand, increasing tryptophan levels will increase psilocybin/in levels because the enzymes catalyze what's there.. the fungi doesn't have to say, "ok, i need to make this enzyme".. it has already made all the enzymes it's going to make, so they are there and simply catalyze the reaction with the excess tryptophan that the fungi hasn't used for other purposes..

Right? That seems to makes sense... You're right that I *was* originally thinking one might be able to boost enzyme levels with other additives but maybe that would be something more strain specific. While probably out of the realm of possibility anywhere in the near future, one might tweak this or that gene and create a potential for less regulation of the production of our enzymes of interest to the point where traditional substrates, while productive, would be less than nominal to boost alkaloids.

Sounds neat on paper at least.. :)

synaptic



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OfflineOpi
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: synaptic]
    #331278 - 06/01/01 10:10 AM (21 years, 4 months ago)

There is one problem we are not going to get around and that is there are rate limiting genes to contend with.

Also adding tryptophan is starting at the wrong end of the pathway.. Let me explain. If you add excess typtophan, you must also supply excess amino acids and nutrition to allow conversion of tryptophan into tryptamine, tryptamine into N-methyltryptamine, N-methyltryptamine into N,N-dimethyltryptamine, and finnally N,N-dimethyltryptamine into gold. With genetically imposed limits at each and every step. Start as close as you can to your final product and you reduce the stress on the fungus by orders of magnatude. As it is, if you want psilocybin as your final goal, expect to add some excess phosphate to the substrate... in the form of KH2PO4 - Potassium dihydrogen phosphate. AKA - Mono Potassium phosphate (MKP).



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Offlinegray1
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #331420 - 06/01/01 05:08 PM (21 years, 4 months ago)

opi
since i have started posting here, especially on this subject, i have tried to stress one ideal, and that is supporting the natural biosynthesis of psilocybin to the organsims full inherrent capacity by optimizing the environment in which it lives.
in this respect, i am not suggesting that there is a direct relationship to the level of tryptophan supplied in the substrate to the level of psilocybin produced, and, in fact i agree that there may in fact be a negative feedback pathway associated with extreme excess of precursor.
i also agree that adding an intermediate that is further toward the end of the completion of the metabolic process as a supplement is theoretically more desirable because it requires fewer enzymatic reactions to reach the final product, however, two factors immediately come to mind that decrease the facility of this:
1. the further intermediates are not readily availiable to the lay person. they must be extracted from plants or are controlled chemicals that are basically impossible for the mushroom hobbyist to come by.
2. those intermediates that are availiable, such as dmt after the process of extraction, maqy not be biologically available to the organism when introduced to the substrate. please see the radiolabel incorporation paper that i referenced earlier in this thread.
because of these two factors, i continue to support the theory that suplementation with tryptophan may increase psilocybin levels, and if tryptophan is unavailiable, 5-htp, though one step further away from the goal, may also serve as a valuable substrate addition.
mono and di phosphates are also good supplementation candidates to promote overall growth and function of the organism, they are often added in liquid culture medium for the culture of other microorganisms such as bacteria(e. coli used in molecular biology research) ans yeast.



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Offlinegray1
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: synaptic]
    #331425 - 06/01/01 05:22 PM (21 years, 4 months ago)

i apologize if that reply was hostile in any way, i find that despite the fact that the majority of the people that post here are good natured and have good intent to learn and /or share info, there are quite a few assholes who are either too negative to experimentation or to insistent that they know everything. because of this, i am generally very defensive of my posts, i'm sorry if i ofended you.
to briefly reiterate by response to opi in relation to the levels of enzymes, i am in favor of supporting psilocybin biosynthesis by attempting to provide the organism with the optimal growth environment, in this case doing so with nutritional supplements, to allow for the full natural functioning of metabolic processes. in this respect, utilizing existing physiological enzyme concentrations.

this is completely beyond the scope of my knowledge and it is purely a speculation of possibility, but the pathways involved in cellular feedback and control are immensly complex, and adding tryptophan could serve to mediate several things, such as a negative feed back loop, or actual indirect transcriptional up-regulation of the involved enzymes. who knows...

however, for my other interest, genetic engineered systems that produce psilocybin/psilocin, the involved genes would definately be put to strong promotion/transcrtiption/translation to provide high levels of enzymes that could then convert high levels of supplemented tryptophan to psilocybin.

this is obviously a rough experiemnt and theory because we lack the information about the true nutritional requirements of the organsim and also the full nutritional information for the substrates that those who cultivate the organism use.



