Cold Exposure & Other Mild Stressors for Increased Health & Longevity

Dean Pomerleau · May 13, 2016

Marijuana (Component) Promotes Browning of White Fat Too!

I'm not sure if Sthira (or anyone but Kenton and Tom?) are still reading this thread, but I suspect he'll be pleased to hear about this new research study [1].

Background: Cannabidiol (CBD), is a major medicinal, non-psychoactive component of marijuana, accounting for up to 40% of the plant's extract.

Results: Researchers in [1] treated human white adipose tissue with various concentrations of CBD for 72 hours and then tested to see if they were expressing the usual markers of fat browning/beiging. They found them in spades, in a linear dose-response relationship, as can be seen in this graph of protein expression:

Conclusion: It looks like we can add the cannabidiol component of the "wacky weed" ☺ to the list of brown / beige fat promoters.

Discussion: I could be in luck, since my home state of Pennsylvania legalized marijuana just last month for medical use. Now I just have to convince a judge that "promoting fat browning for health and longevity" is a legitimate medical use, to be added to the list of 17 medical conditions for which medicinal marijuana has been approved in PA ☺.

It's not entirely clear to me where cannabidiol oil (refined from hemp seed oil) fits into the existing marijuana laws. In fact, it's looks like very much of a gray area, both in terms of it's legality and product quality (as with almost all supplements...). Plus the stuff is damn expensive. Here is where the researchers in [1] bought theirs - $38 for 5 mg!

It's sold to consumers elsewhere on the web as "High CBD hemp oil". You can supposedly get 165mg of CBD as high CBD hemp oil for $65, but my advice is "buyer beware". It's not even clear what the oral dosage you'd need to gain this WAT-browning effect, since [1] was an in vitro study, as are most of the studies investigating CBD for medical conditions like cancer (as opposed marijuana use for palliative care). Self-described "authorities" suggest starting with a dosage of 1-3 mg of CBD per day, which would put it at somewhere around $1/day (not that bad), if the dosage recommendation is relevant and the CBD quality can be trusted - which are by no means a given...

I've added cannabidiol to the list of brown / beige fat inducers below, but I'll be sticking with cold exposure, capsaicin, and the other foods & practices on the list. If I were going to experiment with marijuana, it would be for very occasional recreational purposes, although it goes without saying that I'm not endorsing any such practice...

--Dean

----------------

Here is the latest full list of modifiable and [nonmodifiable] factors associated with increased brown/beige adipose tissue and/or thermogenesis:

Cold exposure - by far the best BAT inducer/activator
Spicy / pungent foods, herbs & supplements - capsaicin / chilli peppers, curcumin / turmeric root, menthol/mint/camphor, oregano, cloves, mustard, horseradish/wasabi, garlic, onions
Arginine-rich foods - Good vegan sources include seeds (esp. sesame, sunflower & pumpkin), nuts (esp. almonds and walnuts) and legumes (esp. soy, lupin & fava beans and peas)
Healthy Fats - Olive Oil / MUFA-rich diet, DHA / EPA / fish-oil
Nitrate-rich foods - beets, celery, arugula, and spinach
Other foods - Apples / Apple Peels / Ursolic Acid
Beverages - green tea, roasted coffee, red wine, cacao beans / chocolate
Drugs / Supplements - metformin, caffeine, creatine, nicotinamide riboside (NAD), resveratrol, ginseng, cannabidiol / hemp oil / medicinal marijuana
Low gluten diet
Methionine restriction - Reduce animal protein. Soy is low in methionine and high in arginine.
Low protein diet
Fasting
Exercise
Acupuncture - locations Zusanli (foot - ST36) and Neiting (lower leg - ST44)
Whole body vibration therapy
Avoid obesity/overweight
[being naturally thin - high metabolic rate]
[being younger]
[being female]
[Ethnicity - having cold-climate ancestors]

-----

[1] Mol Cell Biochem. 2016 May;416(1-2):131-9. doi: 10.1007/s11010-016-2702-5. Epub

2016 Apr 11.

Cannabidiol promotes browning in 3T3-L1 adipocytes.

Parray HA(1), Yun JW(2).

Author information:

(1)Department of Biotechnology, Daegu University, Kyungsan, Kyungbuk, 712-714,

Republic of Korea. (2)Department of Biotechnology, Daegu University, Kyungsan,

Kyungbuk, 712-714, Republic of Korea. jwyun@daegu.ac.kr.

Recruitment of the brown-like phenotype in white adipocytes (browning) and

activation of existing brown adipocytes are currently being investigated as a

means to combat obesity. Thus, a wide variety of dietary agents that contribute

to browning of white adipocytes have been identified. The present study was

designed to investigate the effects of cannabidiol (CBD), a major nonpsychotropic

phytocannabinoid of Cannabis sativa, on induction of browning in 3T3-L1

adipocytes. CBD enhanced expression of a core set of brown fat-specific marker

genes (Ucp1, Cited1, Tmem26, Prdm16, Cidea, Tbx1, Fgf21, and Pgc-1α) and proteins

(UCP1, PRDM16, and PGC-1α). Increased expression of UCP1 and other brown

fat-specific markers contributed to the browning of 3T3-L1 adipocytes possibly

via activation of PPARγ and PI3K. In addition, CBD increased protein expression

levels of CPT1, ACSL, SIRT1, and PLIN while down-regulating JNK2, SREBP1, and

LPL. These data suggest possible roles for CBD in browning of white adipocytes,

augmentation of lipolysis, thermogenesis, and reduction of lipogenesis. In

conclusion, the current data suggest that CBD plays dual modulatory roles in the

form of inducing the brown-like phenotype as well as promoting lipid metabolism.

Thus, CBD may be explored as a potentially promising therapeutic agent for the

prevention of obesity.

PMID: 27067870

TomBAvoider · May 13, 2016

What always makes me nervous about popping something because it's been shown to be good for x, y, or z, is that you have basically no idea as to what else that clearly bioactive substance does. I don't want to end up with an "well arsenic definitely cures a headache - guaranteed not to have one" effect. For that matter, this even applies to things like capsaicin. Which is why the only surefire way of assessing it would be all-cause mortality outcomes against controls, but obviously that's not practical in humans, and of dubious translatability from animal models. At least with capsaicin we can weakly re-assure ourselves that it's been a part of human diet for generations and if large numbers of people were dropping dead, we'd at least know something was off. It may brown you fat, but what good is it, if it cuts your life or health in other ways. What are its other effects? How safe is it?

