Wellllll, my dissertation is finally handed in, so I thought I’d write a bit of a long post about Kratom today. Kratom is probably my favourite entheogen, so I’m surprised I haven’t written about it before. Actually, it did get a brief mention in JWH-018, Spice & Me (to the disappointment of a couple of kratom vendors more interested in profit than helping to spread factual information), but I think it’s worthy of a post of its own. I also recently added a tonne of kratom products to Coffeesh0p that I want to talk about too, so a post about kratom is about due.

It’s a shame that I don’t have any more presentations to give (well, kind of, because I hate giving presentations) because Kratom would definitely be the subject of the next one. So…

What is Kratom?

Kratom refers to the leaves of the Mitragyna speciosa tree that’s native to Southeast Asia. A member of the Rubiaceae family, Kratom is also related to the Coffee plant (Coffea spp.) and Psychotria viridis, a hallucinogenic shrub used throughout the Amazon. It’s effects include analgesia (ie, it’s a pain killer), stimulation at low doses and sedation at higher doses accompanied by euphoria. Sounds awesome, right?

Kratom Use

So, how is kratom used? Traditionally, workers of Southeast Asia would chew on the fresh leaves for a mild stimulatory effect and also as a pain killer. Sounds like a perfect natural remedy to get you through a long day, but unfortunately, fresh leaves are pretty scarce outside their country of origin. Luckily for us though, dried leaf retains its potency, so can be shipped worldwide – just don’t try and chew it. Dried leaf can be be brewed into a tea, refined into an extract (which can also be brewed up or eaten) or smoked, although the effects of smoking it are limited.

Kratom tea is generally prepared by boiling your leaf (see below for dosage info) in water and lemon juice for about 15 minutes. This is then strained and more water/lemon juice added to the leaves and the process repeated. The lemon juice is important, as the citric acid present in it acidifies your tea, meaning more of the alkaloids present in kratom will be extracted into solution. Once the leaves are strained for a second time, they can be chucked away, leaving you with a murky brown liquid that doesn’t taste amazing. This large quantity of liquid can be boiled down further to reduce the volume, but it’s not essential. You might want to add some sugar or honey for taste though. My lab rat and his gerbil friend tend to make their tea up to the nearest 100 ml, so it can be taken as 50 ml shots throughout the evening. Turning your particular dosage of kratom leaves into approximately 200 ml of tea is enough for 4 shots. If you’re new to kratom, try spacing these shots out every half an hour to an hour. This way, you won’t take too much and you can experience all of the subtleties kratom has to offer instead of diving straight in at the deep end. Alternatively, some people just throw all of their crushed leaf into a glass of juice, stir it up and just knock it back. This can be just as effective if you’ve got the stomach for it.

As far as smoking goes, it’s really not that great. If you’re going to smoke kratom, it’s far more rewarding to smoke some alongside a cup of kratom tea than just smoking it exclusively.

Kratom Dosage & Effects

An effective dosage for one person may be completely different for another person, as a tolerance can develop, especially with daily use. Other factors can also affect how much you might need (more on that later), so the best thing to do is start off with a low dose and increase it by small increments each time until you arrive at a dosage you’re comfortable with. This advice should be followed whenever you try something new, not just for kratom.

Here’s a rough dosage guide:

Dosages greater than 15g can be too much for some people, while dosages above 25g will be too much for most people. It’s statistically likely that you are included in the “most people” group, so please be careful!

Kratom Pharmacology

Kratom contains a number of active alkaloids, including mitragynine, 7-hydroxymitragynine & mitraphylline, which are structurally similar to the hallucinogenic tryptamines (eg LSD, psilocybin), as if this plant wasn’t interesting enough! At low doses, these alkaloids are thought to act on the adrenergic system (the same sets of receptors that respond to adrenaline & noradrenaline), responsible for the plant’s stimulatory effects.

At higher doses, however, these compounds start to activate your opioid receptors. These are the same receptors that opiates such as codeine, morphine and heroin bind to which cause the same sedation, analgesia and euphoria associated with opiate use. 7-hydroxymitragynine actually appears to be 17x more potent than morphine at these receptors. Powerful stuff! It’s no surprise then that:

1. Kratom can be used to help opiate addicts overcome their addiction
2. An effective kratom dosage depends on any pre-existing opiate tolerance
3. Kratom itself can be habit forming, just like opiates (so don’t use it every day!)

Growing Kratom

Unfortunately, growing kratom will be more hassle that it’s worth. Mitragyna Speciosa prefers the warm, swampy conditions of Southeast Asia, which makes growing it outside impossible in most areas of the world. Even a green house won’t be enough. If you’re going to grow it, you need to be serious about it. You’ll need an area of your house set aside for climate control, lamps, etc. Here’s the final blow: the kratom you grow yourself will likely be of inferior quality compared to that grown natively. Sorry to disappoint.

If you still want to go ahead, you’ll need to find a rooted clone, as you won’t find any seeds commercially available, or if you do, they might be fake, and cuttings tend to be vulnerable to infection.

Kratom Products

There are a number of kratom products available for purchase. I’m always on the lookout for more because I think this plant is truly amazing, so if you find something you think I should be stocking, let me know! Anyway, here’s a quick rundown of what we’ve got so far:

Kratom Leaf This is just bog-standard, dried, crushed kratom leaf. This is also available in a prerolled joint to enjoy in conjunction with other kratom products.

Thai Dragonfly Liquid Kratom Extract Each bottle of liquid kratom extract contains 20 ml. I’m not sure what strength this extract is, but it suggests a dose of 5 to 10 ml will be sufficient.