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OfflineOpi
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #331616 - 06/01/01 10:01 PM (21 years, 4 months ago)

I guess my basic feeling is not so much favoring one precursor over another, but simply illustrating that the further you are from the end of the path, the more work you must do to take care that you also have an excess of all building blocks from that point on, plus enough to take care of the fungus, as it is doing more work than normal.

This might require more work than it is worth. I don't mean overall. I think these experiments should be attempted (UNDER CONDITIONS THAT DON'T VIOLATE ANY LAWS). I just mean that it might be more work to try and use tryptophan (even for an extreme amature) than to try and obtain, extract, and prepare DMT as a precursor.

I speculate that early atempts at adding tryptamine, which resulted in gnarled stems, and only excess psilocin suffered mainly from lack of this sort of thinking. The fungus was probably doing everything it could to get rid of this excess tryptamine, at expense of its own healthy functioning, and without the benifit of excess phosphate to create psilocybin.

I also want to thank you all for participating in what I feel is one of the most productive threads in a long time. And we really haven't accomplished anything yet! But we are straightening out a lot of muddled thinking on the subject, and I think we are close to a reasonably easy approach to boosting the levels of the desired alkaloids.

OPI



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Offlinegray1
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #331662 - 06/01/01 11:01 PM (21 years, 4 months ago)

"I just mean that it might be more work to try and use tryptophan (even for an extreme amature) than to try and obtain, extract, and prepare DMT as a precursor. "


again, i feel that i need to stress that DMT is NOT a good substrate addition
although it is an intermediate in the biological process, studies have shown that it is not incorporated into the organism from the substrate. if it doesn't get into the cells it can't be a part of any biosynthetic pathways, regardless of how good a potentiator it may or may not be. period.
this is why i continue to suggest tryptophan as the only realistic substrate addition for those who are so inclined to carry out experiments. it is readilly availiable, of reasonable expense, and can be incorporated into the organism.

whether the addition of tryptophan will give rise to any noticable difference (increase) in potency and psilocin/psilocybin remains to be seen, and even so will be highly subjective.

also, i am not trying to suggest that the fungi should do "more work than normal" or function abnormally,
just attempting to creat a theoretical optimal environment for the NATURAL function of the organism to it's full inherrent capabilities.

gray1

additionally, i don't see how adding tryptophan to the substrate during regular substrate preparation is more work than growing/ordering, extracting and adding DMT to the substrate, in fact, the reverse seems true to me. i think that any amature could pop open a few capsules and mix it inwith the vermiculite/brf...

Edited by gray1 on 06/01/01 04:06 PM.



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Offlinejonnyshaggs420
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #331790 - 06/02/01 02:01 AM (21 years, 4 months ago)

In the "Tryptophan" post, also in this forum, it is stated that primrose seeds and sunflower seeds carry an abundance of biologically produced tryptophan more than any other organic source.

If this info. is accurate than why not just add a little crushed primrose seed or sunflower seed to brf/verm or birdseed substrates.  This help boost levels of alkaloids, but still provide other nutrients needed by the fungus, perhaps avoiding the gnarled fruits.

4 out of 5 inmates agree.  The FSR is better than being raped by your overweight roomate!;)
Support the FSR.


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OfflineOpi
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #332198 - 06/02/01 02:45 PM (21 years, 4 months ago)

I guess you only read half of what people say... I won't explain why it is less work in this post, as I did more than an adaquate job in my last post.

And actually DMT works fine as a substrate addition.
"studies have shown that it is not incorporated into the organism from the substrate"

Just plain not true. There is a difference between not AS effective, and NOT effective.

It HAS been shown to increase psilo levels, just is not AS effectivly as tryptamine (probably due to solubility).

And if you are elevating the levels of precursors above what you will ever find in nature, I would hesitate to call it natural. It certainly is more work for the fungus than it would encounter in nature..

I know this is something you are investing a lot of thought into, but please don't get defensive over ideas.



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OfflineOpi
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #332199 - 06/02/01 02:51 PM (21 years, 4 months ago)

OK.. I can't resist RE-explaining myself...
Tryptophan is plain NOT AN OPTION. By itself it does not work. That HAS been demonstrated. You might be able to "WITH A LOT OF WORK" get it happen. But you would have to take so many things into consideration (i.e. Every step of the biosynthetic process from tryptophan to psilo) to get it to happen, that it would be harder to accomplish than using DMT. And the organism would have a chance to limit you at SEVERAL steps instead of just at the final one.