Dean Pomerleau · May 13, 2016

Tom,

Very good point about unknown side effects. The more you look at human metabolism and the effects of particular substances (like fish oil), the more you see just how complicated things are. Fish oil may protect the brain, make your arteries more pliable, but also make your mitochondria more peroxidizable, or not, as you and I discussed yesterday.

For that matter, this even applies to things like capsaicin. Which is why the only surefire way of assessing it would be all-cause mortality outcomes against controls, but obviously that's not practical in humans... At least with capsaicin we can weakly re-assure ourselves that it's been a part of human diet for generations and if large numbers of people were dropping dead, we'd at least know something was off.

As your say, capsaicin can't be that bad since people have been consuming it in pretty large quantities for generations. But even better, there is evidence [1] it is associated with lower all-cause mortality, as discussed by Michael here. While [1] is not a double-blind, placebo controlled intervention trial, it does seem like promising evidence in favor of hot peppers, if not capsaicin directly. Good enough, apparently, that Michael has added hot peppers to his diet...

--Dean

-------------

[1] Lv J, Qi L, Yu C, Yang L, Guo Y, Chen Y, Bian Z, Sun D, Du J, Ge P, Tang Z, Hou W, Li Y, Chen J, Chen Z, Li L; China Kadoorie Biobank Collaborative Group. Consumption of spicy foods and total and cause specific mortality: population based cohort study. BMJ. 2015 Aug 4;351:h3942. doi: 10.1136/bmj.h3942. PubMed PMID: 26242395; PubMed Central PMCID: PMC4525189. (See also the Rapid Response by Prof. Nicholas D Moore).

Gordo · May 14, 2016

In a prior post I showed a log of CE and blood glucose and mentioned I was going to post again about an even higher GI meal (despite the fact that I made the last one intentionally pretty high GI by including foods like rice and bananas). Here is a new log to give a sense of the phenomenal glucose control that can be achieved with CE/BAT/Thermogenesis (I don't even know what to call this anymore, we need to come up with our own name). This time I used different cooling methods compared to the last log posted.

LOG

Started morning with max cold shower (as I now do every morning) at 7:10AM

Had large glass of crushed ice water at 7:45AM

Went out, came back. 9:30AM had another large glass of crushed ice water.

House was 70 degrees all day. No food or exercise all morning, just mental work (iPad app development). Focus and mental clarity are stellar, I believe the cold showers help with this.

1:55PM fasting blood glucose (BG)=58 mg/dl

(I've seen readings as low as 54, while still feeling great, never saw such readings before I started cold exposure)

Put on techkewl cooling vest and started making lunch at 2:30PM.

(I blame Dean for this)

3:01PM (final fasted reading) BG=68 mg/dl

Finished preparing nice high GI meal consisting of a carefully measured and weighed:

The barley was first milled into "flour" using a 30 second blend in my blendtec blender (for maximum blood sugar spiking capacity) along with the flax and chia then combined with the almond milk (plus a cup of water) and oats and pressure cooked for 10 minutes at 15PSI. Everything else was added after cooking.

That's 219g of carbs and 82 grams of sugar, including pure liquid sugar in the form of maple syrup.

Breakdown of the macros:

Pic of meal - can't tell by the pic, but a 1352 calorie bowl of porridge is pretty huge and heavy and would fill almost anyone up:

Started eating at 3:03PM

Drank one cup of crushed ice and water with lemon squeeze while eating.

Finished eating at 3:21PM

3:23PM (2 minutes postprandial) BG=68 mg/dl

3:40PM Had another cup of crushed ice water.

3:43PM (22 minutes postprandial) BG=64 mg/dl

(wow, I was expecting a 20 minute postprandial spike with a high GI meal like this, on the contrary, my BG actually went DOWN)

4:09PM (48 minutes postprandial) BG=70 mg/dl

4:35PM (1 hour 14 minutes postprandial) BG=62 mg/dl

5:09PM (1 hour 48 minutes postprandial) BG=76 mg/dl

5:30PM Cooling vest is spent. Removed. Had another large glass of crushed ice water.

6:00PM (2 hours 39 minutes postprandial) BG=77 mg/dl (final reading)

I am impressed by the BAT induced BG stability. Again I never saw results like this before I started doing CE.

Refs:

http://diabetes.diabetesjournals.org/content/early/2014/07/15/db14-0746.short

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3533266/

I continue to be humored by all of the "things that activate BAT" that just happen to be part of my regular routine even before I knew there was a link. For example I eat beets almost daily (they may be the most affordable superfood on the planet, I get a giant bag of them for $1 from my local produce store and they have them year round). I've been eating most of the items from the Dean list on a regular basis for a long time. I am excited to see apples added to the list -- as I've mentioned before, I grow my own organic apples and eat them daily when in season (3 more months to go). A new Dr. Greger video also mentions that apples can repair DNA damage. Speaking of which, if any of you are in the market for it, this year surprisingly I've found fantastic quality, low cost ($20) fruit trees at Walmart and Lowes (but not Home Depot). These trees are still available in my area, have good branching/branch angles, and fantastic roots, I believe they were grown in pots for a full year, some will produce fruit in the first year, and are far ahead of any bare root trees you could buy (I saw honeycrisp and fugi apple trees, belle of Georgia peach trees at Walmart, good varieties, lowes had fantastic stanley plum trees, red haven and hale peach trees, and moonglow pear trees as well as winesap apple trees). I added 7 new fruit trees this year, all are doing well so far.

Another coincidence - for years my wife has been using that ovuview software Dean mentioned recently (great software, but not sure it has any application related to this thread, haha).

Regards,

Gordo

Edited June 3, 2016 by Gordo

Dean Pomerleau · May 14, 2016

Gordo,

That is another amazingly good glucose result - thanks for sharing it! But I must say, your previous test meal looked a lot more appetizing to me ☺. But the well-known insulin boosting effects of dairy (ice cream) makes this a much better, though less appealing, test of postprandial glucose control. I wonder if Paul McGlothin is following any of this... I'm sure even he would be impressed.

