X-Message-Number: 33258
Date: Thu, 20 Jan 2011 18:57:12 -0800 (PST)
From: 
Subject: upgrade for glycerol cryopreservation?

http://www.cryonics.org/perfusion/Glycerol.html

CI now suggests cephalic cryoprotectant perfusion for their dry ice
"customers".  The solutions recommended are saline (0.91 w/v%
NaCl) supplemented with 2 to 8 M glycerol. I wondered whether
there might not be a low cost upgrade for such solutions. The following
suggestion is what I came up with.

Suggestion: Add a dash of filtered red wine, and replace a pinch of salt
with some sodium bicarbonate.

The rationale: The cost is negligible. The only safety concern derives
from the very small amount of acid, which can easily be neutralized by a
pinch of sodium bicarbonate. The main reason for adding a "dash" of red
wine is because this offers powerful protection against glycerol toxicity
in live bodies. The main drawback is that it is not known what the
magnitude of this protection would be in dead bodies. However since both
cost and risks are negligible, there appears to be no reason not to
gamble with this addition.

Nephrol Dial Transplant. 2004 Sep;19(9):2237-44. Epub 2004 Jul 6.

Amelioration of myoglobinuric renal damage in rats by chronic exposure to 
flavonol-rich red wine.

Rodrigo R, Bosco C, Herrera P, Rivera G. Laboratorio de Fisiopatologia Renal, 
Programa de Farmacologia Molecular y Clinica, ICBM, Facultad de Medicina, 
Universidad de Chile, Independencia 1027, Casilla 70058, Santiago 7, Chile.
Abstract

  BACKGROUND: Myoglobinuric acute renal failure causes increased oxidative 
  stress. Since ethanol upregulates renal antioxidant enzymes and wine 
  polyphenols behave as antioxidants, we tested the hypothesis that red wine 
  components would ameliorate the renal damage caused by rhabdomyolysis.

METHODS: Adult rats received water (control), alcohol-free red wine, ethanol 
12.5% (v/v) or red wine for 10 weeks. Rhabdomyolysis was induced by glycerol 
injection (50%, 10 ml/kg, i.m.), and urine and blood samples were collected 6 h 
later to measure renal function parameters, creatine kinase (CK) activity, free 
F(2)-isoprostanes and total antioxidant capacity. Kidneys were then harvested 
for morphological studies and determinations of lipid peroxidation, protein 
carbonylation, (Na + K)-ATPase and antioxidant enzyme activities.

RESULTS: In the control group, myoglobinuria was associated with a 68% decrease 
in creatinine clearance and increases in plasma creatinine and blood urea 
nitrogen of 3.2 and 1.8 times above baseline, respectively. Controls also showed
increases in plasma free F(2)-isoprostanes levels and CK activity, together 
with enhanced renal expression of the antioxidant enzymes catalase, glutathione 
peroxidase and superoxide dismutase, as well as increased production of 
malondialdehyde and carbonyls. Rhabdomolysis reduced renal (Na + K)-ATPase 
activity and this reduction was associated with a 5-fold increase in fractional 
sodium excretion as well as morphological damage to the kidney. These changes 
were significantly attenuated by pretreatment with chronic red wine exposure 
prior to glycerol injection. A less marked degree of functional and biochemical 
protection was also observed in response to the administration of alcohol-free 
red wine and ethanol.

CONCLUSIONS: The present data suggest that red wine protects against functional,
biochemical and morphological damage caused by rhabdomyolysis in the rat, and 
this protection may be due to the synergistic effects of ethanol and 
non-alcoholic red wine components.
PMID: 15238628
Free text>
http://ndt.oxfordjournals.org/content/19/9/2237.full.pdf+html

Atherosclerosis. 2010 Oct;212(2):426-35. Epub 2010 Jun 25.

Moderate intake of red wine improves ischemia-induced neovascularization in 
diabetic mice--roles of endothelial progenitor cells and nitric oxide.

Huang PH, Tsai HY, Wang CH, Chen YH, Chen JS, Lin FY, Lin CP, Wu TC, Sata M, 
Chen JW, Lin SJ. Division of Cardiology, Department of Medical Research and 
Education, Taipei Veterans General Hospital, Institute of Clinical Medicine, 
Cardiovascular Research Center, National Yang-Ming University, Taipei, Taiwan.
Abstract

  OBJECTIVE: Circulating endothelial progenitor cells (EPCs) play a significant 
  role in postnatal neovascularization. Patients with diabetes have attenuated 
  EPC functions and impaired angiogenic response after tissue ischemia. We 
  investigated whether moderate red wine consumption can enhance blood flow 
  recovery in response to tissue ischemia by enhancement of EPC functions in 
  diabetic mice.

