The Entourage: One Of The Many Missing Pieces To The Puzzle

[in vivo]

That's only sixteen profiles with less quantified terpenes than each of us have access to. Already patterns begin to emerge:

 

mostly indica’ strains were characterized by dominancy of β-myrcene, present in high relative contents, with limonene or α-pinene as second most abundant terpenoid,

 

^^ The lemon skunk is similar to a couple of these listed indicas. High in myrcene and alpha-pinene. I'm surprised there wasn't more limonene, but after learning that alpha-pinene is responsible for much of the aroma associated with mangos, I kind of figured that all bets were off. I posted about "Juniper Berry - a-Pinene 43%, b-myrcene 12%" back on the second page of this thread, and we've been using it in topicals. Alpha-pinene and myrcene might be associated with one another in nature.  

 

while ‘mostly sativa’ strains were characterized by more complex terpene profiles, with some strains having α-terpinolene or α-pinene as dominant terpenoid, and some strains having β-myrcene as dominant terpenoid with α-terpinolene or trans-β-ocimene as second most abundant terpenoid.

 

In the Amnesia chemotype the main terpene is alpha-terpinolene, but none of those listed have the same type of beta-caryophyllene content, which was second most prominent in that cut. 

[in vivo]

Isn't legal use of medical cannabis in Michigan just to relieve symptoms of qualifying conditions?

 

I have a long history of documented migraines. As many might be aware migraines can be quite debilitating, and there isn't much in terms of successful rescue meds, or preventative treatments. Cannabis not only helps relieving the symptoms of a migraine when it sets in, but can also be used as a preventative treatment.

 

I've got pretty sever anxiety as well, and I prefer not to take benzos (too forgetful), so while it's not a qualified condition I also get a great deal of therapeutic value in that regard. Maybe I should remove that question. This thread is enough liability as is.

[+washtenaut]

I like the prevention explanation regarding migraines.  Very plausible

[Norby]

I think you just have to think outside the box.  Your not really seeking to relieve anxiety your finding a med with the side effect of not exacerbating the anxiety.  And hey, if it relieves it then it's a bonus.  Your just trying to find the meds that relieve your qualifying condition with side effects that don't make other symptoms any worse.

 

I've often thought of this because anxiety makes IBS symptoms occur, you can't really separate the 2 or your not treating the condition for IBS.

[in vivo]

I think people build up a tolerance to a point that strains that might make one person anxious or have a panic attack, can be quite therapeutic and enjoyable for others. It seems likely that the level of expression of receptors plays into this. The more receptors, the more an agonist will work. This was my first run with the sour og. I love it, but I've got a patient who didn't like it at all. They wanted to go to sleep, but were unable and felt very anxious. It tested at over 22% THC. I also find that I build a tolerance to it quickly if I don't rotate it. 

 

Treating anxiety is possible with cannabis. That being said, the anxiolityc effects do happen to be an added benefit of my migraine treatment.    

[+washtenaut]

I also find that I build a tolerance to it quickly if I don't rotate it. 

 

I find I quickly build tolerances too.  I like to have a few strains on hand for that very reason.  I have been fearful over the years though that having several containers of cannabis looked more like a cannabis dealer as opposed to just a user.

 

Anyway, I digress  

 

Why do we build up these non-permanent tolerances? 

[trichcycler]

the people who take rx often have a variety of different ones, to treat various ailments at different times of the day. I don't feel so bad with 8 small jars....

[in vivo]

That's a big question. I don't know if anyone is entirely certain, I know that I'm not. There are some seemingly obvious factors, but I doubt they offer a complete explanation.

 

The use of agonists lowering the level of expression of receptors seems to be a common issue in a variety of treatments. Opiates, benzos, and I was just reading about how it's also an issue in epilepsy treatment with GABA agonists. While there is more to tolerance than the level of expression of receptors, it appears to come into play in a significant way.

 

First let's think about the cannabinoids. If the majority of the psychoactive effects are associated with CB1 receptors in the brain then the greater the efficacy of cannabinoids to activate those CB1 receptors, the lower the level of expression of CB1 should become as a result.  

 

While I build a tolerance to any strain, I find that I build one more quickly to some when compared to others. Why is this? One explanation can be found in the way that cannabinoid receptors modulate one another, and other GPCRs. CB1 receptors form heteromic (this can all be found in the pdf named 'Cannabinoid Receptors Beyond CB1 and CB2' in my first post of the ECS thread) complexes with other receptors. Here are some definitions of those mechanisms:    

 

 

 

VI. Glossary

 

• Allosteric modulator. An exogenous or endogenous
molecule that binds to a distinct and nonoverlapping
site to influence binding or signaling at
another, usually orthosteric, site.