Kratom 15x Extract This extract is 15x stronger the plain leaf. Each pack contains 2g of powered extract. Unfortunately, this is a little expensive compared to the rest of the kratom products available, because it comes in fancy packaging and I have to import it.

Freeze Dried Kratom 20x Extract This brittle, resinous extract is 20x stronger than plain leaf.

Kratom Gold 35x Extract Capsules Each pack contains 3 capsules of 500mg 35x extract. A convenient way to consume your kratom!

Krypton Kratom 50x Extract This is without a doubt my lab rat’s favourite kratom product. Being 50x stronger than plain leaf, 0.5g is plenty for an entire evening. Comes in a 3g and 1g pack.

Since my last post about the spice behind Spice, it has been brought to my attention that some initial toxicology testing has been done on the synthetic cannabinoid JWH-018. Before we get down to the details however, here’s some pretty weird background information – the sponsor and provider of these studies wishes to remain anonymous! Unfortunately, this makes the whole thing a lot less credible, but since this is the only information we have right now, let’s hope someone else can verify these things at a later date. So far, one professor (who also wishes to remain anonymous) thinks these are real, but as of yet, no one is willing to put their name down on any kind of formal statement. If you, or anyone you know, has the relevant expertise to look over these studies, please drop me a line!

Feel free to invent your own conspiracy theories, but for now, let’s take a look at the data. You can download the PDF documents in this Zip file [2.04 MB]

CYP450 Inhibition Assay

This first assay looks at the effect of a drug on specific enzymes in your liver. These Cytochrome P450 enzymes are responsible for metabolising the vast majority of drugs you might put in your body, so if you’ve got too much of one drug in your system (ie paracetamol/acetaminophen), then other drugs that are also metabolised by these enzymes (ie alcohol) may compete for these enzymes and so hang around in your system for longer. As you can imagine, it’s important to understand how one drug may affect the metabolism of another, in case of any disasterous drug-drug interactions.

Results: JWH-018 will probably interact with the metabolism of other drugs, so more in vivo work is necessary.

hERG Binding Assay

hERG stands for human Ether-à-go-go Related Gene. This gene codes for a particular type of potassium channel found on heart tissue. This channel pumps potassium ions out of the heart muscle cells and are critical in coordinating the heart’s electrical activity. Unfortunately, these channels are a prime target for drugs to bind to, disrupting their function. This can lead to “Long QT Syndrome”, associated with fainting and can lead to sudden death, so you can see why these kinds of tests are important. Here’s a typical ECG recording showing what’s called the “QT interval” shown in blue, which lasts for longer than it should do if these channels are disrupted.

Results: JWH-018 does not interfere with these channels. That’s a good thing.

Cytotoxicity Assay

This simple test essentially looks at how many cells die when you perfuse them with a drug. The more cells that die, the more toxic the drug.

Results: JWH-018 is not cytotoxic at low concentrations.

GreenScreen HC Genotoxicity Assay

This assay looks at how much a drug will interfere with our DNA. Typically, anything that damages DNA is bad news, being potentially carcinogenic, making the rationale behind this test glaringly obvious. This test was also performed in the presence of a fraction taken from liver cells, which will break down the drug. This not only checks if the drug will damage DNA, but also its breakdown products.

Results: JWH-018 does not damage DNA, so shouldn’t give you cancer.

Rat Repeat Toxicity Assay

Guess what happens in this experiment. A number of renagade lab rats looking for a bad time are rounded up and promised free drugs (kind of like Pleasure Island from Pinocchio; that shit was scary!). The rats are then dosed up and observed. Initially, they appear lethargic (read: totally baked) but a few of them died at higher doses. This appears to be down to problems breathing rather than organ toxicity, but only affected the male rats, who appeared more sensitive to the compound. The drug didn’t appear to accumulate in their systems either, but they did lose some weight, probably because they couldn’t be arsed to eat. JWH-018 showed a huge potency and was found to be tachyphylactic (my new favourite word – it means that more of a drug is required to reach the same state following an initial dosage).

Results: According to FDA guidelines, the human equivalent dose is 0.016 mg/kg but it should be tested in other species before this can be seen as reliable!

Rat Pharmacokinetics

Data is collected on a number of different “pharmacokinetic” aspects of the drug, such as how it is absorbed, distributed throughout the body, metabolised and excreted, which can help with the design of future clinical trials.

Results: JWH-018 is distributed well throughout the rat’s tissues. Metabolism and excretion are normal, with a plasma half-life of approximately 2 hours

Summary

Well, from the looks of these tests, JWH-018 seems to be pretty safe, but unless you want to piss off Ben Goldacre, it would be wise not to rely on this “test tube data” entirely. Also, like I said before, we don’t know where this data has come from, clouding the issue even further.

Feel free to ask any questions in the comments.

Big thanks to Alfa @ Drugs-Forum.com for letting me know about these studies.

Today, I thought I’d share another one of my essays I had to do recently. This one looks at animal testing, problems concerning species differences and what we can do to avoid them. This essay is a little more sciency than my other one on living forever, so I’ll include the references this time. Here goes:

The use of non-human animals in the drug development process can attract criticism due to the issue of species differences. How significant is this problem and what strategies can be employed to minimise the impact of species differences?

Animal testing is a major tool in the drug development process, required by law before any new drug can enter the market. Animal models are set up to not only test the efficacy of a compound for its intended effect, but also to observe any potential side effects, to calculate a safe dosage for humans and to check for any addiction potential. Although animal testing is a legal requirement, implemented for our own safety, it is still only a model; a substitute for human physiology, whose results could be completely erroneous if they were derived from a poorly planned experiment. Differences between species are always a concern when setting up an appropriate animal model, and a lot of time is spent agonising over them to ensure any results obtained are both accurate and applicable to humans. When it comes to experimental design, species differences can be broadly classified into the following categories: anatomical/physiological differences, differences in metabolism and subsequent toxicity, pharmacological differences and behaviour.

