CryoNet #32267 - #32271

CryoNet - Wed 30 Dec 2009

    #32267: Re: temp without LN2 [yvan Bozzonetti]
    #32268: Yet another dark story [[email protected]]
    #32269: The Decade of Improgression? [Mark Plus]
    #32270: flavonoids protect against freezing and freeze-drying damage [oberon]
    #32271: Re: Rebranding cryonics [David Stodolsky]

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Message #32267
Date: Tue, 29 Dec 2009 14:40:33 +0100
Subject: Re: temp without LN2
From: yvan Bozzonetti <[email protected]>

--001485f87d2a9147f9047bde2ace

> From: [email protected]
> Subject: temp without LN2
>
> Yvan mentioned the potential usefulness of transporting
> at liquid nitrogen temperature without using liquid nitrogen.
> I and others have mentioned this in the past, and in
> principle it isn't difficult. You just supercool dry ice
> (for example) to liquid nitrogen temperature and insulate
> well. The experiments to determine parameters have not
> yet been done as far as I know.
>
> Robert Ettinger
>

 What I was writing about is far simpler:
It is a LN2 cryostat lined with a kind of mineral "sponge".
To use it, you fill it up with LN2, a large part of it cool the "sponge" and
get absorbed by it.
After some hours, the equilibrium temperature is reached.
You then drain the excess LN2 or let it evaporate.
You have a dry cryostat with a capability to remain LN2 cool for at least
two week.
I have experimented with such a system, it work well.
You can put it in any position, there is no liquid spill, an important
feature for transportation.
The system I have seen is loaded with near 8 gallons of LN2 and has a free
volume of 6 gal.
I don't know if there are larger systems on the market.
Yet, even if there is no one, they could be built simply with open cell foam
in an ordinary LN2 cryostat.

Yvan Bozzonetti.

--001485f87d2a9147f9047bde2ace

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Message #32268
From: "[email protected]" <[email protected]>
Date: Tue, 29 Dec 2009 23:42:33 GMT
Subject: Yet another dark story

http://www.365tomorrows.com/04/28/is-hard-to-find/

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Message #32269
From: Mark Plus <[email protected]>
Subject: The Decade of Improgression?
Date: Tue, 29 Dec 2009 18:23:27 -0800

Starting with the arbitrary date of January 1, 2000, and ending with December 31, 2009, can someone defend the case that cryonics on the whole has made progress over that ten year period?

Because when I think back on it, I just remember one organization's series of ineffectual CEO's, sabotage-minded employees and episodes of bad publicity.

Mark Plus


 		 	   		  
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Message #32270
Date: Wed, 30 Dec 2009 00:05:02 -0800 (PST)
From: [email protected]
Subject: flavonoids protect against freezing and freeze-drying damage

[Although the results are impressive, flavonoid additions are still surprisingly rare. The reason is not cost. For example, the distilled water in cryoprotectant solutions could be replaced with filtered green tea at a negligible cost.]

[Genistein reduces cryopreservation induced DNA damage.]

Hum Reprod. 2009 Sep;24(9):2061-70. Epub 2009 Jun 12.
Cryopreservation-induced human sperm DNA damage is predominantly mediated by oxidative stress rather than apoptosis.
Thomson LK, Fleming SD, Aitken RJ, De Iuliis GN, Zieschang JA, Clark AM. Fertility First, PO Box 807, Hurstville, NSW 2220, Australia.
    BACKGROUND: Whereas studies have revealed that the cryopreservation of human semen increases sperm DNA fragmentation, the mechanisms involved in this type of cryo-injury are largely unknown. Elucidation of these mechanisms may provide insight into preventing such injury. METHODS: We obtained 60 semen samples from 60 men and conducted experiments to determine the cause of cryopreservation-induced DNA fragmentation using 8-oxo-7,8-dihydro-2'deoxyguanosine (8OHdG) as a biomarker of oxidative stress, percentage caspase positive cells as an indicator of apoptosis, the potential antioxidant genistein and the caspase inhibitor Z-VAD(OMe)-FMK. RESULTS: Cryopreservation led to a significant increase in percentage DNA fragmentation, percentage 8OHdG and percentage caspase positive cells (P < 0.0
 01). Percentage DNA fragmentation was positively correlated with percentage 8OHdG before (r = 0.756, P < 0.001) and after cryopreservation (r = 0.528, P = 0.017). The addition of 50 and 100 microM genistein to the cryoprotectant had a significant protective effect on sperm DNA (P < 0.001) although the caspase inhibitor demonstrated no difference to the control. CONCLUSIONS: Human sperm DNA fragmentation is associated with an increase in oxidative stress during cryopreservation, rather than the activation of caspases and apoptosis. The estrogenic compound genistein may be useful in reducing this effect but larger trials are needed to confirm this.
PMID: 19525298