 

• Cooperativity. The effect(s) of multiple equivalents
of the same ligand binding to multiple (generally)
identical sites.

 

• Functional selectivity. Selective activation of a
subset of the signaling pathways available to a receptor
by a ligand.

 

• GPCR allosterism. The reciprocated effect(s) of
binding two (or more) distinct ligands at different
sites on a receptor monomer, homomer or heteromer.
Such effects can be positive or negative.

 

• Heteromeric receptor. A signaling unit composed
of two or more GPCR protomers that by themselves
are nonfunctional.

 

• Negative allosteric modulator. Reduces binding or
activity.

 

• Positive allosteric modulator. Enhances binding
or activity.

 

• Receptor heteromers. Two or more molecularly distinct
and individually functional GPCRs that combine to
form a molecular entity with distinct pharmacology.

 

• Receptor homomers. Two or more molecularly
equivalent and functional GPCRs that combine to
form a molecular entity with distinct pharmacology.

 

CB1 and CB2 form functional heteromers in the brain. That means that activation of one automatically 'turns down' the other. So, if you've got a strain with cannabinoids hitting CB2 (CBG, CBN), or blocking CB1 (CBD), that will lower the overall efficacy of CB1 agonists to activate those receptors. I think it might take longer to build a tolerance to those strains.

 

It's interesting that CBC can increase THC levels in the brain. That seems like it would increase the rate of tolerance.

 

Okay, so now that I've built up a tolerance to one, why does another one 'work better'?

 

I have a picture of this in my head, but I'm not sure how to convey it, and I'm not sure how accurate it is.

 

First the issue of novelty seems appropriate to bring up. I was watching a Brain Games episode where they recently discussed this. They showed that our bodies have a greater reaction to novelty, be it thrill seeking, sexual, and possibly pharmaceutical. But that explanation leaves a lot to be desired.  

 

Maybe a music analogy.

 

There's more going on than CB1 and CB2. Cannabinoids activate a number of receptors. Terpenes and flavonoids activate a number of receptors. Some of these can modulate one another similar to the CB1 and CB2 example. They can also share signal transduction pathways.

 

Look at the various cannabinoids and terpenes like the keys on a piano. Alone each key has its own vibration/tone (activity). Some combinations of keys make up chords (I think, unless that's just guitars). Think of a chord as a subjectively pleasant strain. Think of the piano as your brain. Each time you 'chord' (cannabinoids/terpenes) you're activating the receptors in your brain in a very similar fashion. The novelty soon diminishes, and you're activating the same combination of receptors (which should lead to specific receptor tolerance) over and over. By playing in a new 'chord' you hit a different combination of receptors and signal transduction pathways. if you are able to rotate in strains that vary in terms of CB1 activation, by incorporating one that is less efficient (at activating CB1 not treatment) you allow your CB1 receptors to build back up. The same is likely true for the combination of receptors being activated by a particular chemotype. 
 

I don't consider that to be anywhere near a complete picture, or a good analogy, but I'll continue to work on it.

Edited March 9, 2014 by in vivo

[in vivo]

This thread has been updated. I unpacked some info from Taming THC and added some visual aids to the first page. I also plan to add a list of terpenes that we're currently able to quantify in MI with information and links about each one in the remaining edited posts on the first page.

 

The cannabinoid and terpene content percentages has been added to each of the chemotypes listed so far.

 

Sour OG Review:

 

Method of consumption being reviewed: Smoked

 

The period of the day you use this strain the most: Day or Evening (won't help to sleep)

 

List of some of the effects that I feel feel describe this strain:

 

Anxious, Creative, Dizzy, Energetic, Euphoric, Expectorant, Focused, Giggly, Happy, Paranoid, Positive, Soothing, Talkative, Therapeutic, Uplifted

 

List of aromas that I associate with this strain:

 

Flowery, Earthy, Pungent, Nutty, Chemical, Diesel, Astringent, Pepper

 

 

Here's a Sour OG chemotype (diesel pheno/74 days, plan to go 84 next round):

 

 

 

Edited March 12, 2014 by in vivo

[in vivo]

Elemene:

 

This is the main terpene in this sour og chemotype. There are over 20k hits on a elemene in relation to its anticancer characteristics.  If you look at other 'sour' or 'diesel' lines you'll see that elemene is the highest in some, and is relatively high in almost all of them. It looks like most of them have elemene, myrcene, limonene, and beta-caryophyllene as the four most prominent terpenes. I suspect that elemene might have a role in the 'diesel' aroma. Someone else has had a terpene profile done on a sour og here in MI. I notice that the main terpene in theirs is myrcene, then limonene, then beta-caryophyllene, then elemene. I wish I could ask them if their chemotype is associated with sedation, which wouldn't describe this one at all.    

