Scientists (Yoshihiro Matsushima, et al) investigated the potential for 日陰痺茸 - 'Hikageshibiretake' (sun-shadow-numbing-mushroom, formerly Psilocybe argentipes, now P. subcaerulipes, Fig 1) as a treatment for obsessive compulsive behaviors by utilizing previously established mouse models. Their research showed that P. subcaerulipes mycelium outperformed both SSRI and pure Psilocybin in improving responses to marble burying and locomotor activity.
Project Proposal and Author Note
Here, the group of scientists springboard off of previously established work in the lat 90's to compare the effectiveness of entheogenic "mushrooms on marble-burying behavior as compared with the effects of anti-psychotic agents such as fluvoxamine...[and] authentic psilocybin." Literature notes that there has been a previous case of entheogenic mushroom therapy that was "maintained as a treatment for OCD symptoms for 4 years." [Moreno 1997]; which was the inspiration for this investigation. The marble burying mouse model proposed as tests for repetitive, compulsive-like behaviors; specifically number of marbles buried and spontaneous movements. While not without its flaws, the marble burying model was used to show effectiveness of the prescription drug fluvoxamine, and thus should provide a similar set of insights into entheogen effects on same or similar behaviors.
The importance of this paper to me has many levels, it was looking at endemic Japanese entheogenic mushrooms, it utilized mycelia and not fruit bodies, and further, investigated natural products versus isolated constituents. Each of these points, and more, pushed this paper to the forefront of my list. Taking time to work with and understand the unique properties of individual species is important to helping preserve their biodiversity. Noted later on, there is a large influx of non-native Psilocybe cultivation that has been occurring in Japan since before the 21st century; this inherently puts the native species at risk as they compete with their cousins over natural habitats. Also taking the investigation into the space of mycelia versus fruit bodies was also very interesting. As previously noted in this blog, mycelia have been shown to contain different metabolites than fruiting bodies, especially when it comes to psychoactive tryptamines. This is an important distinction as the mycelia is likely to act differently due to varying metabolites when compared to fruiting bodies. Furthermore, it is these distinct chemical profiles that are likely the cause for different results when compared to isolated the compounds psilocybin and psilocin.
History of entheogenic mushrooms in Japan
As of available research in 2021, there are no no historical records of intentional use of entheogenic mushrooms in Japan. The oldest record of "poisoning by psychoactive mushrooms" is “a story of nuns who climbed a mountain, ate a mushrooms, and danced” in 今昔物語集 - 'Konjaku Monogatari Shuu', a Heian period collection of stories compiled near the end of the in the 12th century that emphasizes the path of Buddhism from India to China and then Japan; and while 'dancing' is mentioned no other psychedelic or hallucinatory descriptions are. [Musha 1988] That being said, the presence and knowledge of these mushrooms were not foreign to the Japanese; this can be seen in the etymology of the mushrooms' names. For instance: 踊い茸 - 'Odoritake' (dance-inducing mushroom); sometimes confused for the edible 舞い茸 - 'Maitake' (Grifola frondosa, hen-of-the-woods) get their names from "that joy of finding delicious mushrooms" inducing dancing. Taxonomical differences between entheogenic maitake and the edible maitake are unclear based on observable features [Matsushima 2009], if they even are different organisms. Also consider 笑い茸 - 'Waraitake' (Panaeolus papilionaceus, laugh-inducing mushroom), while considered inedible and found to cohabit dung piles with engheogenic fungi, they are not known themselves to contain psilocybin. [Allen 2013] That being said, P. subcaerulipes are only one of many entheogenic fungi that inhabit the Japanese archipelago; interestingly the National Institute of Health Sciences reported that two mushroom species from European culture kits, P. cubensis and P. cyanescens, were what comprised 90% of all entheogenic mushrooms distributed for intentional ingestion in Japan, not the endemic P. subcaerulipes. [Musha 1988]
Before the turn of the century, the availability of entheogenic mushrooms to the average individual was staggering; street vendors existed and head shops sold packages of cultured or foraged fruiting bodies for around 10 USD. While cultivation and collection of entheogenic 'vegetables' was widely overlooked in the drug enforcement policies in Japan, near the turn of the century, the Japan Poison Information Center noticed relatively sharp increases in 'psychedelic mushroom poisoning': from to10 in 1998, 19 in 1999, 32 in 2000, and 55 in 2001. [Maruyama 2003] This surge of interest spurred the revision of the Japanese“Law for the Control of Narcotic Drugs, Psychotropic Substances, and their Raw Materials” in June 2002. [Maruyama 2003] The revision included regulations against the culture, possession, trade, and ingestion of psilocybin, psilocin and mushrooms that produce them.