[Green tea flavonoid ECGC comes to the rescue here.]

Cell Transplant. 2009;18(5):513-9.
Long-term preservation of rat skin tissue by epigallocatechin-3-o-gallate.
Kim H, Kawazoe T, Matsumura K, Suzuki S, Hyon SH. Department of Medical Simulation Engineering, Research Center for Nano Medical Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.
    Skin grafts can be preserved by cryopreservation and refrigerated storage at 4 degrees C. Epigallocatechin-3-O-gallate (EGCG) enhances the viability of stored skin grafts and also extends the storage time up to 7 weeks at 4 degrees C. EGCG, the major polyphenolic constituent present in green tea, has potent antioxidant, antimicrobial, antiproliferative, and free radical scavenging effects. This study examined the effects of EGCG on skin cryopreservation. Skin sample biopsy specimens from GFP rats were previously treated with/without EGCG then moved to -196 degrees C. Skin samples were transplanted to nude mice after 2, 8, and 24 weeks of preservation. Glucose consumption was measured after thawing to assess the metabolic activity. Two weeks later the transplanted skin grafts were excis
 ed and histologically analyzed. Histological examinations revealed the degeneration of the epidermal and dermal layers in all groups. In the EGCG groups, the grafts showed higher integrity in the epidermal layer and dermal matrix. The present findings suggest the future clinical usefulness of EGCG for skin preservation; however, the mechanism by which EGCG promotes skin preservation still remains unclear.
PMID: 19775511

[ECGC enables cells to survive freeze-drying. Its benefit is much greater than that of trehalose, and a trehalose/ECGC combination offers a synergistic effect. Before washing, cellular viability with this combination is over 90% after both freeze-thawing and freeze-drying.]

PLoS One. 2009;4(4):e5240. Epub 2009 Apr 21.
Freeze-drying of mononuclear cells derived from umbilical cord blood followed by colony formation.
Natan D, Nagler A, Arav A. Core Dynamics Ltd, Ness-Ziona, Israel.
    BACKGROUND: We recently showed that freeze-dried cells stored for 3 years at room temperature can direct embryonic development following cloning. However, viability, as evaluated by membrane integrity of the cells after freeze-drying, was very low; and it was mainly the DNA integrity that was preserved. In the present study, we improved the cells' viability and functionality after freeze-drying. METHODOLOGY/PRINCIPAL FINDINGS: We optimized the conditions of directional freezing, i.e. interface velocity and cell concentration, and we added the antioxidant EGCG to the freezing solution. The study was performed on mononuclear cells (MNCs) derived from human umbilical cord blood. After freeze-drying, we tested the viability, number of CD34(+)-presenting cells and ability of the rehydrated 
 hematopoietic stem cells to differentiate into different blood cells in culture. The viability of the MNCs after freeze-drying and rehydration with pure water was 88%-91%. The total number of CD34(+)-presenting cells and the number of colonies did not change significantly when evaluated before freezing, after freeze-thawing, and after freeze-drying (5.4 x 10(4)+/-4.7, 3.49 x 10(4)+/-6 and 6.31 x 10(4)+/-12.27 cells, respectively, and 31+/-25.15, 47+/-45.8 and 23.44+/-13.3 colonies, respectively). CONCLUSIONS: This is the first report of nucleated cells which have been dried and then rehydrated with double-distilled water remaining viable, and of hematopoietic stem cells retaining their ability to differentiate into different blood cells.
PMID: 19381290
Free text>
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2667668/pdf/pone.0005240.pdf


[Kaempferol-7-O-glucoside reduces vitrification solution toxicity.]

