CryoNet #32518 - #32521

CryoNet - Tue 30 Mar 2010

    #32518: Do we live in a manufactured or virtual world? [Raphael T. Haftka]
    #32519: Uploading [Keith Henson]
    #32520: 32516 [doctor Robert]
    #32521: vitrification - the little beetle larvae that could [oberon]

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Message #32518
Date: Mon, 29 Mar 2010 09:45:34 -0400
From: "Raphael T. Haftka" <[email protected]>
Subject: Do we live in a manufactured or virtual world?
References: <[email protected]>

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Message #32519
Date: Mon, 29 Mar 2010 08:40:11 -0700
Subject: Uploading
From: Keith Henson <[email protected]>

To completely muddle the concept of uploading, consider your brain
being infiltrated by nano devices on the same scale as the number of
cells in the brain.  The devices are small enough so there is plenty
of room for them without expanding your skull.  The devices can both
read and stimulate nerve cell activity.  There are no physical
barriers I can think of that prohibit building such nano machines.

In monitoring mode, the devices map out all the signal pathways and
learn to predict what inputs result in specific outputs.  They build a
parallel brain and operate it long enough to be confident the brain in
the nano devices duplicates the running brain, including the formation
of long and short term memory.

Now we cool the brain or otherwise shut down the firing of the nerve
cells, but let the parallel brain continue to operate.  Activity
continues in the parallel brain which now takes over forming long and
short term memory.  After hours to days, we warm up the natural brain
and synch it to the nano devices.

There is no reason I can think of that subjective consciousness would
not persist across the cool down and warming up of the natural brain.

I used this in "The Clinic Seed" to painlessly seduce people out of
the physical world.

Keith

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Message #32520
Date: Mon, 29 Mar 2010 13:01:03 -0700
Subject: 32516
From: doctor Robert <[email protected]>

--0016e64c0b000b67510482f5f9d8

Hi all.  This may just be a small point, but it is one that I think that we
all should discipline ourselves to pay attention to; e.g. no guarantee of
eternal life. Not only is there no guarantee of *eternal life, but there is
not even a mention or thought of it by any rational cryonicist. Noe of us
should use this terminology, we do NOT have the data to support it, hell, we
don't even know what it means. We are hoping for a very long life in which
death becomes optional. Please do not make us look  ridiculous  by trying to
compete with a God that most  of us do not even believe in. Dr. Bob Newport*

-- 
Dr Robert's Fine Art Studio
www.doctorobertsart.com
[email protected]

--0016e64c0b000b67510482f5f9d8

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Message #32521
Date: Mon, 29 Mar 2010 19:41:21 -0800 (PST)
From: [email protected]
Subject: vitrification - the little beetle larvae that could

[Glycerol is one of the "miracle" ingredients responsible for an
amazing feat of natural bioengineering (vitrification) by the Alaskan
beetle larvae Cucujus clavipes. Glycerol is interesting because its
toxicity profile does not include protein denaturation, which is a
limiting factor for many cryoprotectants (eg: DMSO), which are commonly
used in human engineered vitrification solutions. Glycerol itself has seen
relatively little use in vitrification solutions, possiblely due to its
higher risk of osmotic toxicity. Also the higher viscosity of glycerol
would limit the concentration that can be used with organ
cryopreservation. As with beetles, partial slow dehydration would be
required to achieve organ vitrification.]

[Note: Authors include Wowk B, and Fahy GM. Beetle survival was low at
-100C, but this was believed to be due to manhandling of the
specimens causing physical damage. Freezing did not occur in specimens
cooled to -150 C, which confirmed vitrification had occurred.]

J Exp Biol. 2010 Feb;213(Pt 3):502-9.
Deep supercooling, vitrification and limited survival to -100{degrees}C in the Alaskan beetle Cucujus clavipes puniceus (Coleoptera: Cucujidae) larvae.
Sformo T, Walters K, Jeannet K, Wowk B, Fahy GM, Barnes BM, Duman JG. Institute of Arctic Biology, University of Alaska, Fairbanks, AK 99775, USA.
    Larvae of the freeze-avoiding beetle Cucujus clavipes puniceus (Coleoptera: Cucujidae) in Alaska have mean supercooling points in winter of -35 to -42 degrees C, with the lowest supercooling point recorded for an individual of -58 degrees C. We previously noted that some larvae did not freeze when cooled to -80 degrees C, and we speculated that these larvae vitrified. Here we present evidence through differential scanning calorimetry that C. c. puniceus larvae transition into a glass-like state at temperatures<-58 degrees C and can avoid freezing to at least -150 degrees C. This novel finding adds vitrification to the list of insect overwintering strategies. While overwintering beneath the bark of fallen trees, C. c. puniceus larvae may experience low ambient temperatures of around -40
  degrees C (and lower) when microhabitat is un-insulated because of low snow cover. Decreasing temperatures in winter are correlated with loss of body water from summer high levels near 2.0 to winter lows near 0.4 mg mg(-1) dry mass and concomitant increases in glycerol concentrations (4-6 mol l(-1)) and thermal hysteresis. Finally, we provide direct evidence that Cucujus from Wiseman, Alaska, survive temperatures to -100 degrees C.
PMID: 20086136 [PubMed - in process]

[Glycerol has been successfully incorporated into a few human engineered
vitrification solutions.]

