X-Message-Number: 29421 Date: Sat, 14 Apr 2007 16:23:12 -0700 (PDT) From: Subject: engineered negligible senescence Part IV [Effective intervention in human aging may not be as far off as one might suppose, provided information is acted on in a timely manner.] J Gerontol A Biol Sci Med Sci. 2005 Nov;60(11):1386-93. Prevention of accelerated cell aging in Werner syndrome using a p38 mitogen-activated protein kinase inhibitor. Davis T, Baird DM, Haughton MF, Jones CJ, Kipling D. D.Phil, Department of Pathology, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, United Kingdom. We investigated the role of p38 mitogen-activated protein kinase (MAPK) signalling in the accelerated aging of Werner Syndrome (WS) fibroblasts by use of SB203580, a cytokine-suppressive anti-inflammatory drug that targets p38 activity. SB203580 treatment reverts the aged morphology of young WS fibroblasts to that seen in young normal fibroblasts. In addition, SB203580 increases the life span and growth rate of WS fibroblasts to within the normal range. In young WS cells, p38 is activated coincident with an up-regulation of p21(WAF1), and a reduction in the levels of both activated p38 and p21(WAF1) are seen following treatment with SB203580. As these effects are not seen in young normal cells, our data suggest that the abbreviated replicative life span of WS cells is due to a stress-induced, p38-mediated growth arrest that is independent of telomere erosion. With some p38 inhibitors already in clinical trials, our data suggest a potential route to drug intervention in a premature aging syndrome. PMID: 16339323 Biogerontology. 2005;6(5):313-23. Effect of S-adenosylmethionine on age-induced hepatocyte damage in old Wistar rats. Castillo C, Salazar V, Ariznavarreta C, Fossati M, Tresguerres JA, Vara E. Laboratory of Experimental Endocrinology, Department of Physiology, School of Medicine, Complutense University, Avda. Complutense s/n, 28040, Madrid, Spain. Aging is accompanied by changes in the morphology and physiology of organs and tissues, such as the liver. This process might be due to the accumulation of oxidative damage induced by reactive oxygen (ROS) and reactive nitrogen species (RNS). Hepatocytes are very rich in mitochondria and have a high respiratory rate, so they are exposed to large amounts of ROS and permanent oxidative stress. S-Adenosylmethionine (SAMe) is an endogenous metabolite that has shown to exert protective effects on different experimental pathological models in which free radicals are involved. The aim of this study was to investigate the effect of SAMe on age-induced damage in hepatocytes. For this purpose, male and female Wistar rats of 18 and 2 months of age were used. Cells were isolated and, after incubation in the presence or in the absence of SAMe, different parameters were measured. Aging induced a significant increase in nitric oxide, carbon monoxide and cGMP, and a reduction in reduced glutathione, ATP and phosphatidylcholine synthesis, as well as in methionine- adenosyl-transferase and methyl-transferase activities. Incubation of old cells with SAMe prevented all these age-related changes, reaching values in some of the parameters similar to those found in young animals. In conclusion, SAMe seems to have beneficial effects against age-induced damage in hepatocytes. PMID: 16463108 Proc Natl Acad Sci U S A. 2005 Sep 6;102(36):12873-8. Epub 2005 Aug 29. Blocking protein farnesyltransferase improves nuclear shape in fibroblasts from humans with progeroid syndromes. Toth JI, Yang SH, Qiao X, Beigneux AP, Gelb MH, Moulson CL, Miner JH, Young SG, Fong LG. Department of Medicine and Division of Cardiology, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA. Defects in the biogenesis of lamin A from its farnesylated precursor, prelamin A, lead to the accumulation of prelamin A at the nuclear envelope, cause misshapen nuclei, and result in progeroid syndromes. A deficiency in ZMPSTE24, a protease involved in prelamin A processing, leads to prelamin A accumulation, an absence of mature lamin A, misshapen nuclei, and a lethal perinatal progeroid syndrome: restrictive dermopathy (RD). Hutchinson-Gilford progeria syndrome (HGPS) is caused by a mutant prelamin A that cannot be processed to lamin A. The hallmark cellular abnormality in RD and HGPS is misshapen nuclei. We hypothesized that the farnesylation of prelamin A is important for its targeting to the nuclear envelope in RD and HGPS and that blocking farnesylation would ameliorate the nuclear shape abnormalities. Indeed, when RD fibroblasts were treated with a farnesyltransferase inhibitor (FTI), prelamin A was partially mislocalized away from the nuclear envelope, and the frequency of nuclear shape abnormalities was reduced (P < 0.0001). A FTI also mislocalized prelamin A and improved nuclear shape in Zmpste24-deficient mouse embryonic fibroblasts (P < 0.0001) and improved nuclear shape in