 

Antitumor effect of β-elemene in non-small-cell lung cancer cells is mediated via induction of cell cycle arrest and apoptotic cell death

 

Beta-elemene inhibits melanoma growth and metastasis via suppressing vascular endothelial growth factor-mediated angiogenesis

 

Anti-tumor effect of β-elemene in glioblastoma cells depends on p38 MAPK activation

 

Elemene displays anti-cancer ability on laryngeal cancer cells in vitro and in vivo

[in vivo]

I had to post this in the grow section, but this is definitely something I'd like to learn more about:

 

 

Abstract

We studied the effect of ethephon on levels of the major cannabinoids (tetrahydrocannabinol and cannabidiol) and chlorophyll, carotenoids and α-tocopherol in Cannabis sativa at productive stage. Results revealed that ethephon increased THC content of leaf in male and female plants and of male flowers. However, ethephon unable to enhancing THC content in female flowers. Treatment with etheohon increased CBD content in male and female leaf and female flowers. The treatment of male flowers with low ethephon concentration caused an increase, and those treated with high ethephon concentration resulted in a decrease in CBD content. The lowest level of ethephon (1 μM) enhanced chlorophyll a, b and total chlorophyll in male and female plants. Both sexes treated with ethephon showed an increase in carotenoids content, but 1 μM ethephon had the stronger effect in this regards. Male and female plants had a higher content of α-tocopherol when treated with ethephon. These results showed ethephon is a suitable treatment for increasing cannabinoids and α-tocopherol in productive stage of cannabis and there was not a relation between primary and secondary terpenoids.

Highlights

► Ethephon treatment has considerable effects on increasing of cannabinoids in male and female cannabis. ► There is no correlation between cannabinoids and other plastidial terpenoids. ► Using of ethephon treatment in flowering stage do not adverse effect on plant growth.

 

Ethephon application stimulats cannabinoids and plastidic terpenoids production in< i> Cannabis sativa</i> at flowering stage

 

 

Levying more control via the grow process to promote sought after cannabinoids and terpenes is interesting to me. They mention in Taming THC that nutrient rich mediums produce plants with fewer cannabinoids, and cutting off the nitrogen and putting the plant through oxidative stress is beneficial. Most growers are already doing that, but I'm always eager to learn new tricks. I'll have to do some serious due diligence before I'd actually try something like this, but learning about the mechanisms involved might be beneficial. I wish I had full access. 

[Norby]

Interesting, it seems to be converted into ethylene quickly upon uptake and force matures a plant.  I wonder if you'd get the same effect just letting the plant ripen longer?

 

I've always equated wild ginseng, in that, the wild roots are higher in ginsenoids? as they experience adverse conditions which produces more chemicals to help the plant and not as much starch or in this case bud material as opposed to cannabinoids. Which is why I look for signs of deficiency and fertilize accordingly to get rid of it and then I don't have to flush and I don't produce as much weight and more cannabinoids.  I can't wait to start testing.

[in vivo]

This is an interesting topic that seems worthy of discussion. The following quote is from Taming THC: 

 

As observed for cannabinoids, terpenoid production increases with light exposure, but decreases with soil fertility (Langenheim, 1994), and this is supported by the glasshouse experience that demonstrates higher yields if plants experience relative nitrogen lack just prior to harvest (Potter, 2004), favouring floral over foliar growth.

 

I feel as though the nitrogen comment putting more energy into leaf production rings to true. I've seen colas not quite come together due to this. The soil fertility decreasing terpenoid production is new to me, but I have read about oxidative stress raising cannabinoids, I'm not sure where. Here are the citations from that quote:

 

Growth and morphology of medicinal cannabis

 

Higher plant terpenoids: a phytocentric overview of their ecological roles

 

Unfortunately I can't find free versions anywhere.

[hempsci]

You might want to add the 20 odd flavonoids to what could be modifying the effects of Cannabis.