Cryobiology. 2007 Oct;55(2):148-57. Epub 2007 Jul 4.
Cryopreservation of Radopholus similis, a tropical plant-parasitic nematode.
Elsen A, Vallterra SF, Van Wauwe T, Thuy TT, Swennen R, De Waele D, Panis B. Laboratory of Tropical Crop Improvement, Department of Biosystems, Faculty of Bioscience Engineering, KU Leuven, Kasteelpark Arenberg 13, 3001, Leuven, Belgium.
    For obligate plant-parasitic nematodes, cryopreservation has advantages over the usual preservation methods on whole plants or axenic culture systems, because the latter two are labourious and time and space consuming. In addition, cross contamination among different isolates can occur easily. Moreover, specific genetic studies require maintenance of the original population. The nematode under investigation, Radopholus similis, is a plant-parasitic nematode from the humid tropics. Therefore, any treatment at low temperatures is likely to add extra stress to the nematode, making the development of a cryopreservation protocol extremely difficult. In this paper, we describe experiments to achieve a successful cryopreservation protocol for the tropical nematode R. similis using vitrificati
 on solution-based methods based on a well defined mixture of cryoprotectants in combination with ultra-rapid cooling and thawing rates. A two-step treatment was used consisting of an incubation in glycerol followed by the application of a vitrifying mixture of methanol, glycerol and glucose. After cryopreservation, the pathogenicity of the nematodes was not altered, since they could infect and reproduce on carrot discs after recovery in the Ringer solution. The cryopreservation method described can be used for routine cryopreservation of R. similis lines from different origins.
PMID: 17707790

[A glycerol/methanol combination gave the best results for  flounder
embryos. Methanol probably acted to reduce the osmotic toxicity of
glycerol.]

Theriogenology. 2005 Feb;63(3):763-73.
Toxicity and protective efficiency of cryoprotectants to flounder (Paralichthys olivaceus) embryos.
Zhang YZ, Zhang SC, Liu XZ, Xu YJ, Hu JH, Xu YY, Li J, Chen SL. Department of Marine Biology, Ocean University of China, Qingdao 266003, PR China.
    With the purpose of finding an ideal cryoprotectant or combination of cryoprotectants in a suitable concentration for flounder (Paralichthys olivaceus) embryo cryopreservation, we tested the toxicities, at culture temperature (16 degrees C), of five most commonly used cryoprotectants-dimethyl sulfoxide (Me2SO), glycerol, methanol (MeOH), 1,2-propylene glycol (PG) and ethylene glycol (EG). In addition, cryoprotective efficiency to flounder embryos of individual and combined cryoprotectants were tested at -15 degrees C for 60 min. Five different concentrations of each of the five cryoprotectants and 20 different combinations of these cryoprotectants were tested for their protective efficiency. The results showed that the toxicity to flounder embryos of the five cryoprotectants are in the
  following sequence: PG < MeOH < Me2SO < glycerol < EG (P < 0.05); whereas the protective efficiency of each cryoprotectant, at -15 degrees C for a period of 60 min, are in the following sequence: PG > Me2SO approximately MeOH approximately glycerol > EG (greater symbols mean P < 0.05, and approximate symbols mean P > 0.05). Methanol combined with any one of the other cryoprotectants gave the best protection, while ethylene glycol combined with any one of the other cryoprotectants gave the poorest protection at -15 degrees C. Toxicity effect was concentration dependent with the lowest concentration being the least toxic for all five cryoprotectants at 16 degrees C. For PG, MeOH and glycerol, 20% solutions gave the best protection at -15 degrees C; whereas a 15% solution of Me2SO, and a 1
 0% solution of EG, gave the best protection at -15 degrees C.
PMID: 15629795

[Glycerol is never used by itself in vitrification solutions.]

J Reprod Fertil. 1993 Nov;99(2):471-7.
Design of vitrification solutions for the cryopreservation of embryos.
Ali J, Shelton JN. John Curtin School of Medical Research, Australian National University, Canberra.
    A series of experiments was performed to determine the concentrations at which ten cryoprotectants singly and in pairs would vitrify on plunging into liquid nitrogen and remain vitreous when warmed by plunging into a water bath at 25 degrees C. From these tests eight solutions (VS) were selected for testing of toxicity to mouse morulae in vitro. One of these (VS1) was modified as a further five VS of which one (VS11) was tested for toxicity to all stages of mouse embryos and to sheep compacted morulae. The concentrations at which the cryoprotectants vitrified on cooling were: butylene glycol, 3.0 mol l-1; propylene glycol, 4.0 mol l-1; dimethyl sulfoxide (DMSO) and glycerol 5.0 mol l-1; ethylene glycol, 6.5 mol l-1. None of these, at the highest concentration tested, remained vitreous 
 during warming. Methanol and the high molecular weight polymers, dextran, Ficoll, polyethylene glycol and polyvinylpyrrolidone, did not vitrify at the concentrations tested. Toxicity studies showed the order of increasing toxicity to be ethylene glycol, methanol, DMSO, glycerol, propylene glycol and butylene glycol. Of the mixtures composed of two cryoprotectants, those containing ethylene glycol and glycerol were the least toxic at vitrifying concentrations. VS11 (6.0 mol ethylene glycol l-1 and 1.8 mol glycerol l-1) was well tolerated by mouse morulae, less well by eight- and one-cell embryos and poorly by two-cell embryos. Dilution of the VS11 from mouse embryos by exposure to 1.0 mol sucrose l-1 for 10 min did not enhance their survival.(ABSTRACT TRUNCATED AT 250 WORDS)
PMID: 8107029