Anatomical/physiological Differences

This is perhaps the most obvious class of species difference. It is no good testing a drug on an animal and looking for effects that are physically impossible for the animal to manifest. Any tests carried out on one species with implications for another must only test parts of the physiology common to both species, or identify an analogous symptom that corresponds to the effect you are looking for.

A prime example of this kind of difference crops up when investigating the emetogenic potential of a drug – unfortunately, evolution has not provided rats with a vomiting reflex, so an different model would have to be devised looking for an alternative behaviour or using another species with a physiology closer to ours.

Metabolism & Toxicity Differences

Different species also metabolise drugs differently – either via different metabolic pathways or with different kinetics. As such, a drug toxic to one species may have little effect on another, which is particularly important when trying to determine the toxicity in humans. A drug’s LD50, the amount required to kill 50% of subjects in a particular sample, is usually given in mg/kg of body mass, scaled up from animal experiments. If a drug’s toxicity or pharmokinetics are only determined from one animal species and extrapolated for the average human, the data would not take into account any differences in metabolism that may be present, resulting in potentially extreme inaccuracies.

For example, dogs should never be given coffee or chocolate, as they are poor metabolisers of theobromine¹, a xanthine alkaloid occurring naturally in both, as well as being a metabolite of caffeine. As little as 50g of chocolate can result in theobromine poisoning for small dogs, while humans can metabolise it fast enough without issue.
Similarly, metabolism of NSAIDs shows a huge variation across different species. The plasma half-life of aspirin ranges from 1 hour in ponies up to 37 hours in cats², due to their poor glucuronidation ability, while dogs are more susceptible to aspirin’s gastrointestinal side effects³.

One final example would be the varying MPTP toxicity between species. MPTP can be formed as an unintended byproduct in the manufacture of MPPP, a synthetic opioid with great potential for abuse. MPTP on its own is not harmful, but MPP+, the natural metabolite of MPTP, is a potent neurotoxin. MPP+ is produced via MonoAmine Oxidase B in neuroglia and the capillary endothelia comprising the blood-brain barrier, and results in rapid-onset Parkinsonian symptoms barely indistinguishable from typical Parkinson’s disease⁴. These symptoms are also reduced by L-DOPA, a drug commonly used in Parkinson’s disease. Rats, however, are almost entirely immune to MPTP toxicity, most likely due to a different level of expression of MAO B⁵. Mice, on the other hand, do produce MPP+, but clear it from their brain in a matter of hours, unlike the primate brain, in which clearance can take days.

Pharmacological Differences

The chemical pathways and their associated protein machinery will not necessarily be structurally identical, or indeed act in the same way. Pathways may be more or less complex, depending on the species, with more or less scope for modulation by other factors. Receptors too may also differ in structure, ligand affinity and the type of G proteins they may couple with. All of these factors may be of huge importance when designing a drug with a particular molecular target in mind.

A few interesting cases have resulted from these types of differences. For a while, Leptin was theorised to suppress hunger, as knockout mice that did not express leptin or its associated receptor got fat. Giving leptin to those that could not express it themselves, but still possessed the appropriate receptor, caused them to lose weight⁶ – a potential gold mine if the results were also applicable to humans. Unfortunately, they were not. Leptin showed little effect in humans, as weight problems tended to concern signal transduction rather than a lack of leptin⁷, in much the same way as insulin-resistant diabetes.

Another, rather more serious example is that of TGN1412, a monoclonal antibody with not only a high affinity for the human CD28 receptor, but a strong agonist ability too. Originally intended to help patients with rheumatoid arthritis and B cell chronic lymphocytic leukaemia, TGN1412 was initially tested on animals and an apparently safe dosage calculated. Of the 6 volunteers hospitalised, each given a dose 500 times smaller than that given to their animal counterparts, 4 developed multiple organ failure as a result of cytokine storm⁸. Hopefully, this example highlights the importance of species difference; that it is a real issue and not just a theoretical concern.

Behavioural Differences

The final category, and perhaps least obvious, is that concerning animal behaviour. Unfortunately for us, animals are not able to clearly express their feelings, so we are left to try and interpret that behaviour, which can be particularly difficult. Humans seem to have an intrinsic penchant for anthropomorphism – we are always unconsciously trying to attribute characteristics that are uniquely human, such as complex emotions or intention, onto animals and even non-living objects. Children are especially guilty of this, smacking a rock, perhaps, as a punishment because it tripped them up. It is only as we grow older and put in a little more thought that we realise that perhaps the rock was not to blame. With animal models, we must also put in that extra thought when it comes to interpreting an animal’s behaviour, instead of opting for the instinctive, humanised interpretation.

Other problems are encountered when we assume a particular behaviour is a result of a particular effect. For example, in the tail flick assay, designed to measure effects on nociception, analgesia is associated with an increased latency in moving the tail away from a heat source. Approving a new drug as an analgesic based on only this interpretation could be disastrous if the increased tail flick latency was instead due to a loss of muscle control or paralysis.

One final thought concerning animal behaviour, is that some behavioural responses may be unique to the species in question. For example, a hedgehog might curl up into a ball as a typical fear response. While this may be easy to interpret, other idiosyncratic responses may not.