For anyone who has forgotten, here are Gordo's tips for implementing cold exposure effectively and cheaply - including his cold shower protocol. They certainly seem to be working like a charm for your Gordo!

--Dean

drewab · May 14, 2016

Paul is aware of Gordo's CE experiments. See these two threads over at his website:

http://www.livingthecrway.com/home/forum/diabetes---glucose-control/a-daily-log-example-of-the-phenomenal-glucose-control-that-is-possible-with-cold-exposure

http://www.livingthecrway.com/home/forum/cr-way-support-group-general-/getting-enough-calories-in-just-two-meals

And BTW, I am not Drew Lawton over there. It's a bizarre occurrence that their is another Drew active their. My tag is Drew_ab there.

Gordo,

That is another amazingly good glucose result - thanks for sharing it! But I must say, your previous test meal looked a lot more appetizing to me ☺. But the well-known insulin boosting effects of dairy (ice cream) makes this a much better, though less appealing, test of postprandial glucose control. I wonder if Paul McGlothin is following any of this... I'm sure even he would be impressed.

For anyone who has forgotten, here are Gordo's tips for implementing cold exposure effectively and cheaply - including his cold shower protocol. They certainly seem to be working like a charm for your Gordo!

--Dean

Edited May 14, 2016 by Dean Pomerleau

Dean Pomerleau · May 14, 2016

Hey - that's wonderful!

Thanks for those pointers Drew, and thanks to you Gordo for carrying the torch over on the LivingTheCRWay™ forums! Glad to see Paul is aware of what we've been doing/researching, and seems to be interested in it. It seems like it would be right up his alley.

--Dean

Kenton · May 14, 2016

When I was in grade school a classmate "taught" me how to not feel pain in my injured left leg by kicking my right leg (thus getting my mind off the left one). Funny how I never seem to feel or mind being weak or hungry any more, being preoccupied with feeling cold ! !

Dean Pomerleau · May 14, 2016

Ray Cronise and the "Metabolic Winter" Hypothesis

All,

I just finished watching this video (embedded below) of an interview of Ray Cronise by Rhonda Patrick of FoundMyFitness.com.

Ray was on day 23 of a 24-day supervised water fast at the time of this interview. Ray looks good, and says he feels great. He acknowledges though that clear thinking isn't always easy at that point in a fast, and I have to admit he is a bit rambly at times in the interview.

But the reason Ray and his interview is relevant for this thread is that Ray is a pioneer in human application of mild cold exposure. He has a paper [1] in which he co-author David Sinclair (of SIRT1 fame) outline their "Metabolic Winter" Hypothesis.

In short, they think modern society coddles us too much; we aren't exposure to cold temperatures or significant time away from food, and as a result our health suffers. In other words, we're killing ourselves by being unnaturally warm and well-fed. Interestingly, Ray goes so far as to suggest that intentional exercise is simply leveraging the health-promoting response the body developed originally to cope with cold exposure via shivering at first, and then non-shivering thermogenesis once cold-adapted. Exercise triggers a similar response to increase thermogenic capacity by upregulating irisin (as discussed here and here). From [1]:

Thus, it might be reasonable to consider that many of the health benefits of physical activity are actually adaptive responses related to times of cold stress and shivering. In nature, animals do not intentionally participate in high levels of activity to mitigate excess calorie ingestion—available calories are limited and animals conserve activity. In fact, the main factors influencing energy expenditure are body mass and ambient temperature, not activity.

This idea sort of turns Michael's "jiggling pecs" observation on it's head - i.e. exercise is piggybacking on the adaptive cold response rather than the cold exposure piggybacking on the beneficial response to exercise.

Ray also suggests that CR and CE share many of the same beneficial metabolic responses. The spend the first hour or so of the video talking about fasting. Interesting stuff, but not as relevant for this thread.

He starts talking about CE at 1:10:00 in the video. Here are some highlights:

1:11:00 - The link between CR and CE via the "Metabolic Winter" hypothesis

1:13:00 - CE and improved sleep

1:15:15 - His (and Wim Hof's) cold shower protocol - (10-sec warm → 20-sec cold) repeated 10 times, end on cold.

1:21:00 - CE for treating depression via elevated epinephrine

1:28:50 - More on the relationship between CE and exercise

1:34:15 - "We've engineered winter out of our lives in the last century"

1:37:30 - Ray's "trichotomy" of cold stress, dietary restriction and sleep

1:46:00 - Sleeping in cold bedroom. No need for blanket or sheet once you're adapted (I can confirm!)

1:48:00 - Melatonin and how it decreases body temperature. He does cold shower then melatonin (30mg!) before bed

1:52:45 - Synergy between melatonin, resveratrol and SIRT1

--Dean

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[1] Metab Syndr Relat Disord. 2014 Sep;12(7):355-61. doi: 10.1089/met.2014.0027. Epub

2014 Jun 11.

The "metabolic winter" hypothesis: a cause of the current epidemics of obesity

and cardiometabolic disease.

Cronise RJ(1), Sinclair DA, Bremer AA.

Free full text: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4209489/

The concept of the "Calorie" originated in the 1800 s in an environment with

limited food availability, primarily as a means to define economic equivalencies

in the energy density of food substrates. Soon thereafter, the energy densities

of the major macronutrients-fat, protein, and carbohydrates-were defined.

However, within a few decades of its inception, the "Calorie" became a commercial

tool for industries to promote specific food products, regardless of health

benefit. Modern technology has altered our living conditions and has changed our

relationship with food from one of survival to palatability. Advances in

agriculture, food manufacturing, and processing have ensured that calorie

scarcity is less prevalent than calorie excess in the modern world. Yet, many

still approach dietary macronutrients in a reductionist manner and assume that

isocalorie foodstuffs are isometabolic. Herein, we discuss a novel way to view

the major food macronutrients and human diet in this era of excessive caloric

consumption, along with a novel relationship among calorie scarcity, mild cold

stress, and sleep that may explain the increasing prevalence of nutritionally

related diseases.

