
Persistence of Long-Term Memory in Vitrified and Revived C. elegans worms - gwern
http://online.liebertpub.com/doi/pdf/10.1089/rej.2014.1636
======
reasonattlm
Cryonics is the low-temperature preservation of the deceased in order to grant
them a chance at a renewed life when technologies for restoration are
developed. One of the big outstanding questions is the degree to which a well-
conducted cryopreservation successfully preserves the fine structure of the
brain and thus the data of the mind. The fact that cold water drowning victims
can live again after an hour or more of brain death tells us that memory is
encoded in physical structures, with the current consensus (not undisputed)
suggesting it is located in synaptic connections between neurons.

Scanning technologies have been used to show that cryopreservation via
vitrification of cryoprotectant-infused tissue does indeed preserve the fine
structure of brain cell connections. E.g.:

[http://www.brainpreservation.org/content/competitors](http://www.brainpreservation.org/content/competitors)

So it seems reasonable to move forward with initiatives based on the theory
that yes, cryonics is going to preserve the mind. Certainly all too many
people are left with no other viable option for a shot at a longer life in the
future. There is always the need for more and better proof, however. At the
present time studies involving restoration of vitrified individuals must be
carried out in lower animals, unfortunately, as researchers are still far from
the point at which they can safely restore vitrified mammals. That said,
restoration of a vitrified mammalian organs has been demonstrated, though even
that is very early stage work in the grand scheme of things.

~~~
stefantalpalaru
> cold water drowning victims can live again after an hour or more of brain
> death

They don't reach brain death. The cold causes a drop in the oxygen
requirements, but the brain keeps on living.

~~~
ciphergoth
Indeed. If by "brain death" you mean "massive ischemic damage", then
cryopreservation under good circumstances also takes place before brain death.

------
ejstronge
Take this with a huge block of salt.

This PDF is of a paper recently accepted by an unremarkable journal. Further,
the research is sponsored by a company that performs cryonics. If this is
true, it may be exciting but we should wait for others to replicate these
findings.

~~~
gwern
I don't see why we should be skeptical. It seems to lack the usual warning
signs like small samples or questionable statistics, and, for all the
skepticism most laymen have ('but how do we know vitrification does preserve
memory even if it preserves everything else?!') this is what one should have
expected, if only from related instances like comas and drownings - whatever
memory is physically encoded as, exactly, it's clearly not ultra-evanescent.

~~~
ejstronge
I think there are a few causes for concern regarding this paper:

1\. Rejuvenation Research isn't a widely-recognized biology journal. If this
is the first time memories have been recovered from a cryopreserved organism,
this should be in a top journal as it's great news.

2\. The article was accepted on the day it was received[ref 1]. This doesn't
inspire confidence in the strength of the associated peer review as such
minimal review is unusual in biology. Indeed, the average review time for the
journal is about 4 weeks[ref 2] (which itself is a short time).

3\. This is pretty concerning - the key protocol used in the paper hasn't been
published (see reference 14 in the PDF). While it's described in passing in
this paper, there's no way to assess the full protocol.

4\. There's a juxtaposition between the authors' new technique and an
established protocol for freezing _C. elegans_. However, the authors only
freeze _C. elegans_ for 30 minutes in their protocol whereas they freeze
organisms in the established protocol testing group for 2 weeks. There seem to
be appropriate controls but this difference in protocol is stated very
unclearly.

 _References_

1\.
[http://online.liebertpub.com/doi/10.1089/rej.2014.1636](http://online.liebertpub.com/doi/10.1089/rej.2014.1636)