Strategies

A number of strategies have been devised for combating the issues species difference brings up, ranging from simple common sense to the rather more complex. An in-depth knowledge of the species under investigation is a good start. Experience and familiarity with a particular species will naturally lead to a better ability to read an animal’s behaviour, just as we become better at reading the people around us the longer we spend in their company. Someone new to animal work will be more likely to anthropomorphise, drawing instead from their experience with other people, whereas someone with ample experience could make a more accurate judgement. Another benefit from experience is that any of the more subtle differences between that species and us is more likely to spring to mind, reducing the risk of something important being overlooked. For example, rat models are a useful tool when studying the intestinal bioavailability of drugs, but are a poor choice when it comes to intestinal metabolism⁹.

Another strategy to reduce the risks imposed by any unknown or overlooked differences, and one that is required by law, is to test on more than one species. Doing so greatly reduces the chances that any observed response is unique to one species in particular, and is therefore likely to be exhibited by humans too.

Although there are an incredible number of individual species, some proteins remain relatively conserved. Working with these specific proteins that share a great deal of similarity between their human counterparts will likely lead to more reliable results. For example, the muscarinic receptor family has remained much the same throughout evolution such that the human and rat receptors share a very similar agonist/antagonist profile¹⁰. It is very likely that something acting on rat muscarinic receptors will elicit the same response in humans, making this an accurate model.

More recently, the latest tools and techniques of the genetic engineer promise to make animal models even more relevant. Genetic manipulation has already delivered knockout animals, not expressing particular genes, and transgenic animals, expressing genes belonging to another species, but in 2008 a chimeric mouse with 90% human hepatocytes (liver cells) was produced¹¹. Until now, the best tool for studying the effects of drugs on the liver would be to use actual human liver (another strategy for overcoming species differences is to use human cells if possible), but the chimeric mouse has already shown great potential. The liver is mainly responsible for the pharmacokinetics of a drug, as it is the primary place that drugs are metabolised, which has subsequent effects on the toxicity and efficacy of that drug. The chimeric mouse has shown a similar pharmacokinetic profile to the human donor, as well as human-specific metabolites not ordinarily found in mice, making this an excellent model with which to study pharmacokinetics and toxicity. This advancement brings with it all the benefits of testing drugs on an actual human target, without any of the ethical considerations raised with human testing.

We humans are an animal species like any other, and we may have our own species-specific responses that are impossible to capture or anticipate with any animal model. It is important to remember that an animal model is just that – a model. Species differences will always be an issue; there are even idiosyncratic reactions to drugs within the same species, such as some humans being allergic to penicillin, so we can never eliminate these differences completely. Increasing research, awareness, criticisms from the animal rights campaigners and new genetic techniques will continue to help us reduce the severity of these issues until they can be reduced no further.

References

1. Kahn CM, editor. The Merck Veterinary Manual. 9th Ed. New Jersey: Merck & Co., Inc; 2008.
2. Boothe DM. The Analgesic, Antipyretic and Anti-inflammatory Drugs. In: Adams HR, editor. Veterinary Pharmacology and Therapeutics. 8th Ed. Iowa: Iwoa University Press; 2001. p. 433-454
3. Crosby JT. Veterinary Questions and Answers – Can you give a dog or cat aspirin? [cited: 2008 Sept 02] About.com: Veterinary Medicine. Available from: http://vetmedicine.about.com/cs/altvetmedgeneral/a/dogcataspirin.htm
4. Langston JW, Ballard P. Parkinsonism induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP): implications for treatment and the pathogenesis of Parkinson’s disease. Can J Neurol Sci. 1984 Feb;11(1 Suppl):160-165.
5. William Langston JW. The Impact of MPTP on Parkinson’s Disease Research: Past, Present, and Future. In: Factor SA, Weiner WJ, editors. Parkinson’s Disease: Diagnosis and Clinical Management, New York: Demos Medical Publishing, 2002. p. 407-436
6. Pelleymounter MA, Cullen MJ, Baker MB, et al. Effects of the obese gene product on body weight regulation in ob/ob mice. Science. 1995 Jul 28;269(5223):540-543
7. Considine RV, Sinha MK, Heiman ML, et al. Serum immunoreactive-leptin concentrations in normal-weight and obese humans. N Engl J Med. 1996 Feb 1;334(5):292-295
8. Suntharalingam G, Perry MR, Ward S, et al. Cytokine storm in a phase 1 trial of the anti-CD28 monoclonal antibody TGN1412. N Engl J Med. 2006 Sep 7;355(10):1018-1028
9. Hurst S, Loi CM, Brodfuehrer J, El-Kattan A. Impact of physiological, physicochemical and biopharmaceutical factors in absorption and metabolism mechanisms on the drug oralbioavailability of rats and humans. Expert Opin Drug Metab Toxicol. 2007 Aug;3(4):469-489
10. Venter JC, Eddy B, Hall LM, Fraser CM. Monoclonal antibodies detect the conservation of muscarinic cholinergic receptor structure from Drosophila to human brain and detect possible structural homology with alpha 1-adrenergic receptors. Proc Natl Acad Sci USA. 1984 Jan;81(1):272-276
11. Katoh M, Tateno C, Yoshizato K, Yokoi T. Chimeric mouse with humanized liver. Toxicology. 2008 Apr 3;246(1):9-17

What with the holidays and the decision to move all the articles from Coffeesh0p over here, it’s about time I posted something with a bit more meat. Having said that, this post is also suitable for vegetarians, so read on!

As I mentioned briefly before, I had to give another presentation to my neuropharmacology class in a similar vein to the one on Salvia divinorum I published earlier. In the end, I chose to talk about psychoactive mushrooms, so here’s the slides and a bit of bloggified talking along with each. Before we begin though, I’ll just say this was the worst presentation I’ve ever given – I (probably) had the most severe case of flu ever recorded and only managed to summon the courage to deliver it with Beechams flu plus, aspirin and a cheeky dihydrocodeine. Without these unsung heroes, this talk would not have been possible!

