Accepted Abstracts 2004

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American Aging=20 Association - 33rd Annual Meeting

Molecular = Mechanisms of=20 Aging: As Modulated by Genes, Hormones and Oxidative=20 Events

June = 4-7,=20 2004 St Petersburg,=20 = Florida

Meeting=20 Report Speakers Program-at-a-= Glance=20 Program=20 Details Abstracts = ; =20 Sponsors &= nbsp;=20 Pictures=20

ACCEPTED=20 ABSTRACTS

O R A = L
Brewer, Gregory J.	AGING=20 NEURON MITOCHONDRIAL FUNCTION AND REJUVENATION = WITH=20 ESTROGEN OR MITOSIS=20 =20 G. Brewer,=20 J. Nash, T. Jones, J. Reichensperger, M.=20 Parihar=20 =20 PO Box=20 19626, Springfield, IL 62794-9626=20 =20 The=20 mitochondrial oxyradical theory of aging is = gaining=20 increased support from studies of brain, heart = and=20 muscle tissue. Is this aging caused by = systemic=20 hormones or intrinsic to aging = mitochondria in=20 neurons, for example. The intrinsic = hypothesis=20 is supported by our findings with isolated = neurons of=20 increased susceptibility with age to glutamate = and=20 A-beta toxicity, two endogenous toxins = important to=20 age-related neurodegenerative disease (Brewer, = 1998). For neuron-specific function, we = have=20 established culture techniques that allow us = to=20 compare neurons isolated from embryonic, = middle-age=20 (12 month) and old (24 month) rat hippocampus = (Brewer,=20 1997). For use with these cultured = neurons, we=20 monitored cytochrome oxidase activity (COX),=20 cardiolipin specific staining of mitochondria = with=20 nonyl-acrydine orange (NAO), as well as = labeling with=20 mitotracker-red and immunostain for cytochrome = C,=20 fluorescent NADH, glutathione levels and = respiration.=20 Cytochrome C immunostain and mitotracker red = stained=20 equal numbers of mitochondria per cell for = middle-age=20 and old neurons, compared to higher numbers = per=20 embryonic neuron. In contrast to = mitochondrial=20 numbers, COX activity/cell declined sharply = with age=20 of the neurons: levels of old neurons were = reduced by=20 30% of those of middle-age = neurons. =20 Similarly, NAO stain for mitochondria was 40% = lower=20 for old neurons compared to middle-age = neurons. =20 By addition of FGF2 and low density passage, = old=20 neurons divide in culture (Brewer, = 1999). COX=20 activity in these dividing old neurons was = equivalent=20 to that of middle-age neurons. = Age-related=20 reductions in glutathione levels were = exacerbated by=20 exposure to glutamate. NADH levels and=20 respiration in old neurons were normal, but = showed=20 dramatic deficits in response to = glutamate. In=20 addition to restoring youthful calcium = dynamics,=20 treatment of old neurons with = 17-beta-estradiol=20 restored NAO staining levels/cell to those of=20 middle-age neurons and provided full = neuroprotection=20 from glutamate and A-beta toxicity. = These=20 results demonstrate that mitochondrial = function is=20 impaired in old neurons, which correlates with = increased susceptibility to toxic stress, but = function=20 can be restored by treatment with FGF2 or=20 estradiol.=20 top=20
Carter, Christy S.	ACE=20 inhibition intervention: Implications for = improving=20 age-related declines in physical performance = and=20 longevity.=20 =20 CS Carter,=20 M Cesari, M Pahor=20 =20 Wake Forest=20 Univeristy School of Medicine, J. Paul Sticht = Center=20 on Aging, 1 Medical Center Blvd. Winston-Salem = NC,=20 27157=20 =20 Age-related=20 disregulation of the renin-angiotensin = system=20 (RAS) leads to variety of pathologies = including end=20 organ damage to kidney, heart, blood vessels, = and more=20 recently has been suggested to exacerbate = declining=20 cognitive and physical performance. Reversing = or=20 attenuating these effects could have a = significant=20 impact on maintaining independence and = extending=20 longevity in the elderly. In fact, both=20 pharmacological and genetic studies have = demonstrated=20 that lower circulating levels of angiotensin=20 converting enzyme (ACE) results in enhanced=20 responsiveness to exercise, increased muscle = strength,=20 and lower adiposity. The relationship between = low=20 physical performance in healthy older adults, = shifting=20 body composition (increases in fat and = decreases in=20 fat-free mass) and longevity has been = documented and=20 more recently this same assessment has been = made in=20 aging rodents. Therefore, interventions = that=20 retard declining performance such as ACE = inhibition=20 (ACEi) may also have validity for studies of=20 longevity. We have conducted studies to = assess=20 the effects of ACEi on body composition and = physical=20 performance in aged rats. Animals were = randomized to=20 daily injections of 40 or 80 mg/kg of = enalapril or=20 saline at baseline (24 months of age) and were = followed for 6 months. ACEi attenuated = declining=20 physical performance as measured by both grip = strength=20 and inclined plane tasks, and was most likely=20 explained by a significant reduction in total = fat=20 mass, as no differences in lean mass were = observed=20 between groups (as measured using DXA). A = possible=20 interpretation is that beyond its powerful = hemodynamic=20 effects, ACEi regulates many aspects of = metabolic=20 functioning, decreases oxidative stress in = many=20 tissues, and may act ubiquitously to reduce = age and=20 disease related chronic inflammatory = states. =20 Therefore, our understanding of these results = could be=20 enhanced by a more thorough and mechanistic=20 characterization of the effects of long-term = ACEi on=20 lifespan and declining = performance. =20 top=20
Chang, Sandy	ESSENTIAL ROLE OF LIMITING = TELOMERES IN=20 THE PATHOGENESIS OF WERNER SYNDROME=20 AbstractAuthors: S. Chang, A. = Multani, N.=20 Cabrera, M. Naylor, P. Laud, D. Lombard, S. = Pathak, R.=20 de Pinho MD Anderson Cancer Center, = Houston, TX=20 and Dana Farber Cancer Center, Boston,=20 MA Mutational inactivation of the Wrn = helicase=20 gene causes Werner Syndrome (WS), an autosomal = recessive disease characterized by premature = aging,=20 elevated genomic instability and increased = cancer=20 incidence. The capacity of enforced telomerase = expression to rescue premature senescence of = cultured=20 human WS cells and the lack of a phenotype in = Wrn=20 deficient mice with long telomeres have = implicated=20 telomere attrition in WS pathogenesis. Here, = we show=20 that the varied and complex in vivo and = cellular=20 phenotypes of WS are precipitated upon = exhaustion of=20 telomere reserves in mice. In mice doubly null = for Wrn=20 and the telomerase RNA component (Terc), = telomere=20 dysfunction elicits a classical WS premature = aging=20 syndrome typified by premature death, hair = graying,=20 alopecia, osteoporosis, type II diabetes, and=20 cataracts. This model also exhibited = accelerated=20 replicative senescence and accumulation of = DNA-damage=20 foci in cultured cells as well as increased=20 chromosomal instability and cancer, = particularly=20 non-epithelial malignancies typical of WS. = This=20 faithful mouse model of WS establishes that = the=20 complex and pleiotropic in vivo sequelae of = Wrn=20 deficiency are provoked largely by critical = telomere=20 shortening. top*=20
Heilbronn, Leonie K.	THE=20 EFFECTS OF 6-MONTHS OF CALORIE RESTRICTION ON=20 BIOMARKERS OF AGING IN NON-OBESE = HUMANS.=20 =20 Leonie K=20 Heilbronn, Eric Ravussin and the Pennington = CALERIE=20 group.=20 =20 Pennington=20 Biomdeical Research Center, 6400 Perkins Rd, = Baton=20 Rouge, LA=20 =20 Caloric=20 restriction (CR) extends life span and retards = age-related diseases in rats, mice, flies, = worms and=20 yeast. Several biomarkers of aging (fasting = glucose,=20 insulin, DHEAS and core temperature) have been = identified in human and monkey studies of = longevity.=20 Whether prolonged CR improves biomarkers of = aging in=20 humans is unknown. Forty-eight healthy, = nonsmoking,=20 male (25-50y) and female (25-45y), overweight=20 participants (25 < BMI < 30) were = randomized=20 into one of four groups; group 1 =3D 25% CR, = group 2 =3D=20 12.5% CR + 12.5% increase in physical = activity, group=20 3 =3D low calorie diet until 15% weight = reduction, group=20 4 =3D Control. Subjects were fed by the = metabolic=20 kitchen from Weeks 1-12 whilst they were = taught a=20 calorie counting system to allow them to = prepare their=20 own meals (Weeks 12-22). Subjects returned to=20 infeeding from Weeks 22-24. Fasting glucose, = insulin,=20 DHEAS and 24h core temperature were assessed = at=20 baseline, weeks 12 and 24. At the time of = writing,=20 only 30 subjects had completed the study and = so CR=20 groups (Groups 1-3) were combined for = analysis.=20 Average weight loss was 10% in the CR groups. = Weight=20 was not changed significantly in the controls. = Fasting=20 insulin was reduced 24% in CR groups as = compared to a=20 5% increase in the controls (p<0.05). = Glucose was=20 reduced 1% in CR groups as compared to a 4% = increase=20 in controls (p<0.03). DHEAS and daytime = core=20 temperature were not significantly changed = after 6=20 months. However, nighttime temperature = (recorded from=20 10pm to 6am) was reduced by 0.2=BAC in CR = groups as=20 compared to a 0.3=BAC increase in controls = (p=3D0.06).=20 This study suggests that 6-mo. of calorie = restriction=20 in non-obese humans was sufficient to observe=20 improvements in 3 out of 4 known biomarkers of = aging.=20 Whether these changes are sustained over = longer=20 periods of time is unknown.=20 =20 top*=20
Kitani, Kenichi	THE=20 PRESENCE OF AN OPTIMAL DOSE RANEGE MAY EXPLAIN = DISCREPANCIES IN THE EFFECTS OF (-)DEPRENYL = (D) ON=20 SURVIVALS OF ANIMALS IN PAST = REPORTS=20 =20 K.Kitani1,=20 S.Kanai2, K.Miyasaka2, M.C.Carrillo3 ,GO = Ivy4=20 =20 Tokyo=20 Metropolitan Institute of Gerontology =20 THE=20 PRESENCE OF AN OPTIMAL DOSE RANGE MAY EXPLAIN=20 DISCREPANCIES IN THE EFFECTS OF (-)DEPRENYL = (D) ON=20 SURVIVALS OF ANIMALS IN PAST REPORTS =20 K.Kitani1,=20 S.Kanai2, K.Miyasaka2, M.C.Carrillo3 ,GO = Ivy4=20 1National=20 Institute for Longevity Sciences, 36-3, Gengo, = Moriokacho, Obu-shi; 2Tokyo Metropolitan = Institute of=20 Gerontology, Tokyo, Japan; = 3Univ.Rosario,Rosario,=20 Argentina; 4Univ.Toronto, = Scarborough,Canada=20 =20 To explain=20 discrepancies in the effect of D on life spans = of=20 animals in past reports, we examined the = effect of D=20 at a dose of 0.25 mg/kg/inj s.c. 3x a week = starting=20 from 18 months of age on life spans of = F344Du/Crj rats=20 of both sexes. Average life spans = (days)=20 were significantly increased in both male=20 (895.2=B1109.7,mean =B1S.D., n=3D30; = 967.8=B188.6, n=3D30,=20 control vs. D treated,+8.1%, P<0.01) and = female=20 (924.7=B1132.2, n=3D38; 987.1=B1133.4, = n=3D39,+6.7%,=20 P<0.05) rats that received D injections = compared to=20 control animals given saline. Increases = in life=20 expectancy from 24 months of age were 44% in = males and=20 32% in females. We previously reported = that a=20 dose of 0.5mg/kg/inj significantly increased = the life=20 span of male F344 rats (1). However, a dose of = 1.0=20 mg/kg/inj somewhat shortened the life span of = animals,=20 although not statistically significantly (2).=20 Together,our data indicate that a proper dose = range of=20 D can significantly increase the life spans of = rats of=20 both sexes, however, a greater dose becomes = less=20 effective and may adversely affect the life = span of=20 animals. The presence of this effective = dose=20 range of D may explain discrepancies in the = effect of=20 D on life spans of animals previously = reported. =20 Furthermore, the fact that the effective dose = range=20 observed for the life prolonging effect of D = parallels=20 that for increasing activities of SOD and = catalase=20 strengthens our contention that these two = effects are=20 causally related. Our new observation = that the=20 same dosage of D (0.25mg/kg) increases life = spans of=20 both male and female rats further supports our = contention, since we previously reported that = the=20 optimally effective dose of D to increase = antioxidant=20 enzyme activities becomes closer for male and = female=20 rats as they get older, while at young ages, = the=20 optimal dose is 10 times greater in male rats=20 (3). Ref. 1) Life Sci. 52:281-288, 1993. = 2) Life=20 Sci. 67:577-585, 2000.3)Life Sci.=20 52:1925-1934,1993=20 top*=20
Ladiges, Warren C.	AGE-ASSOCIATED=20 CELLULAR STRESS IN HSP-40 MOLECULAR CHAPERONE = MUTANT=20 MICE=20 =20 S=20 Knoblaugh, J Morton, G Moore-Sanders, A = MacAuley, W=20 Ladiges* =20 Department=20 of Comparative Medicine, University of=20 Washington, Seattle, WA 98195=20 =20 Cellular=20 stress associated with a malfunctioning = endoplasmic=20 reticulum (ER) is causally related to the = inability of=20 molecular chaperones to maintain structural = integrity=20 of proteins. Proteins which are structurally = damaged=20 under conditions of oxidative stress or = aggregation=20 during the aging process place an increased = burden on=20 the processing capacity of the ER. The result = is an=20 exaggerated and extended ER stress response, = which=20 triggers transcriptional and translational = pathways of=20 cell death and decreased protein synthesis. ER = dysfunction has been implicated in a number of = age-related diseases including Parkinsons, = Alzheimers,=20 cardiovascular conditions, and diabetes. The = HSP-40=20 molecular chaperone family member, p58ipk, = functions=20 in the later stages of the ER stress response = as an=20 off switch to prevent excessive cell death and = promote=20 protein synthesis. Mutant mice deficient = in=20 p58ipk have evidence of an exaggerated ER = stress=20 response in several cell types including = pancreatic=20 beta cells, hepatocytes, and plasma cells. = Gene=20 expression, Western immunoblots and=20 immunohistochemistry in cells from p58ipk = mutant mice=20 confirm the upregulation of ER stress response = genes=20 and an enhanced cell death pattern reflective = of an=20 age-dependent onset of diabetes and immune=20 dysfunction, and an average life span of 18=20 months. We conclude that ER resident = proteins,=20 including HSP-40/p58ipk, are targets for = investigating=20 specific aging and age-related disease = questions. The=20 p58ipk mutant mouse model will be of interest = in=20 intervention studies to determine capabilities = of=20 decreased disease and increased life span = mediated by=20 a functional ER. top=20