2\.
[http://www.liebertpub.com/DContent/Files/FastFacts/FastFacts...](http://www.liebertpub.com/DContent/Files/FastFacts/FastFacts_RejuvenationResearch.pdf)

~~~
gwern
> If this is the first time memories have been recovered from a cryopreserved
> organism, this should be in a top journal as it's great news.

Cryonics related research never gets published in top journals no matter its
quality, so this is what you would expect. There is a long-standing war by
cryobiologists on anything to do with cryonics - the old canards like the
exploding lysosomes o' death may have been refuted but last I heard, they were
still explicitly banning from membership any cryonicists.

> 2\. The article was accepted on the day it was received[ref 1].

Look up the journal policies and note the dates: it was posted the day it was
received, as part of Liebert Instant Online/LION
[http://www.liebertpub.com/forauthors/rejuvenation-
research/1...](http://www.liebertpub.com/forauthors/rejuvenation-
research/127/) but that doesn't mean it was reviewed or accepted in a day:

> To enable the release of new scientific findings as quickly as possible, the
> Journal has a policy of pre-publishing all manuscripts in their unedited
> format upon acceptance. The papers will have undergone full peer review but
> will not yet have been copyedited, typeset, or proofread/corrected by the
> authors. All accepted papers appear online within 72 hours of acceptance as
> a part of Liebert Instant Online (LION).

Given the editing gap is at least October 2014 - April 2015...

> 3\. This is pretty concerning - the key protocol used in the paper hasn't
> been published

Reference 14 is to the SafeSpeed cryopreservation protocol, not to
vitrification in general. They _also_ tested out traditional vitrification. If
you distrust SafeSpeed, you can throw out that entire part of the paper and
the results are exactly the same: preservation of olfactory memory. (This was
covered on pg3, I don't know how you missed it if you could look up the
reference number...)

> However, the authors only freeze C. elegans for 30 minutes in their protocol
> whereas they freeze organisms in the established protocol testing group for
> 2 weeks.

? The 30 minutes refers to the traditional vitrification itself (this is why
it's 'slow') where they're immersed in Ln2, but then they were all stored in
some more regular freezers at -80 celsius (much warmer than LN2) for several
weeks (pg11-12).

~~~
ejstronge
> There is a long-standing war by cryobiologists on anything to do with
> cryonics - the old canards like the exploding lysosomes o' death may have
> been refuted but last I heard, they were still explicitly banning from
> membership any cryonicists

I'm not sure what to say here - politics definitely plays into biology
publication but there are multiple well-respected journals to send 'small
results', of which I don't believe Rejuvenation Research is one.

> Look up the journal policies and note the dates

I did exactly this. Unless Rejuvenation Research has an unusual application of
the terms 'Received' and 'Accepted' (see ref. 1), I'm more inclined to believe
the information on the article page than a general journal policy.

Weirdly enough, all the articles in the most recently published edition of
Rejuvenation Research
([http://online.liebertpub.com/toc/rej/18/2](http://online.liebertpub.com/toc/rej/18/2))
had at least some gap between receipt and acceptance. I'm even more inclined
to think something unusual has started to happen with the current batch of
online-before-publication papers. This could, of course, be a bug with the
journal's content management system.

> Reference 14 is to the SafeSpeed cryopreservation protocol

Publishing a reference to an non-peer-reviewed protocol is unusual,
irrespective of the details. This is even more unusual when the protocol forms
the core of the paper.

To be a bit more precise (using details from the paper):

Vitrification and slow-freezing are distinct methodologies for preserving
specimen. The authors _only_ use two methodologies for preservation, which
they explain in the methods section:

    
    
        The cryopreservation methods included worms that were trained and cryopreserved by vitrification and worms that were trained and cryopreserved by slow freezing
    

> If you distrust SafeSpeed, you can throw out that entire part of the paper
> and the results are exactly the same: preservation of olfactory memory

I think this is part of why my original comment said we should wait for
replication and be skeptical. Trust in science is a difficult thing to gain
and an easy thing to lose. The choice of journal and the very-unclear
description of the cryopreservation protocols make me a bit more skeptical of
the results.

The SafeSpeed protocol seems to be under review at Nature Methods. If it's
accepted there, the authors will likely need to supply supporting data which
will only bolster the claims they made here.

Further, the slow-freezing result is a fairly cheap one to replicate; we're
likely to see many people try it and they'll likely cite Alcor's work.

> The 30 minutes refers to the traditional vitrification itself

I think the authors were intentionally vague in comparing the two preservation
methods, especially given how clear the rest of the report is.

First, note that only _one_ vitrification technique was used, SafeSpeed.
Additionally, only _one_ slow-freezing technique was used, a standard approach
that one of your sibling commenters references in passing.

The SafeSpeed-vitrified animals were stored in liquid nitrogen (LN2) for 30
minutes and then recovered. This is stated (unclearly) in the methods section
(pg 8) and alluded to in the discussion.

> 30 minutes refers to the traditional vitrification itself (this is why it's
> 'slow')

This is counter to what the authors state - they explain that the SafeSpeed
method is actually slower than 'slow-freezing'. See page 11.