Oh, you can also click on the slides to enlarge them. Without further ado:

I’ll be talking about both the traditional “Magic Mushrooms” and the fly agaric mushroom, which is less well known, but is actually pretty culturally significant. For both of these, I’ll touch on a bit of history and tradition, pharmacology and a few other interesting bits and pieces.
The typical magic mushrooms are actually many species of the Psilocybe genus with each species having its own subtle differences. There are 60 species of Psilocybe mushrooms growing throughout the united states, of which 25 are hallucinogenic. These mushrooms will grow in nearly any kind of habitat, apart from arid deserts, so are found throughout the world. The greatest species diversity falls within the neotropic climate zones, encompassing much of South America.

These mushrooms were traditionally used by the native peoples of middle America for divination & healing purposes as well as religious communion. In fact, these people referred to the mushrooms as “God flesh” in their native language. Traditional use continued until the Spanish invaded, bringing European culture with them in the 14-1500s which pushed mushroom use underground. In 1955, Robert Gordon Wasson was the first westerner to take the mushrooms, and since then, western interest has exploded.

Some of the positive effects brought on by these mushrooms include a euphoric change in mood accompanied by giggling and laughter, as well as an increased flow of ideas and tendency to think “deep”. Objects and lights also appear more interesting and colourful. The neutral effects include a general shift in consciousness, as with most other psychoactive substances, but also an increased emotional sensitivity, pupil dilation & photosensitivity, lethargy and time dilation – the feeling that time is passing faster than it actually is. The negative effects of mushroom use can include intense fear, a headache as the effects begin to wane, gastrointestinal discomfort such as cramps & nausea, anxiety, confusion and fainting. There has been no evidence of organ damage following use.

The pharmacology – The important constituents are two compounds in the tryptamine family, psilocybin and psilocin. Psilocybin is not actually biologically active – rather, it’s a prodrug that gets dephosphorylated by the body to form psilocin, which is psychoactive. I’ve also put a model of 5-HT on there for comparison. Psilocin is an agonist at 5-HT 2A, 2C and 1A receptors, but it’s hallucinatory effects are due to the binding to 5-HT2A receptors in the brain. Psilocin shows no effect on dopaminergic pathways, and only affects noradrenergic pathways in high doses. It is believed to be the degradation of psilocin into some kind of blue pigment responsible for the characteristic blue/black bruising of these mushrooms following handling. The ease at which they bruise is a good indicator of the mushroom’s potency. One species will even turn blue from just blowing on it.

While there are no recognised medical uses of magic mushrooms, they have been used as an experimental treatment for a number of disorders. There’s significant anecdotal evidence to suggest that mushrooms can abort the period where people with cluster headaches are prone to attacks and also prevent relapses. Cluster headaches are quite a serious condition, being described as more painful than childbirth (by women!), so it’s no wonder people are willing to break the law to treat themselves. There are also currently studies under way on the effect of these mushrooms at easing the psychological suffering associated with cancer.

There’s not a lot more to say about these mushrooms, only that making them illegal naturally hampers research into a potentially useful drug.

The Amanita muscaria mushroom is a whole different kettle of fish. Here’s a few pictures so you know what I’m talking about.

Also known as the Fly agaric, this mushroom is the archetypal toadstool of the fairy tales, and is native to many places throughout the northern hemisphere, where it has been used ceremonially and recreationally for thousands of years. The mushroom, when freshly picked, is poisonous, but with careful preparation, the mushroom loses its toxicity. Unlike its psilocybin containing counterpart, this mushroom is completely legal.

Amanita have a long past, appearing in artwork from as long ago as 3500 BC. They also appear in paintings from the renaissance period, becoming more prominent during the Victorian era. This mushroom is associated in particular with fairies, elves and little people in general. They also began appearing on Christmas cards as a symbol of luck, and models of the mushroom were hung on Christmas trees as decorations. This could be due to the natural association between these mushrooms and pine forests.

It’s also been suggested that Santa Clause himself is modelled after the fly agaric mushroom, with his red ‘n’ white suit. Reindeer have also been observed eating this mushrooms in the wild and becoming intoxicated, so could that be behind the stories of flying reindeer? In fact, here’s another article on Amanita muscaria & Christmas – a very interesting read. Alice in Wonderland by Lewis Carol seemed to draw it’s inspiration from Amanita muscaria too.

Here’s a few images of this mushroom appearing in art through time. The top left one is from Disney’s Fantasia from 1940 – another example of just how widespread this mushroom has become within our culture.

Use of these mushrooms has been as widespread as their geographic distribution, but heavy use has been recorded in Siberia in particular. The Siberian shamans use the fly agaric as an alternative method to drumming and chanting to enter a trance state, but in eastern Siberia, the mushrooms were used by everyone both religiously and recreationally.

These mushrooms have a much more of a sedative effect with less hallucinations than the psilocybin containing counterparts. The positive effects include euphoria, analgesia, trance-like states being achieved, synaesthesia, and seeing “little people”. Maybe that one’s not so positive… The neutral effects include sedation, although some people can feel particularly energetic, along with changes in body perception, blurred vision and such. The most common negative effects associated with fly agaric use are nausea & gastrointestinal discomfort, but a powerful dissociation and delirium can occur at higher doses.

The active compounds in Amanita muscaria are Ibotenic acid and it’s derivative, muscimol. Ibotenic acid is a neurotoxin, which has since found a use in research, being a good inducer of brain lesions. This is the compound responsible for the toxic delirium resulting from ingestion of the fresh mushrooms. When dried in a particular manner, the ibotenic acid is decarboxylated into muscimol, making the mushrooms a lot safer to eat.