Nikolich-Zugich, = Janko	PHENOTYPIC,=20 FUNCTIONAL AND GENETIC PROFILES OF THE AGING = T-=20 LYMPHOCYTES IN PRIMATES AND THEIR ALTERATION = BY=20 CALORIC RESTRICTION=20 =20 Janko=20 Nikolich-Zugich, Jessica Warner, Bree Fisher, = Dragana=20 Nikolich-Zugich and Ilhem=20 Messaoudi=20 =20 Vaccine and=20 Gene Therapy Institute and the Oregon National = Primate=20 Research Center, Oregon Health & Science=20 University, Beaverton, OR, USA=20 =20 Numerous=20 T-cell functions are diminished or = dysregulated in old=20 age, including T-cell population dysbalance = (altered=20 na ve:memory and CD4/CD8 T-cell ratios), = diminished=20 T-cell responsiveness to a variety of signals = and=20 altered cytokine networks. Yet, despite = intense=20 research, numerous gaps in the understanding = of T-cell=20 senescence still exist. Moreover, much of our = current=20 knowledge on T-cell biology and aging comes = from=20 inbred, specific pathogen-free rodents, and it = is=20 unclear which observations translate to human = immune=20 aging. =20 We begun to=20 develop a primate model of human immune = senescence=20 using Rhesus macaques (RM). We evaluated = T-cell=20 surface phenotype, in vivo and in vitro = turnover, the=20 complexity of T-cell receptor (TCR) repertoire = and=20 correlated them to functional studies of = isolated cell=20 subsets and gene expression profile analysis. = We also=20 evaluated the ability of an experimental = manipulation=20 - caloric restriction, the only intervention = that=20 consistently and reproducibly increases = longevity and=20 quality of life in a variety of animal models = - to=20 impact upon these parameters. =20 We showed=20 that sharp differences exist in the phenotypic = and=20 functional T-cell aging between the CD8 and = CD4 T-cell=20 subsets in: (i) cell cycle programs (as = assessed by=20 both in vitro proliferation and in vivo = turnover=20 measurement); (ii) CD28 regulation upon cell = cycle=20 entry; (iii) accumulation of immediate = effector cells=20 amongst the CD28- cells, believed to be close = to or at=20 replicative senescence. These results = underscore poor=20 reliability of CD28 as a marker for = senescence, and=20 further suggest that some of the T-cell aging=20 phenomenology in RM can be ascribed to = accentuation=20 over time of the inherent differences in = activation=20 programs in CD8 and CD4 T-cells. We also found = age-related differences in T-cell turnover, = and=20 modest, but consistent, gene expression = changes in=20 certain resting T-cell subsets. Caloric = restriction=20 was able to modulate most, if not all, of = these=20 parameters, resulting in = young-like =20 characteristics of examined T-cells. We = conclude that=20 the results so far are consistent with = resource=20 conservation induced by caloric = restriction.=20 =20 top=20
Shaikh, Aasef G.	cAMP=20 forms a neurochemical = correlate of=20 tinnitus=20 =20 A, = Shaikh; P,=20 Finlayson=20 =20 Department of = Otolaryngology,=20 Wayne State University, Detroit, MI=20 48201 =20 =20 Cochlear damage that = commonly=20 occurs in elderly is either due to ototoxicity = of=20 anticancer drugs (cisplatin) or a process of = aging.=20 Such hearing loss is often accompanied by = tinnitus. It=20 was suggested that increased spontaneous = neural=20 activity (SA) may be the underlying mechanism = for=20 tinnitus. Neurochemical consequences of = cochlear=20 ablation (CA) have also been widely explored. = A=20 significant synaptic plasticity has been = reported=20 following cochlear insults. Yet a direct = relationship of=20 increased SA with plastic neurochemical = alterations=20 remains to be recognized. In central auditory = neurons,=20 the signals emerging after CA are transduced = by range=20 of mechanisms including extracellular = signal-regulated=20 kinase (ERK) pathway. One of the functional = roles of=20 this pathway is to enhance the inhibition of=20 phosphodisterase E 4 (PDE4), thereby elevating = intracellular cAMP concentrations ([cAMP]i). = We sought=20 to determine if increasing [cAMP]i affects SA, = and if=20 so what possible mechanisms may be involved = and the=20 possible physiological/pathological = ramifications. We=20 also investigated such elevations in SA could = mimic=20 the neural code for the pure-tone. Forskolin = (an agent=20 that systematically increases [cAMP]i) = specifically=20 increases tone-evoked responses and SA of the = SOC=20 neurons in dose-dependent manner. = Interestingly,=20 the increased SA following application = of 50 =B5M=20 forskolin mimics the neural code that is = normally=20 generated by the tonal stimulus. We also = report=20 that effects of forskolin are predominantly = due to PKA=20 independent pathway that involves = hyperpolarization=20 activated inward conductances. These results = provide=20 for the first time, direct evidence that = systematic=20 increase in the [cAMP]i, in the auditory = brainstem=20 neurons, by application of forskolin elevates = the SA,=20 which mimics the neural code for the = pure-tone. Here=20 we suggest, increased levels of [cAMP]i that = could=20 follow cochlear insults, forms the basis of = increased=20 SA and therefore could be the =93neurochemical = correlate=94 of tinnitus. = top=20
Sun, Liou Y.	HIPPOCAMPAL=20 GH AND IGF-1 EXPRESSION IN GH-DEFICIENT=20 MICE=20 =20 Liou Y.=20 Sun, Khalid Al-Regaiey, Michal M. Masternak, = Jian=20 Wang and Andrzej Bartke=20 =20 Geriatrics=20 Research, Department of Medicine and = Department of=20 Physiology=20 Southern=20 Illinois University, Springfield, IL 62794, = USA=20 =20 Liou Y.=20 Sun, Khalid Al-Regaiey, Michal M. Masternak, = Jian=20 Wang and Andrzej Bartke=20 Geriatrics=20 Research, Department of Medicine and = Department of=20 Physiology=20 Southern=20 Illinois University, Springfield, IL 62794, = USA=20 =20 Beneficial=20 effects of GH and IGF-1 on the development and = function of the central nervous system are = well=20 documented. In spite of primary deficiency of = GH and=20 secondary IGF-1 deficiency, Ames dwarf mice = live=20 considerably longer than normal animals, = exhibit=20 apparently normal cognitive functions and = maintain=20 them into advanced age. In an attempt to = reconcile these findings, we have examined = local=20 expression of GH and IGF-1 in the hippocampus, = brain=20 region involved in learning and memory, of = these=20 mice. RNA and protein was extracted from = the=20 hippocampus of dwarf and normal mice and = assayed for=20 the GH and IGF-1 transcripts and their encoded = proteins. With the real-time RT-PCR analyses,=20 hippocampus of Ames dwarf mice was found to = express=20 normal levels of GH and IGF-1 messenger RNA = indicating=20 ectopic GH expression and the integrity of=20 transcription capability in dwarf mice. The = identities=20 of the PCR products were confirmed by = sequencing.=20 Hippocampal levels of GH and IGF-1 were = evaluated by=20 western blotting using the antibodies specific = for the=20 respective proteins. Both GH and IGF-1 protein = levels=20 are significantly increased in the hippocampus = of Ames=20 dwarf compared with normal mice suggesting a=20 compensatory mechanism of peripheral hormonal=20 deficiency. In contrast, IGF-1 expression in = the liver=20 of Ames dwarf mice is profoundly suppressed,=20 consistent with congenital GH deficiency and = lack of=20 detectable GH and IGF-1 in peripheral = circulation. In=20 addition, Increased phosphorylation of Akt and = CREB=20 were also detected in the hippocampus of Ames = dwarf=20 mice. Our results suggest that increase in = hippocampal=20 GH and IGF-1 protein expression and subsequent = activation of PI3K/Akt-CREB signal = transduction=20 cascade might contribute to the maintenance of = cognitive function and is likely to be = responsible for=20 the integrity of neuronal structure, and = maintenance=20 of youthful levels of cognitive function in = these=20 long-lived mice during aging.=20 top=20
Wilcock, Donna M.	PASSIVE=20 AMYLOID IMMUNOTHERAPY CLEARS AMYLOID AND = TRANSIENTLY=20 ACTIVATES MICROGLIA IN A TRANSGENIC MOUSE = MODEL OF=20 AMYLOID DEPOSITION=20 =20 Donna M.=20 Wilcock1, Amyn Rojiani2, Arnon Rosenthal3, = Jennifer=20 Alamed1, David Wilson1, Nedda Wilson1, Melissa = J.=20 Freeman1, Marcia N. Gordon1, Dave = Morgan1.=20 =20 AbstractLocation:=20 1,2: Alzheimer=92s Research Laboratory, = 1Department of=20 Pharmacology, 2Departments of = Interdisciplinary=20 Oncology and Pathology, University of South = Florida,=20 12901 Bruce B Downs Blvd, Tampa, FL 33612, = USA.=20 3: Rinat=20 Neuroscience Corp. 3155 Porter Drive, Palo = Alto,=20 California, 94304, USA.=20 =20 The role of=20 microglia in the removal of amyloid deposits = following=20 systemically administered anti-A=DF antibodies = remains=20 unclear. In the current study we injected = Tg2576 APP=20 transgenic mice weekly with anti-A=DF antibody = for a=20 period of one, two or three months such that = all mice=20 were 22 months at the end of the study. In = mice=20 immunized for three months we found an = improvement in=20 alternation performance in the Y maze. = Histologically,=20 we were able to detect mouse IgG bound to = congophilic=20 amyloid deposits in those mice treated with = anti-A=DF=20 antibody but not in those treated with control = antibody. We found that Fcgamma receptor = expression on=20 microglia was increased following one month of = treatment while CD45 was increased following = two=20 months of treatment. Associated with these = microglial=20 changes was a reduction in both diffuse and = compact=20 amyloid deposits following two months of=20 treatment. Interestingly, the microglia = markers=20 were reduced to control levels following three = months=20 of treatment while amyloid levels remained=20 reduced. Serum A=DF levels and anti A=DF = antibody=20 levels were elevated to similar levels at all = three=20 survival times in mice given anti-A=DF = injections rather=20 than control antibody injections. These data = show that=20 antibody is able to enter the brain and bind = to the=20 amyloid deposits, likely opsonizing the A=DF = and=20 resulting in Fcgamma receptor mediated = phagocytosis.=20 Together with our earlier work, our data argue = that=20 all proposed mechanisms of anti-A=DF antibody = mediated=20 amyloid removal can be simultaneously active. = top=20
P O S T E=20 R
Al-Regaiey,=20 Khalid	43. =20 EFFECTS OF LONG-TERM CALORIC RESTRICTION ON = WILD-TYPE=20 AND GROWTH HORMONE RECEPTOR KNOCKOUT MICE = Khalid = Al-Regaiey,=20 Michal M. Masternak, Michael Bonkowski, Liou = Sun,=20 Andrzej Bartke Southern Illinois = University=20 School of Medicine, Springfield, IL 62794=20 Reduced IGF-1/insulin signaling and = caloric=20 restriction (CR) are known to extend life span = and delay=20 age related diseases. To address the interaction = of=20 these two interventions, we subjected normal (N) = and=20 growth hormone receptor knockout (KO) mice to CR = starting at weaning for 20 months. = Molecules=20 involved in glucose homeostasis were = investigated. =20 CR resulted in decreased plasma glucose levels = in both=20 phenotypes (N-CR and KO-CR). Circulating = IGF-1 was=20 reduced in N-CR and was undetectable in KO=20 animals. Insulin was also reduced in N-CR = to=20 levels comparable to those in KO animals. = Corticosterone=20 and adiponectin levels were higher in KO than in = N=20 animals while leptin was reduced by CR in both=20 phenotypes. We also analyzed hepatic gene and = protein=20 expressions of molecules involved in insulin = signaling=20 using real-time PCR and western blotting, = respectively.=20 Diet and phenotype did not affect Akt gene and = total=20 protein expression while KO mice exhibited = reduced Akt=20 activation with no diet effect. Phosphorylation = of=20 protein kinase C γ/ζ was not = affected=20 by either diet or phenotype. KO mice exhibited = increased=20 CREB activation. Gene expression and protein = levels of=20 Foxo1 and PGC-1 were increased in KO mice with = little=20 diet effects. Genes that are known to be = regulated by=20 CREB, Foxo1, and/or PGC-1including pepck, = g6pase,=20 igfbp-1, and MnSOD had increased expression in = KO mice.=20 Genes encoding proteins that are involved in = lipolysis,=20 including hormone-sensitive lipase (HSL) and=20 lipo-protein lipase (LPL) were upregulated in KO = mice.=20 We conclude that CR and growth hormone = resistance=20 probably affect longevity of mice by different=20 mechanisms and that gluconeogenesis pathway as = well as=20 fatty acids mobilization are more active in KO = mice.=20 These data also suggest that liver of KO mice is = more=20 protected against oxidative stress as indicated = by=20 increased Foxo proteins and MnSOD mRNA. = Supported by=20 NIA.