PMCID: PMC4209489

PMID: 24918620

Dean Pomerleau · May 16, 2016

Cold Exposure and Cold-Shock Protein RBM3

I'm sure all of us have had the experience of forgetting the name of someone who we haven't seen (or thought of) in a few months or years. It's on the tip of your tongue, but you just can't recall it. Memory is one of those "use it or lose it" sort of deals. If the synaptic connections that encode long-term memories aren't refreshed occasionally via recall, they atrophy and disappear. Loss of synapses that encode memories is also the first indication of neurodegenerative diseases like Alzheimer's Disease (AD), happening well before the development of plaques & tangles, and well before the neuronal death that characterizes late-stage AD [7][8].

Animals that hibernate have this problem in spades - since their brains are pretty much shut down for several months during which time they aren't actively rehearsing their memories to maintain them. But when they wake up, they still need to remember where they hid their nuts the previous fall and where the neighborhood predators live.

Fortunately, it appears nature and their own biochemistry have got them covered. In response to cold and a modest (i.e. 1 °C [1]) drop in temperature, cells all over the body, but particularly neurons in the brain, synthesize a 'cold shock' protein called "RNA binding motif protein 3" (RBM3) [2]. From [4]:

In hibernating animals, RBM3 is upregulated in muscle, liver, and heart tissues of black bears [ref, ref], as well as in brain, heart, and liver tissues of squirrels at late torpor [ref].

And it's not just hibernating mammals where RBM3 is found. RBM3 or it's analogs are prevalent across both the plant and animal kingdom, present in plants, fish, amphibians, birds and mammals, including mice, rats and importantly, humans [4]. Interestingly, this cold-induced synthesis of RBM3 is boosted by the presence of either melatonin and life-extending, cold-induced FGF-21 [1]. In addition to cold, RBM3 also induced by UV radiation and hypoxia [5].

What does RBM3 do, particularly in neurons? It appears to play a role in neuronal stem cell maintenance and proliferation [4]:

In mammals, RBM3 level peaks during the early postnatal period and then decreases to very low levels in youth and adulthood in most regions of the brain, except for areas where proliferation remains active, such as the subventricular zone (SVZ) and the hippocampal subgranular zone (SGZ) [ref, ref], indicating a pivotal role of RBM3 in the maintenance of stemness and proliferation in neural stem/progenitor cells. Regardless to this dynamic temporal expression of CIRP and RBM3 with high abundance in early developmental stages and low in mature organisms, many mature cells maintain their ability to overexpress CIRP and RBM3 in response to stressful conditions, such as cold.

So right off the bat we see cold-induced RBM3 plays a role in neurogenesis, particularly in the brain's memory formation hub, the hippocampus. So that's nice to see. But it gets even better. Again from [4]:

RBM3 expression ... correlates with good [cancer] prognosis and reduced risk of disease progression and recurrence...

Conversely [4]:

A large variety of immunohistochemical studies, including many tumor types, have shown consistently that loss of RBM3 expression is associated with clinically more aggressive tumors and an independent factor of poor prognosis.

In particular, high expression of RBM3 is associated with good prognosis in breast, epithelial, ovarian, prostate, testicular, bladder, esophageal, stomach, colorectal, and skin cancer [4].

RBM3 even appears to be beneficial for preventing sarcopenia and bone loss [4]:

RBM3 is involved in the regulation of skeletal muscle size and the prevention of muscle loss, indicating a novel vital function of RBM3 in muscle disease [ref]. A subsequent study has further revealed that RBM3 inhibits both necrosis and apoptosis in muscle myoblasts, consistent with a general cytoprotective function of RBM3 [ref]. Moreover, RBM3 is suggested to mediate hypothermia-induced overexpression of bone protein alkaline phosphatase and osteocalcin [ref].

The above combination of effects is interesting and pretty unusual. Usually anabolic agents that help preserve muscles and bones (i.e. IGF-1) are associated with increased cancer proliferation. But here, we see the best of both worlds - muscle & bone preservation and lower cancer risk associated with expression of RBM3.

In summary, RBM3 appears to be a) induced by mild cold exposure / reduced body temperature, and b) have many beneficial effects all over the body, including preventing cancer proliferation, preventing muscle and bone loss, and boosting growth of new neurons, especially in the hippocampus.

But a closer look at the effects of RBM3 on the brain is warranted. A certain molecular variant of RBM3 (i.e. RBM3 which is 'absent a single arginine residue in the RGG domain' - whatever that means...) is found in heavy concentrations in neural dendrites [6] - which anyone familiar with neuroanatomy will know are the post-synaptic (i.e. the receiving side) of synapses.

What's it doing there? In general, from [3]:

[RBM3] is known to increase local protein synthesis at dendrites [ref] and global protein synthesis through ribosomal subunit binding and/or microRNA biogenesis [ref].

That sounds good, but what exactly does it mean in terms of brain health? That is what this study [3] from Nature last year (popular press account, another one, and a detailed review on an Alzheimer's forum), aimed to find out. Here is where things get really interesting. What prompted the researchers from [3] was the question we started with - how do mammals pull off the previously-observed [9][10] trick of preserving memories & synapses during hibernation and/or reconstruct them upon awakening? By now you won't be surprised to hear that it appears to be RBM3 that mediates this synaptic preservation/reconstruction.

In particular, study [3] investigated the effects of cold exposure and RBM3 on the synapses in both normal mice, and in mice models of two neurodegenerative disease, Alzheimer's Disease (AD) via genetically-mutant 5XFAD mice¹ and a mad-cow-like prion disease, which I'll abbreviate as MCD for "mad cow disease" although it isn't exactly, simply to avoid using PD as an abbreviation for "prion disease". PD would be more accurate, but it is too easily confused with another neurodegenerative disease, Parkinson's Disease.

Basically what they found was that in wild-type mice (w/o the AD or MCD), a short but deep bout of hypothermia (17 °C body temperature for 45 minutes) resulted in the loss of synapses, but upon rewarming the synapses were restored, at least in number, through a process dubbed structural plasticity. I'll talk about whether memory performance was also preserved/restored shortly (preview - it was). This sort of cold-induced structural plasticity is exactly what appears to happen during and after hibernation in mammals.