Muscimol itself is a selective agonist at the GABA-A receptor and a partial agonist at the GABA-C receptor. Muscimol’s effect profile is the sum of its actions at both these receptors, where it binds to the GABA site rather than that of an allosteric modulator, such as benzodiazepines or babituates. These GABAergic effects alter neuronal activity in many regions of the brain including the cerebral cortex, the hippocampus and the cerebellum. Muscimol is not metabolised further by the body, but is excreted in large quantities, as we shall see…

Time for some interesting bits and pieces about muscimol. Alcohol withdrawal can lead to hallucinations of little people much like muscimol. Since alcohol also acts on GABAergic pathways, maybe the effects could be related?

Siberian tribes used to drink the urine of their shaman, as it contains a high concentration of muscimol after ceremonial fly agaric use. I can’t think of any reason someone might find this out in the first place though.

And despite the name, Amanita muscaria have negligible muscarinic effects. They do contain muscarine, but in such tiny quantities to not make a difference.

Muscimol has also found use as a pharmacological tool, being a GABA agonist. GABA itself plays an inhibitory role, so GABA agonists applied to the brain will also have an inhibitory role. This is a useful method of simulating axon-sparing brain lesions, making reversible inactivation of brain areas a great way to study brain-behaviour relationships, such as where and when neuronal events for learning and memory take place.

And that’s that!

At this point I handed round a fly agaric cap for extra cool points.

The slides are available as a PDF here: Psychoactive Mushrooms Presentation [1.79 MB]

Introduction

This guide is by no means a complete reference for any would-be Salvia divinorum user. It simply describes, compares and contrasts the different methods by which one can experience the effects of Salvia Divinorum. There are other factors which must first be taken into consideration before you should embark on your journey with salvia: don’t take too much. Start off with smaller doses first and get comfortable with the experience, then maybe later, increase that dose slightly. Never jump in at the deep end – it could put you off salvia for a long time. Also, especially if it’s your first time with salvia, or the first time you’ve increased your dose, make sure you have someone sober nearby making sure nothing bad happens. With that warning out of the way, here we go!

The “Mazatec Oldskool” Method

Traditionally, the Mazatec people, indigenous to the Oaxaca region of Mexico where Salvia divinorum was first found, used to use salvia as part of their shamanic practices. It is believed they used to grind up large quantities of Salvia divinorum leaf, which was then added to water and drank. This method leaves a lot to be desired. We know now that salvinorin-a, the active chemical in Salvia divinorum is not very readily absorbed through the stomach, so large quantities of leaf must be used. It also doesn’t taste particularly fantastic. These drawbacks are countered by the fact that the effects from the salvia last much longer than any other method outlined here. The Mazatecs also used to chew fresh leaf for long periods of time, which is still quite popular today. See the Quid method for more details.

Pros:

• Safe
• Doesn’t harm the lungs
• Longer lasting effects

Cons:

• Inefficient – lots of leaves required for desired effect
• Tastes horrible

Smoking Leaf

Smoking salvia leaf can be effective, but it’s not ideal for non-smokers. The active chemical in Salvia divinorum, salvinorin-a, requires a high temperature to vaporise, so the leaf should be smoked through a pipe or bong rather than rolled as a cigarette. When smoking the leaf through a pipe or bong, you should try and use a torch lighter if possible. The extra heat generated by the torch flame will vaporise more of the salvinorin-a per hit compared with a regular lighter. That said, many users have reported a more relaxed mood shift when smoked as a cigarette, and a more “trippy” high when using it to replace tobacco in a cannabis joint. It’s generally considered harder to achieve the full effects of Salvia divinorum when smoking only leaf, compared with the stronger extracts. This is because the salvinorin-a from the leaf is metabolised by the body rather quickly, so smoking more over a longer duration will only maintain the level of trip, rather than enhancing it. To get the most from this method, it is advised that you take two to three hits from the pipe or bong, each time holding your breath for as long as you can, exceeding 30 seconds if possible. The effects will be noticeable after about one minute, giving you up to about three hits before you should put the pipe or bong down. The effects will remain for up to about half an hour.

Pros:

• Quick
• Easy
• No bad taste
• Relatively safe – It’s quite hard to get too much salvinorin-a into your body from smoking only leaf

Cons:

• Hard to achieve effects
• Smoking anything is never good for your lungs
• Harsh on the throat/lungs – the smoke is very hot
• Short effect duration

Smoking Extract

Salvia divinorum extracts are quite simply salvia leaf with a lot more kick. Extracts are prepared by taking the salvinorin-a from a large quantity of leaf and depositing it back onto a much smaller quantity of leaf. For example, one gram of 20x extract is, give or take, one gram of salvia leaf, with the salvinorin-a of 20g of leaf added to it. To visualise it, imagine filling your bowl with 20x the amount of normal leaf, and smoking it all. For more information on the extracting process, you might like this article: How To Make Salvia divinorum Extract. This method ensures you get enough salvinorin-a into your body as soon as possible, opening up the deeper levels of the salvia experience. Unfortunately, due to the strength of some extracts, it can be hard to accurately measure out a correct dose, so you could end up taking in far more than you intended. The extract should also be smoked in a pipe or bong.