OPI



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OfflineOpi
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #332202 - 06/02/01 03:00 PM (21 years, 4 months ago)

Damn why couldn't I just put this all in one post 8^)

I speculate that the specific reason tryptophan doesn't seem to elevate alkaloid levels is that the mechanism is already functioning at a level where it is converting as much tryptophan into tryptamine, as is going to happen. Maybe I am wrong about being limited at every step. Maybe this is the only limiting step the fungus happens to have, which would make this a little easier. I certainly hope so.



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Offlinejonnyshaggs420
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #333327 - 06/04/01 02:25 AM (21 years, 3 months ago)

just a little question here.  If the fungus is limited with tryptophan than it may just as well be limited with tryptamine and DMT and psilo's. 
Maybe we should all just sit down and ask the fungus what it wants.

4 out of 5 inmates agree.  The FSR is better than being raped by your overweight roomate!;)
Support the FSR.


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Offlinegray1
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #333875 - 06/04/01 05:04 PM (21 years, 3 months ago)

opi:
you say that DMT is proven to work: where is the scientific proof/literature behind this statement? please post a link or reference.

DMT IS NOT INCORPORATED INTO THE ORGANISM
go back and read through this thread again (or more likely for the first time) and you will see the literature reference to this that used a radiolabel incorporation study. if you give me your email i'll attach the paper as a pdf. (let me remind you again of what i have previously stated, in theory, i think that increased in vivo levels of DMT would cause an increase in end product, but simply adding DMT to the substrate will not increase in vivo levels, therefore no increase in end product will be seen.)

you state that adding tryp alone won't work, while i concede that this may be the true, possibly because the substrates commonly used are already supplying all of the needed tryptophan, but you seem to say so definatively.
please reference to what makes you say this.

i consider supplementing to the full nutritional requirements of the mushroom natural. who is to say that the natural environments in which these mushrooms grow don't provide optimal conditions, including nutritional requirements? and why shouldn't we strive to provide equally optimal environments in our artifical process, but do so as naturally as possible?
obviously the difficulty here is twofold:
1. we don't don't know the precise nutritional value of the substrates used
2. we don't know the true nutritional requirements of the organism.
for humans, don't you think that getting the full RDA ov every vitamin and mineral, correct mg/kg protein, correct carbos, fatty acids, etc... is natural? with proper diet it certainly can be achieved, if you don't think that taking vitamins is natural.
again, through supplementation, i only suggest that we try to create the optimal environment for the organism to grow, and let it do it's thing concerning the production of indole alkaloids.





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Offlinegray1
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #333879 - 06/04/01 05:11 PM (21 years, 3 months ago)

"I guess my basic feeling is not so much favoring one precursor over another, but simply illustrating that the further you are from the end of the path, the more work you must do to take care that you also have an excess of all building blocks from that point on, plus enough to take care of the fungus, as it is doing more work than normal.

This might require more work than it is worth. I don't mean overall. I think these experiments should be attempted (UNDER CONDITIONS THAT DON'T VIOLATE ANY LAWS). I just mean that it might be more work to try and use tryptophan (even for an extreme amature) than to try and obtain, extract, and prepare DMT as a precursor."

you appear to be speaking about two different "works" here:
the work of the mushroom that is escalated because of supplying more precursor
and the work of the cultivator, supplying more precursor.

my response,

additionally, i don't see how adding tryptophan to the substrate during regular substrate preparation is more work than growing/ordering, extracting and adding DMT to the substrate, in fact, the reverse seems true to me. i think that any amature could pop open a few capsules and mix it inwith the vermiculite/brf...

stands true. the excess work you are refering to is the work of the fungus itself, obviously the cultivator is not responsible for this work.