Mice with the AD and MCD models, in the first few weeks after exposure, i.e. before their diseases had time to take hold, showed the same structural plasticity - they lost synapse with cooling but then the synapses were restored after rewarming. And in these mice, they did test memory, via a clever object recognition task you can read about in the full text. The memories that the infected (and wild-type) mice had formed prior to cooling was preserved upon rewarming. Just like in the case of hibernation (now where did I hide those nuts?).

But a few weeks/months later, when the diseases had progressed more in the AD and MCD mice, if they cooled these messed up mice their synapses were not restored after rewarming, and memory performance was impaired after cooling and then rewarming. Importantly, as time (and the diseases) progressed, the AD and MCD mice produced less and less RBM3 in response to cooling, and this drop paralleled their drop in synapse recovery and memory performance upon rewarming - suggesting RBM3 was causal in the synapse and memory preserving process (more on that below).

By now you should be thinking to yourself - so what Dean? So you can cool normal or (early) neurodegenerative mice and rewarm them without trashing their synapses or their memory performance and this may happen as a result of upregulation of RBM3 - what good it that?

Here are three possibilities:

It might bode well for Zeta's hope (I miss Zeta. Wonder where he's at?) that someday we'll develop technology for suspended animation. Brian points out in this post that one of the challenges of suspended animation is preserving the delicate structures of the brain (particularly synapses) during cryo-suspension. RBM3 might play a role in such preservation. BTW, I came across [11], titled "Is Human Hibernation Possible?", which is a good review of the feasibility of this approach to both life extension, and short-term suspended animation (see next point).
Understanding / leveraging how RBM3 preserves the brain during hypothermia / hypoxia may be useful in clinical practice, where temporary cooling of the body is done to given doctors more time, with the hope (and fortunately, the observation) that patients can be temporarily cooled and then rewarmed without significant deleterious effects, particularly on brain function. This clinical application of cooling is discussed in this recent popular press article on a pilot study being done here in Pittsburgh with gunshot victims who come into the hospital in critical condition. The goal is to put them into "suspended animation" for a few hours or days to give doctors more time to stabilize them and treat their injuries. Understanding and leveraging RBM3 could make this sort of short-term, medically-induced cryo-suspension safer and more effective.
These results might help explain the amazing mammalian diving reflex (MDR),which I mentioned back in my CE Albatross post two months ago, and which I promised to discuss someday. The MDR enables animals (and people!) to recover from being submerged in cold water without oxygen for up to two hours without brain damage. It would not be at all surprising to me if scientists discover that RBM3 plays an important role protecting the brain in the MDR, although I looked and couldn't find any papers that addressed this possibility.

But the original question is a good one - If I don't think I'll be shot, fall into a frozen lake, or undergo cryo-suspension for life extension anytime soon, what use is all this RBM3 synapse preservation stuff for me now?

That's where the rest of [3] comes in. It turns out that cold exposure, and specifically boosting RMB3, acts as a hormetic agent - providing long-lasting protection against loss of synapses, and preserving memories.

Specifically, as we saw above, cooling and rewarming the AD and MCD mice once their disease had progressed was worse than useless - i.e. they lost synapses (and memories) during the cooling process which weren't restored upon rewarming. In contrast, cooling and rewarming early in the progress of the disease (i.e. just a couple weeks after the mice had been treated to induced AD or MCD) not only kept their memories from before the cooling intact immediately after rewarming - it also served as a hormetic stressor, which helped preserve their synapses for many weeks to come. Specifically, long after control mice with the AD or MCD conditions had lost their synapses and their ability to form new memories, the AD and MCD mice who had been cooled and rewarmed early on in their diseases were protected - their synapses and memory abilities were preserved much longer. In addition, they lived longer than the control mice who had either the AD or MCD treatment, but no cooling and rewarming. Don't get too excited. Their lives were still greatly shortened relative to wild-type mice who weren't messed up by the AD or MCD treatments.

Further, they found that this preservation of synapses and memory performance was mediated by upregulation of RBM3. In particular, a single episode of cooling raised RBM3 levels in wild-type mice for three days, and in the AD and MCD mice, a single cooling episode "resulted in sustained several-fold increase in RBM3 expression up to 6 weeks later" [3]. When they knocked out the ability of the mice to produce RBM3 in response to cooling, it erased all the benefits - without elevated RBM3, the AD and MCD mice lost synapses and memories after cooling/rewarming at the same rate as control AD and MCD mice who hadn't been cooled/rewarmed, and they didn't live any longer either. In short, RBM3 was critical for cold-induced preservation of synapses and memories.

Finally, to get the real smoking gun, they skipped the cooling/rewarming and simply injected the AD and MCD mice with a virus that expresses RBM3. Yup - sure enough. Injecting this RBM3-expressing virus into the hippocampi of the AD and MCD increased hippocampal RBM3 by a factor of 3 and in the MCD mice, this boost to RBM3 preserved synapses, neurons, memories and extended survival relative MCD mice who got placebo virus injection into their hippocampi. The AD mice injected with the RBM3-expressing virus showed similar (although not identical) benefits. Finally, to cap it off:

RBM3 knockdown [in the absence of cooling] also reduced synapse number and

novel object memory in wild-type mice (Extended Data Fig. 10b), thus it is

likely to be involved in synaptic maintenance under normal physiological conditions.

Here is that figure 10b they mentioned, showing the number of synapses (left graph) and memory performance (right graph) of wild-type mice with normal levels of RBM3 (white bars) and suppressed levels of RBM3 (yellowish bars):

In short, when RBM3 was cut to 70% below its usual level, normal mice kept in normal conditions (i.e. no cooling) were 50% worse at retaining memories!

Here is the paper's optimistic final paragraph:

In conclusion, we have shown that early synapse loss in mouse models

of neurodegenerative disease results, at least in part, from defective

synaptic repair processes associated with failure to induce the cold-shock

RNA-binding protein, RBM3. This results in impaired synaptic

reassembly after cooling, but also appears to be important in the context

of protecting against ongoing synaptic toxicity during disease, and

in synaptic maintenance in wild-type mice. Our data suggest that further

understanding the mechanisms of action of cold-shock proteins

such as RBM3 may yield insights into endogenous repair processes and

bring new therapeutic targets for neuroprotection in neurodegenerative

disease.