Pros:

• Effects are very easy to achieve
• Less material needs to be smoked compared to leaf for the same effects

Cons:

• Short effect duration
• Easy to take too much
• Smoking anything is never good for your lungs
• Harsh on the throat/lungs – the smoke is very hot

The Quid Method

A “quid” is basically a big wad of leaves. Fresh leaves, if possible, but dry leaves can be used too. If the leaves are dry, immerse them in a cup of warm water for about a minute before you wish to begin – this step is essential, otherwise you’ll be chewing on dry leaf, which will taste just plain disgusting. Take about ten to fifteen fresh or soaked leaves, roll them up into a ball and pop the ball, or quid, into your mouth. Now all you have to do is chew those awful tasting leaves for a good fifteen to thirty minutes. Sounds easy? Well, you have to do it swallowing as little saliva as possible. This method works by a process called “sublingual absorption”: the salvinorin-a is absorbed into your blood through the mouth, so the quid needs to stay in your mouth for as long as possible. After the time is up, you should begin to feel the effects, although rather subtly compared to smoking the leaf or extract. You can now either spit out the contents of your mouth, or swallow it. Swallowing it can be a ghastly experience, but it’s recommended, since any remaining salvinorin-a in your saliva or the leaf will eventually get absorbed through your stomach. Just like the “Mazatec Oldskool” method, the effects last longer than smoking.

Pros:

• Safe
• Doesn’t harm the lungs

Cons:

• Tastes horrible
• Takes longer for the effects to set in
• Can be harder to obtain full effects

Salvinorin Tincture

Salvinorin tincture works in the same way as the quid method: sublingual absorption. That is, you let the liquid sit in your mouth for 15 to 30 minutes, allowing the salvinorin-a to diffuse into your blood through your mouth. The tincture itself is an alcohol-based solution of salvinorin-a, meaning doses can be measured more accurately by diluting it, and it doesn’t taste nearly as bad. The effects are also brought about much faster.

Pros:

• Quick
• Easy
• Relatively safe
• Tastes nicer than quid method
• Doesn’t harm the lungs

Cons:

• If the tincture is not diluted enough, it can burn the mouth

Conclusions

Many people report a variety of different effects from each method, from nothing at all, to a full-blown shit-your-pants trip. Non-smokers will naturally prefer the oral methods, while smokers would naturally be more comfortable smoking the leaf or extract. It’s also not unheard of for people to combine two or more of the above methods to achieve a greater effect than either on their own.

I’m very excited! We’ve just bought quite a few new things to pad out our Legal Highs and Entheogen categories, which happen to be our favourite. Over the next few weeks, we’ll be trickling new products on here and there, including more kratom, herbal teas and other delicious extracts, as well as popular smoking mixtures in the same league as Spice Diamond. It’s going to take forever to write all the descriptions and produce a nice set of images for each. We’re going to try and put more on there about traditional usage, and maybe some pharmacology too. I just wish I was working on it full time!

Pyramidal neurones in the Cerebral Cortex

As it happens, I’m pretty busy with my second module of the semester: neuropharmacology. In a few weeks, I’ll have to do another presentation in the same vein as my one on salvia. I’m thinking I’ll talk about either kratom, ayahuasca or psychoactive mushrooms. Kratom has some very interesting properties, acting as an opioid, ayahuasca highlights the importance of drug-drug interactions, containing DMT and an MAO inhibitor, while psychoactive mushrooms are just really interesting. The last option also allows me to bring in some whole Amanita muscaria caps (another exciting new product!) in to pass around – everyone would love that, right? As if that wasn’t reason enough, it’s active constituent, muscimol, is also of use pharmacologically. During some preliminary research, I found this great paper on hallucinogens and dissociative agents naturally growing in the United States I thought I’d share with you. Not hard going at all, and totally worth a read.

I’ve also got to pick a 10 week research project to do after Christmas. One of the options, and no doubt the one with the most competition, is on cannabinoids, eating behaviour and GABA signalling. Another one that stands out is the effects of ketamine on the visual cortex. Looks like a busy few weeks ahead.

As I mentioned before, I got to give a 10 minute presentation on Salvia divinorum to my pharmacology class. It went down pretty well, so I thought I’d write it up for you guys! Clicking on any slide will open it full size in a new window/tab.

The first thing you should know about Salvia Divinorum is that it’s a very potent hallucinogen. I mean, realllly potent. It’s name directly translates to “Sage of the Seers” or “Diviner’s Sage”, so already we know it’s gonna be cool. Salvia Divinorum is a member of the mint family, along with other common herbs, such as basil, rosemary and garden sage. In fact, Salvia is the Sage genus, which includes the common sage as well as S. Divinorum. This plant is native to one region only – the Oaxaca ["wahaka"] province of Mexico, where it grows best in a moist, shady environment.

Salvia Divinorum found extensive use among the Mazatec, the indigenous people of this region. Their religion is a blend of traditional superstition and a flavour of Christianity (also superstition!) brought over by the Spanish conquistadors. They make extensive use of the natural psychoactive plants and fungi in their rituals, including Salvia Divinorum, Morning Glory seeds and psilocybin mushrooms. Salvia in particular was used very much as a learning tool to facilitate visions, particularly in the context of healing or divination (hence the name).

Salvia was also a common medicine, prescribed for such ailments as diarrhoea, headaches and rheumatism. It was also the number one cure for a semi-magical disease known as “panzón de borrego“, or swollen belly.It’s no surprise to see a difference in use between the Mazatec and the “west”. The majority of use in the USA, UK and other developed nations is for recreation, while the Mazatec adopt a somewhat more “respectful” approach. Traditionally, salvia leaves were chewed, or an extract was prepared by crushing the leaves and consuming the liquid. There is no indication the plant was ever smoked by these people, which makes sense – the active compound has a very high vaporisation temperature. It is only the western world who smoke high powered extracts though a bong with a turboflame lighter!