Vet Med (Praha). 1991 Oct;36(10):585-92.
[Comparison of various cryopreservation media for vitrification of 7-day bovine embryos]
[Article in Czech]
Liehman P. Fyziologicky ustav CSAV, Praha.
    A vitrification medium, attested for cryopreservation of mouse eight-blastomere embryos (6.85 mol/l glycerol as a cryoprotective agent), was checked up with respect to preservation of bovine seven-day morulas and blastocysts. As this medium was not found to be convenient either as to its technical parameters or the reached embryo survival, its composition was modified. The glycerol content was complemented or partly replaced by other cryoprotectives (methanol, saccharose, L-proline). A comparison of technical parameters and viability of rewarmed embryos shows that our requirements (technically simple technique of freezing and rewarming and good survival) are met in the best way by a cryoprotective combination of 4.11 mol/l glycerol and 1.0 mol/l saccharose. The total of 54.5% embryos d
 eveloped in-vitro conditions following vitrification in the medium of this composition and after rewarming.
PMID: 1807015

[Glycerol induced damage can also be reduced by some antioxidants.]

Pharmacology. 2005 Jan;73(1):49-56. Epub 2004 Sep 27.
Reversal of experimental myoglobinuric acute renal failure in rats by quercetin, a bioflavonoid.
Chander V, Singh D, Chopra K. Division of Pharmacology, University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh, India.
    The occurrence of acute renal failure (ARF) following rhabdomyolysis has been put at between 10 and 40% of cases, and accounts for between 3 and 15% of all cases of ARF. Reactive oxygen intermediates have been demonstrated to play an etiological role in myoglobinuric renal failure. This study was performed to explore the protective effect of quercetin, a bioflavonoid, in an experimental model of myoglobinuric ARF in rats. Four groups of rats were employed in this study: group 1 served as control, group 2 was given 50% glycerol (8 ml/kg, i.m.), group 3 was given glycerol + quercetin (2 mg/kg, i.p.), and group 4 was given glycerol + DMSO (the solvent for quercetin, 5 ml/kg, i.p.). Renal injury was assessed by measuring serum creatinine, blood urea nitrogen, creatinine and urea clearance.
  The oxidative stress was measured by renal malondialdehyde levels, reduced glutathione levels and by enzymatic activity of catalase, glutathione reductase, and superoxide dismutase. Glycerol administration resulted in a marked renal oxidative stress, significantly deranged the renal functions as well as renal cytoarchitecture. All these factors were significantly improved by quercetin treatment. Because of its radical-scavenging and iron-chelating properties, quercetin protected the kidney against the glycerol-induced oxidative stress and resultant renal dysfunction. Based on these results, this study confirms the role of oxidative stress and demonstrates the renoprotective potential of quercetin in this rhabdomyolysis-mimicking model. 2005 S. Karger AG, Basel.
PMID: 15452363

Folia Microbiol (Praha). 2009;54(3):230-2. Epub 2009 Aug 2.
Viability of commercial wine yeasts during freezer storage in glycerol-based media.
Sidari R, Caridi A. Dipartimento di Scienze e Tecnologie Agro-Forestali e Ambientali, Universita degli Studi Mediterranea di Reggio Calabria, Reggio Calabria, Italy.
    Glycerol-based medium (BM) with and without the addition of 1 g/L ascorbic acid (Asc) and/or 100 mg/L (+/-)-catechin (Cat) was tested for the storage of three commercial wine yeasts at -20 degrees C. The medium supplemented with Asc was also used to store 706 strains to verify the maintenance of the liquid state. A decline in survival throughout the storage period was observed. The media containing Asc maintained viability better than the other three. The BM caused a loss of viability of 7 orders for one strain and of 6 orders for the other two. All three strains exhibited a loss of viability of 4 orders when stored in BM+Asc. Two strains decreased viability by 5 orders while one strain by 4 orders, when stored in BM+Cat. Two strains decreased viability by 6 orders while one strain by 
 5 orders, when stored in BM+Asc+Cat. Regarding the physical state of the medium tested on 706 yeast strains, three cases were observed: completely liquid (56.5 %), liquid with only the upper part frozen (40.4 %) without involving the yeast biomass settled at the bottom, and completely frozen (3.12 %). It is practicable to prepare a BM that remains liquid at -20 degrees C enhancing yeast viability when Asc is added as cryoprotectant.
PMID: 19649740

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