Anderson, Rozalyn M.	44. =20 AGING RETARDATION BY CALORIE RESTRICTION: ROLE = OF=20 REGULATORS OF ENERGY METABOLISM. =20 R. Anderson=20 (1), J.Barger (1), T. Pugh (1), S. Park (2), = T. Prolla=20 (2) and R. Weindruch (1).=20 =20 (1)Department=20 of Medicine, University of Wisconsin and = GRECC, VA=20 Hospital, Madison WI 53705 USA. (2) Department = of=20 Genetics and Medical Genetics, University of=20 Wisconsin, Madison WI 53706 USA.=20 =20 University=20 of Wisconsin Madison, GRECC, VA Hospital, = Madison WI=20 53705, USA.=20 =20 =20 Caloric restriction (CR) extends lifespan in a = broad=20 spectrum of organisms and retards the = progression of a=20 wide range of age-associated biological = changes;=20 however, the underlying mechanisms are = unclear. =20 In mice, there is an inverse linear = relationship=20 between calorie intake and lifespan extension=20 suggesting that factors central to energy = metabolism=20 may underlie aging retardation by CR. = One=20 hypothesis is that CR triggers an active = response=20 involving a reprogramming of energy metabolism = that=20 reduces the rate of aging and the development = of=20 several age-related diseases. = Transcription=20 profiling of tissues from control and CR = animals=20 points to mitochondrial function as a central = feature=20 in this process. We have used a = combination of=20 techniques to identify factors that may be = involved in=20 the mechanism of lifespan extension by = CR. We=20 then investigated how these potential = effectors of CR=20 are regulated and examined their relationship = to=20 elements of known longevity pathways, = including=20 members of the FOXO and Sirtuin = families.=20 top=20
Armbrecht, Harvey J.	45. =20 REGULATION OF THE RENAL VITAMIN D = 1ALPHA-HYDROXYLASE=20 CYTOCHROME P450(CYP27B1) BY DIETARY PHOSPHORUS = CHANGES=20 WITH AGE=20 =20 H.J.=20 Armbrecht, M.A. Boltz=20 =20 Geriatric=20 Center, St. Louis VA Medical Center, St. = Louis,=20 MO 63125=20 =20 Young rats=20 adapt to a low phosphorus diet by increasing = plasma=20 levels of 1,25-dihydroxyvitamin D (1,25D), the = biologically active metabolite of vitamin D, = which=20 then increases intestinal phosphate = absorption. =20 Previous studies have shown that the capacity = of rats=20 to adapt to low dietary phosphorus declines = with=20 age. The purpose of this study was to = determine=20 whether this decreased adaptation was due to = decreased=20 expression of the renal 1alpha-hydroxylase = (1-OHase),=20 which makes 1,25D. The 1-OHase consists = of=20 ferredoxin reductase, ferredoxin, and a = terminal=20 cytochrome P450 =96 CYP27B1. Young (2 = months) and=20 adult (12 months) F344 rats were placed on a = low=20 phosphorus (0.1%) or high phosphorus (1.0%) = diet for 2=20 weeks. Plasma 1,25D was markedly = increased by=20 the low phosphorus diet in young animals but = not in=20 adults. To determine whether this = difference was=20 due to decreased 1-OHase expression, mRNA = levels of=20 CYP27B1 were measured by ribonuclease = protection=20 assay. In young animals, the low = phosphorus diet=20 increased renal CYP27B1 mRNA levels 8-fold = compared to=20 the high phosphorus diet, but there was no = significant=20 effect in adults. The effect of low = dietary=20 phosphorus on plasma calcium, phosphate, and=20 parathyroid hormone levels was similar in both = age=20 groups. These results suggest that the = decreased=20 CYP27B1 expression in the adult in response to = dietary=20 phosphorus depravation is not due to changes = in the=20 major regulators of 1-OHase=20 activity. =20 top*=20
Ball, Sheldon S.	46. =20 SENEX: INTEGRATING GERIATRICS AND MOLECULAR=20 GERONTOLOGY=20 =20 S=20 Ball, A Nhalil and V Mah=20 =20 Department=20 of Veterans Affairs, Greater Los Angeles = Healthcare,=20 11301 Wilshire Boulevard, GRECC 11G, Los = Angeles CA=20 90073=20 =20 Senex is a=20 computer application 15 years in development=20 integrating geriatrics and molecular = gerontology (from=20 bench to bedside). It includes information = (and tools=20 for interpretation of data) that span internal = medicine, molecular pathology (biochemistry, = molecular=20 biology, pathology), laboratory medicine, = radiology,=20 pharmacology, anatomy, and statistics. = Clinical=20 and molecular information is needed from = laboratory=20 bench to patient bedside, during contact = between=20 physician and patient, educator and student, = scientist=20 and technician. The information needs to = be=20 structured so that specific and real-time = retrieval is=20 achievable. The information needs to be = presented in a=20 manner that facilitates a conceptual = understanding of=20 the details presented. Additionally, an=20 information system should display a certain = degree of=20 intelligence, including flexibility in = accepting input=20 from the user, the capacity to reason with = structured=20 information, and the esthetic display of=20 context-specific information. Implementation = of such=20 an information system should allow use to = become=20 better physicians, scientists, educators, = students=20 and/or informed citizens. Senex functions=20 independently as a stand-alone application = (i.e. not=20 dependent upon internet access). Thus it is = fast,=20 reliable, and mobile. Senex also provides = direct links=20 into the world-wide-web, to molecular and = clinical=20 databases and to the original literature when = access=20 to the internet is available. Senex is a = large=20 application with: 1514 organisms, 2417 = anatomic=20 structures, 222 cells, 131 cellular = compartments,=20 13,138 molecules, 9607 proteins, 628 genes, = 795=20 motifs, 67 molecular pathways, 1580=20 diseases, 540 clinical laboratory tests, = 18,680=20 database links (6140 Swiss Prot, 6280 = OMIM, 870=20 Prosite, 2000 PIR, 3390 Locus Link). = Senex=20 allows a user to add proprietary information = on top of=20 the core Senex knowledge base, thus = customizing the=20 application for a user=92s own information = needs. =20 Senex runs on Macintosh and Windows computers. = The=20 Macintosh version has a microarray data = analysis and=20 data mining module. Reference:=20 www.senex-medical-software.com=20 top=20
Bergamini, Ettore	47. =20 THE AGE-RELATED ACCUMULATION OF DOLICHOL IN = TISSUES=20 SATISFIES ALL CRITERIA TO BE QUALIFIED A = BIOMARKER OF=20 AGING=20 =20 E.=20 Bergamini, G. Cavallini, A. Donati, Z. Gori, = A.=20 Manfrini, I. Parentini=20 =20 Centro di=20 Ricerca di Biologia e Patologia = dell=92Invecchiamento=20 dell=92Universita=92 di Pisa, Dipartimento di = Patologia=20 sperimentale, Via Roma 55, 56126 PISA (Italy); = Tel=20 0039-050-2218584 Fax=20 0039-050-2218581 e-mail=20 ebergami@med.unipi.it=20 =20 It was=20 stated that the identification of specific = biomarkers=20 of aging would be an important milestone in=20 gerontological research, and criteria for = defining=20 biomarkers were suggested (Mooradian, 1990). = In this=20 communication evidence on male and female = Sprague=20 Dawley and Lewis rats fed ad-libitum or on = regimens of=20 food restriction and on human tissues are = reviewed,=20 showing that accumulation of dolichol in = tissues of=20 older animals meet these criteria: levels of = dolichol=20 exhibit a quantitative correlation with age in = all=20 tissues; the biological parameter is not = altered by=20 several age-dependent diseases in the same = direction=20 as that of aging; data with transplanted = organs show=20 that the age-related accumulation is not = secondary to=20 metabolic changes of aging; factors that = modulate the=20 aging rate like caloric restriction and = physical=20 exercise appropriately alter the accumulation = of=20 dolichol; this putative biomarker of aging = appears to=20 be applicable to different tissues across = mammalian=20 species including human; biomarker is = applicable to=20 the hypothalamic digoxin-mediated model for = trisomy 21=20 (Kurup & Kurup, 2003). Reliable changes in = tissue=20 dolichol levels are seen in relatively short = intervals=20 of time compared to life-time, and levels can = be=20 tested on a small amount of tissue without = causing=20 death of the animal, which are desirable = features of a=20 biomarker of aging. The age-associated = alteration in=20 dolichol levels is likely to reflect an = age-dependent=20 derangement of free radical metabolism in = membranes.=20 Useful applications are shown, which include a = study=20 of the effect of the donor-recipient = age-mismatch on=20 the biological age of the graft tissues, and = the=20 detection of gender-related differences in=20 aging.=20 Research=20 was supported in part by a grant of MIUR = (Ministero=20 Istruzione, Universita=92 e Ricerca)=20 Mooradian,=20 A.D. (1990) J Gerontol 45(8) B183-186.=20 Kurup R.K.,=20 Kurup, P.A. (2004) Pediatr Pathol Mol Med = 22(5)=20 411-422.=20 top=20
Bowen,=20 Richard	107.=20 LIVING AND DYING FOR SEX: A THEORY OF AGING = BASED ON THE=20 MODULATION OF CELL CYCLE SIGNALING OF = REPRODUCTIVE=20 HORMONES R. Bowen, C.=20 Atwood* Voyager Pharmaceutical = Corporation,=20 Raleigh, North Carolina, USA * Department of=20 Pathology, Case Western Reserve University, = Cleveland,=20 Ohio and School of Medicine, Section of = Geriatrics and=20 Gerontology, Department of Medicine, University = of=20 Wisconsin, Madison, Wisconsin, USA We = put forth=20 a new theory based on a novel definition of = aging - any=20 change in an organism over time. This definition = includes not only the changes associated with = the loss=20 of function (i.e. senescence), but also the = changes=20 associated with the gain of function (growth and = development). Using this definition, the rate of = aging=20 would be synonymous with the rate of change. The = rate of=20 change/aging is most rapid during the fetal = period when=20 organisms develop from a single cell at = conception to a=20 multicellular organism at birth. Therefore, = =93fetal=20 aging=94 would be determined by factors = regulating the=20 rate of mitogenesis, differentiation, and cell = death. We=20 suggest these factors also are responsible for=20 regulating aging throughout life. Thus, whatever = controls mitogenesis, differentiation and cell = death=20 must also control aging. Since life-extending = modalities=20 consistently affect reproduction, and = reproductive=20 hormones are known to regulate mitogenesis and=20 differentiation, we propose that aging is = primarily=20 regulated by the hormones that control = reproduction=20 (hence, the Reproductive-Cell Cycle Theory of = Aging). In=20 mammals, reproduction is controlled by the=20 hypothalamic-pituitary-gonadal (HPG) axis = hormones.=20 Longevity inducing interventions, including = caloric=20 restriction, decrease fertility by suppressing = HPG axis=20 hormones and HPG hormones are known to affect = signaling=20 through the well-documented longevity regulating = GH/IGF-1/PI3K/Akt/Forkhead pathway. This is = exemplified=20 by genetic alterations in C. elegans where = homologues of=20 the HPG axis pathways, as well as the daf-2 and = daf-9=20 pathways, all converge on daf-16, the homologue = of human=20 Forkhead that functions in the regulation of = cell cycle=20 events. In summary, we propose that the hormones = that=20 regulate reproduction act in an antagonistic=20 pleiotrophic manner to control aging via cell = cycle=20 signaling; promoting growth and development = early in=20 life in order to achieve reproduction, but later = in=20 life, in a futile attempt to maintain = reproduction,=20 become dysregulated top=20
Chacon,=20 M.A.	108.=20 UP-REGULATION OF FREE FATTY ACIDS - A POTENTIAL = MASTER=20 SWITCH OF CALORIC RESTRICTION = PROTECTION P.=20 Wills¹, H. Brown-Borg²,and = M.A.=20 Chacon¹(P) = ¹Irazu=20 Biodiscovery, LLC, Baltimore, MD 21224;=20 ²Department of Pharmacology, = Physiology and=20 Therapeutics, University of North Dakota School = of=20 Medicine and Health Sciences, Grand Forks, ND=20 58203. Seventy years=20 of scientific investigation has demonstrated = that=20 caloric restriction (CR) without malnutrition is = the=20 only experimental intervention that can extend = life=20 span, and decrease the incidence and delay the = onset of=20 pathologies associated with aging. = Nevertheless,=20 the precise mechanism by which caloric = restriction=20 induces its life-extending effect remains poorly = understood. One consequence of caloric = restriction=20 observed in experimental rats, which has not = been=20 previously investigated, is an elevation in = serum levels=20 of free fatty acids (FFA). In our studies, we = found that=20 calorie restricted rats, mice, and dwarf mice = all have=20 elevated free fatty acids. In addition to = their=20 well-characterized role as an energy substrate, = recent=20 scientific studies have demonstrated that these=20 molecules possess signaling properties. = Therefore,=20 we tested the hypothesis that free fatty acids = may=20 induce (mimic) some of the protective effects = seen in=20 calorie restricted animals. Using an in vitro,=20 cell-based ROS assay, we demonstrated that=20 supplementation of serum from ad lib animals = with free=20 fatty acids to levels comparable to those found = in the=20 serum of caloric restricted animals conferred = protection=20 from the cytotoxic effects of H2O2. We = contend=20 that the up-regulation of FFA and their CoA = derivatives,=20 through their effects on ion channels, oxygen=20 utilization, and gene expression, offer a = plausible=20 mechanism for the protective phenotype = associated with=20 CR. top=20