The effects are many and varied, depending greatly on the amount consumed. Uncontrollable laughter is perhaps the most obvious effect, but it doesn’t happen to everyone. Other effects include remembering past memories, dissociation of the body and mind, a sensation of a force or pressure pushing or pulling on the body, usually in a particular direction, perceiving membranes or films or multiple small tiles covering surfaces and merging with, or becoming other objects. This is in no way comparable to any of the classic hallucinogens, such as LSD or Mescalin in effect or duration, as the Salvia experience usually lasts 15 to 60 minutes.

The active compound of Salvia Divinorum is Salvinorin-A, a diterpine compound. I know what you’re thinking – “why should I care?” – well, you should! Salvinorin-a is the only known psychoactive diterpene AND the first non-alkaloidal (or non-nitrogenous) hallucinogen to be discovered. It acts as an agonist at the kappa opioid receptor, which is also unusual. The other, classical hallucinogens work at the 5-HT2a receptor, and the other opioid receptor ligands tend to be alkaloids.

Shown here is a receptor selectivity profile, comparing the LSD in red with salvinorin-a in green. As you can see, the salvinorin-a is very selective for the kappa opioid receptor and not a lot else, while LSD shows activity across multiple receptors.

Shown here is the proposed kappa receptor:salvinorin-a binding complex, produced from various mutagenesis studies.An active dose of salvinorin-a can be as low as 200 micrograms, around the same as LSD, making it one of the most potent hallucinogens. But, as I’ve already mentioned, the experience usually lasts under an hour. Salvinorin-A does not remain in the body for long, with a half life of between 20 to 80 minutes in nonhuman primates.

So, an interesting drug, but is it also interesting clinically? Definitely! First off, salvinorin-a has shown promise in analgesia (pain relieving) studies in mice. Salvinorin-a, when injected intraperitoneally, produced an increased tail flick latency in these mice. The tail flick test is designed to measure the pain threshold – the mouses tail is laid out flat on a plate, and at one point along the plate, a beam of light is focused on the plate from underneath, creating a hot spot underneath the end of the mouses tail. As soon as the mouse begins to notice any pain, it flicks its tail to the side, so an increase in this amount of time shows an increased pain threshold. This antinociceptive effect is abolished if the mice are first pre-treated with a kappa antagonist, or are genetically engineered to lack kappa receptors, which proves salvinorin-a acts on these receptors in vivo as well as in vitro, shown by the previous graph. To make sure this effect was consistent and really did show an increased resistance to pain, other assays, such as the hotplate and chemo-nociceptive acetic acid abdominal constriction assays were done and produced results concordant with analgesia.

As I mentioned before, Salvia was administered by the Mazatec for diarrhoea, but it has now been shown to prevent myenteric cholinergic transmission in the small intestines of a guinea pig, effectively stopping muscle contraction.

Salvia may also shed some light on depression. Other kappa selective agonists typically produce depressive like behaviour in animal models, and salvinorin-a seems to produce a similar response. This supports the hypothesis that kappa opioid receptor signalling plays a role in depressive behaviours, but there has been at least one case report where salvia divinorum was used to treat refractory depression – depression that responds to nothing else. Before we can conclude anything from this, further work, including clinical trials must be undertaken. Either way, interesting stuff!

As we’ve seen, salvinorin-a is a bit strange, offering us an exciting new molecule to play around with. Already, chemical tweaking of the molecular structure has given us a selective agonist for the mu receptor and further research might lead to many novel, receptor-specific compounds.

Salvia has also shed some light on kappa receptors and their role in hallucinatory diseases. If the kappa agonist, salvinorin-a is able to produce such intense hallucinations (proving the involvement of kappa receptors in modulating our perception), could a kappa antagonist help reduce hallucinations in diseases with prominent perceptual disturbances, such as Alzheimer’s or Schizophrenia? There are many avenues Salvia Divinorum could lead us down, but if one thing’s for certain, more research is needed!

There are some great papers here, all of which I’d recommend, but if you can’t be bothered, there’s a brilliant TV documentary on there, Sacred Weeds. Definitely worth a watch. Thanks!

The slides are available as a PDF here: Salvia Divinorum Presentation [836kB]

Salvia Divinorum

It’s been a week now since I started back at university, and everything’s going great – we’ve dived head first into my first module, Pharmacology: from Molecules to Man. This comprises of 25% coursework and a 75% exam right at the end. The coursework is further divided up into essays, lab write-ups and …a 10 minute presentation on the topic of my choice! Right now, I’m pretty certain I’ll be talking about Salvia Divinorum, as it’s just plain more interesting than anything else, both for me to talk about for ten minutes and for everyone else to listen to it. Soooo, what to talk about for ten minutes?

Although this will be a serious pharmacological talk, I think I can get away with a bit of history of it’s use and what the plant represented to the ancient civilization that used it. If anyone has any interesting facts, now’s the time to share them! But what about the serious pharmacology stuff? Well, these papers are pretty good.

The first talks about the extremely selective action of Salvinorin-A on kappa opioid receptors. The

Salvinorin-A

hallucinogenic properties of this compound, mediated by activation of these receptors could be useful clinically. If activation of these receptors is capable of bringing about hallucinations, turning them off (with a selective antagonist) could help diseases in which hallucinations are promenant, such as Alzheimer’s and schizophrenia.

The second paper is much more easy going. This one focuses on the method of absorption and its effect on the experience. By comparing what happens when the compound is vaporised and inhaled, absorbed sublingually (under the tongue) and eaten, it proves that absorption through the gut has little or no effect.

But what else is there to talk about? Any other papers worth a read? I might write up a transcript of the presentation here, along with the PowerPoint presentation itself.