 _Conclusion_

I think that this will be exciting if SafeSpeed gets published somewhere more
reputable. As you alluded to earlier, if there is a physical correlate to
benzaldehyde training that isn't damaged upon freezing, we'd definitely expect
it to be recovered after freezing.

This paper just isn't compelling enough to say that we've seen this occur
definitively.

 _References_

1\.
[http://online.liebertpub.com/doi/10.1089/rej.2014.1636](http://online.liebertpub.com/doi/10.1089/rej.2014.1636)

    
    
        'Received' usually indicates receipt of a manuscript;
        'Accepted' usually indicates the end of scientific edits.
        Often, articles are held up (i.e., not published online) to coordinate 
        publishing with other labs/projects within a lab.

~~~
gwern
> I'm not sure what to say here - politics definitely plays into biology
> publication

This is well beyond regular scientific politics; the attacks are now older
than a lot of people working in the field. I don't know why you would expect
random journals to be unaffected (where do you get cryobiologist reviewers who
are unaffected or fearless?).

> I'm more inclined to believe the information on the article page than a
> general journal policy.

The journal policy which explains what the dates mean?

> Weirdly enough, all the articles in the most recently published edition of
> Rejuvenation Research
> ([http://online.liebertpub.com/toc/rej/18/2](http://online.liebertpub.com/toc/rej/18/2))
> had at least some gap between receipt and acceptance.

So are you even objecting to anything besides a software bug?

> The authors only use two methodologies for preservation, which they explain
> in the methods section:

I don't understand your objection here. OK, so one protocol is still upcoming.
I don't think this is that unusual since I see plenty of references to
publications in press in papers I read - econ papers spend years as preprints,
I regularly comment on drafts, fast moving areas often do this, work which has
been turned into multiple publications does this all the time because
different journals move at different rates etc. I don't think this is a big
problem but _even if it was_ , they _also_ tested out the standard well-
understood procedure you cannot possibly have any objection to, so what's with
this 'only' business? That's the method one wants them to test - and they did.
And it worked the same for preserving memories. Which is the point.

> Further, the slow-freezing result is a fairly cheap one to replicate; we're
> likely to see many people try it and they'll likely cite Alcor's work.

That seems unlikely. No one wants to go near cryonics for the political
reasons I mentioned and there's no funding for it. I have been saying for at
least 5 years now that I really hoped someone would run exactly this
experiment with seeing if training survived, and it took that long for any
study to be done - despite always being cheap to run. Sometimes being cheap to
replicate doesn't matter.

> This is counter to what the authors state - they explain that the SafeSpeed
> method is actually slower than 'slow-freezing'. See page 11.

I don't see any such claim. I think you are again confusing different stages
of storage and revival.

> This paper just isn't compelling enough to say that we've seen this occur
> definitively.

I find it compelling. None of your arguments goes to the root, and most are
tangential: it's not a prominent journal because of the very well known
politics involved, the dates are due to quirks of the website or acceptance
process, the new protocol is not well-described but seems to work as well as
the older methods and the older methods suffice to prove the point, and some
confusion about the details doesn't obviate the core claim either. The worms
were trained, were frozen, were revived, and remembered. Which of your
criticisms defeats this? This isn't some mushy social psychology paper with
n=15 and the authors could have tried a thousand different analyses to pull
out a p<0.05; the worms either remembered or they didn't, there's not many
ways to slice the data.

~~~
ejstronge
I don't think we're going to agree on the quality of this paper, which is
perfectly fine and a normal part of science.

Two things:

> I don't think this is that unusual since I see plenty of references to
> publications in press in papers I read - econ papers spend years as
> preprints

This isn't normal in biology. It's a stance that researchers are trying to
change (see [http://biorxiv.org/](http://biorxiv.org/), notably), however.

> I don't see any such claim. I think you are again confusing different stages
> of storage and revival.

At a molecular level, the two processes may work differently (we don't, of
course, have the SafeSpeed protocol). However, the article states:

"During this time, it is necessary to allow 30-45 minutes for vitrification
processes, 15-30 minutes for slow freezing processes, and 24 hours for all
processes to safely recover and verify the survival of the worms"

------
dcminter
Not directly related, but it reminded me of this:
[http://everything2.com/title/The+ability+of+planarian+worms+...](http://everything2.com/title/The+ability+of+planarian+worms+to+run+a+maze+more+successfully+after+being+fed+the+remains+of+a+successful+worm)

------
trhway
well, today we can restore information - sound - from a photo of the
gramophone recording media
([https://mediapreservation.wordpress.com/2012/06/20/extractin...](https://mediapreservation.wordpress.com/2012/06/20/extracting-
audio-from-pictures/)). Tomorrow - from a frozen brain.

