@TheWillBilly Very interesting. Did you find they only crystalize in very wet solution in a cool place? Less than 68?
And by don’t sugar out you mean they don’t chalk up after sitting around or it doesn’t crash into the sugar version of that pollymorph? I had some that lasted almost 4 months before they chalked.
They definitely start crashing well before the normal saturation point, the oldest are from July so there’s a chance it could still chalk out, but now that we can grow them repeatedly on demand and feel confident about the cause of the polymorphism, we don’t think these will see the same issue
Good point! Thank you, that’s definitely not the way I thought about it.
Feels like, if you using 13x for reasons beyond being a dessicant, you should toss it out more frequently.
Since you’ve stated you believe you know the cause of this polymorphism, could you share your hypothesis? Curious if it’s similar to @Dred_pirate or not.
Were waiting until we get a Crystallography report back to finalize and realease the study
Looking forward to it dude. I’ve not experienced this issue, but it has me utterly desperately curious.
@TheWillBilly We are hoping to get it done as soon as possible. Apparently, very few of the crystallographers want to touch THCa, but we may have found one who will! @DocKnott
I hope to hear back very soon.
I am not sure whether that is correct.
There are reports that the THC synthase enzyme and CBGA
are excreted into the vacuole. Topologically exterior to the cell.
( proteomic and transcriptosome work?)
there are people who have used micro capillaries to remove contents of capitate heads and have done analyses.
enzymatic synthesis of THCA synthase takes place in cytoplasmic side of (disc cell. ?) and the enzyme is then secreted to the vacuole…where THCA synthesis takes place. Subtle difference
from interior. This is the whole issue we are trying to define in
“Butane works but no theory”. You understand that cytoplasmic pH is like 6.8 to 7.0 or something like that. The pK of the salicylic acid like moiety that makes up the COOH part of the THCA is
about 3.0. That means there is only about one in 1000 to 10000
molecules of CBGA (substrate) in COOH form, the xray diffraction studies on the synthase co crystalized with THCA show the binding requires the COO- (minus) form. So it is thought that
that the substrate binding and release of THCA as product
is the ionized form. How is it that the COO- ion is protonated
in the vacuole…in order for butane to work as a solvent?
a number of theories are being entertained.
the protonated (undefined mechanism) seems likely.
best regards
Interesting, @moronnabis … although I’m not sure if the vacuole of which you speak is the same as the space inside the trichome head membrane (which is composed of a single membrane over the combined set of disc cells).
Btw, @AlexSiegel , this is the thread I was talking about.
I refer to the “storage space” as a vacuole…
the one above the disc cell set…particular to the
capitate stemmed trichome. Perhaps the term is incorrect.
when I click the alex siegel thread…I just get an overview…
but I don’t know how to reference the thread…?
Please advise.
I was surprised to read the synthase was excreted.
I think the researchers cloned the enzyme into tabacco cells (?)
fed it CBGA and it killed the cells. there is a limited number of papers on the synthase…I am sure you can find it …if it interests you…
also: this is very interesting:
https://doi.org/10.1016/j.plantsci.2019.04.008
a lot seems to go on in storage area…I am sure you can appreciate the complexities.
Ah, yes, thank you for clarifying your use of the term “vacuole”. I see what you mean, and I can certainly agree with the notion that the “balloon” ( @cyclopath ) or “vacuole” ( @moronnabis ) is probably a cauldron of synthetic activity… especially any syntheses which can make use of light energy!
I didn’t mean to confuse anyone with my mention of @AlexSiegel . I only did that for his sake, since we were discussing this matter outside the forum… I was just “pinging” his Future4200 notifications, so he knew the thread to which I was referring in our conversation. I also just copied the url from my browser address field and truncated it to remove the last numeric value (which is the number of the specific post in the thread).
By the way, @moronnabis , that doi link is faulty.
Thanks, @Franklin ! 
probably not the correct term. although using “vacuole” and “storage compartment” interchangeably is certainly well supported in most cases.
this particular storage compartment is (apparently) weird.
the vacuole (by definition) is NOT topologically exterior to the cell.
the storage space in question IS considered extracellular.
the cannabinoid synthases are secreted into said extra-cellular space, and imo that would not happen if there was no synthesis occurring there.
that would be a damn good reason for shipping the synthase “outside” before turning it on.
https://www.jbc.org/article/S0021-9258(20)72782-1/fulltext
does not indicate that CBGA kills tobacco expressing THCA synthase. Nor exporting the synthase by tobacco. it does state that baculovirus based expression in insect cells DID lead to secretion of most of the THCAS
Thank you…
but to be clear the interior if a vacuole in not topologically
exterior to the plant cell…?