METHODS AND RESULTS: Starting at 4 weeks after diabetes onset, red wine (4 
ml/kg/day) or ethanol were administered to streptozotocin (STZ)-induced (type 1)
diabetic mice and KKAy-Ta (type 2) mice. Unilateral hind limb ischemia surgery 
was conducted after 2 weeks of red wine or ethanol ingestion. Type 1 and type 2 
diabetic mice given red wine, but not ethanol, had significantly increased 
collateral flow about 30% and augmented capillary density in ischemic tissues. 
These beneficial effects were markedly abolished by an eNOS inhibitor (L-NAME). 
Flow cytometry analysis showed impaired EPC-like cells (Sca-1+/Flk-1+) 
mobilization after ischemia surgery in diabetic mice, but augmented mobilization
in red wine group (baseline vs. 2 days after operation: 0.88 0.06% vs. 1.73 
0.29%, p=0.010). C-kit positive bone marrow cells isolated from diabetic mice 
given red wine had enhanced adhesion and migration compared to mice given 
vehicle. By in-vitro studies, incubation with red wine in high-glucose medium 
significantly reduced H2O2 production, and improved high glucose-suppressed EPC 
functions by nitric oxide-related mechanisms.

CONCLUSIONS: Our findings demonstrate that red wine consumption enhances blood 
flow recovery after tissue ischemia in diabetic mice. These effects may partly 
derive from enhanced EPC functions by upregulation of eNOS activity.
Copyright C 2010 Elsevier Ireland Ltd. All rights reserved.
PMID: 20637466

Lipids. 2000 Feb;35(2):143-8.

A high-fat diet induces and red wine counteracts endothelial dysfunction in 
human volunteers.

Cuevas AM, Guasch V, Castillo O, Irribarra V, Mizon C, San Martin A, Strobel P, 
Perez D, Germain AM, Leighton F. Department of Nutrition, Metabolism & Diabetes,
Faculty of Biological Sciences, Pontificia Universidad Catolica de Chile, 
Santiago.
Abstract

  Endothelial dysfunction is associated with atherogenesis and oxidative stress 
  in humans. In rat and rabbit blood vessels, wine polyphenol antioxidants 
  induce vascular relaxation in vitro through the NO-cGMP pathway. To assess the
  effect of a regular high-fat diet (HFD) and moderate red wine consumption on 
  endothelial function (EF), a study was performed in healthy male volunteers. 
  EF was measured as flow-mediated dilatation of the brachial artery, employing 
  high-resolution ultrasound after an overnight fast. Other clinical and 
  biochemical parameters related to EF were also measured. Six volunteers 
  received a control diet, rich in fruits and vegetables (27% calories as fat) 
  and five volunteers received an HFD (39.5% calories as fat). Measurements were
  done twice on each volunteer: after a period of 30 d with diet plus 240 mL of
  red wine/d, and after a period of 30 d with diet, without wine. In the 
  absence of wine, there is a reduction of EF with HFD when compared to the 
  control diet (P = 0.014). This loss of EF is not seen when both diets are 
  supplemented with wine for 30 d (P = 0.001). Plasma levels of n-3 fatty acids 
  (R2 = 0.232, P = 0.023) and lycopene (R2 = 0.223, P = 0.020) show a positive 
  correlation with individual EF measurements, but they do not account for the 
  significant differences observed among dietary groups or after wine 
  supplementation. These results help elucidate the deleterious effect of a 
  high-fat diet and the protective role of wine, n-3 fatty acids and dietary 
  antioxidants in cardiovascular disease.
PMID: 10757544

Ann Neurol. 2000 Oct;48(4):686-7.

Lyophilized red wine administration prolongs survival in an animal model of 
amyotrophic lateral sclerosis.

Esposito E, Rossi C, Amodio R, Di Castelnuovo A, Bendotti C, Rotondo T, Algeri 
S, Rotilio D.
PMID: 11026458 [PubMed - indexed for MEDLINE]

Brain Res. 2010 Jul 30;1346:247-50. Epub 2010 May 27.

Red wine but not ethanol at low doses can protect against the toxicity of 
methamphetamine.