The effects of THC are pretty well known. The effects of 100% pure THC smoked alone are not so well known.

Same with the other Cannabinoids that have been shown to modify THC, like CBD, CBN, THCV, and maybe CBC a bit, they are pretty well known. The 130+ terpenes effects to modify THC and other Cannabinoids are being discovered now and will be known in the next decade or so.

The first test of pure THC spiked with a terpene was only a little over a decade ago.

I never got around to testing the flavonoids, I had hoped to after the terpene study, but I did not ever do it.

If this is only about terpenes maybe you need to change the title of the thread?

My focus was how terpenes, Cannabinoids, and maybe flavonoids and how they effected the "high".

I understand medical users have other priorities but I assumed they could use my info to help themselves.

-Hempsci

Edited March 17, 2014 by hempsci

[hempsci]

Whoops

Edited March 17, 2014 by hempsci

[in vivo]

darn right we can use your insight! I'm only aware of one person online that's publicly speaking about having done terpene research. When I saw those few relevant questions I had to pop my head in. I love it when a plan might have come together. 

 

Didn't mean to bait and switch on the title. When I started this thread there was no terpene quantification available at all in MI. I'm not sure if this thread had anything do with the change, or the more likely scenario that I just rowed in on the 'terpene wave'. Regardless, I'm stoked to be able to start out by quantifying around 36 of them. That seems to be more than most labs are currently offering. There's no doubt that a more complete profile will always be preferred.  I'd like to see:

 

Apigenin
Boiling point: 178°C / 352.4° Fahrenheit
Properties: Anxiolytic, Anti Inflammatory, Estrogenic

Quercetin
Boiling point: 250°C / 482° Fahrenheit
Properties: Antioxidant, Antimutagenic, Antiviral, Antineoplastic

Cannflavin A
Boiling point: 182°C / 359.6° Fahrenheit
Properties: COX inhibitor, LO inhibitor

β-Sitosterol
Boiling point: 134°C / 273.2° Fahrenheit
Properties: Anti Inflammatory, 5-α-reductase, inhibitor

 

But that's only because those are the few that I've read about being some of the most prominent. I'll ask about them the next time I speak with the analyst. He might be able to at least let me know if he identifies something worthy of mention, even if he can't quantify it. 

 

What I've been pushing for is getting THCV and CBDV quantified. I'd like to work them into breeding lines. I've also got them playing around with an magnolia officinalis extract to see if they can quantify magnolol and honokiol (two phytocannabinoids). 

 

When I wasn't quantifying terpenes and was limited to THC/CBD/CBN I had no ability to account for variance. For my purposes there was no point paying for that type of profile. If I feel I reach that point again I'll be the first one in here whining about how more people need to ask for an even more complete profile. At this point it seems as though there might be quite a bit to learn from what we can quantify.

 

Don't sweat any political correctness. Psychoactivity or getting 'high' is therapeutic. I know I made some statements that were kinda prickish about it, but I was only attempting to stress the importance of what this type of research has to offer. There's a bit of an aversion in MI to testing and labs in general (but we won't get into that). 

 

Something I didn't state publicly but should is that if you buy into the 'Botany of Desire' it's really no coincidence at all that psychoactive strains have so much therapeutic value in some of the most common ailments in our society (pain, nausea, stress, etc). Feeling good is therapeutic. That being said any and all information about the effects of these compounds is highly appreciated.

 

How many were you quantifying? How many profiles were completed? What associations and patterns were identified? 

 

I want to say I also read something about having developed a questionnaire. That's something I'm working on, but seem to be struggling with. I'd love to see the types of questions others were asking, particularly those that provided the most insight. 

 

I wish I had held onto my synthetics. I didn't care enough for them to keep them after their legal status changed. It seems like that would be a helpful way of systematically approaching this. I've only experimented with terpenes from essential oils (with GC/MS), and I think they offer economic value in some applications (like topicals), but the taste is not all that great on top of flowers. Now that this research can finally get started I plan to distill and capture essential oils from chemotypes and vaporize them, once I get my fun tickets up I also plan to start investigating individual terpenes.

 

I appreciate you coming in here to offer your insight!

Edited March 17, 2014 by in vivo

[Norby]

So does this mean that those, are they flavenoids, will stay intact during decarb at 250f?  Is that what boiling point means?

Edited March 17, 2014 by Norby

[hempsci]

You need to order a bunch of analytical terpene's found in Cannabis samples from SIGMA.