Chai, = Weihang	48. =20 SENESCENT NORMAL HUMAN FIBROBLASTS MAINTAIN = THEIR 3'=20 TELOMERIC OVERHANG LENGTH W. Chai, J.W. = Shay, and=20 W.E. Wright Department of = Cell=20 Biology, University of Texas Southwestern = Medical=20 Center, 5323 Harry Hines Blvd., Dallas, TX=20 75390-9039 Replicative senescence = involves=20 progressive telomere shortening but the exact = molecular=20 mechanisms triggering the growth arrest are = poorly=20 understood. Recently, using a = telomere-oligonucleotide=20 ligation assay (T-OLA), it was reported1 that = the 3=92=20 G-rich overhang was eroded at senescence, = leading to the=20 hypothesis that loss of the 3=92 G-rich overhang = is the=20 molecular signal that triggers senescence. To = better=20 understand the molecular mechanisms triggering=20 replicative senescence, we have developed a = quantitative=20 assay to measure the length of the G-rich 3=92 = telomeric=20 overhangs from cultured cells. By using this = assay and=20 the conventional non-denaturing hybridization = assay for=20 measuring G-rich overhangs, we show that normal = human=20 fibroblasts maintain their 3=92 G-rich overhangs = at=20 senescence. Furthermore, cells maintain their = G-rich=20 overhangs when they bypass senescence after the=20 inactivation of p53 and retinoblastoma proteins = by human=20 papillomavirus type 16 (HPV) oncoproteins E6 and = E7=20 (E6/E7). Interestingly, cells expressing just = HPV E7=20 exhibit longer overhangs when they growth = arrest. We=20 also found that expression of simian virus 40 = large=20 T-antigen induces some telomeric overhang = shortening.=20 These results demonstrate that a significant = reduction=20 in overhang length is not the molecular signal = that=20 triggers senescence. Reference: 1. = Stewart,=20 S.A., I. Ben-Porath, V.J. Carey, B.F. O'Connor, = W.C.=20 Hahn, and R.A. Weinberg. 2003. Erosion of the = telomeric=20 single-strand overhang at replicative = senescence. Nat=20 Genet 33: 492-6. top=20
Chow, Vivian W.H.	49. =20 APOPTOTIC DNA FRAGMENTATION IN THE BRAINS OF = YOUNG AND=20 AGED eNOS-, iNOS- and nNOS-KNOCKOUT MICE: = MEASUREMENT=20 BY NEW ULTRASENSITIVE CE-LIF = TECHNIQUE=20 =20 VW Chow,=20 RR Fiscus, SB Chew=20 =20 The=20 Department of Physiology, The Chinese = University of=20 Hong Kong, Shatin, N.T. Hong Kong.=20 =20 Previously,=20 we showed that elevation of cGMP levels in = PC12 cells=20 exposed to atrial and brain natriuretic = peptides (ANP=20 & BNP) inhibits onset of apoptosis and = promotes=20 survival [1]. We further showed that = 24h-preincubation=20 of NG108-15 cells with ANP protects against=20 pro-apoptotic effects of nitric oxide (NO) = donor (high=20 concentrations) [2]. At lower physiological=20 concentrations, NO, via cGMP elevations, = inhibits=20 apoptosis and improves neural survival [3]. = Depending=20 on the source of NO, whether from nNOS, iNOS = or eNOS,=20 and depending on age, NO may promote or = protect=20 against apoptosis. The present study = determines if=20 aging-induced-apoptosis in brain is affected = by=20 genetic deletion of nNOS, iNOS and eNOS using = nNOS-,=20 iNOS-, eNOS-knockout mice. Young = (2-4-month-old) and=20 aged (12-22-month-old) nNOS-/-, iNOS-/-, = eNOS-/-=20 knockout and control mice (C57BL/6J & = B6129SF2/J)=20 were used to determine levels of apoptosis in=20 half-brain and different brain regions by a = new=20 ultrasensitive technique using capillary=20 electrophoresis with laser-induced fluorescent = detector (CE-LIF) to accurately quantify = apoptotic-DNA=20 fragmentation. Using half-brain, amounts of=20 apoptotic-DNA-fragments were 11-fold higher in = aged=20 than in young B6129SF2/J (nNOS-/-control) mice = (p<0.0001). Apoptotic-DNA-fragments in = young=20 nNOS-/- were 23-fold higher than in young = B6129SF2/J=20 (p<0.0001). Apoptotic levels in aged = B6129SF2/J=20 were 2.5-fold higher than in aged nNOS-/-=20 (p<0.001). These data suggest that nNOS = contributes=20 to aging-dependent-increase in brain apoptosis = of=20 B6129SF2/J, but provide neuroprotective effect = in=20 young adults. Neuroprotective effects of nNOS = were=20 especially large in young hippocampus.=20 Apoptotic-DNA-fragments in young iNOS-/- were = 30-fold=20 higher than in young C57 controls = (p<0.001). Aged=20 iNOS-/- showed 6.5-fold lower level of = apoptosis=20 compared to young adult (p<0.001). iNOS had = anti-apoptotic effect in young C57 but not = aged C57.=20 eNOS showed significant neuroprotective effect = in=20 medulla oblongata of both young and aged C57. = All NOS=20 isoforms provided neuroprotective function in = young=20 adult brain, but this neuroprotection appeared = change=20 during aging.=20 1. =20 Fiscus RR, Tu AW, Chew SB. Neuroreport=20 2001;12:185-189.=20 2. =20 Cheng Chew SB, Leung PY, Fiscus RR. Histochem = Cell=20 Biol. 2003;120:163-171.=20 3. =20 Fiscus RR. Neurosignals = 2002;11:175-190.=20 Support:=20 Competitive Earmarked Grant from RGC of Hong = Kong to=20 RRF. (#CUHK 4169/02M)=20 =20 top*=20
Citron, Bruce	50. GENE EXPRESSION = CHANGES IN=20 NEURODEGENERATIVE MICE AND CULTURED=20 CELLS =20 B.A.=20 Citron^1,2, A. = Surguchov^3,4, I.=20 Surgucheva^3,4, and B. W.=20 Festoff^3,4,5 ¹Lab = of=20 Molecular Biology, VA Medical Center, Bay = Pines, FL,=20 ²Dept. of Biochemistry & = Molecular=20 Biology, USF College of Medicine, Tampa, FL,=20 ³Neurobiology Research Lab, VA = Medical=20 Center, Kansas City, MO, Depts. of=20 ⁴Neurology and = ⁵Pharmacology,=20 University of Kansas Medical School, Kansas = City,=20 KS A variety of genes have been = implicated in=20 late onset neurodegenerative disorders. = Several=20 reports linked synucleins to motor neuron = diseases=20 including ALS. Present in aggregates, = α-synuclein is=20 important for the long term survival of = neurons. The=20 role of γ-synuclein is less clear but = recent studies=20 indicate that it regulates other = neurodegenerative=20 factors. Synucleins modulate certain = proteases,=20 however their own proteolytic degradation may = be=20 essential in the process of formation of = inclusion=20 bodies. We have been studying cultured neurons = and=20 wobbler mice, which undergo an age dependent = loss of=20 motor neurons in the cervical region of the = spinal=20 cord and model sporadic ALS. We have measured=20 activation of transcription factors (with=20 electrophoretic mobility shift assays) in = wobbler=20 spinal cords and cultured neurons exposed to = insults.=20 Specifically, we found that activated = transcription=20 factors SP1 and NF-kB, but not AP1 are = increased in=20 wobbler mice and in motor neurons exposed to = apoptosis=20 inducing insults (thrombin or mechanical = injury). We=20 have also examined mRNAs levels with = Affymetrix gene=20 arrays. Genes responsible for CNS maintenance = (e.g.=20 myelin-associated oligodendrocytic basic = protein) are=20 downregulated in the wobbler spinal cord. = Ubiquitin=20 C-terminal hydrolase is also expressed at = lower levels=20 in the wobbler cord and this could affect the=20 ubiquitination of proteins that aggregate in = the=20 degenerating CNS. α-synuclein is = upregulated in the=20 wobbler compared to the wild-type littermate = spinal=20 cord. Directed elevation of α-synuclein, = γ-synuclein=20 or both, in transfected neurons resulted in a = marked=20 dysregulation of gene expression in several = functional=20 categories. For example, of the genes that are = significantly expressed in these cells, more = genes=20 involved in cell adhesion are upregulated = compared to=20 genes in several other categories. The over = and=20 underrepresented functional classes in gene = groups=20 upregulated and downregulated by α- = and/or γ-synuclein=20 introduction have been determined to obtain a = picture=20 of synuclein regulatory influences.=20 top*=20
Cohen, Haim=20 Y.	51. The = SIRT1=20 NAD+-dependent deacetylase regulates = Bax-mediated=20 apoptosis Haim Y. Cohen, Christine = Miller,=20 Kevin Bitterman and David A.=20 Sinclair. Department of Pathology, = Harvard=20 Medical School. Boston MA USA = 02115. Decreased=20 apoptosis and increased cell survival is = observed in=20 organisms whose lifespan has been extended by = calorie=20 restriction or genetic manipulation. A crucial = but=20 poorly understood step in the stress-induced = apoptotic=20 pathway is activation of the pro-apoptotic = factor=20 Bax. In yeast, lifespan extension by = calorie=20 restriction requires Sir2, a = stress-responsive,=20 NAD+-dependent deacetylase. Here we show that = the=20 human Sir2 homolog, SIRT1, controls cell = survival in=20 response to stress by inhibiting Bax-mediated=20 apoptosis. SIRT1 deacetylates specific lysines = in the=20 Ku70 C-terminus, which increases the Ku70-Bax=20 interaction and prevents Bax from relocalizing = to=20 mitochondria. We show that this pathway can be = manipulated using small molecules and discuss = the=20 implications of this finding to the general = decline in=20 physiological function with age.=20 top*=20
Coumoul, Xavier	52. Modulation of IGF = members=20 expression in Brca1-knockout Mice X = Coumoul, L Cao and C Deng National = Institute=20 of Health, National Institute of Diabetes and=20 Digestive and Kidney Diseases, 9000 Rockville = Pike,=20 Bethesda MD 20892, USA Breast cancer = affects=20 200.000 women in the United States which = results in=20 40.000 deaths every year. Heterozygous = carriers of=20 Breast Cancer Associated Protein 1 (Brca1) = mutations=20 are at great risk to develop breast or ovarian = cancers=20 through their lifetime. Our mice models of = Brca1=20 deletions have shown that indeed lack of this = gene=20 plays a role in mammary tumorigenesis but also = interestingly in aging process. Here, we shown = that=20 deletion of Brca1 modulates expression of = several=20 Insulin-like Growth Factor (IGF) members in = mice=20 liver. IGF members expression has previously = been=20 linked to tumorigenesis and aging process.=20 Interestinlgy, IGF1 serum levels but also mRNA = liver=20 levels are increased as shown by microarrays = and=20 RT-PCR analysis. Other members like = IGF1-Receptor,=20 Insulin Receptor Substrate 1 or IGF-Binding = Protein 1=20 and 2 also appears to be modulated. We also = use a=20 human cell culture system UBR60, allowing = modification=20 of Brca1 expression, to shown that those = results are=20 reproducible with a different model. Those = results=20 allow us to hypothesize that modulation of IGF = members=20 expression by Brca1 could be one of the = mechanism that=20 links lack of this protein to aging and = tumorigenesis=20 process.=20 top*=20
de Grey, Aubrey D.	53. =20 THE=20 UNFORTUNATE INFLUENCE OF THE WEATHER ON THE = RATE OF=20 AGING=20 =20 A. de=20 Grey=20 =20 Department=20 of Genetics, University of Cambridge, Downing = Street,=20 Cambridge CB2 3EH, UK=20 =20 Much=20 research interest, and recently even = commercial=20 interest, has been predicated on the = assumption that=20 reasonably closely-related species -- humans = and mice,=20 for example -- should in principle respond to=20 aging-retarding interventions with an increase = in life=20 expectancy roughly proportional to their = control=20 lifespan (life expectancy without the = intervention).=20 Here it is argued that the best-studied = life-extending=20 manipulations of mice are examples of a = category that=20 is highly unlikely to follow this rule, and = more=20 likely to exhibit only a similar absolute = increase in=20 lifespan from one species to the next, = independent of=20 the species' control lifespan. That category = --=20 deprivation of dietary calories or of the = organism's=20 ability to metabolise or sense them -- is = widely=20 recognised to extend lifespan as an = evolutionary=20 adaptation to transient starvation in the = wild, a=20 situation which alters the organism's optimal=20 partitioning of resources between maintenance = and=20 reproduction. What has been generally = overlooked is=20 that the extent of the evolutionary pressure = to=20 maintain adaptability to a given duration of=20 starvation varies with the frequency of that = duration,=20 something which is -- certainly for = terrestrial=20 animals, and less directly for others -- = determined=20 principally by the weather. The pattern of = starvation=20 that the weather imposes is suggested here to = be of a=20 sort that will tend to cause all terrestrial = animals,=20 even those as far apart phylogenetically as = nematodes=20 and mice, to possess the ability to live a = similar=20 maximum absolute (rather than proportional) = amount=20 longer when food is short than when it is = plentiful.=20 This generalisation seems to be strikingly in = line=20 with available data, leading to the = biomedically and=20 commercially sobering conclusion that human=20 interventions which manipulate caloric intake = or its=20 sensing are unlikely ever to confer more than = two or=20 three years' increase in lifespan at the = most.=20 =20 top=20
Ding, Shi-Ying	54. =20 CRP LEVELS ARE STRONGLY ASSOCIATED WITH = AGING =20 =20 SY. Ding,=20 X.T. Tigno, and B.C. Hansen=20 =20 Obesity,=20 Diabetes, and Aging Animal Resource Center, = Department=20 of Physiology, School of Medicine, University = of=20 Maryland at Baltimore, 10 S. Pine St, = Baltimore MD=20 21201=20 =20 Epidemiological=20 studies have consistently shown that the = plasma level=20 of C-Reactive Protein (CRP), a marker of = inflammation,=20 is a strong, independent predictor of future=20 cardiovascular events, both in patients with a = history=20 of coronary heart disease (CHD) and in = apparently=20 healthy subjects. Inflammatory markers have = also=20 lately been implicated in aging and in type 2=20 diabetes. The plasma concentrations of CRP = were=20 determined by ELISA in a cohort of 33 diabetic = and=20 non-diabetic rhesus monkeys: 13 young normal = (Y) with=20 a mean age of 9.1=B12.9 yr, 11 old normal (O, = 25.9=B11.5=20 yr), 8 old type 2 diabetic monkeys (DM, = 26.2=B12.3 yr)=20 and 4 calorie-restricted (for more than 20 = years)=20 monkeys (CR, 30.1=B1 2.8 yr). Results show = that CRP=20 levels were positively correlated to age = (r=3D0.46,=20 p<0.01) and body weight (r=3D0.33, = p<0.02). The=20 mean levels of CRP were found to be highest = among the=20 O (246=B116=B5g/L), a value significantly = greater than=20 that among the Y (184=B114=B5g/L). However, = mean CRP=20 values did not differ significantly between O, = DM=20 (221.4=B118=B5g/L) and CR (214=B126=B5g/L), = all of whom were=20 elderly. The data suggests that the proportion = of the=20 variation in CRP that can be accounted for by=20 variation in age is 0.21, and that age is a = stronger=20 predictor of CRP than weight (R2=3D 0.18) or = metabolic=20 status, as reflected by circulating levels of = CRP in=20 the non-human primate.=20 top=20