Ali SF, Bondy SC. Neurochemistry Laboratory, Division of Neurotoxicology, 
National Center for Toxicological Research, Jefferson, AR 72079-9502, USA.
Abstract

  The goal of this study was twofold: (a) to search for possible interactive 
  effects between two common drugs of abuse, ethanol and methamphetamine. b) To 
  inquire whether any effects of ethanol could be replicated using an equivalent
  amount of ethanol in the form of red wine. Adult male C57/6N mice received 2%
  ethanol for 8 weeks in drinking water or red wine diluted to yield the same 
  ethanol content. On the 9th week animals received multiple injections of 
  methamphetamine (4 x 10 mg/kg, ip, every 2 h). They were then sacrificed 72 h 
  after treatment. Methamphetamine produced a significant depletion of dopamine 
  and DOPAC in the striatum. Treatment with both ethanol and methamphetamine led
  to a reduction of striatal dopamine and DOPAC that were both 
  non-significantly greater than that observed with methamphetamine alone. 
  Alcohol alone produced no changes in the striatal content of dopamine or its 
  metabolite, DOPAC. These data suggest that low doses of alcohol potentiate 
  methamphetamine-induced neurotoxicity in mice and that this combination may be
  especially detrimental to the brain. However, an equivalent dose of ethanol 
  in the form of red wine actually partially protected against 
  methamphetamine-induced depletion of dopamine and DOPAC in red wine treated 
  mice. This implies the presence of other agents in red wine, which may 
  mitigate the toxicity of methamphetamine.
Copyright 2010 Elsevier B.V. All rights reserved.
PMID: 20510887

Biochem Biophys Res Commun. 2011 Jan 14;404(2):743-9. Epub 2010 Dec 16.

Chronic intake of red wine polyphenols by young rats prevents aging-induced 
endothelial dysfunction and decline in physical performance: Role of NADPH 
oxidase.

Dal-Ros S, Zoll J, Lang AL, Auger C, Keller N, Bronner C, Geny B, Schini-Kerth 
VB. UMR CNRS 7213, Laboratoire de Biophotonique et de Pharmacologie, Faculte de 
Pharmacie, Universite de Strasbourg, Illkirch, France.
Abstract

  Aging is associated with oxidative stress-mediated endothelial dysfunction and
  decline in physical performance, which promote cardiovascular diseases. This 
  study examined whether chronic intake of red wine polyphenols (RWPs), a rich 
  source of natural antioxidants, prevents aging-related impairment of vascular 
  function and physical exercise capacity. Vascular reactivity from 12, 20 and 
  40week-old rats was assessed in organ chambers. Rats received from week 16 to 
  40 either solvent, RWPs or the antioxidant and NADPH oxidase inhibitor, 
  apocynin. Aging was associated with blunted endothelium-dependent relaxations,
  oxidative stress (dihydroethidine staining), and an upregulation of eNOS, 
  arginase I, NADPH oxidase p22phox and nox1 subunits, and AT1 and AT2 receptors
  (assessed by immunohistochemistry) in the mesenteric artery. RWPs and 
  apocynin improved the endothelial dysfunction, normalized oxidative stress and
  the expression of the different proteins. RWPs also improved aging-related 
  decline in physical exercise. Thus, intake of RWPs protects against 
  aging-induced endothelial dysfunction and decline in physical performance. 
  These effects likely involve the ability of RWPs to normalize oxidative stress
  and the expression of proteins involved in the formation of NO and the 
  angiotensin II pathway.
Copyright AC 2010 Elsevier Inc. All rights reserved.
PMID: 21167817

Ethanol has been tested as a cryoprotectant. Unfortunately, red wine
itself has never been tested, despite its much lower toxicity
profile. The only reason that has occurred to me to account for this
omission alludes to psychological limitations of scientists.

Cryobiology. 2009 Apr;58(2):166-9. Epub 2008 Dec 11.

Influence of cryoprotectants on abnormality and motility of baung (Mystus 
nemurus) spermatozoa after long-term cryopreservation.

Muchlisin ZA, Azizah MN. Department of Marine Sciences, Faculty of Sciences 
Syiah Kuala University, Kopelma Darussalam, Banda Aceh, NAD 23111, Indonesia.
Abstract

  Study on the effect of cryoprotectants on abnormality and motility of baung, 
  Mystus nemurus spermatozoa were evaluated using transmission and scanning 
  electron microscopy. Four cryoprotectants, dmso, ethanol, methanol and 
  glycerol at concentration of 10% were tested in triplicates. Three ml of fresh
  sperm which was diluted with 60 ml of ringer solution was added to each of 
  twelve 5-ml vials containing of 0.50-ml of the cryoprotectants. The vials were
  placed in an icebox containing dry ice 5 min and then storage into container 
  containing liquid nitrogen for 13 months. The effect of cryoprotectants on the
  spermatozoa abnormality and motility were significant (P<0.05). The 
  spermatozoa abnormality was significantly lower in methanol (62.65%) compared 
  with the other cryoprotectants. The spermatozoa motility was higher in 
  methanol, but not significantly different with ethanol (P>0.05). It is a 
  negative correlation between sperm motility and abnormality. Generally, higher
  abnormalities of spermatozoa resulted low motility.
PMID: 19114036

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