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OfflineBleuboxo
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MIMOSA BARK IN MAiLBOX!!!!!
    #321571 - 05/20/01 01:43 PM (16 years, 2 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 03:18 PM (16 years, 2 months ago)

excellent! *sinister look*

peace, -=To0dLeS=-
TM


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OfflineoOjonahOo
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: The Messiah]
    #321718 - 05/20/01 04:30 PM (16 years, 2 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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Offlinehubertd8
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: oOjonahOo]
    #323641 - 05/22/01 08:39 PM (16 years, 2 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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OfflineDarK_SavioR
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Bleuboxo]
    #323716 - 05/22/01 10:13 PM (16 years, 2 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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InvisibleTripsAreForKids
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: DarK_SavioR]
    #324349 - 05/23/01 06:03 PM (16 years, 2 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 - 05/23/01 10:01 PM (16 years, 2 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 03:07 PM (16 years, 2 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 06:55 AM (16 years, 1 month 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 12:46 AM (16 years, 1 month 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 03:18 PM (16 years, 1 month 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 02:22 AM (16 years, 1 month 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


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Invisiblesynaptic
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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: the universe]
    #330313 - 05/31/01 03:14 AM (16 years, 1 month 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 08:50 AM (16 years, 1 month 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 12:11 PM (16 years, 1 month 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 04:20 PM (16 years, 1 month 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 - 05/31/01 08:02 PM (16 years, 1 month 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 03:10 AM (16 years, 1 month 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 10:08 AM (16 years, 1 month 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 10:22 AM (16 years, 1 month 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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