In short, it appears that the cold-shock protein RBM3 can be induced by cold exposure in mice and humans. It helps to prevent cancer proliferation, sarcopenia and bone loss, boosts growth of new (hippocampal) neurons, preserves synapses and memories, and perhaps can even help prevent the onset and/or slow the progression of neurodegenerative diseases, including Alzheimer's disease.

Yet another pathway by which cold exposure appears good for your health!

--Dean

------

¹The 5XFAD mice model of Alzheimer's disease involved genetically messed up mice who express 5 human genes associated with familial, early-onset AD, which causes them to rapidly develop amyloid-beta plaques. They lose synapses and memories first, then neurons and eventually die, just like in human AD.

----------

[1] Neuroscience. 2015 Oct 1;305:268-78. doi: 10.1016/j.neuroscience.2015.08.012.

Epub 2015 Aug 8.

Cold stress protein RBM3 responds to temperature change in an ultra-sensitive

manner in young neurons.

Jackson TC(1), Manole MD(2), Kotermanski SE(3), Jackson EK(3), Clark RS(4),

Kochanek PM(4).

Extremely mild hypothermia to 36.0 °C is not thought to appreciably differ

clinically from 37.0 °C. However, it is possible that 36.0 °C stimulates highly

sensitive hypothermic signaling mechanism(s) and alters biochemistry. To the best

of our knowledge, no such ultra-sensitive pathway/mechanisms have been described.

Here we show that cold stress protein RNA binding motif 3 (RBM3) increases in

neuron and astrocyte cultures maintained at 33 °C or 36 °C for 24 or 48 h,

compared to 37 °C controls. Neurons cultured at 36 °C also had increased global

protein synthesis (GPS). Finally, we found that melatonin or fibroblast growth

factor 21 (FGF21) augmented RBM3 upregulation in young neurons cooled to 36 °C.

Our results show that a 1 °C reduction in temperature can induce pleiotropic

biochemical changes by upregulating GPS in neurons which may be mediated by RBM3

and that this process can be pharmacologically mimicked and enhanced with

melatonin or FGF21.

PMCID: PMC4570027 [Available on 2016-10-01]

PMID: 26265550

------------

[2] Williams DR, Epperson LE, Li W, Hughes MA, Taylor R, Rogers J, Martin SL, Cossins AR, Gracey AY (2005) Seasonally hibernating phenotype assessed through transcript screening. Physiol Genomics 24(1):13–22. doi:10.1152/physiolgenomics.00301.2004 PubMedCrossRef

------------

[3] Nature. 2015 Feb 12;518(7538):236-9. doi: 10.1038/nature14142. Epub 2015 Jan 14.

RBM3 mediates structural plasticity and protective effects of cooling in

neurodegeneration.

Peretti D(1), Bastide A(1), Radford H(1), Verity N(1), Molloy C(1), Martin MG(1),

Moreno JA(1), Steinert JR(1), Smith T(1), Dinsdale D(1), Willis AE(1), Mallucci

GR(2).

Full text: http://sci-hub.cc/10.1038/nature14142#

Comment in

Nat Rev Neurosci. 2015 Mar;16(3):124.

Nature. 2015 Feb 12;518(7538):177-8.

Nat Rev Neurol. 2015 Mar;11(3):124.

In the healthy adult brain synapses are continuously remodelled through a process

of elimination and formation known as structural plasticity. Reduction in synapse

number is a consistent early feature of neurodegenerative diseases, suggesting

deficient compensatory mechanisms. Although much is known about toxic processes

leading to synaptic dysfunction and loss in these disorders, how synaptic

regeneration is affected is unknown. In hibernating mammals, cooling induces loss

of synaptic contacts, which are reformed on rewarming, a form of structural

plasticity. We have found that similar changes occur in artificially cooled

laboratory rodents. Cooling and hibernation also induce a number of cold-shock

proteins in the brain, including the RNA binding protein, RBM3 (ref. 6). The

relationship of such proteins to structural plasticity is unknown. Here we show

that synapse regeneration is impaired in mouse models of neurodegenerative

disease, in association with the failure to induce RBM3. In both prion-infected

and 5XFAD (Alzheimer-type) mice, the capacity to regenerate synapses after

cooling declined in parallel with the loss of induction of RBM3. Enhanced

expression of RBM3 in the hippocampus prevented this deficit and restored the

capacity for synapse reassembly after cooling. RBM3 overexpression, achieved

either by boosting endogenous levels through hypothermia before the loss of the

RBM3 response or by lentiviral delivery, resulted in sustained synaptic

protection in 5XFAD mice and throughout the course of prion disease, preventing

behavioural deficits and neuronal loss and significantly prolonging survival. In

contrast, knockdown of RBM3 exacerbated synapse loss in both models and

accelerated disease and prevented the neuroprotective effects of cooling. Thus,

deficient synapse regeneration, mediated at least in part by failure of the RBM3

stress response, contributes to synapse loss throughout the course of

neurodegenerative disease. The data support enhancing cold-shock pathways as

potential protective therapies in neurodegenerative disorders.

PMCID: PMC4338605

PMID: 25607368

----------

[4] Cell Mol Life Sci. 2016 May 4. [Epub ahead of print]

Cold-inducible proteins CIRP and RBM3, a unique couple with activities far beyond

the cold.

Zhu X(1), Bührer C(2), Wellmann S(3,)(4).

Free full text: http://link.springer.com/article/10.1007%2Fs00018-016-2253-7

Cold-inducible RNA-binding protein (CIRP) and RNA-binding motif protein 3 (RBM3)

are two evolutionarily conserved RNA-binding proteins that are transcriptionally

upregulated in response to low temperature. Featuring an RNA-recognition

motif (RRM) and an arginine-glycine-rich (RGG) domain, these proteins display

many similarities and specific disparities in the regulation of numerous

molecular and cellular events. The resistance to serum withdrawal, endoplasmic

reticulum stress, or other harsh conditions conferred by RBM3 has led to its

reputation as a survival gene. Once CIRP protein is released from cells, it

appears to bolster inflammation, contributing to poor prognosis in septic

patients. A variety of human tumor specimens have been analyzed for CIRP and RBM3

expression. Surprisingly, RBM3 expression was primarily found to be positively

associated with the survival of chemotherapy-treated patients, while CIRP

expression was inversely linked to patient survival. In this comprehensive

review, we summarize the evolutionary conservation of CIRP and RBM3 across

species as well as their molecular interactions, cellular functions, and roles in

diverse physiological and pathological processes, including circadian rhythm,

inflammation, neural plasticity, stem cell properties, and cancer development.