human HGPS fibroblasts (P < 0.0001). Most remarkably, a FTI significantly improved nuclear shape in two fibroblast cell lines from atypical progeria patients with lamin A missense mutations in the absence of prelamin A accumulation (P = 0.0003 and P < 0.0001). These findings establish a paradigm for ameliorating the most obvious cellular pathology in lamin-related progeroid syndromes and suggest a potential strategy for treating these diseases. PMID: 16129834 [Some examples of divergence between human aging and rodent aging are with repect to telomeres, amyloid accumulation, and p66(shc).] Mech Ageing Dev. 2005 Aug;126(8):839-44. Epub 2005 Apr 8. p66(shc) is highly expressed in fibroblasts from centenarians. Pandolfi S, Bonafe M, Di Tella L, Tiberi L, Salvioli S, Monti D, Sorbi S, Franceschi C. Department of Experimental Pathology, University of Bologna, via S. Giacomo 12, 40126 Bologna, Italy. p66(shc-/-) mice exhibit prolonged lifespan and increased resistance to oxidative and hypoxic stress. To investigate p66(shc) involvement in human longevity, p66(shc) mRNA and protein were evaluated in fibroblasts from young people, elderly and centenarians, exposed to oxidative or hypoxic stress. Unexpectedly, centenarians showed the highest basal levels of p66(shc). Oxidative stress induced p66(shc) in all samples. At variance, hypoxic stress caused p66(shc) reduction only in cells from centenarians. These changes occurred in absence of any modification of p66(shc) promoter methylation pattern. Intriguingly, in cells from centenarians, p66(shc) induction was affected by p53 codon 72 polymorphism. Thus, cells from centenarians present a peculiar regulation of p66(shc), suggesting that its role in mammalian longevity is more complex than previously thought. PMID: 15992607 [Engineered Reversible Cryostasis is not yet available. A precursor called cryonics is currently used as a processing option for the treatment of corpses. By comparison, ERC would typically be used to place still living human beings into an ultra-low temperature "hibernation". Although ERC subjects would be suffering from a terminal disease, they would still be both medically and legally alive both before and after treatment. If there was any doubt on this matter, any ERC subject could be resusitated, and show up in court to prove this legally. After technical development of ERC, it is anticipated that there may be a further 10 year delay before legal hurtles are overcome to allow still living human beings to become ERC subjects.] Ann N Y Acad Sci. 2004 Jun;1019:559-63. The arrest of biological time as a bridge to engineered negligible senescence. Lemler J, Harris SB, Platt C, Huffman TM. Alcor Life Extension Foundation, 7895 E. Acoma Drive, Scottsdale, AZ 85260, USA. Biological systems can remain unchanged for several hundred years at cryogenic temperatures. In several hundred years, current rapid scientific and technical progress should lead to the ability to reverse any biological damage whose reversal is not forbidden by physical law. We therefore explore whether contemporary people facing terminal conditions might be preserved well enough today for their eventual recovery to be compatible with physical law. The ultrastructure of the brain can now be excellently preserved by vitrification, and solutions needed for vitrification can now be distributed through organs with retention of organ viability after transplantation. Current law requires a few minutes of cardiac arrest before cryopreservation of terminal patients, but dogs and cats have recovered excellent brain function after 16-60 min of complete cerebral ischemia. The arrest of biological time as a bridge to engineered negligible senescence, therefore, appears consistent with current scientific and medical knowledge. PMID: 15247086 [Quote from the paper listed below: "Approaches to reduce chemical toxicity of a CPA solution without dimishing its glass-forming tendancy include using a combination of CPAs with different mechanisms of toxicity, using agents that antagonize each other's toxicity and using reduced temperatures of exposure". Note that pretreatment of cells to modulate cryoprotectant toxicity is an avenue which has NOT been significantly explored in published papers. I will shortly be pretreating brine shrimp to see if this might be a viable approach to reducing cryoprotectant toxicity. Various additives to the cryoprotectant solutions will also be tried.] Cryobiology. 