The importance of being specific when discussing the therapeutic potential of cannabis and cannabinoids

When it comes to discussing the therapeutic potential of cannabis, it’s really important to differentiate between the artisanal preparations that are not regulated, the pharmaceutical-grade botanical drug substances that are regulated and the isolated cannabinoids that have been purified (ie. not synthetic). Being able to easily understand and differentiate between all three (and assume that others can too) is something I take for granted.

Some people are quick to dismiss all claims that cannabis can have any therapeutic potential whatsoever. They assume that these are the proclamations of a bunch of stoners seeking validation. They ignore the hype because they believe that there can be no real science behind it all. Seriously, how can smoking a herbal preparation grown in someone’s backyard benefit anyone’s health? Thing is, I used to be one those sceptics until I realised there’s so much more to it than just smoking weed.

What I’m trying to say is that there is real science being conducted using cannabis. However, it may not resemble what you would usually associate with the plant or drug. In order to accurately study it, scientists need to know exactly what they’re dealing with. For the vast majority of research, this means breaking down the entire cannabis plant into its individual constitutive components (of which there are more than 500!); the most obvious ones being the cannabinoids (of which there are more than 100). Each cannabinoid is rigorously studied in isolation in order to have its pharmacological effects properly characterised.

Sometimes cannabinoids work very well in combination with other cannabinoids. Research like this requires the pharmacological characterisation of each cannabinoid in isolation and then further characterisation when they are combined. As this can get quite complicated, most combined preparations usually contain 2 cannabinoids at a specific ratio.

This is a slightly different story when it comes to studying botanical drug substances, which are well-defined whole-plant extracts. The main cannabinoids (usually just 2) are present at a specific ratio, but there are tiny amounts of other cannabinoids present (less than 1%) in addition to non-cannabinoid compounds too (at low concentrations). These preparations are pharmaceutical grade because they are highly reproducible. This means that these types of cannabis preparations can be used in randomised clinical trials and the results can be taken seriously by the scientific community and clinicians. I should also point out here that isolated cannabinoids can (and have been) used in randomised clinical trials too.

The sad truth is that people’s misconceptions about cannabinoids has massively delayed the research and development of pharmaceutical agents that can be used to treat billions of people worldwide.

Collective noun for a group of electrophysiologists?

My husband and I have had an ongoing discussion for quite a while now about what the collective noun for a group of electrophysiologists should be. This discussion came up again a few nights ago, where he firmly believes that the collective noun should be a ‘superstition‘. I, on the other hand, think it should be a ‘paranoid‘, or maybe an ‘expletive‘.

I started my first career-related job recently. I’m now working as a manual (conventional) patch clamp electrophysiologist for a contract research organisation. It’s definitely a completely different world to academia and I’m having to pick up a lot of new concepts very quickly, but happily, I’m enjoying myself very much (despite having to write my thesis in the evenings and weekends).

Although it’s a whole new world for me, there’s definitely something oddly familiar about the environment and the people I work with.

I find myself smiling whenever I think back to my uni days, when I would sit at my patch clamp rig and fantasise about electrophysiology equipment that wasn’t held together with dog-eared autoclave tape or covered in aluminium foil that was torn in several places. I also used to dream about what it would be like to have a comfortable chair, a responsive computer and enough workspace to accommodate a keyboard, a mouse, an open lab book and a monitor (or 2 – if I was doing MEA experiments). On my first day at work it came as a little bit of a surprise to find that some things really don’t change.

I operate two patch clamp rigs now (not at the same time!) and although they are pretty much identical, they each have their own foibles. Whether it’s the optics of the microscope, or the amount of patience (and swearing) it requires to get the perfusion flow rate just right, or the extra stability a few centimetres of tape gives to the manipulation of the patch electrode, or the aluminium foil that was once wrapped around a cable but is now half hanging off and catches on the stage of the microscope. There are even uncomfortable chairs and very limited workspace! But thankfully I still get to work beside a window (yay daylight!) and at least the computers (and IT support) are much better now.

I sometimes used to think that maybe I’m a little bit too particular when it comes to conducting electrophysiological experiments. I have a certain way of doing things, often in a certain order, and I like my work environment to be organised and tidy. I also like to have particular (and in my opinion, essential) bits of equipment to hand in case I need it (ie., plastic syringes, forceps, super glue, copper wire, silver wire, tubing, foil, soldering iron, hot air gun, pipette glass, heat-shrink, blu tac, tape, lab roll, pipette tips etc etc). Having now found myself surrounded by highly skilled and very experienced electrophysiologists, I’ve discovered that maybe I’m not so weird after all. I may have a certain way of doing things, but I’m definitely not the only one. It also turns out that I’m not the only one who gets paranoid about flushing the perfusion tubing sufficiently, or the particular sequence of events that need to be followed in order to successfully patch a cell, or having too many people in the room when you’re trying to do an experiment.

There’s even a hint of slightly irrational superstition. One of my colleagues has said that their two best days, in terms of the success rate of patching a certain cell line, was when they were listening to music by a certain band. As we’re currently having our patience tested by this really troublesome cell line, it is now a firmly held belief that our chances of getting data will be massively improved by listening to this particular band. The only caveat being that my colleague was the only person in the lab at the time, so it may not work if the rest of us are there to watch/try it out. We all solemnly agreed that the conditions should be kept the same as last time, otherwise it will never work.