So, i wonder, if i had enough money - may be instead of whole body or head
cryonics, i should go for sub-micron thick slicing of my brain and making
photo and various other wavelength scans of it and storing this digitized info
somewhere (like Glacier :) Or may be i should just start a start-up offering
just such a service to make money for my future cryonics :)

~~~
gwern
This is a good question. There's a lot of active research into chemical
fixation and then doing brain scanning on thin slices to reconstruct the
connectome and other features, with regular progress like "High-resolution
whole-brain staining for electron microscopic circuit reconstruction", Mikula
& Denk 2015
[http://connectomes.org/pdf/mikula2015_.pdf](http://connectomes.org/pdf/mikula2015_.pdf)
. I have some more links in
[http://www.gwern.net/plastination](http://www.gwern.net/plastination)

The upsides to chemical fixation or plastination is that you avoid issues with
temporary interruptions in cooling (as far as we know, Alcor and CI have a
nearly perfect record in maintaining LN2 coldness, but one could wonder how
well that will work for the next century) since a fixated brain will be stable
at room temperature, and you can have a much more quantifiable idea how much
progress has been made in fixation and scanning through things like the BPF's
prizes
([http://www.brainpreservation.org/](http://www.brainpreservation.org/)). So
that makes this attractive.

The downside is that there is no infrastructure setup (there's no one you can
go to and say "here's $10k, please wait by my bedside and as soon as I'm
declared legally dead, dunk my head in osmium"), and the concerns about
cryonics being too slow are magnified massively - people are worried about
vitrification taking several hours and cryonics possibly being too late, but
the current fixation solutions take more on the order of a _month_ and don't
necessarily fixate the entire brain volume. So you may get a great
preservation which can be easily stored and imaged at high quality without any
concerns about how to heat it back up... but what will you be imaging?

If I died tomorrow, I would go with regular cryonics hands-down no contest.
But in 30 years? If Mikula, Hayworth, the BPF, Seung, etc keep on pushing
forward this area, maybe then plastination will be equal or superior.

------
jacquesm
Frogs can do this too (and more in fact), so I wouldn't be surprised if it was
true. That said it's a bit of a (ok, not such a bit) stretch to extrapolate
from being able to freeze a worm which has absolutely minimal nervous tissue
to how this finding will apply to the non-science of cryonics, though I can
see why those with a stake in the game would want to make such connections as
strongly as possible.

Not only are there differences in scale between frogs, worms and humans, there
are also differences in complexity and structure.

------
dang
Also
[https://news.ycombinator.com/item?id=9593988](https://news.ycombinator.com/item?id=9593988).

------
teddyh
The viability (or not) (and the incentives of different parties in) the field
of cryonics was previously discussed here:

[https://news.ycombinator.com/item?id=8241374](https://news.ycombinator.com/item?id=8241374)

------
pavlov
Cryonics is fascinating because it's the closest thing to a technological
religion so far. (Scientology does not count, it's more like a pulp sci-fi
religion.)

St. Paul eloquently described the upcoming Singularity and the raising of the
vitrified minds:

 _" Listen, I tell you a mystery: We will not all sleep, but we will all be
changed -- in a flash, in the twinkling of an eye, at the last trumpet. For
the trumpet will sound, the dead will be raised imperishable, and we will be
changed."_

~~~
ggreer
I often hear the "cryonics is a religion" claim, but a closer examination
refutes this. Unlike religions, cryonics is not exclusive. A person can
practice Christianity or Islam and be signed up for cryo. Cryonics makes no
claims about morality or politics. There's no dogma about how to live one's
life, or the origins of the cosmos, or what happens to consciousness after the
brain is destroyed.

The most important difference between cryonics and religions is that religious
people are typically convinced of the afterlife. In contrast, cryonicists
rarely assign high probability to being revived. Most think of it like an
experimental cancer treatment: Unlikely to work, but better than the
alternative.