Dolle,=20 Martijn	55. =20 AGE-RELATED GENOME INSTABILITY IN DNA = REPAIR-DEFICIENT=20 MICE M. Doll=E91, R. Busuttil2, L.=20 Niedernhofer3, G. van der Horst3, J. = Hoeijmakers3, S.=20 Wijnhoven1, P. Lohman1, H. van Steeg1, J.=20 Vijg2 1National Institute of Public = Health and=20 the Environment, Department of Toxicology, = Pathology and=20 Genetics, Bilthoven, the Netherlands; = 2University of=20 Texas Health Science Center, Department of = Physiology,=20 San Antonio, TX, USA; 3Erasmus University, = Department of=20 Cell Biology and Genetics, Rotterdam, the=20 Netherlands Genomic instability has been=20 implicated as a major cause of both cancer and = aging.=20 Previously we have shown that mutations = accumulate with=20 age in an organ-specific manner, using a = transgenic=20 mouse model with a chromosomally integrated lacZ = mutational reporter gene. To examine the = correlation=20 between age and mutation load, we are = investigating=20 genomic instability in aging DNA = repair-deficient mice.=20 Here we present our preliminary findings on = different=20 mouse mutants with deficiencies in nucleotide = excision=20 repair (NER) and DNA interstrand crosslink (ICL) = repair.=20 NER removes a broad range of lesions, such as = UV-induced=20 damage, other bulky adducts and some forms of = oxidative=20 damage. Life span studies and mutant frequency=20 determinations in liver and kidney were = performed on=20 male mice with defects in global and/or=20 transcription-coupled repair (Xpa, Csb or = Xpd(ttd)). We=20 observed a reduced life span for Xpa and = Xpd(ttd), as=20 compared to Csb and wild type control animals. = Conform=20 to our initial studies with just wild type mice, = all=20 NER-deficient models showed an age-related = increase of=20 mutant frequencies in liver and kidney. However, = only=20 the Xpa-mice showed elevated mutant frequencies = over=20 control animals in both organs. In addition, we = examined=20 mutant frequencies in liver of the very = short-lived=20 Ercc1(292) mouse. This mouse model lacks the = last 7=20 amino acids of Ercc1 and is deficient in both = NER and=20 DNA ICL repair. We observed a two-fold increase = in=20 mutant frequency over control mice at 23 weeks, = which is=20 close to their maximum life span. A comparison = of=20 mutation spectra in liver revealed that the = general=20 NER-deficiency of XPA-mice resulted in point = mutations,=20 predominantly consisting of one base pair = deletions. The=20 increased mutant frequency in the Ercc1(292) = mice was=20 due to both point mutations and translocations, = the=20 latter presumably reflecting the additional = cross-link=20 repair deficiency compared to = Xpa-mice. top=20
Falcon,=20 Alaric A.	56. DNA = EPISOMES AND=20 REPLICATIVE SENESCENCE IN S. = CEREVISIAE=20 =20 Alaric A.=20 Falc=F3n, Diego M. Ayo, Natalie Rios, and = John P.=20 Aris=20 =20 Department=20 of Anatomy and Cell Biology, Health Science = Center,=20 University of Florida, Gainesville, FL =20 32610-0235=20 =20 We=20 have shown that plasmids containing an = autonomously=20 replicating sequence (ARS; yeast DNA = replication=20 origin) reduce yeast replicative life span, = most=20 likely due to their accumulation during = replicative=20 aging (Plasmid accumulation reduces life span = in=20 Saccharomyces cerevisiae(Falc=F3n and Aris, = 2003). =20 This raises the question: how is plasmid = accumulation mechanistically linked to = replicative=20 senescence in yeast? One possibility is = that=20 accumulated DNA exerts an effect on other = cellular=20 DNA, perhaps by influencing DNA replication = and/or=20 inheritance. To explore this, we have = studied=20 =93interactions=94 between different types of = episomes in=20 yeast: recombinant plasmids, = extrachromosomal=20 rDNA circle (ERCs), and the = naturally-occurring 2=20 micron plasmid. A short-lived = sir2∆=20 strain that contains elevated ERC levels = transmits=20 recombinant ARS1-plasmids to daughter = cells=20 more often than a SIR2* control = strain. =20 However, an ARS1-CEN4-plasmid is = transmitted to daughter cells equally well in=20 sir2∆ and SIR2 = strains. Levels of=20 ARS1-plasmids are reduced in old = sir2∆=20 cells, compared to old SIR2 = cells. =20 Unexpectedly, an ARS1-plasmid, but not = an=20 ARS1-CEN4-plasmid, significantly = increased the frequency of loss of the 2 = micron=20 plasmid (i.e., curing of a cir+ strain to = cir0). Consistent = with this,=20 levels of the 2 micron plasmid are reduced in = old=20 cells compared to young cells. Thus,=20 accumulation of different types of nuclear = episomal=20 DNAs can have significant and unexpected = effects on=20 other nuclear DNAs. These findings = support a=20 relationship between episomal DNA = accumulation,=20 effects on cellular DNA metabolism, and cell=20 senescence in S. cerevisiae. =20 top=20
Fiscus, Ronald R.	57. =20 INHIBITOR=20 OF APOPTOSIS PROTEIN-2 (IAP-2) EXPRESSION = LEVELS ARE=20 DOWN-REGULATED BY CYCLIC GMP DEPLETION AND=20 UP-REGULATED BY CYCLIC GMP ELEVATIONS INDUCED = BY=20 NATRIURETIC PEPTIDES OR ANALOGS OF = CYCLIC GMP IN=20 NG108-15 CELLS: MOLECLULAR MECHANISM OF = CYCLIC=20 GMP/PROTEIN KINASE G (PKG)-MEDIATED=20 NEUROPROTECTION=20 =20 R.R.=20 Fiscus* and J.P. Yuen=20 =20 Dept.=20 Physiology (Fac. Medicine), Epithelial Cell = Biology=20 Res. Ctr. and Ctr. for Gerontology & = Geriatrics,=20 The Chinese University of Hong Kong, Shatin, = NT, Hong=20 Kong=20 =20 Previously,=20 we showed that cGMP elevations protect = hippocampal=20 neurons against glutamate toxicity [1], = inhibit=20 apoptosis and prolong cell survival in = stressed PC12=20 cells [2] and protect NG108-15 cells against = the=20 pro-apoptotic effects of nitric oxide (NO, = used at=20 high/neurotoxic concentrations mimicking = pathogenesis=20 of Alzheimer=92s and Parkinson=92s disease) = [3]. =20 Activation of cGMP/PKG signaling pathway in = neurons=20 may represent an important physiological = mechanism=20 protecting against neurotoxicity and = neurodegeneration=20 (reviewed [4]). Even basal levels of = cGMP appear=20 sufficient (and necessary) to prevent = spontaneous=20 development of apoptosis in some neural cells = (e.g.=20 NG108-15 cells), because depletion of cGMP by = ODQ=20 triggers apoptosis [4]. Recently, cAMP = was shown=20 to inhibit apoptosis in intestinal epithelial = cells=20 via protein kinase A (PKA)-mediated = phosphorylation of=20 CREB and increased IAP-2 expression [5]. = Because=20 PKG phosphorylates CREB at the same site as = PKA [4],=20 we hypothesized that cGMP/PKG-mediated = neuroprotection=20 may involve IAP-2 expression. The = present study=20 found that ODQ (10-100 micromolar, 24 h), = which=20 lowered basal cGMP levels, greatly reduced = protein=20 content of IAP-2 (Western blot) in a=20 concentration-dependent manner in NG108-15=20 cells. Elevation of cGMP levels with = either=20 atrial or brain natriuretic peptides (ANP or = BNP,=20 10-100 nM) or with 8-pCPT-cGMP (1-100 = micromolar, a=20 cell-permeable direct activator of PKG), all = of which=20 by-pass the ODQ block, completely reversed the = ODQ-induced down-regulation of IAP-2. = Indeed,=20 ANP and BNP up-regulated IAP-2 levels above = normal=20 control. The results suggest that = up-regulation=20 of IAP-2 expression in neural cells may = represent one=20 of the molecular mechanisms that mediates=20 cGMP/PKG-induced protection against = apoptosis. =20 1. Barger et al (1995) J Neurochem = 64:2087-2096, =20 2. Fiscus et al (2001) NeuroReport = 12:185-189, =20 3. Chew et al (2003), Histochem Cell Biol=20 120:163-171, 4. Fiscus (2002) = NeuroSignals=20 11:175-190, 5. Nishihara et al (2003) = PNAS=20 100:8921-8926. (Support: Grant # = CUHK4169/02M=20 from Research Grants Council of Hong Kong to=20 RRF)=20 top=20
Gemma, = C	58. =20 nINHIBITION OF CASPASE 1 IMPROVES CONTEXTUAL = FEAR=20 CONDITIONING IN AGED RATS C. = Gemma=20 (P), M. Fister, C. Hudson, P.C. = Bickford=20 James A Haley=20 VAH, Tampa Florida and Center for Aging and = Brain=20 Repair, USF, 12901 Bruce B. Downs BLVD, Tampa, = FL=20 33612 An = interplay=20 between the immune system and the central = nervous system=20 may underlie the neuropsychological changes = occurring=20 with aging. It has been long accepted that = brain=20 levels of certain cytokines increase as a = function of=20 age, even in absence of a pathologic = stimulus. In=20 aged rats there is an increase in IL1b that has = been=20 implicated in declines of synaptic plasticity in = the=20 hippocampus and performance on cognitive tasks. = IL-1b=20 receptor expression is high in the hippocampus, = an area=20 of the brain, which plays a pivotal role in = memory and=20 learning, suggesting that the effects of IL-1b = may be=20 specific to hippocampal-dependent memory=20 processes. However, although at = pathophysiological=20 levels IL-1b produces detrimental effects on = learning=20 and memory processes, under physiological = circumstances=20 IL-1b seems to be required for normal learning = and=20 memory processes. IL-1b is a = proinflammatory=20 cytokine initially synthesized in an inactive = precursor=20 form that is cleaved to generate the biological = mature=20 17kDa form by a protease named caspase-1. = In the=20 present study we chronically inhibit the = cleavage of=20 IL-1b using a specific inhibitor of=20 Caspase-1(Ac-YVAD-CMK, 10 pmol), both in old (22 = months)=20 and young (4 months) rats, and observe the = effect on=20 contextual fear conditioning paradigm. = Ac-YVAD-CMK=20 was delivered for 28 days icv through a brain = infusion=20 cannula connected to an osmotic minipump (Alzet, = Model=20 2004 pumping rate, 0.25ml/h; total volume 200ml) = implanted subcutaneously. On day 20 the = animals=20 were exposed on contextual fear conditioning and = the=20 memory for context was tested on day 22. = Chronic=20 infusion of a specific Caspase-1 inhibitor in = aged rats=20 resulted in an improvement in the memory for=20 context. top*=20
Gorbunova,=20 Vera	59. =20 EXPRESSION OF HUMAN TELOMERASE (hTERT) DOES = NOT=20 PREVENT STRESS-INDUCED SENESCENCE IN NORMAL = HUMAN=20 FIBROBLASTS BUT PROTECTS THE CELLS FROM = STRESS-INDUCED=20 APOPTOSIS AND NECROSIS Vera=20 Gorbunova¹(P), Andrei = Seluanov¹,=20 and Olivia M. = Pereira-Smith² ¹Verna and=20 Marrs McLean Department of Biochemistry and = Molecular=20 Biology, Baylor College of Medicine, Houston, TX = 77030 ²Department of=20 Cellular and Structural Biology, Sam and Ann = Barshop=20 Center for Longevity and Aging Studies, = University of=20 Texas Health Science Center, San Antonio, TX=20 78229-3900 Cells=20 subjected to sub-lethal doses of stress such as=20 irradiation or oxidative damage enter a state = that=20 closely resembles replicative senescence. = What=20 triggers stress-induced premature senescence = (SIPS) and=20 how similar this mechanism is to replicative = senescence=20 are not well understood. It has been = suggested=20 that stress-induced senescence is caused by = rapid=20 telomere shortening resulting from DNA = damage. In=20 order to test this hypothesis directly, we = examined=20 whether overexpression of the catalytic subunit = of human=20 telomerase (hTERT) can protect cells from = SIPS. We=20 therefore analyzed the response of four = different lines=20 of normal human fibroblasts with and without = hTERT to=20 stress induced by UV, -irradiation, and=20 H2O2. SIPS was induced with the same = efficiency in=20 normal and hTERT-immortalized cells. This = suggests=20 that SIPS is not triggered by telomere = shortening and=20 that nonspecific DNA damage serves as a signal = for=20 induction of SIPS. Although telomerase did = not=20 protect cells from SIPS, fibroblasts expressing = hTERT=20 were more resistant to stress-induced apoptosis = and=20 necrosis. We hypothesize that healing of = DNA=20 breaks by telomerase inhibits the induction of = cell=20 death, but because healing does not provide = legitimate=20 DNA repair, it does not protect cells from=20 SIPS. top=20
Guerin, John C.	60. =20 ROCKFISH: RETARDING AGING WITH "NEGLIGIBLE=20 SENESCENCE"=20 =20 J.=20 Guerin=20 =20 Portland,=20 OR=20 =20 Field=20 observations have suggested for quite some = time that=20 certain fish, turtles and invertebrates have = extremely=20 long maximum lifespan potential. Age = validation=20 techniques have since confirmed these=20 observations. Negligible senescence is = defined=20 in part as no observable age-related increase = in=20 mortality rate or decrease in reproduction = rate after=20 maturity, and no observable age-related = decline in=20 physiological capacity or disease = resistance. =20 Recent data compiled on rockfish (genus = Sebastes) have=20 documented ages exceeding 200 years. = However,=20 many rockfish species are reported under 30 = years=20 maximum observed lifespan, raising the = intriguing=20 possibility of intra-species lifespan=20 comparison. The Centenarian Species and = Rockfish=20 Project has 14 total pilot studies. The = three=20 studies reported in this poster are: = =93Rockfish liver=20 microarrays using existing 16,000+ zebrafish = gene=20 chips=94, a collaboration between Glenn S. = Gerhard,=20 Dartmouth Medical Center, and Renee Malek, = TIGR (The=20 Institute for Genomic Research); =93Heat Shock = Protein=20 comparison between younger and older = rockfish=94,=20 Marcelle Morrison-Bogorad, Associate Director, = Neuroscience and Neuropsychology of Aging = Program,=20 NIA; and =93Electron transport abnormalities = and=20 mitochondrial DNA mutations in rockfish heart = tissue=94,=20 Judd Aiken, University of Wisconsin-Madison. = top=20
Halaschek-Wiener, = Julius	61. ANALYSIS OF LONG = LIVED C.=20 ELEGANS DAF-2 MUTANTS USING SERIAL ANALYSIS OF = GENE=20 EXPRESSION J Halaschek-Wiener, S = McKay, S=20 Jones, M Marra, D Riddle, A=20 Brooks-Wilson Genome Sciences Centre, = BC Cancer=20 Agency, 600 West 10th Avenue, Vancouver, BC = V5Z 4E6,=20 Canada Serial Analysis of Gene = Expression=20 (SAGE) was used to identify = longevity-associated genes=20 in a long-lived daf-2 C. elegans mutant. The = daf-2=20 gene encodes an insulin/IGF-1 receptor-like = protein=20 and mutations therein lead to a 100% increase = in mean=20 life span. SAGE is a method to efficiently = count large=20 numbers of mRNA transcripts by sequencing = short tags.=20 We prepared adult C. elegans SAGE libraries of = days 1,=20 6 and 10 for daf-2 and days 1 and 6 for = control worms.=20 Analyses of gene expression profiles within = daf-2=20 libraries and between daf-2 versus control = SAGE=20 libraries (day 6 vs. 6 and day 10 vs. 6) = identified=20 not only single genes but also whole gene = families=20 that were differentially regulated. = Furthermore, daf-2=20 mutants at day 6 show a strong hypometabolic = phenotype=20 when compared to equally aged control worms = that=20 diminishes at advanced age. Identified gene = families=20 regulate important metabolic processes = including=20 stress response, lipid-, DNA/RNA-, protein, = and energy=20 metabolisms as well as intracellular signaling = and=20 cell structure. Identical expression patterns = of=20 various members of several gene families = emphasize the=20 importance of these types of genes in=20 longevity-related processes. Our results = suggest that=20 long-lived daf-2 mutants are severely = hypometabolic in=20 mid-life and we present evidence that specific = gene=20 families are involved in these metabolic=20 changes.=20 top*=20