Obsessive compulsive disorder (OCD), selective serotonin reuptake inhibitors (SSRIs), and entheogenic tryptamines
The power of the subconscious mind over one's actions cannot and should not be understated. Compulsive actions, those actions when performed persistently and repetitively without any apparent reward or pleasure are experienced by a wide number of individual during their lifetime. However, when those compulsions become excessive, all-consuming, and begin to disrupt daily living; some individual seek to
gain more autonomy back from their obsessive behaviors.
Modern medicine has found that increasing
the amount of extracellular serotonin (serotonin outside of and around neurons and other cells) which can double patient response to psychological treatment. [Busko 2008] This was typically done through chemically preventing serotonin being taken back in by some neurons through the use of compounds like fluvoxamine (Tbl 1); which is currently prescribed as the first line of treatment for OCD in both Canada and the UK. [Katzman 2014] While the mechanisms of psilocybin are understood well enough to point towards their serotonin receptor activity, deeper mechanisms for action will be saved for later posts. Suffice to say, the structural similarities between serotonin and psilocybin-like tryptamines depict a shared origin for both in the amino acid tryptophan (Tbl 1).
Marble Burying to test repetitive/compulsive-like behaviors
Establishing baseline parallels between one animal's behavior to another is one of the key hallmarks of psychological animal models. It is fairly well understood that "when rodents are put in a cage with marbles they will bury the marbles." In the marble burying model, 20-25 marbles are arrayed on the surface of clean bedding. The number of marbles buried in a 30 min session is scored by investigators blind to the treatment or status of the subjects (Fig 2). Generally, "this behavior is seen as anxiety related or OCD behavior." Based on previous studies reductions in the number of marbles buried were observed when mice were injected with drugs used to treat anxiety or OCD; and the scientist note that "marble burying doesn't show any difference between anti-compulsive or anxiolytic (anxiety-breaking) activity." Notably, the test tends to be sensitive to two major classes of drugs: selective serotonin reuptake inhibitors (SSRIs) and benzodiazepines (muscle relaxants). Benzodiazepines are commonly used to treat anxiety and SSRIs are used to treat both OCD and anxiety. [Borsini 2002]
In researching I have come across mixed opinions over the validity of the marble burying test. One asks: if marbles cause anxiety, rodents would most likely remain immobile or avoid the side containing marbles. Furthermore they point towards marble burying test being sensitive to two types of drugs: SSRIs and muscle relaxers. Benzodiazepines have been shown to have no effect on patients suffering from OCD. If the test measured OCD then benzodiazepines ought not affect observable mouse behavior. [Borsini 2002]
I feel these criticisms are fairly weak and can be answered with a little logical thinking. First the idea that the marbles are the 'cause of anxiety' is incorrect, the marbles represent neutral data points to be monitored by the scientist, not as a source of anxiety for the mouse. The anxiety of the experiment comes from the transferring to a new cage and how the animal handles that transition. Furthermore, on the argument of benzodiazepines sensitivity of the test: benzodiazepines are muscle relaxants, and thus reduce locomotor activity generally and would prevent any mouse from scoring high on the test (marbles buried and anxious movements).