And like experimental treatments, research is ongoing. Since cryonics
organizations were first founded, there have been many improvements to
preservation protocols. Modern cryopreservation methods allow a rabbit kidney
to function after thawing, albeit with some damage.[1] This is very promising,
as parts of the kidney have poor vasculature. Brains are much easier to
saturate with cryoprotectants.

1\.
[http://www.21cmpublications.com/PubFiles/11/12FahyORG5-3[1]....](http://www.21cmpublications.com/PubFiles/11/12FahyORG5-3\[1\].pdf)

~~~
kansface
I will eschew Wittgenstein's advice and take the bait- I don't think the
techno-religion claim is too far off the mark. Of course, the entirety of our
argument is over the definition of religion.

| Unlike religions, cryonics is not exclusive.

I think we'd both agree that exclusivity isn't the principle quality of
religion, though many religions are exclusive.

Religions typically have some core set of beliefs - some of which may be
untestable, ceremony, dress, and often attempt to explain the mysteries of the
universe. I'd also throw in the quality that religions advocate their
followers either do something they wouldn't otherwise do or abstain from
something they would otherwise enjoy. Its possible some religion lacks
proscriptions for life, but I certainly haven't met it.

Cyronics certainly has a core set of beliefs (faith!?) that lead to action. I
very much doubt that even if we were to develop sufficient technology to
unfreeze all those heads, future humans would do so. Perhaps we'd unfreeze
some heads for science and then a few more for the novelty. We both agree
cyronics is unlikely to work and that there is no way to run this experiment
within our lifetime - cryonics requires faith (by any other name) in humanity
and technology.

|In contrast, cryonicists rarely assign high probability to being revived.
Most think of it like an experimental cancer treatment: Unlikely to work, but
better than the alternative.

I know Christians that would describe their experience in similar terms.

| The most important difference between cryonics and religions is that
religious people are typically convinced of the afterlife.

Christians believe in an afterlife, but not all religions do, or even weigh in
on the matter - Judaism immediately comes to mind. Apropos, cryonics proposes
its own cycle of rebirth! Well, possible rebirth until the great robot wars,
the sun explodes, or the heat death of our local cluster.

As for the other attributes I mentioned - dress, ceremony, and the like,
cryonics offers bracelets, (non)ceremonial decapitation with ensuing
vitrification, and requires its devotees to die somewhere convenient! OK, I
agree, this doesn't exactly fit the bill.

Precisely defining religion and faith is tricky. At the very least, I would
say cyronics is uncomfortably close to a religion.

~~~
nshepperd
> Cyronics certainly has a core set of beliefs (faith!?) that lead to action.

You could describe basically every mundane activity as having a core set of
beliefs that lead to action, but ok.

> I very much doubt that even if we were to develop sufficient technology to
> unfreeze all those heads, future humans would do so. Perhaps we'd unfreeze
> some heads for science and then a few more for the novelty.

Not reviving cryopreserved people is equivalent to leaving them to die...
Unless you're counting on future society to be completely 100% morally
bankrupt, I would bet on there being eventually at least one person with both
enough of a conscience and enough money to fund the revival of people in
suspended animation (as the cost of revival should decrease over time with
advancing technology). Not that this really seems relevant to any connection
to religion.

> Apropos, cryonics proposes its own cycle of rebirth! Well, possible rebirth
> until the great robot wars, the sun explodes, or the heat death of our local
> cluster.

FWIW, unlike religion, cryonics does not really claim you can come back from
death. Legal death, yes, but legal death is merely a convenient criterion that
captures "the point at which we give up on treating a person with current
technology". If you set a cryonicist on fire and destroy their brain's
information, they are permanently dead, just like anyone else.

(ETA: Since physics is reversible, destroying information is technically
impossible, but capturing and calculating reversal of the trajectories of
smoke, soot, air and photons back into a whole brain seems thermodynamically
infeasible, even for a civilization with Big Angelic Powers.)

~~~
kolinko
iirc physics is non-reversible. Something to do with entrophy and possibly
quantum physics.

~~~
FeepingCreature
Quantum Physics is reversible; see
[http://en.wikipedia.org/wiki/Delayed_choice_quantum_eraser](http://en.wikipedia.org/wiki/Delayed_choice_quantum_eraser)
. (Warning: amateur, may be misunderstanding.) You do have to be able to
capture the complete environment of the quantum process, which is why
decoherence is usually considered one-way.

------
crablar
Cold storage