Harper, James M.	62. =20 SERUM IGF-I AND LIFESPAN IN CROSSES BETWEEN = WILD AND=20 LABORATORY STOCKS OF MICE=20 J.M. Harper=20 (P)¹, S.N. Austad¹, R.A. = Miller¹, R.C.=20 Dysko² ¹Department=20 of Pathology and Geriatrics Center, University = of=20 Michigan School of Medicine,²Unit = for=20 Laboratory Animal Medicine, University of=20 Michigan =20 Using F2=20 hybrid stocks constructed as a cross between = two=20 stocks of wild-derived mice (Id and Ma) and = the=20 C57BL/6J inbred strain, we have assessed the = ability=20 of a suite of life-history and physiological = traits to=20 act as predictors of lifespan, as a prelude to = mapping=20 genes that may regulate lifespan in = mice. The Id=20 parental strain, the Ma parental strain, and = the (Id x=20 B6)F2 hybrid stock are all significantly = longer-lived=20 than a laboratory-derived control stock (DC),=20 generated as the offspring of (BALB/cJ x = C57BL/6J)F1=20 females and (C3H/HeJ x DBA/2J)F1 males = (log-rank test=20 p =A3 0.02). The lifespan of the (Ma x B6)F2 = hybrid=20 stock is indistinguishable from the control = (log-rank,=20 p =3D 0.84). Because serum IGF-I levels = measured=20 in 6-month old mice are lower in both the Id = and Ma=20 stocks than in the DC control (p < 0.001), = we=20 tested to see if IGF-I levels could predict = life=20 expectancy in mice of the two segregating F2=20 stocks. We found in each case that low = IGF-I=20 levels at 6 months of age were associated with = longer=20 life span (r2 =A3 0.19, p =3D 0.02). = IGF-I levels=20 did not predict life span in either of the=20 wild-derived parental stocks Ma and Id, nor in = the=20 laboratory control DC mice (p > 0.2 for = all). =20 These data suggest that endogenous levels of = early=20 life IGF-I may play a key role in the = regulation of=20 lifespan, and that there are polymorphisms = between at=20 least the Id and C57BL/6J stocks of mice that=20 influence lifespan. The QTL mapping of = the loci=20 responsible for these effects will provide = valuable=20 insight into the hormonal and genetic control = of=20 mammalian aging.=20 top=20
Kirchman, Paul A.	63. =20 ANALYSIS OF THE INFLUENCE OF MITOCHONDRIAL DNA = POINT=20 MUTATIONS ON LONGEVITY IN SACCHAROMYCES=20 CEREVISIAE=20 =20 P.=20 Kirchman=20 =20 Harriet L.=20 Wilkes Honors College, Florida Atlantic = University,=20 5353 Parkside Dr., Jupiter, FL 33458=20 =20 Evidence=20 from several species points to mitochondrial = DNA=20 (mtDNA) as one determinant of longevity. = Interpretation of these studies is hampered by = nuclear=20 DNA heterogeneity of the test subjects and, in = some=20 cases, by the variable environments in which = subjects=20 live(d). To eliminate variability in = both the=20 nuclear genome and the environment, the = budding yeast,=20 Saccharomyces cerevisiae, was used to examine = the=20 influence of mtDNA variation on=20 longevity. Mitochondrial = DNA=92s=20 containing point mutations in the COX1, COX3, = or CYTB=20 gene were transferred to a yeast strain that = lacks=20 mtDNA (rho0). The created strains vary = only in=20 the point mutations on the mtDNA. These = strains=20 were analyzed for variation in longevity = relative to a=20 control strain containing the non-mutated = mtDNA. =20 No statistically significant variation in = longevity=20 has been found between any of the strains=20 tested. To influence longevity, = variation in=20 mitochondrial DNA may need to be more = extensive than=20 the single nucleotide mutations tested. = Analysis=20 of additional mitochondrial mutant strains is = ongoing=20 and additional results will be = reported.=20 top=20
Lee, Yongwoo	64. =20 MICROARRAY ANALYSIS OF GENE EXPRESSION CHANGES = IN=20 INTERLEUKIN-4-STIMULATED HUMAN VASCULAR = ENDOTHELIAL=20 CELLS=20 =20 YW Lee, M=20 Toborek=20 =20 Department=20 of Surgery/Division of Neurosurgery, = University of=20 Kentucky College of Medicine, Lexington, = Kentucky=20 40536=20 =20 Oxidative=20 stress-mediated inflammatory reactions within = vascular=20 endothelium have been implicated in the = development of=20 age-related human diseases including = atherosclerosis.=20 We have reported that interleukin-4 (IL-4) can = induce=20 the pro-oxidative and pro-inflammatory = pathways in=20 human vascular endothelial cells. The cellular = and=20 molecular regulatory mechanisms underlying = this=20 process, however, are not fully understood. In = the=20 present study, we performed GeneChip = microarray=20 analysis to investigate global gene expression = patterns in human vascular endothelial cells = after=20 treatment with IL-4 using the Affymetrix = GeneChip=AE=20 Human Genome U133A Arrays, which contain more = than=20 22,000 human genes. Our results showed that = mRNA=20 levels of a total of 106 genes were = significantly=20 up-regulated and 41 genes significantly = down-regulated=20 with more than a twofold change. Majority of = these=20 genes are critically involved in the = regulation of=20 inflammatory responses, apoptosis, signal=20 transduction, transcription factors, = metabolism;=20 functions of the remaining genes are unknown. = The=20 changes in gene expression of selected genes = related=20 to inflammatory reactions such as VCAM-1, = E-selectin,=20 MCP-1 and IL-6 were verified by quantitative = real-time=20 RT-PCR and ELISA, respectively. IL-4 treatment = also=20 significantly increased the adherence of = inflammatory=20 cells to endothelial cell monolayers in a=20 dose-dependent manner. These results may help=20 determine the molecular mechanisms of action = of IL-4=20 in human vascular endothelium. In addition, a = better=20 understanding of IL-4-induced vascular injury = at the=20 level of gene expression could lead to the=20 identification of new therapeutic strategies = for=20 atherosclerosis. (This work was supported by = the=20 American Heart Association, Ohio Valley = Affiliate and=20 University of Kentucky Microarray = Facility =20 Program)=20 top*=20
Li, Hong	65. =20 GENDER DIFFERENCES IN NEURONAL CELL DEATH = AFTER OGD=20 INJURY AND NMDA-MEDIATED EXCITOTOXICITY IN = ORGANOTYPIC=20 HIPPOCAMPAL CULTURES=20 =20 Hong Li,=20 MD, Katrin Andreasson, MD, Louise McCullough, = MD,=20 PhD=20 =20 Department=20 of Neurology, Johns Hopkins University, = Baltimore,=20 Maryland, 21287=20 =20 Background=20 and Objective: Increasing evidence has = demonstrated=20 striking sex differences in outcomes after = acute=20 neurological injury. Females are less = vulnerable to=20 acute insults associated with experimental = cerebral=20 ischemia. It is believed that the greater=20 neuroprotection seen in females is due to = circulating=20 estrogens. However, this endogenous female=20 neuroprotection may not be due solely to = hormonal=20 influences. The objective of this study was to = determine if there are gender differences in = neuronal=20 cell death in organotypic hippocampal cultures = after=20 oxygen glucose deprivation (OGD) injury and=20 M-Methyl-d-Aspartic Acid (NMDA)-mediated=20 excitotoxicity. Methods: Organotypic = hippocampal slice=20 cultures were prepared from individual = postnatal day 8=20 Sprague=96Dawley rat pups. Gender genotyping = was=20 accomplished by PCR analysis from pup tail = DNA. The=20 cultures were exposed to 10 mM NMDA for 1 h or = subjected to OGD for 45 min, in presence or = absence of=20 30 mM 7NI (7 Nitroindazole; a selective nNOS=20 inhibitor), or 10nm 17b-estradiol (E2) for 7 = days=20 prior to OGD. Neuronal death was quantified = with=20 propidium iodide (PI). Data were analyzed by = one-way=20 ANOVA followed by Newman=96Keuls post hoc = test. Results:=20 Baseline neuronal cell death in control, 7NI, = and E2=20 treated groups was equivalent in males and = females.=20 Cell death markedly increased after = stimulation with=20 NMDA or OGD (P < 0.001) in both groups, but = there=20 was a significant increase in cell death in = male=20 cultures (P < 0.001 ~ 0.05) after both OGD = and NMDA=20 treatment. Treatment with7NI and E2 reduced = neuronal=20 damage induced by NMDA and OGD (P < 0.001 ~ = 0.05).=20 Interestingly, this effect was more notable in = males=20 than in females after OGD injury (P <0.01). = Conclusions: There is a consistent gender = difference=20 after OGD and NMDA toxicity, demonstrating=20 neuroprotection in female cells. Dimorphisms = in cell=20 survival may underlie enhanced neuronal = survival (or=20 decreased apoptosis) in female brain. top*=20
Lopez-Cruzan, Marisa	66. =20 ROLE OF MITOCHONDRIAL CASPASE-2 ACTIVITY IN = OXIDATIVE=20 STRESS-INDUCED APOPTOSIS=20 =20 Marisa=20 Lopez-Cruzan, Victoria Centonze, and Brian=20 Herman=20 =20 Cellular=20 and Structural Biology Department, University = of Texas=20 Health Science Center at San San Antonio. 7703 = Floyd=20 Curl Drive, San Antonio, Texas = 78229-3900=20 =20 The role of=20 mitochondrial caspase-2 in oxidative = stress-induced=20 apoptosis was examined by first determining if = mitochondria contain oxidative = stress-inducible=20 capase-2 activity and second, what the effect = of loss=20 of mitochondrial caspase-2 would be on=20 mitochondrial-specific oxidative = stress-induced=20 apoptosis. Oxidative stress-induced=20 mitochondrial-specific caspase-2 activation = and=20 apoptosis were assayed in murine NIH-3T3 = fibroblasts=20 or fibroblasts obtained from caspase-2 = knockout mice=20 using Florescence Resonance Energy Transfer=20 (FRET)/optical microscopy (mitochondrial = caspase-2=20 activity) and uptake of propidium iodide = (apoptosis).=20 Mitochondrially targeted FRET caspase-2 fusion = protein=20 containing the preferential caspase-2 = substrate=20 flanked by CFP and YFP, as donor and acceptor=20 fluorophores respectively, were used as an = in-situ=20 sensor of mitochondrial caspase-2 activity. = FRET=20 efficiency, measured using acceptor = photobleaching,=20 demonstrated a time-dependent loss of FRET = after=20 exposure of cells to tert-butyl hydroperoxide, = indicative of mitochondrial specific caspase-2 = activation. Inhibition of complex I or III of = the=20 mitochondrial electron transport chain is = known to=20 induce reactive oxygen species (ROS) within = the=20 mitochondria. Treatment with the complex I = inhibitor=20 rotenone, the complex III antimycin A or = staurosporine=20 (STS), a non-ROS inducer of apoptosis, = produced a=20 significant amount of cell death in = fibroblasts=20 isolated from wild type mice. However, cells = isolated=20 from caspase-2 deficient mice were highly = resistant to=20 rotenone and antimycin A, but not STS-induced=20 apoptosis. These results indicate that = mitochondria=20 contain oxidative stress-inducible caspase-2 = and that=20 caspase-2 activation is required for = mitochondrial=20 oxidative stress-induced apoptosis.=20 top=20
Mamczarz, Jacek A.	67. =20 PATTERN OF=20 GENE EXPRESSION IN LIVER OF YOUNG AND AGED = RATS AFTER=20 SHORT-TERM DIET RESTRICTION OR = 2-DEOXY-D-GLUCOSE=20 INJECTION: cDNA=20 =20 Jacek=20 Mamczarz, Min Zhu, Jonna Bowker, Kara Duffy, = Donald=20 Ingram=20 =20 Lab. Exp.=20 Gerontology, National Institute on Aging, NIH, = 5600=20 Nathan Shock Drive, Baltimore, MD 21224 = =20 Diet=20 restriction (DR) in rodents increases = lifespan,=20 reduces age-related disease and pathology, = increases=20 stress responses, and maintains better = function later=20 into life compared to conventional ad libitum = (AL)=20 feeding. We have been investigating DR regimen = and=20 also DR mimetics that stimulate stress = response=20 pathways that are activated by DR. By = inhibiting=20 glycolysis, feeding or injection of = 2-deoxy-D-glucose=20 (2DG) has been proposed as a DR mimetic. In = the=20 current study we investigated the pattern of = gene=20 expression using cDNA microarray in liver of = young (4=20 mo) and aged (26 mo) male Fischer-344 rats = subjected=20 to 3 weeks of 40% DR or 2DG injections once a = day. 40%=20 DR reduced initial body weight (BW) around 15% = in=20 young and 14% in old rats, respectively. 2DG = in the=20 dose of 125 mg/kg attenuated food intake and=20 developmental gains in BW in young rats, while = in aged=20 rats 2DG decreased BW and food intake below = initial=20 levels. 2DG at a dose of 250 mg/kg also = decreased BW=20 and food intake in both young and aged rats; = however,=20 in aged rats the decrease was at the level of=20 DR. In general, DR and 2DG treatment=20 up-regulated more genes in aged rats than = recorded in=20 young rats (around twice in DR, 6 times in = 2DG-125,=20 and 3.5 times more in 2DG-250). = Down-regulation of=20 gene expression was similar in young and old = DR rats,=20 twice higher in young 2DG-125, and twice lower = in=20 young 2DG-250, respectively, compared to aged = rats. DR=20 and 2DG-125 shared much more common patterns = of gene=20 expression in aged rats than in young, but for = 2DG-250=20 there was a generally common pattern shared in = both=20 age groups. Analysis of gene expression = patterns can=20 be used to assess potential effectiveness of = DR=20 mimetics.=20 top*=20