PMID: 27147467

----------

[5] Wellmann S, Buhrer C, Moderegger E, Zelmer A, Kirschner R, Koehne P, Fujita J, Seeger K (2004) Oxygen-regulated expression of the RNA-binding proteins RBM3 and CIRP by a HIF-1-independent mechanism. J Cell Sci 117(Pt 9):1785–1794. doi:10.1242/jcs.01026 PubMedCrossRef

---------

[6] Smart F, Aschrafi A, Atkins A, Owens GC, Pilotte J, Cunningham BA, Vanderklish PW (2007) Two isoforms of the cold-inducible mRNA-binding protein RBM3 localize to dendrites and promote translation. J Neurochem 101(5):1367–1379. doi:10.1111/j.1471-4159.2007.04521.x PubMedCrossRef

----------

[7] Selkoe, D. J. Alzheimer’s disease is a synaptic failure. Science 298, 789–791 (2002).

-------------

[8] Mallucci, G. R. Prion neurodegeneration: starts and stops at the synapse. Prion 3, 195–201 (2009).

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[9] Magarin˜ os, A. M., McEwen, B. S., Saboureau, M. & Pevet, P. Rapid and reversible

changes in intrahippocampal connectivity during the course of hibernation in

European hamsters. Proc. Natl Acad. Sci. USA 103, 18775–18780 (2006).

---------

[10] Popov, V. I. & Bocharova, L. S. Hibernation-induced structural changes in synaptic

contacts between mossy fibres and hippocampal pyramidal neurons.

Neuroscience 48, 53–62 (1992).

-----------

[11] Annu Rev Med. 2008;59:177-86. doi: 10.1146/annurev.med.59.061506.110403.

Is human hibernation possible?

Lee CC(1).

Full text: 10.1146/annurev.med.59.061506.110403

The induction of hypometabolism in cells and organs to reduce ischemia damage
holds enormous clinical promise in diverse fields, including treatment of stroke
and heart attack. However, the thought that humans can undergo a severe
hypometabolic state analogous to hibernation borders on science fiction. Some
mammals can enter a severe hypothermic state during hibernation in which
metabolic activity is extremely low, and yet full viability is restored when the
animal arouses from such a state. To date, the underlying mechanism for
hibernation or similar behaviors remains an enigma. The beneficial effect of
hypothermia, which reduces cellular metabolic demands, has many well-established
clinical applications. However, severe hypothermia induced by clinical drugs is
extremely difficult and is associated with dramatically increased rates of
cardiac arrest for nonhibernators. The recent discovery of a biomolecule, 5'-AMP,
which allows nonhibernating mammals to rapidly and safely enter severe
hypothermia could remove this impediment and enable the wide adoption of
hypothermia as a routine clinical tool.

PMID: 18186703

Sthira · May 16, 2016

Marijuana (Component) Promotes Browning of White Fat Too!

I'm not sure if Sthira (or anyone but Kenton and Tom?) are still reading this thread, but I suspect he'll be pleased to hear about this ....

Jah mon.. I know I'm completely alone here, but personally I don't like these crazy high thc sativa strains. I prefer the mellow indicas -- admittedly without much BAT consideration -- rich cbd indica strains like Stephen Hawking kush, Darkstar, Remedy... Hey, no leaf expert; but I do know some sweet and sour widow will definitely brown-down some white fat, well, anecdotally of course since science likes mice studies. That's great news that Pennsylvania of all places is legalizing -- did I read that correctly?

AlPater · May 16, 2016

So cold-inducible RNA-binding protein (CIRP) and RBM3 are the Dr/ Jekyll and Mr. Hyde of' of cold exposure?

Heat shock proteins have some nice things to be said of them too, I believe

Khurram quoted earlier in the thread:

"Connie - So you found this protein, RBM3 - where do you go now?

Giovanna - So, we didn’t find the protein; I mean the proteins a known cold shock protein. What we’ve done is associate it with the failure of structural plasticity in neurodegenerative disease in Alzheimer type mouse models and what we now want to do is understand the relevance for human disease. Because what we found in the mice is that if you put the protein back in it’s incredibly protective, it gives them new synapses, it stops them getting neurodegeneration, it stops them getting memory loss, and it protects them in the long term and you can do that by either cooling the mice early to boost their indodgenous or their own RBM3 levels, or by putting it in artificially. So now, obviously, this is a way in for neuroprotection for human disease but cooling itself is not realistic or practical in the long term. It is used medically; it’s used in newborn babies that have had hypoxic damage; it’s used in post-stroke and it’s used in cardiac surgery, and in many forms of neurosurgery. So we thinks that that’s acting through RBM3 and our ideal would be to be able to manipulate RBM3 levels for protection without having to cool."

Dean Pomerleau · May 16, 2016

Thanks Al,

So cold-inducible RNA-binding protein (CIRP) and RBM3 are the Dr/ Jekyll and Mr. Hyde of' of cold exposure?

Yes - CIRP doesn't look as friendly as RBM3 - e.g. CIRP is associated with worse cancer prognosis. Fortunately RBM3 is induced by mild cold exposure, while CIRP appears to kick in more for quite severe cold, and not much at all in the brain, as this Western Blot analysis from [1] shows:

As you can see, RBM3 kicks in strongly in the brain even from a modest drop in body temperature from normal (37 °C) to 36 °C, While CIRP (= CIRBP) is nowhere to be seen.

Khurram quoted earlier in the thread: ...