So this has provided my husband with more proof that a group of electrophysiologists should be called a ‘superstition‘, which is a good call, but I personally think the collective noun should still be a ‘paranoid‘ or an ‘expletive‘. Funnily enough though, I get really anxious when my husband puts new shoes on the table. I know it’s completely irrational, but I just can’t help myself. Maybe electrophysiology appeals to a certain kind of person.

attempted murder of crows

Never underestimate the power of a thorough literature review

As someone who is currently writing their thesis, I have a major piece of advice for PhD students embarking on their studies…

Never underestimate the power of a thorough literature review!

Always, always, always read as much as you can about the field of research you are working in. Especially if there are numerous inconsistencies and unresolved controversies surrounding the expression/function of proteins/receptors and the pharmacology of their ligands. I’m specifically talking about the field of cannabinoid research and its relatives.

I carried out one of my early experiments* under strict instructions from my project manager and supervisor. Their rationale at the time sounded ok to me: look at the effects of drug A in the presence of drug B, which happens to be a selective antagonist for receptor X. My knowledge of the literature at the time (which was quite substantial, with over 100 references for the literature review I wrote 9 months after starting the PhD) concurred that this seemed like a legitimate thing to do: If drug B, which has no effect on its own, blocks the effects of drug A, then drug A must bind to receptor X.

Seems legit

Recently however, while doing a bit of extra research on the drugs I used**, I came across a few research articles (written by one of the most famous scientists in the field) that described both drug A and drug B to be inverse agonists of receptor X. Basically, that means both drugs should produce an effect by themselves. The same effect. At the same receptor.

*face palm*

Great. How on earth am I supposed to explain that in my thesis, let alone in my viva?

Luckily, my trump card was the fact that neither drug did the same thing when applied individually, which kinda goes against the ‘inverse agonists at the same receptor’ theory. Also, drug A appeared to be antagonised by drug B afterall. To top it all off, I found more research articles with evidence to suggest that not only do drug A and B bind to receptor X, but they also bind to receptor Y!

Hurrah for the inexplicable and frustrating weirdness of cannabinoid research!

Seriously though, do lots of reading. Read the reviews written by the most famous and respected scientists. Read the original articles that they refer to. Read articles about similar research conducted at other laboratories. Even read the letters and commentaries on these articles written by other scientists in the same field – they might dispute the findings, or highlight problem areas. Even attending conferences can open your eyes to what other scientists really think.

However, try to stay open minded as science is subject to fashion and popularity. Established scientists can denounce the results of perfectly good work just because it challenges the dogma. But sometimes it can take a fresh pair of eyes and a different way of thinking to make real progress.

Never make the excuse that you don’t have enough time to read research papers. There is always time to read papers. Your supervisors/advisors may be an expert in a particular field of research, but you are the expert in your PhD. Try to make sure that every experiment is properly thought through because you don’t want to be struggling to find reasons and explanations when it comes to your thesis/viva. Saying ‘My supervisor told me to do it’ is simply not good enough.

*This was supposed to be a simple control experiment to show what the majority of the literature has already described. My ‘proper’ experiments involved trying to figure out how drug C works.

**Drug A and drug C are similar, but there’s so much more written about drug A that I use this to help me interpret what drug C does.

A typical day – In the beginning

A typical day at the beginning of my PhD was very different to a typical day towards the end of my third year.

When starting a PhD (at least in Life Sciences), before anyone is let loose in a lab, there’s a lot of form-filling, health and safety inductions and even occupational health assessments that need to be done first. Sometimes this can take a while and usually depends on how organised and attentive your supervisor is.

In the meantime, there’s a huge amount of background literature that needs to be thoroughly read, understood and reviewed. This part can be quite tedious, as sitting at a desk reading doesn’t feel like you’re doing much at all, especially when you’re surrounded by busy PhD students and post-doctoral researchers.

In the beginning I used to turn up at around 9.30am, to find most PhD students and post-docs already at their desks or in the lab doing experiments. It used to make me feel a bit guilty, especially as almost all of them would still be there when I left the office at 5pm. But in actual fact, it’s entirely normal to feel like you’re at a loose end when everyone around you seems to be working incredibly hard.

After all the necessary paperwork and admin stuff was out the way, I was able to get into the lab and start learning how to perform my first experiment. Again, this was a little tedious because it mainly involved shadowing other people while they tried to explain what they’re doing and why they’re doing it. For me, this was learning how to ‘patch’ (patch clamp) cells in fresh brain slices and it took about 3 months to go from absolute beginner to first successful attempt. I think it was another month or so before I was confident enough to do a ‘proper’ experiment.

Once I’d found my feet and got into a bit more of a routine, I started to feel less guilty about not working as hard as some of the more seasoned researchers in the lab. I suppose it’s difficult to make yourself work hard when you’re still trying to fathom exactly what you’re PhD is about. I know numerous people, myself included, who didn’t include any data from the first year in their thesis. For some, it was simply due to the direction that the science took them in and for others it was because their experiments didn’t work or weren’t optimised enough.

My advice to people who have just started a PhD; Be patient. The stress will come later.

Guilty feelings

The feelings of a PhD student just starting out