Martin, Rolf J.	68. =20 BLUEBERRY STUDY WEB SITES FOR COGNITIVE = PERFORMANCE=20 MEASUREMENT APPEAR SUFFICIENTLY PRECISE TO = DETECT A=20 0.1% CHANGE IN PERFORMANCE DURING = AGING=20 =20 R. Martin,=20 MMT Corp., Sherman, CT 06784, R.J. Coppings, = Lane=20 College, Jackson, TN 38301, K.E. Gerstmann, = NY, NY=20 10014, William Holme, Bethel, CT 06801, K. = Hull, New=20 Fairfield Senior Center, New Fairfield, CT = 06812, J.A.=20 Joseph, Human Nutrition Research Center on = Aging at=20 Tufts University, Boston, MA 02111, A.C. = Kokesh,=20 Charleston, WV 25301, B. Kristal, Weill = Medical=20 College-Cornell University Medical Center, NY, = NY=20 10021 and Burke Medical Research Institute, = White=20 Plains, NY 10605, M. Luzi and T. Millard, = Sherman IGA=20 Supermarket and B. Sachs, HR Herbs, Sherman, = CT 06784,=20 H.A. Raphaelson, Mansfield Senior Center, = Mansfield,=20 CT 06268, A. Pruchnicki, Mount Sinai Medical = Center,=20 NY, NY 10029, R. Schnoll, Brooklyn College, = Brooklyn,=20 NY 11210, and A. Wetherell, Defence Science = &=20 Technology Laboratory, UK. Contact email:=20 BlueberryStudy@aol.com=20 =20 1=20 Barlow Farm Road, Sherman, Connecticut = 06784=20 =20 To evaluate=20 methods for reducing age-associated cognitive = decline=20 during 3-year study periods, measurement = precision=20 should detect with 95% confidence differences = of 0.1%=20 or more between treatment and control groups = having=20 ~1% average annual decline. To develop such = high=20 precision for our 2000, 2002 and 2004 = blueberry=20 studies, self-calibrating web sites were = developed for=20 online measurement of 4-choice and 2-choice = decision=20 speed and immediate and delayed word recall = (patent=20 no. 6,712,615). These sites enable = participants to=20 measure their performance repeatedly and = thereby=20 obtain very precise average measures of these = aspects=20 of cognition. Annual test-retest reliability = values=20 for decision speed during 2000-2004 ranged = from 0.95=20 to over 0.99. Results collected from these = sites were=20 used to develop computer simulations which = indicate=20 that both our blueberry study design, with two = major=20 types of performance measurement (decision = speed and=20 recall), and Alex Comfort's (1970) approach to = measuring human aging, with 60 different types = of=20 physiological and performance measurement, are = capable=20 of identifying agents that can reduce a 1% = rate of=20 annual decline to 0.95% for studies involving = 10,000=20 or more participants. The major problem of = accurately=20 monitoring food and supplement intake over = long study=20 periods can be solved in part with supermarket = purchase logs and weekly food sharing and = spoilage=20 reports. Purchase logs received to date = indicate that=20 18 participants purchased 2,845 items at the = Sherman=20 IGA Supermarket with electronic discount cards = that=20 provide automatic "health discounts" of ~40% = on=20 blueberries and 35 other health-related foods. = Automatic discount cards can significantly aid = recruitment and retention and provide a = high-density=20 data stream to help examine whether local ad = campaigns=20 or health news reports cause unbalanced diet = or health=20 supplement shifts in either treatment or = control=20 groups. Blueberrystudy.com measurement and=20 diet-tracking software are expected to be = available to=20 other research groups later this year. =20 top*=20
Masternak, Michal M.	69. =20 THE EFFECT OF GROWTH HORMONE RECEPTOR KNOCKOUT = AND=20 CALORIC RESTRICTION ON EXPRESSION OF GENES = RELATED TO=20 INSULIN SIGNALING IN MICE=20 =20 Michal M=20 Masternak, Khalid A. Al-Regaiey, Michael S. = Bonkowski,=20 Andrzej Bartke=20 =20 Departments=20 of Internal Medicine and Physiology, = Geriatrics=20 Research, Southern Illinois University School = of=20 Medicine, 801 N. Rutledge, Springfield IL = 62794=20 =20 Growth=20 hormone receptor knockout (GHR-KO) mice are=20 long-lived, hypoinsulinemic and hypoglycemic = and=20 exhibit enhanced sensitivity to injected = insulin=20 (Ins). Using Real-time PCR (RT-PCR) we = analyzed=20 hepatic and muscle levels of insulin receptor = (IR),=20 insulin receptor substrate 1 (IRS1), IRS2, = peroxisome=20 proliferators-activated receptor gamma (PPARg) = glucose=20 transporter 4 (GLUT4), insulin-like growth = factor 1=20 (IGF1) and IGF1 receptor (IGF1R) mRNA = expression in=20 GHR-KO and in normal (N) animals from the same = strain.=20 We have also analyzed gene expression in = GHR-KO (KO)=20 and normal mice subjected to 30 % caloric = restriction=20 (CR). In the liver, gene expression was = affected by=20 genotype but not diet. The expression of IR, = IRS1 and=20 IRS2 mRNA was increased (P<0.0044, = P<0.024,=20 P<0.0003, respectively) in GHR-KO animals. = PPARg=20 mRNA expression was also increased in KO mice=20 (P<0.0042). However, IGF1 mRNA expression = in the=20 liver was decreased in GHR-KO mice as expected = (P<0.0001). Different effects were detected = in the=20 skeletal muscles of these animals. The mRNA = expression=20 was not affected by genotype but not by CR. = The=20 expression of IR, IRS1 and IRS2 was decreased = by CR in=20 N (P<0.02, P<0.0034, P<0.025, = respectively)=20 and in KO mice (P<0.0005, P<0.0003 and=20 P<0002, respectively). PPARg mRNA was = decreased by=20 CR in normal and KO animals (P<0.0096 and=20 P<0.0009, respectively). The expression of = GLUT4=20 mRNA in muscle showed significant down = regulated by CR=20 in both N and KO mice (P<0.0044 and = P<0.0003=20 respectively). The expression of IGF1 mRNA was = decreased by CR in normal and KO animals = (P<0.0053=20 and P<0.0001). Moreover, the expression of = IGF1=20 mRNA was reduced in KO vs. normal mice = (P<0.0001).=20 In contrast, the expression of IGF1R mRNA was = not=20 affected by CR in N mice, but was up regulated = in KO=20 in comparison to normal mice (P<0.014) and = reduced=20 by CR in KO mice (P<0.0005). Supported=20 by NIA=20 top=20
Mattison, Julie A.	70. THE INFLUENCE OF = SODIUM INTAKE=20 ON CARDIOVASCULAR RESPONSES IN RHESUS=20 MONKEYS J. A. Mattison, A. Bagrov, = H.=20 Spurgeon, P. Pullen, M. A. Lane, G. S. Roth, = D. K.=20 Ingram, E. G. Lakatta NIA, NIH Animal = Center,=20 Poolesville, MD 20837 Although there is = considerable evidence linking salt intake to=20 hypertension, how this dietary variable is = involved in=20 remodeling of the vascular wall to affect = arterial=20 stiffness is still unknown. The effect of an=20 incrementally increased salt load on vascular=20 stiffness and modulation of this response by=20 production of an endogenous ligand was studied = in nine=20 old normotensive male rhesus macaques (mean = age at=20 start =3D 19.1 +/- 0.67 years). Serial = measurements of=20 aortic pulse wave velocity (PWV), blood = pressure (BP),=20 urine and sodium output, and marinobufagenin = (MBG)=20 were made before and following each stepwise = increase=20 in dietary sodium chloride (NaCl) from 0.8% to = 6.0% of=20 daily intake. Sodium excretion significantly = increased=20 with each incremental increase in dietary = intake. PWV=20 significantly increased as a direct nonlinear = function=20 of Na intake independent of changes in = BP. Although=20 there was considerable variability among = monkeys in BP=20 response, there was an immediate pressure = increase=20 that was attenuated at higher dietary Na = loads. MBG=20 excretion was positively correlated to Na = excretion at=20 baseline and with a 1.7% and 2.6% NaCl diet. = This=20 direct relationship was not evident at intakes = of 3.5%=20 and 6% NaCl. These results suggest that old = monkeys=20 are susceptible to NaCl dependent vascular = changes=20 that can be partially compensated by increased = MBG to=20 enhance Na excretion.=20 top=20
Mounkes, Leslie C.	71. =20 ABNORMAL SPLICING OF LMNA CAUSES PROLIFERATIVE = DEFECTS=20 LEADING TO A HUTCHINSON GILFORD PROGERIA = SYNDROME=20 PHENOTYPE IN MICE=20 =20 Leslie C.=20 Mounkes, Lidia Hernandez, Serguei Kozlov, = Colin L.=20 Stewart=20 =20 Cancer=20 & Developmental Biology Laboratory, = National=20 Cancer Institute=97Frederick, MD =20 Hutchinson-Gilford=20 Progeria Syndrome (HGPS) is a rare genetic = disorder=20 resulting in the acceleration of age-related=20 phenotypes, including shortened stature, = craniofacial=20 disproportion, parchment thin skin, alopecia, = and=20 osteoporosis, with death predominantly due to=20 premature atherosclerosis in the early = teens1. =20 We derived mice carrying a splicing defect in = the Lmna=20 gene and found that mice homozygous for the = mutation=20 display defects remarkably similar to = progeria1. =20 Lmna encodes the A-type lamins, major = components of=20 the nuclear lamina with putative functions in = nuclear=20 structure, chromatin organization, gene = regulation,=20 and intracellular signaling. Mutant mice = (LmnaHGPS) showed marked reduction in growth = rate and=20 died by 4 weeks of age. Pathology of the = heart,=20 bone, muscle and skin are consistent with = phenotypes=20 associated with aging of these tissues. = The Lmna=20 mutation resulted in nuclear morphology = defects and=20 decreased lifespan of homozygous fibroblasts = from=20 postnatal, but not embryonic tissues, = suggesting a=20 developmentally regulated mechanism of = premature cell=20 death. Furthermore, in a background = deficient=20 for p16/p19ARF, the premature death of the = mutant=20 progeric cells is rescued in vitro, suggesting = the=20 nuclear lamina plays a role in the control of = cell=20 growth, possibly through pathways involving = pRB and/or=20 p53. Muscle myoblast cultures from = progeric mice=20 displayed a slower response to differentiation = cues=20 and made myotubes, which were shorter with=20 disorganized nuclei compared to myotube=20 differentiation of wild type myoblasts. = Defects=20 in proliferation and differentiation could = contribute=20 to the incomplete development of muscle and = other=20 mesenchymal tissues in progeria.=20 =20 1LC=20 Mounkes, S Kozlov, L Hernandez, T Sullivan = & CL=20 Stewart. A progeroid syndrome in = mice is=20 caused by defects in A-type lamins. = Nature 425:=20 298-301 (2003). top*=20
Overton, James M.	72. =20 PHYSIOLOGICAL RESPONSES TO VERY MILD AND = STANDARD=20 LEVELS OF CALORIC RESTRICTION IN = RATS=20 =20 J. M.=20 Overton, A.D. Parsons, S.A. Evans=20 =20 Department=20 of Nutrition, Food and Exercise Sci., Florida = State=20 Univ., Tallahassee, FL 32306-4340=20 =20 Very mild=20 caloric restriction (CR) is commonly used as a = control=20 intervention to assess the effects of more = substantive=20 CR on lifespan. The purpose of this = study was to=20 compare the physiological effects of = short-term, very=20 mild CR to standard levels of CR in = rats. Male=20 FBNF1 rats weighing about 360-380 g were = instrumented=20 with telemetry devices for measurement of mean = arterial pressure (MAP) and heart rate (HR) = and=20 continuously housed in room calorimeters for=20 assessment of oxygen consumption (VO2) and = respiratory=20 quotient (RQ). Rats were acclimated to=20 thermoneutral (TMN; 30=B0C) conditions prior = to imposing=20 very mild (5-10%) or standard levels of = caloric=20 restriction (CR; 40% restriction of ad lib) = for two=20 weeks. Controls continued to consume = food ad=20 libitum (5-6 rats/group). Over the = two-week=20 period, controls gained 15 grams, while mild = CR=20 reduced weight by about 3 grams and 40% CR = reduced=20 weight by 30 grams. As expected, 40% CR=20 significantly reduced body weight, VO2, HR and = RQ;=20 while increasing HR variability. = Unexpectedly,=20 mild CR, which had no effect on body weight, = RQ and HR=20 variability, produced significant reductions = in HR and=20 VO2. Weight maintenance produced 50% of = the=20 physiologic effect associated with = 40%CR. The=20 results suggest that in rats housed at=20 thermonuetrality, the cardiovascular and = metabolic=20 responses to CR are engaged by very mild = stimulus that=20 does not require weight loss.=20 top=20
Paik, David C	73. =20 DETECTION OF NOVEL CROSS-LINK COMPOUNDS FROM THE = NITRITE/COLLAGEN REACTION DC=20 Paik Department of Ophthalmology, = Columbia=20 University, New York, New York and Department of = Surgery, North General Hospital. New York, New=20 York Introduction: Increased collagen=20 cross-linking is one of the hallmarks of human = aging and=20 can contribute to the functional decline of = aging organ=20 systems. Nitrite ion, whose sources include = inflammation=20 and smoking, can mediate damaging nitration,=20 nitrosation, and hydroxylation reactions. The in = vitro=20 neutral pH reaction of nitrite with collagen = produces=20 changes that mimic collagen aging and include = increased=20 cross-linking, UV absorbance, and fluorescence. = The=20 current study was undertaken in order to = identify=20 covalent collagen cross-links produced = specifically by=20 reaction with nitrite. Methods: Model studies = were=20 performed using gelatin. Samples (10mg/ml) were=20 incubated with 0 to 200mM sodium nitrite at = neutral pH=20 for up to 10 days. The reaction was stopped with = ammonium sulphamate. UV/vis and fluorescence = spectra=20 were obtained on the protein mixtures. = Cross-link=20 enrichment was performed on acid hydrolysates = using=20 Skinner=92s (1982) cellulose mini-column method. = Isodesmosine was added as an internal standard=20 (0.5ug/mg) pre-hydrolysis. The water fraction = was=20 collected and analyzed by HPLC (C18) with diode = array=20 and fluorescence detection. Gradient conditions = using=20 20mM heptafluorobutyric acid and 2-20% = acetonitrile over=20 60 min were employed. Pump speed was 1ml/min. = Results:=20 Nitrite modification of gelatin produced = spectroscopic=20 changes similar to those observed previously for = collagen types I and IV and include increased UV = absorbance ca. 350nm and increased fluorescence = with=20 excitation peaks ca. 290 and 340, and an = emission peak=20 ca. 430. Cross-link analysis revealed 6 unique = products=20 of the reaction as detected by UV at 280nm. Five = of the=20 compounds also showed fluorescence = (ex290/em410). The=20 unknowns elute prior to pyridinoline and = isodesmosine.=20 Conclusions: This is the first identification of = cross-link compounds specific to the = nitrite/collagen=20 reaction. The detected compounds may serve as = biomarkers=20 of the reaction in future studies. top=20
Paredes, Daniel	74. NOREPINEPHRINE = RELEASE IN=20 CEREBELLUM DURING DELAY CLASSICAL EYEBLINK=20 CONDITIONING DECLINES IN AGED = RATS D. A.=20 Paredes, A.N. Samec, M. Fister and P.C.=20 Bickford Center for Aging and Brain = Repair,=20 USF, 12901 Bruce B Downs Blvd, Tampa, Fl=20 33612 Delay Classical Eyeblink = Conditioning is=20 an important model of associative, cerebellar=20 dependent learning. Aged animals and humans = show=20 deficits in learning on this paradigm. Our = laboratory=20 has been investigating the role of = norepinephrine (NE)=20 in this paradigm. We have demonstrated that = blocking=20 either the β-noradrenergic receptor = or=20 protein kinase A with local infusions into = cerebellar=20 lobule HVI and interpositus nucleus can = interfere with=20 learning. The goal of this study was to = determine=20 whether NE release is observed in HVI during = delay=20 eyeblink conditioning and if this is altered = in aged=20 rats. In vivo microdialysis coupled to = eyeblink=20 conditioning was performed on three month old = or=20 twenty month old F344 rats. Dialysate samples = were=20 collected every 10 minutes. After 5 baseline = samples=20 were collected rats received 50 training = trials (45=20 paired with a 12 psi airpuff and 5 tone only = [3kHz 85=20 dB] trials), or Rescorla type pseudo = conditioning.=20 Rats showed a significant release of NE during = and=20 after training, for young rats with 47.3 =B1 = 5.0 nM of=20 NE released as a max value reached 30 min = after=20 training started; while aged rats showed a max = of NE=20 release of 23.2 nM at 80 min after training = started.=20 This data clearly shows a decline on NE = release in=20 aged rats compared with young, this could = explain some=20 of the deficiencies of aged animals to learn = new motor=20 tasks which could be linked with the NE = reuptake=20 system deficiencies. Support Contributed = By: NSF=20 019674=20 top=20