If you can gain access to 100% pure THC everything else is easy.

The synthetic THC like compounds can be dangerous, my advice is do not touch them.

WERC SHOP in pasadena tests for 35 terpenes.

Good luck,

-Hempsci

[in vivo]

Does that mean you're not giving up the goods?

 

I don't plan to play with the synthetics, and I'm not prepared to buy the equipment to separate individual cannabinoids. My plan is to clean up some oil a number of times until the terpene content is next to nothing. This might even be a preferred method over isolated cannabinoids. There are sources of isolated terpenes stateside. There are a few hoops to jump through for the Responsible Care requirements but nothing serious for the proper entity. 

[in vivo]

So does this mean that those, are they flavenoids, will stay intact during decarb at 250f?  Is that what boiling point means?

 

Yea, those are flavonoids. When I think of boiling point I think of the point at which the compound vaporizes. I'd don't have a good handle on the ways in which the chemical structure and the volatility of each compound plays into their stability. I feel confident saying that they don't need to come anywhere near their boiling point in order to be broken down or lost to heat. Most of these compounds don't appear to be very static in nature and they're are constantly changing.

 

The study on the solvents believed that the nonvolatile nonpolar solvents were able to retain the largest amount of terpenes and flavonoids by bonding with and trapping them.   

[Norby]

So if there are 160 types of terpenes found in MJ and test sites can't test for all of tehm, will you know if there are unid'd terpenes in your sample or do we just assume there is even more in our sample than we get back in the results?  This could explain why a lemon skunk sample may show no limonene but still smell very lemony?  A different terpene that is similar but not registering?

[in vivo]

It's possible, but I imagine they're continually adding those that are most prominent. Many are likely only present in very minimal amounts.
I would imagine that if there's an anomaly worthy of mention in the results that they would comment on it. 

[Norby]

Kewl.

[in vivo]

seems like an easy thing to find, right? not really.

 

this page mentions 66 cannabinoids

http://www.medicinal...of-cannabis.pdf

Chemical constituents of cannabis

"So far, 66 cannabinoids have been identified."

 

http://www.greenhous...of_cannabis.pdf

"With about 538 known constituents, Cannabis is one of the chemically best studied plants."

 

http://adai.uw.edu/m...annabinoids.htm

"There are over 480 natural components found within the Cannabis sativa plant, of which 66 have been classified as "cannabinoids;" chemicals unique to the plant."

 

you can see, the numbers are all over the map so far. some papers reference other papers and studies and no one has the definitive list!

 

 

I think the point that's attempting to be made is that you can't know how to base the pharmacology of a delivery system if you don't know the totality of the chemical constituent that make it up ie all cannabis species. 

 

One issue surrounding the confusion is in regards to whether some identified compounds are in fact present in cannabis or are created via the process of analysis.

 

Another issue would be that any plant is made up of a number chemicals. That doesn't mean that they're all active. Even those that might be active like THCV or CBDV won't show up unless a plant that's high in either THC or CBD respectively is pushed to its capacity. Even then you'd be lucky to hit 2% as far as I know. That means that unless current methods for quantification are insufficient and falling short of proper separation that by in large we know which cannabinoids we're working with. I believe that the rest are currently believed to primarily be found in miniscule amounts.

 

THC

CBD

CBN (older the sample or higher the heat)

CBG (normally under 2%)

CBC (normally under 2%)

CBDV (normally under 2% in strains over 20% CBD)

THCV (normally under 2% in strains over 20% THC)

 

Are there other cannabinoids that can be found less often and in even lower levels? Sure. The question is whether they're destroyed via combustion or present in significant levels.

 

Then you've got about 200 other terpenoids to contend with. Again like the cannabinoids some are believed to be much more prevalent than others They're claimed to have pharmacological relevance down to 0.05%, but that's an issue of much debate. Less debatable is whether many of them are destroyed via combustion, or significant when found in lower levels.  

 

I'm not exactly certain how the wise men came up with the list of terpenoids that they're currently quantifying. I think Iron Labs and one of the big ones out west are running the same 38 or so. It's more than the majority of studies that I've read, and seems to consists of all of the terpenoids that are believed to be most prominent. More will always be better than less, but the lower the level of any individual constituent the lower the likelihood of its relevance. We might very well require a more complete profile. If we're not yet certain if we do or not then we've got plenty on our plates as is.

[+Malamute]

Honestly, I haven't read all the way through this, but have you covered how some cannabinoids are the precursors for other cannabinoids?