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Offlinegray1
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #333921 - 06/04/01 06:31 PM (21 years, 3 months ago)

bleuboxo:
how's the experiment coming?

gray1



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OfflineOpi
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #336184 - 06/06/01 10:13 PM (21 years, 3 months ago)

Believe me gray1, I have read the thread. I guess my mistake was believing Gartz, Shulgin, or YOU when you said:

' i would reccomend extracting the dmt and including it/exposing it to the fungi organism as early as possible. in order to be effective in increasing the psilocybin content (which we have shown numerous times as theoretically possible based on the simple cellular metabolic conversions) it must be bioavailable to the organism. i recently read a pare concerning incorporation of substances into the fungi based on radiolabelling, and it appears as if dmt is poorly absorbed/transported into the organism. therefore, make it as easy as possible by providing it in a pure form and exposing the organism to it as long as possible.

i also mention this in the doubters of the dmt method post
"tryptamine, whcih is readily formed from tryptophan by P. cubensis, serves as a better precursor of psilocybin than tryptophan. N-methyltryptamine is a still better progenitor of psilocybin, but n, n dimethyltryptamine is rather poorly incorporated as judged from the dilution factors, however, if the poor absorbtion of this compound by the fungus, less than 5%, is taken into account, the high dilution factor does not make it an unlikely intermediate." '

8^)

In reference to tryptophan, feel free to search the shroomery for the numerous posts about people trying to use tryptophan with NO PERCEIVEABLE increase in potency (Ryche Hawk for one). I will assume that perceivable is what counts here. I also understand that you can't trust everything posted on the shroomery, but judging the direction that people tend to lie around here, I would assume that if they were going to lie about this, they would claim it did work, not the other way around. And trust me, certain people would thrust themselves forward to write up a tek and claim credit for their discovery (sorry, that just amuses me to no end). I'm sorry, but I won't admit to having tried it, so I have no first hand experience with it. If you trust that I am not going to lie about it, I will say that I have spoken to a two organic chemists, and one biochemist who HAVE tried this (where it is legal to do so) and they had no luck. So again, if you trust me... Tryptophan (under normal conditions) will not boost psilo levels.

Ok, now I will hold your hand now and explain the "work" I was talking about. 8^)

I have theorized that there are genetic limitations in the biosynthetic process of P. cubensis that will try to hinder the production of excess psilo alkaloids. Do some reading on allosteric enzymes... The problem is end product inhibition. Basically when the amount of psilocin gets high, it starts binding to the enzyme converting tryptophan into tryptamine and prevents that step from occuring any further. The good news (and this explains the known behavior of adding precursors to P. cubensis) is that this usually only happens at the first step of the biosynthetic process. This partially explains why tryptamine results in massive psilocin, but tryptophan does not. Getting psilocybin, would probably require that work I was talking about. My theory WAS that you would have to supply not only enough nutrition for regular metabolic processes, but also EXTRA nutrition (maybe nutrition is not the best word here) to supply the building blocks of the excess: tryptamine, N-methyltryptamine, n, n dimethyltryptamine,psilocin, and psilocybin. We can be reasonable sure this is true for the psilocybin step. It probably doesn't have enough phosphate to create the psilocybin from the psilocin. My suggestion that starting with DMT would be easier meant that you would only have to supply this kind of extra 'nutrition' for the last two steps of the biosynthetic process since simply adding tryptophan isn't going to do it. So I was saying it would be more work to get tryptophan to work than to extract DMT. Looking at it now, we can see I was only partially right.. No matter what you do, you probably can't use tryptophan, but it still would require "something" extra to get tryptamine, or n-methyltryptamine to produce excess psilocybin and still be healthy enough to fruit normally.
That is all I was trying to say, and it really is a while lot of arguing over a petty issue. I think we have a lot more to offer each other working together on this. And even arguing we are comming up with some amazing ideas! The subject seems to have taken over the advanced forum !!!

Back to work:
Tryptamine results in few, gnarled, unhealthy looking mushrooms. Again probably not enough nutrition to carry out this much alkaloid production and maintain healthy growth, sporulation, and fruiting ability.

Now here is the BIG DEAL!!! I think I realized WHY tryptamine works so well and why dmt works only slightly

WATER SOLUBILITY!!!!

You might not be able to get the mushroom to take up what is in the substrate, but if it is in the water that that composes 92% of the fungus, then it will appear to be part of the mushrooms normal biosynthetic process, as it will be absorbed into the fungus..

Tryptamine is soluble in water, trytophan isn't, DMT is slightly, n-methyltryptamine is slightly...

What do you think?