Cryobiology. 2008 Dec;57(3):242-5. Epub 2008 Sep 15.
Improved cryopreservation by diluted vitrification solution with supercooling-facilitating flavonol glycoside.
Kami D, Kasuga J, Arakawa K, Fujikawa S. National Agricultural Research Center for Hokkaido Region, Sapporo 062-8555, Japan.
    The effect of kaempferol-7-O-glucoside (KF7G), one of the supercooling-facilitating flavonol glycosides which was originally found in deep supercooling xylem parenchyma cells of the katsura tree and was found to exhibit the highest level of supercooling-facilitating activity among reported substances, was examined for successful cryopreservation by vitrification procedures, with the aim of determining the possibility of using diluted vitrification solution (VS) to reduce cryoprotectant toxicity and also to inhibit nucleation at practical cooling and rewarming by the effect of supplemental KF7G. Examination was performed using shoot apices of cranberry and plant vitrification solution 2 (PVS2) with dilution. Vitrification procedures using the original concentration (100%) of PVS2 caused
  serious injury during treatment with PVS2 and resulted in no regrowth after cooling and rewarming (cryopreservation). Dilution of the concentration of PVS2 to 75% or 50% (with the same proportions of constituents) significantly reduced injury by PVS2 treatment, but regrowth was poor after cryopreservation. It is thought that dilution of PVS2 reduced injury by cryoprotectant toxicity, but such dilution caused nucleation during cooling and/or rewarming, resulting in poor survival. On the other hand, addition of 0.5mg/ml (0.05% w/v) KF7G to the diluted PVS2 resulted in significantly (p<0.05) higher regrowth rates after cryopreservation. It is thought that addition of supercooling-facilitating KF7G induced vitrification even in diluted PVS2 probably due to inhibition of ice nucleation durin
 g cooling and rewarming and consequently resulted in higher regrowth. The results of the present study indicate the possibility that concentrations of routinely used VSs can be reduced by adding supercooling-facilitating KF7G, by which more successful cryopreservation might be achieved for a wide variety of biological materials.
PMID: 18824164

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Message #32271
From: David Stodolsky <[email protected]>
Subject: Re: Rebranding cryonics
Date: Wed, 30 Dec 2009 10:37:32 +0100
References: <[email protected]>

On 29 Dec 2009, at 11:00 AM, CryoNet wrote:

> A number of cryonicists, myself included, have begun to question  
> whether this association has resulted in "positive" results for  
> cryonics, given that enough time has passed to falsify early  
> transhumanist predictions with dates about medical & technological  
> breakthroughs, including ones in cryonics.

Regardless of this, probability of sign-ups from the Transhumanist  
'church' are higher then they are for the Christian Church.


> And as you point out, some schools of transhumanist thinking have  
> led to passivity or terror management,

According to Terror Management Theory, terror management will always  
be present.


> like Ray Kurzweil's consumption of "life extension" quackery.  
> Kurzweil might have arrangements for his cryonic suspension (a  
> recent "Popular Science" article states that he does); but for an  
> inventor and engineer obsessed with his personal survival, he seems  
> incurious and unmotivated about trying to improve cryonics as a  
> technological product, probably because he believes that the bowls  
> of pills he swallows every day have put him on the path to "living  
> well forever." Even if he knows he lacks the expertise to help  
> cryonics, he has the money, status and connections to attract people  
> who do.

The problem is, that it isn't clear how to 'help' cryonics. While  
there is always another advance in biotech, it isn't clear that that  
is the main problem the movement faces. Kurzweil has certainly made a  
positive contribution as a high profile figure publicly signed up for  
cryonics.


>
> I've also run across transhumanists who dismiss cryonics, even if is  
> succeeds, because they believe that AI's will make humans "obsolete"  
> and human life not worth preserving in a few decades. This sort of  
> transhumanism sets itself up as explicitly hostile to cryonicists'  
> goals.

My question was what approach to repackaging has produced the best  
results. Clearly, Transhumanism as a whole, produces both positive and  
negative effects for cryonics. However, the organizations that aim to  
be positive, like Venturism, haven't gained much of a following.


dss

David Stodolsky
[email protected]  Skype: davidstodolsky

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