2007 Feb;54(1):1-12. Epub 2006 Dec 12. Cryopreservation of rat precision-cut liver and kidney slices by rapid freezing and vitrification. de Graaf IA, Draaisma AL, Schoeman O, Fahy GM, Groothuis GM, Koster HJ. Pharmacokinetics and Drug Delivery, Groningen University Institute for Drug Exploration, A. Deusinglaan 1, 9713 AV, Groningen, The Netherlands. Precision-cut tissue slices of both hepatic and extra-hepatic origin are extensively used as an in vitro model to predict in vivo drug metabolism and toxicity. Cryopreservation would greatly facilitate their use. In the present study, we aimed to improve (1) rapid freezing and warming (200 degrees C/min) using 18% Me(2)SO as cryoprotectant and (2) vitrification with high molarity mixtures of cryoprotectants, VM3 and VS4, as methods to cryopreserve precision-cut rat liver and kidney slices. Viability after cryopreservation and subsequent 3-4h of incubation at 37 degrees C was determined by measuring ATP content and by microscopical evaluation of histological integrity. Confirming earlier studies, viability of rat liver slices was maintained at high levels by rapid freezing and thawing with 18% Me(2)SO. However, vitrification of liver slices with VS4 resulted in cryopreservation damage despite the fact that cryoprotectant toxicity was low, no ice was formed during cooling and devitrification was prevented. Viability of liver slices was not improved by using VM3 for vitrification. Kidney slices were found not to survive cryopreservation by rapid freezing. In contrast, viability of renal medullary slices was almost completely maintained after vitrification with VS4, however vitrification of renal cortex slices with VS4 was not successful, partly due to cryoprotectant toxicity. Both kidney cortex and medullary slices were vitrified successfully with VM3 (maintaining viability at 50-80% of fresh slice levels), using an optimised pre-incubation protocol and cooling and warming rates that prevented both visible ice-formation and cracking of the formed glass. In conclusion, vitrification is a promising approach to cryopreserve precision-cut (kidney) slices. PMID: 17166492 [Here's another example of the extremely slow pace of transmission of information to scientists. Back in 1988 the Gynaephora groenlandica larva were discovered to be nature's most freeze resistant animal. By all rights, cryobiologists should have bumped into each other researching this animal to discover its chilly secret. However for the next two decades nothing happened. There are still currently a grand total of zero references to this animal in the journal Cryobiology. I talked to one of the world's premier cryobiologists recently about these "Woollybear caterpillars". He admitted he had never heard of them! Yes, he was very interested, and thankful for the information. This slow pace of information transmission is partly the reason I agree with conservative cryobiologists, that it may take over 50 years to achieve ERC. Cryobiologists have had a few decades to advance their art, while Mother Nature has had billions of years to evolve the exceptionally high freeze resistance of the woollybear caterpillar.] J Comp Physiol [B]. 1988;158(2):175-83. Glycerol metabolism in a freeze-tolerant arctic insect: an in vivo 13C NMR study.Kukal O, Serianni AS, Duman JG. Department of Biological Sciences, University of Notre Dame, Indiana 46556. Freeze-tolerance in larvae of Gynaephora groenlandica is enhanced by the accumulation of glycerol in the winter. Since summer larvae remain freeze-tolerant despite the lack of glycerol, we investigated glycerol metabolism as a function of acclimation and body temperature using noninvasive 13C NMR spectroscopy. Major constituents of hemolymph isolated from cold- and warm-acclimated larvae were identified with the aid of standard NMR spectra and confirmed by TLC and GLC. Spectra obtained on live, warm-acclimated larvae showed the presence of lipids, glycogen, glucose, trehalose and amino acids. Similar spectra of cold-acclimated or previously frozen larvae showed the additional presence of glycerol. In vitro time-lapse 13C spectra of D-[1-13C]glucose added separately to hemolymph or extracted fat body tissue showed that glycerol is synthesized from glucose in the fat body tissue and distributed to the peripheral tissue via hemolymph. In vivo time-lapse 13C spectra of cold- and warm-acclimated larvae were obtained after injection with D-[1-13C]glucose to monitor the production of labeled metabolic intermediates and end-products. [13C]Glycerol was produced between -30 degrees C and 30 degrees C but accumulated only below 5 degrees C. Above 5 degrees C glycerol was degraded and the 13C label incorporated mainly into glycogen. The mechanism underlying temperature control of glycerol biosynthesis and degradation may provide a clue to the role of glycerol in enhancing freeze-tolerance in these insects. PMID: 3170824 [Further quote from the above paper: "However, cold-acclimation (-15 C, -30 C) enhanced cold tolerance in the larvae so they were able to tolerate temperatures lower than -70 C (the lowest temperature tested, no mortality resulted;"] Guess how many citations there are for "Gynaephora groenlandica" on Pubmed. 600? 60? 6? Check out the answer with this link>> http://www.ncbi.nlm.nih.gov/entrez/query.fcgi Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=29421