OPI




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OfflineHumidity
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #336266 - 06/06/01 11:53 PM (21 years, 3 months ago)

Opi great post I am no chemist but I would think that water solubility would be a important factor in the success of the experiment. I think that is why in the tryptamine cubinses tek they used tryptamine HCL instead of just plain tryptaime because the tryptamine HCL is water soluble and tryptamine alone is not.

"I have theorized that there are genetic limitations in the biosynthetic process of P. cubensis that will try to hinder the production of excess psilo alkaloids. Do some reading on allosteric enzymes... The problem is end product inhibition. Basically when the amount of psilocin gets high, it starts binding to the enzyme converting tryptophan into tryptamine and prevents that step from occuring any further. " Opi

I agree with you 100% here. I was reading some books on secondary metabolies and many of the pathways are limited by the end product. This allows the organism to spend energy one other things.

I posted something not to long ago about secondary metabolite overproduction take a look at it. Instead of trying to add some of these chemicals that are very hard to get and some may not even work we can have the fungi overproduce them for us. Industry does this all of the time. Trace metals play a very important role in many of these processes also industry limits certain nutrients to cause overproduction. Read that post it goes into more detail.





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_____________________________________________________________________________________
"The greatest enemy of knowledge is not ignorance, it is the illusion of knowledge." -Stephen Hawking


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InvisibleTripsAreForKids
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #336562 - 06/07/01 06:01 AM (21 years, 3 months ago)

very good point opi......

Keep Trippin' and Blazin'

-=SporeWorks=-


--------------------
Everything posted above was out of fun, none should be taken serious. I am currently under the influence so take that into consideration.


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Offlinegray1
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #336910 - 06/07/01 05:30 PM (21 years, 3 months ago)

opi- while i still don't aggree with your personal attack on me and your constant disregard for my maturity and intelligence (let me hold your hand... is that bullshit really necessary in the dialogue between two intelligent people?) i do aggree with (and have always comprehended) what you are saying scientifically, i still think that the extra work involved in trying a tryp experiment is minimal and more associated with creating a correct nutritional environment (including proper phosphate levels as that seems to be a concern for you) for the fungi, which is probably already created by a brf type substrate ---however, at this point i'm ready to chalk it up as a miscommunication of ideas and a difference of opioion rather than incorrect information by either of us, and as you suggested, we have more positive potential theorizing and experiemnting together than we do bickering about petty issues. i aggree with the solubility of DMT and the fact that this may play a role in it's poor incorporation into the organism, and was thinking of this when i suggested/posted the method for potency comparrison a few days ago. by liquid cultivating/allowing colonization, DMT may be more readily assimilated into the organism. -i also had an interesting thought earlier this morning when i remembered that for certain natural alkaloid biosynthesis in other organisms that certain stresses actually induces biosynthetic pathways (such as for phalaris and the prodcution of DMT) and should be part of the cultivation process. is it possible that this is also true for the biosynthesis of psilocybin/psilocin in fungi and that attempting to create and optimal environment for growth is actually counterproductive? it seems that tryp or another intermediate would still be necessary for the process, but as has been suggested, tryp probably already exists in sufficient amounts in typical growth substrates such as brf... any thoughts on this?



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OfflineOpi
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #337046 - 06/07/01 08:59 PM (21 years, 3 months ago)

Please, let me COMPLETELY appologize for insulting you. I honestly was trying to be funny (hence the smile faces) about the fact that I said I wouldn't explain it again, and felt drawn into doing it anyway. It wasn't a judgement on your intellegence or maturity. I just found it funny that there was a miscommunication (or inability on my part to communicate clearly) and I still felt a need to explain and re-explain. That was the joke. That was why it was funny to "pretend" it had to do with your intellegence. Regarless of whether I was trying to be funny or not, you were insulted and I am sorry.

"(including proper phosphate levels as that seems to be a concern for you)

Ha ha ha, I will never give up my quest for boosted psilocybin...

I don't think that BRF alone is enough, otherwise you would see the full genetic possibility of the species, huge fruit, max alkaloid content, etc. If we are going to elevate the concentration of one part of BRF we need supplement everything else the fungus is going to need to deal with that excess.

There are a couple good threads going on this subject right now, one of them dealing with adding zinc to the substrate. This sounds very promising. Bringing all of these ideas together might be a good idea. Maybe we should come up with a plan to test different ideas as well as combinations of ideas, to see what works. We ask for competent cultivators to act as virtual lab assistance and report their findings... Its rather subjective, but it might get us somewhere faster..

Opi



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