I'd totally forgotten about that post from Khurram back in January. I guess that's what happens when a thread stretches to over 450 8.5x11 pages! The "Giovanna" person in interview transcript he posted is Giovanna Mallucci one of the authors of PMID 25607368, which was the main reference [3] from my RBM3 post above. Here is the full text of the transcript Khurram posted:

Though the idea of a long sleep may sound pretty tempting, animals actually put themselves through an awful lot. They are continually cooling and reheating their bodies, putting huge stress on their organs, and some even make themselves diabetic. Hibernation is clearly no picnic, and things get even Neuronsworse as, in an attempt to save energy, animals will dismantle the synapses in their brains. These are the parts of the neuron that send and receive signals and without them we’re all pretty useless. But what’s even more amazing is that when it’s time to “wake up” they’ll put them back together again, just as they were. Professor Giovanna Mallucci is a clinical neuroscientist at Cambridge University and she explains to Connie Orbach how this actually works.

Giovanna - I think you’ve heard already from our other speakers, that there’s lots of processes that slow down and are shut down for hibernation including metabolism. And one way to save energy is to stop the brain using its energy and the dismantling of synaptic connections between brain cells is a way of doing that. What happens is, on cooling there is a retraction of what we call the “dendritic arbour”, you know all the connections and branches of a brain cell that’s connecting to another and the actual contacts - it’s like unplugging a plug from its socket, they are just removed so that no energy flows. When they rewarm there’s a signal to reconnect these structures; how that exactly happens is absolutely not known and very, very interesting to us but we do know a lot about the processes that drive that regenerative capacity.

Connie - Is this happening all over the brain?

Giovanna - Yes, it’s happening all over the brain and all of us all the time. So there’s a balance between pruning and generation or regeneration to maintain a sort of status quo and learning and memory need new synapses and then you prune and get rid of all your excess synapses when you sleep and other conditions. But the capacity for regenerating synapses and refreshing them is part of repair and it’s called “structural synaptic plasticity.”

Connie - So let me just get this right. So what’s happening with animals in hibernation is a much more extreme version of actually something that’s happening all the time in humans and animals?

Giovanna - Correct, that’s exactly right.

Connie - So how have you been using this then in your work?

Giovanna - So we know that in neurodegenerative diseases like Alzheimer’s, which is the prototypical disease but also many of the others. The earliest thing that happens, before you get the brain cell degeneration, is that synapses are lost and as synapses are lost memory goes down - what we call cognitive function goes down, and it’s just not clear why this is early loss of synapses which is such an important stage in these diseases, and it’s important a) because it give you symptoms and b) because it's reversible. So that’s the stage before the brain cells have died, before the neurons have died when, actually, if you can increase synapse number you can restore memory so it’s a very attractive, targetable point of intervention. And our starting hypothesis was that the reason that synapses are lost early in Alzheimer’s disease and early in Parkinson’s disease and other disorders is because there’s a failure of this regenerative capacity that is part of our normal structural plasticity. We used hibernation or induced laboratory hibernation in mice to test the ability of synapses to regenerate themselves in mouse neurodegeneration models.

Connie - And what did you find out - what’s happening?

Giovanna - So first of all we found very interestingly that mice which don’t normally hibernate, can hibernate in all the ways that you would normally expect. So if you cool them: they’ll drop their body temperature, they’ll dismantle their synapses and they’ll go into torpor and then, when you re-warm them, they come completely back to normal again. And what we found out was that normal mice dismantle and reassemble their synapses but the mice that we used that had neurodegeneration models - that’s Alzheimer type mice, and mice with prion disease - that’s another neurodegenerative disease. They failed to reassemble their synapses so they could unplug the plugs but they couldn’t put them back in again and this lack of degenerative capacity gives us a good idea of why there’s such an early loss in synapses.

Connie - Did you get a bit deeper into this? Did you get to see the protein that’s involved - is that right?

Giovanna - Yes we did. So hibernating and cooling does two things to you: it shuts down metabolism and it shuts down protein synthesis, but there’s a group of proteins that are upregulated and these are called “cold-shock proteins,” and they’re a relatively new family of proteins. And one of these which is called “RBM3”, which is RNA Binding Motif Protein 3, is highly expressed in the brain and by being upregulated during hibernation that protein keeps a number of really important critical Messenger RNAs, that you need for survival, ready to make into proteins when you wake up. And we found out that RBM3 is failing in the Alzheimer’s brain, and if we put it back in, we can rescue them.

Connie - So you found this protein, RBM3 - where do you go now?

Giovanna - So, we didn’t find the protein; I mean the proteins a known cold shock protein. What we’ve done is associate it with the failure of structural plasticity in neurodegenerative disease in Alzheimer type mouse models and what we now want to do is understand the relevance for human disease. Because what we found in the mice is that if you put the protein back in it’s incredibly protective, it gives them new synapses, it stops them getting neurodegeneration, it stops them getting memory loss, and it protects them in the long term and you can do that by either cooling the mice early to boost their indodgenous or their own RBM3 levels, or by putting it in artificially. So now, obviously, this is a way in for neuroprotection for human disease but cooling itself is not realistic or practical in the long term. It is used medically; it’s used in newborn babies that have had hypoxic damage; it’s used in post-stroke and it’s used in cardiac surgery, and in many forms of neurosurgery. So we thinks that that’s acting through RBM3 and our ideal would be to be able to manipulate RBM3 levels for protection without having to cool.

Thanks Al and Khurram!

--Dean

-------

[1] Neuroscience. 2015 Oct 1;305:268-78. doi: 10.1016/j.neuroscience.2015.08.012.

Epub 2015 Aug 8.

Cold stress protein RBM3 responds to temperature change in an ultra-sensitive

manner in young neurons.

Jackson TC(1), Manole MD(2), Kotermanski SE(3), Jackson EK(3), Clark RS(4),

Kochanek PM(4).

Author information:

(1)University of Pittsburgh School of Medicine, Safar Center for Resuscitation

Research, 200 Hill Building, 3434 Fifth Avenue, Pittsburgh, PA 15260, United

States; University of Pittsburgh School of Medicine, Department of Critical Care

Medicine, 3550 Terrace Street, Pittsburgh, PA 15261, United States. Electronic

address: jacksontc@upmc.edu.

Full text: http://www.sciencedirect.com/science/article/pii/S0306452215007356