In order not to confuse anyone, “storage cavity” or “secretory cavity of the glandular trichome”. since there are multiple types of trichomes,
and the specific studies refer to the capitate heads of the stalked glandular trichome, the latter term may be more correct.
the toxicity studies were done with 50 mM cannabinoic acid
in the extra cellular media… suspension of Tabacco BY-2 cells.
They did express the fusion protein in Tabacco cells…but
toxicity studies were NOT done with expression vector, simply bath application in suspension. “I think.”
Same group as you reference different paper? I think everyone should read both.
Plant Cell Physiol. 46(9): 1578–1582 (2005)
doi:10.1093/pcp/pci166, available online at www.pcp.oupjournals.org
JSPP © 2005
“Thus, we
next prepared transgenic tobacco plants expressing a green fluorescent protein (GFP)-tagged THCA synthase (THCA
synthase–GFP) to visualize the expression of THCA synthase
in the glandular trichome. …The finding that THCA synthase is secreted into the storage cavity raises an interesting question: why must this enzyme
be secreted for the biosynthesis of THCA? In general, secondary metabolites are toxic and must be expelled from the cytoplasm after their biosynthesis. For example, anthocyanin is
imported into the vacuole by a specific transporter to prevent
cellular damage (Goodman et al. 2004). In the case of cannabinoids, it has been reported that THC is toxic and induces the
death of mammalian cells in vitro (Guzman et al. 2001). However, there is no report about other cannabinoids. Since THCA
is biosynthesized outside the cell, it is possible that acidic
cannabinoids, such as THCA, are also cytotoxic. We, therefore,
examined the toxicity of CBGA and THCA, in suspensioncultured C. sativa cells and tobacco BY-2 cells, and compared
it with that of olivetolic acid, a phenolic moiety in cannabinoids. In 10-day-old suspension-cultured cells of C. sativa,
24 h treatment with CBGA and THCA at 50 µM caused 100%
cell death as demonstrated by trypan blue staining, whereas
olivetolic acid did not have any effect on the cells (Fig. 4A).
The same result was observed in 7-day-old tobacco BY-2 cells
treated with these two cannabinoids and olivetolic acid. Moreover, we confirmed that CBGA and THCA induce cell deathvia apoptosis since DNA laddering was detected by gel analysis of genomic DNA from treated plant cells (Fig. 4B). These
results clearly showed that cannabinoids are toxic substances.
We also confirmed that both CBGA and THCA induced
apoptosis not only in plant cells but also in insect (Spodoptera
frugiperda, Sf9) cells (data not shown), suggesting that cannabinoids act as plant defense compounds, like many secondary
metabolites biosynthesized in the glandular trichome. Since
cannabinoid-producing glandular trichomes are distributed in
physically fragile young tissues of the Cannabis plant, THCA
and CBGA would protect these tissues from predators such as
insects. This is the first report suggesting the physiological
importance of THCA and CBGA as apoptosis-inducing defense
compounds.As previously reported, the THCA synthase reaction produces hydrogen peroxide as well as THCA during the oxidation of CBGA (Sirikantaramas et al. 2004). Since the Cannabis
plant produces a very large amount (∼2% of dried tissue) of
THCA (Shoyama et al. 1975), a toxic amount of hydrogen peroxide might also be accumulated in the storage cavity of the
glandular trichome as a result of the THCA synthase reaction.
Thus, THCA synthase might contribute to the self-defense of
Cannabis plants by producing both THCA and hydrogen peroxide. In addition, since these reaction products are toxic to C.
sativa itself, THCA synthase must be secreted from secretory
cells into the storage cavity to avoid cellular damage.”
fixed it, thanks
here it is again in correct form:https://doi.org/10.1016/j.plantsci.2019.04.008
Everyone should read this paper…
it clearly demonstrates that the THCA synthase enzyme, isolated from
the capitate glandular trichome, is functional in hydrophobic solvent,
Hexane.
When I used hexane to re-x it turned into these same shapes. It didn’t Medusa however
I don’t use butane either hexane in it however as the case me be here
?
I am not sure exactly what you mean.
are you saying you redissolved chalked crystals in Hexane and recrystallized. And the recrystallized material chalked up again?
No I’m saying long ago before Medusa stones I was trying different solvents to recrash thca
The Medusa stone grow fast and have a peculiar shape similar to what I saw when I used hexane