Stronger criticisms of the model lie in it's testing of one acute dose of anti-anxiety agent for thirty minutes. Some therapists report weeks before a noticeable change is observed in patients. [Albelda 2012] There is also a latent genetic component that influences this test; some breeds of mice show decreased burying behavior generally. This (and the previous neutral state of the marbles) can be counteracted through Tabasco coating marbles which will increase burying. [Albelda 2012] The best and final response to any criticism of a single model is to then utilize a batter of anxiety tests that have high predictive validity. Through these combined data a direction of efficacy will arise, depicting the therapeutic value (if any) the novel agent might have. [Albelda 2012]
Conclusions and Caveats
The results of this test are quite remarkable in their clarity and potential for deeper research. Two major conclusions can be drawn from the four graphs provided: 1) Psilocybin, by itself provides some effect on reducing compulsive behaviors, but does not out perform the control SSRI fluvoxamine and 2) saline suspensions of powdered mycelium outperformed SSRIs and pure Psilocybin in reduction of compulsive behaviors.
The later statement is observed in Fig. 3, where the dose of 0.1-1.0g of powdered mycelium per kg of mouse weight that administered through in saline suspension was show to reduce marble burying better than both fluvoxamine at typical dose rations (administered by abdominal injection), but saw a loss of that benefit at higher doses. Furthermore, the reduction in locomotor activity was noticeable, but not statistically significant. The scientists go on to note that "the relationship between [doses of psilocybin or P. subcaerulipes and the] number of buried marbles showed an inverted bell curve. This represents a very unusual result for the marble-burying behavior test,since many chemicals show a proportional relationship, high doses reducing both the number of buried marbles and loco motor activity."
The former statement is supported in Fig. 4, where the most effective dose mycelium extract (1.0g/kg) was noted through HPLC analysis to provide about 0.24 mg/kg of psilocybin; was further compared to pure psilocybin. It took, relative to the mycelium, up to six times the amount of pure psilocybin to produce a statistically significant impact on marble burying, which even at the high doses, still did not perform as well at reducing marble burying or locomotor activity as the median dose of psilocybe mycelium.
As previously noted, a point that should not go overlooked is the importance of the choice to use mycelia versus fruiting bodies for the experiment. They might likely end up being the source of future treatments as compared to the fruiting bodies, mycelium is both easier to cultivate and denser in production relative the mushrooms themselves a are likely to provide just as important a product as the fruit bodies themselves.
While administration of fluvoxamine was performed via subdermal injection near the abdomen of mice as compared to psilocybin and P. subcaerulipes via suspension of powdered compound or mycelium in saline and administered by oral ingestion. The difference in administration is likely due to the process of previous testing with fluvoxamine; which when compared to the lack of data on psilocybin and powdered mycelial injections is likely the reason for this disconnect and arguably represents a notable difference in procedure. Are the blood-serum levels of active tryptamines for the mycelial ingestion comparable to those of injected fluvoxamine? A question further worth investigating.
Also noted by the scientists is the difference between pure psilocybin and mycelial suspension, going so far as to note that the improvement to effect is likely due to a concert of other tryptamines present in the suspensions that work together in altering observable behaviors. I would even go so far as to argue that the compounds, when not excreted, are metabolized by the body, become building blocks for serotonin and other tryptamine based neurotransmitters, thus increasing the body's available levels generally. They also note the importance of the "inverted bell curve" of the mycelial data. They argue "this represents a very unusual result for the marble-burying behavior test, since many chemicals [including the tested pure Psilocybin] show a proportional relationship, high doses reducing both the number of buried marbles and loco motor activity."
Project Proposal Success or Failure?
The response to proper dosing of Psilocybin out performed Fluvoxamine; and, saline suspensions of Psilocybe subcaerulipes mycelium outperformed both in reducing marbles buried and anxious movements of mice during testing; both seen as signs of compulsive behavior reduction.
Reviewing the data and the conclusions put forth, the hypothesis proposed by the Scientists was supported. The reason(s) for acceptance are quoted below (emphasis and parentheticals mine):
"The relationship between dose [of psilocybin or P. subcaerulipes] and number of buried marbles showed an inverted bell curve."
"[P. subcaerulipes] at a dose of 0.05–2 g/kg showed a trend toward inhibited marble-burying behavior, while a dose of 0.1–1 g/kg significantly reduced the number of buried marbles (Fig. 2)"
References
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