Pendergrass, Bill R.	75. Mitochondrial = function,=20 organelle degradation, and cataract in lenses = from=20 aging: normal mice, GPX-1 KO mice, and Ghr-KO=20 mice. B Pendergrass,P Penn, A = Bartke, H Van=20 Remmen, and N Wolf 1Department of = Pathology,=20 University of Washington School of Medicine,=20 Department of Pathology, University of = Washington=20 School of Medicine, Seattle, WA. 98195.=20 USA Cataract incidence was increased=20 dramatically in old mice of each genotype = relative to=20 genotype matched young mice. Old Glutathione=20 Peroxidase-1 knock out mice (GPX-1 KO), had an = increased cataract incidence relative to = age-matched=20 controls. Older long lived growth hormone = receptor=20 knock out mice (GHr-KO mice) had significantly = less=20 cataract than age-matched controls. = Mitochondrial=20 oxidative cycling (a measure of O2 use) in = lens=20 epithelial cells (LECs) on the surface of = mouse lenses=20 was determined from the rate of oxidation of = reduced=20 Mitotracker red (H2MTR) to Mitotracker Red = (MTR). This=20 was significantly reduced in old compared to = young=20 control mice lenses. It was even more reduced = in old=20 GPX-1 KO mice lenses compared to age-matched = controls.=20 The reduction in oxidation was apparent = whether=20 measured relative to cell surface area, or to = DNA in=20 the cells. We also found a parallel decrease = in total=20 mitochondrial mass in old mouse LECs. The = relative=20 mitochondrial membrane potential (MMP) in the = LECs,=20 was determined from the fluorescence ratio of = the=20 vital stains Mitotracker Red (sensitive to = MMP) to=20 Mitotracker Green (sensitive to mitochondrial = mass).=20 This MMP ratio was slightly higher in older = LECs than=20 young LECs. Young GHr-KO mice had = significantly=20 reduced MMP relative to young controls, = possibly=20 indicating mild uncoupling of oxidative=20 phosphorylation. We also monitored the = degradation of=20 DNA and mitochondrial proteins using specific=20 fluorescent stains in internalized LEC fiber = cells.=20 The lens fiber cells in the interior of the = lens from=20 old mice were deficient in the ability to = degrade=20 nuclear and mitochondrial organelles. = Cataractous=20 areas of the lenses were especially heavily = stained=20 for DNA and mitochondrial debris. = Dichlorofluorescein=20 (DCF) staining of ROS was also significantly = increased=20 in lens fibers from old mice and this was = frequently=20 associated with cataractous foci in old = lenses.=20 top=20
Pepper,=20 Evan	76. =20 SOS-INDUCED DNA POLYMERASES ENHANCE LONG-TERM = SURVIVAL=20 AND EVOLUTIONARY FITNESS E. Pepper, B. = Yeiser, MF=20 Goodman, and SE Finkel Department of = Biological=20 Sciences, SHS 172, University of Southern = California,=20 Los Angeles, CA. 90089-1340 Escherichia = coli=20 encodes three SOS-induced DNA polymerases: pol = II, pol=20 IV, and pol V. We show here that each of these=20 polymerases confers a competitive fitness = advantage=20 during the stationary phase of the bacterial = life cycle,=20 in the absence of external DNA-damaging agents = known to=20 induce the SOS response. When grown = individually,=20 wild-type and SOS pol mutants exhibit = indistinguishable=20 temporal growth and death patterns. In contrast, = when=20 grown in competition with wild-type E. coli, = mutants=20 lacking one or more SOS polymerase suffer a = severe=20 reduction in fitness. These mutants also fail to = express=20 the =93growth advantage in stationary phase=94 = (GASP)=20 phenotype as do wild-type strains, instead = expressing=20 two additional new classes of GASP. These = polymerases=20 contribute to survival by providing essential = functions=20 to ensure replication of the chromosome and by=20 generating genetic diversity. top=20
Quanjian,=20 Yan	77. =20 NANOG KEEP EARLY EMBRYO AND TROPHOBLASTIC = INVINCIBILITY=20 FROM AGING Quanjian Yan=20 (P), Xiangmei Chen, Yumei Zhang, Daqiang Li, = Quan Hong,=20 Bo Fu. State Key Laboratory of Nephrology, = Kidney=20 Department of 301 Hospital Fuxing Road 28. = Haidian,=20 100853, Beijing. P.R.China. In = our=20 co-culture system, we found that early embryo = and=20 trophoblastic cell of KM mouse can invade = malignant=20 tumor cells and keep youth. Nanog is a newly = identified=20 homeodomain gene that functions to sustain the=20 pluripotency of embryonic stem cells. So we = investigate=20 whether Nanog is necessary for early embryo and=20 trophoblastic cell keep invincibility invasion = and=20 youth. Morphological analysis showed that = mouse=20 blastocysts have the invincibility of adhesion,=20 migration and invasion after implant in many = normal=20 cells or tumor cells. The normal cell strains = include=20 L929 and NIH3T3 mouse fibroblasts, 9HTE human = tracheal=20 epithelial cells and the ECV-304 human umbilical = vein=20 endothelial cells. The malignant tumor cell = strains=20 include differently histological origins, the = HNE1 human=20 nasopharyngeal carcinoma cells include BGC-823 = and=20 SGC-7901, human kidney carcinoma cells 786-0, = human=20 bladder carcinoma cells BIU-87 and BADM-60, = human=20 osteoblastoma cells Ros17/2.8 and Saos-2, human=20 endometrial carcinoma cells RL95-2, rat breast = carcinoma=20 cells SHZ-88, human hepatocarcinoma cell strains = HepG2,=20 SMMC-7721, QGY-7703 and MHCC97-H; human breast = carcinoma=20 cell strains T47D, MCF-7, ZR75-30, Bcap-37, = MDA-MB-231=20 and MDA-MB-435s. After 72 h of co-culture, = integrin=20 alpha V, FAK, MMP-9, Cyclin D1, CD44 protein and = Ki-67=20 mRNA were lower expressed in the trophoblastic = cells in=20 the group without the inner cell cluster or = Nanog=20 expression embryonic stem (ES) cell cluster with = immunocytochemical and in situ hybridization = than that=20 with Nanog expression ES cell cluster. = And the=20 outgrowth area in the group without the inner = cell=20 cluster or Nanog null ES cell cluster is smaller = than=20 that with the inner cell or Nanog = expression ES=20 cell cluster (P>0.05). These results=20 strongly suggest that Nanog activation keep the = inner=20 cell or ES cell cluster from aging, promote=20 trophoblastic adhesion, migration and invasion=20 (etc).
Rikke, Brad A.	78. =20 GENETIC=20 DISSECTION OF DIETARY RESTRICTION=20 =20 Brad A.=20 Rikke, Thomas E. Johnson=20 =20 University=20 of Colorado at Boulder=20 =20 Dietary=20 restriction (DR) is the most validated method = of=20 extending health and longevity n mammals, = extending=20 the life span of rodents by as much as = 50%. Many=20 physiological responses to DR have been put = forth as=20 causal factors underlying DR=92s = benefits. As a=20 first step towards testing some of these = factors,=20 we=92ve examined 83 strains of mice for = significant=20 differences in early responses to DR. = These=20 responses include lowered body temperature, = lowered=20 body weight, reduced growth rates (tail and = hair), and=20 reduced motor activity (runwheel and home = cage). =20 We=92ve also examined the extension of female = fertility=20 after extended DR. For each response, we = find=20 that there is a phenomenal amount of strain = variation,=20 with surprisingly little covariation among=20 responses. These findings could be used = to=20 critically test whether any of these responses = covary=20 with DR-induced longevity. Responses = validated=20 to covary with life extension could be used as = a tool=20 to screen for mutations that are likely to = affect=20 DR-induced longevity. In addition, we = are=20 mapping quantitative trait loci (QTLs) = specifying each=20 response by studying strains from the largest = murine=20 panel of recombinant inbred strains ever = developed,=20 the LXS panel of 77 strains. This = mapping makes=20 it possible to subsequently identify the genes = underlying these QTLs by positional cloning,=20 ultimately leading to a better understanding = of the=20 molecular mechanism by which DR extends = mammalian life=20 span. (Supported by the Ellison Medical=20 Foundation and NIH R01 AA11984)=20 top*=20
Perez-Perez, E.=20 M.	79. =20 ANTIOXIDANT CAPACITY OF HONEY TYPES ACCORDING TO = BOTANICAL AND ENTOMOLOGICAL = ORIGIN E. = M.=20 P=E9rez-P=E9rez¹, A. J.=20 Rodriguez-Malaver¹(P), P.=20 Vit²^{=20 1}Lab. de=20 Bioqu=EDmica Adaptativa, Dep. de Bioqu=EDmica, = Fac. de=20 Medicina; ²Dep. Ciencia de los = Alimentos,=20 Fac. de Farmacia y Bioan=E1lisis, Universidad de = Los=20 Andes, M=E9rida 5101, Venezuela. At = the=20 present, there is overwhelming evidence that = free=20 radicals cause oxidative damage to lipids, = proteins,=20 carbohydrates and nucleic acids. = Therefore,=20 reactive oxygen species (ROS) such as = O2-●,=20 OH●, or lipid peroxyl radical = (LOO●)=20 might lead to many biological complications = including=20 carcinogenesis, mutagenesis, = aging, atherosclerosis=20 and neurodegenerative diseases. Honey has been = known to=20 exert a significant in vitro antioxidant = activity, in=20 part due to its phenolic content. In this work, = we=20 studied the antioxidant capacity of 12 types of = honeys=20 from different places and floral origins, from = Apinae=20 and Meliponinae=20 bee subfamilies, on superoxide anion and = hydroxyl=20 radical. We found that honey samples inhibited = the=20 formation of superoxide anion and hydroxyl = radical,=20 under a range from 86 to 25% depending on the = sample.=20 This inhibition capacity was, for some samples, = higher=20 than quercetin and melatonin. On the other hand, = we=20 measured the antioxidant activity (AOA) values = of honey=20 samples in comparison to uric acid at 1 mM. In = this=20 case, honey samples had AOA values between 0.32 = and 0.84=20 Mm which were considerably high as compared to = melatonin=20 and quercetin. top=20
Perez-Perez, E.=20 M.	80. =20 ANTIOXIDANT CAPACITY OF VENEZUELAN BLACKBERRY = (RUBUS=20 BOGOTENSIS KUNTH) WINE P=E9rez-P=E9rez=20 EM¹, Rodr=EDguez-Malaver = AJ¹(P), Vit=20 P² = ¹Laboratorio=20 de Bioqu=EDmica Adaptativa, Departamento de = Bioqu=EDmica,=20 Facultad de Medicina, ²Departamento = Ciencia=20 de los Alimentos, Facultad de Farmacia y = Bioan=E1lisis;=20 Universidad de Los Andes, M=E9rida 5101,=20 Venezuela The = reported=20 health benefits of moderated wine drinking, = especially=20 in theprevention of chronic diseases associated = with=20 oxidative stress, may be related to the = antioxidant=20 activity of flavonols; quercetin and myricetin, = which=20 can readily reduce free radical by donating an = unpaired=20 electron. Rubus Bogotensis Kunth = (Rosaceae) is the=20 blackberry cultivar in the Venezuelan Andes, = where a=20 fermented product of the berries is marketed as=20 blackberry wine. We studied the antioxidant = capacity of=20 12 samples of blackberry wine produced in = M=E9rida state,=20 on the formation of superoxide anion and = hydroxyl=20 radical. Wine samples inhibited the formation of = superoxide anion and hydroxyl radical under a = range from=20 17% to 53%. For some wine samples, this = inhibition=20 capacity was close to those of quercetin (71%, = for=20 superoxide anion, 53% for hydroxyl radical) and=20 melatonin (79%, for superoxide anion, 54%, for = hydroxyl=20 radical). We also measured the antioxidant = activity=20 (AOA) value of wine samples in comparison to 1 = mM uric=20 acid. Wine samples had AOA values between 0.95 = and 0.80=20 mM, which were, for some samples, actually high = if were=20 compared to those of melatonin (0.83 mM) and = quercetin=20 (0.86 mM), used as reference in this study. This = antioxidant activity can be due to the phenolic = content=20 of wine samples, which are very efficient = scavengers of=20 free radicals because its molecular structure = includes=20 an aromatic ring with hydroxyl groups containing = mobile=20 hydrogens, and they have the capacity to reduce = or=20 chelate divalent ions which catalyze redox=20 reactions. top=20
Rodriguez-Malaver,=20 A.	81. =20 HONEY AND LUTEOLIN EFFECT ON STREPTOZOTOCIN, = SELENITE=20 AND OSMOTIC INDUCED CATARACTS P.=20 Vit¹,5, R. De Jesus², M.=20 Gudi=F1o³, E. = Perez⁴, A.=20 Rodriguez-Malaver⁴(P), A.=20 Melendez⁵ ¹Dep. Food=20 Science, Fac. Pharmacy, ² Dep. = Biology, Fac.=20 Sciences, ³BIOULA Animal House,=20 ⁴Dep. Biochemistry, Fac. Medicine,=20 Universidad de Los Andes, M=E9rida, Venezuela,=20 ⁵Dep. Physiology, Fac. Medicine, = National=20 University of Singapore, Singapore Ocular=20 cataracts are of epidemiological interest in = human=20 populations. The intraocular lens implantation = is a=20 successful technique to improve vision by = removing the=20 damaged ocular lens. However, a medical therapy = to=20 prevent, retard or reverse lens opacity is not=20 available. In the Neotropics, stingless bee=20 (Meliponinae) honey eyedrops are instilled to = treat=20 cataracts. Trigona Tetragonisca angustula=20 angustula honey is currently used for this = purpose,=20 therefore this honey type was harvested from a = hive and=20 refrigerated until use. Antioxidant capacity, = luteolin=20 and luteolin derivatives were detected in = previous=20 studies with honey extracts. From a screening = with=20 commercial flavonoids, luteolin tetra-methyl = ether,=20 luteolin 4=92-glucosyde, luteolin = 3=92-7-diglucosyde reduced=20 osmotic induced opacification in an ovine lens = culture=20 model assessed by digital image analysis. We = further=20 investigated two in vivo rat models of cataracts = induced=20 by streptozotocin and selenite, with slit lamp=20 evaluations. Luteolin tetra-methyl ether and = honey=20 eyedrops were applied before and after cataract = onset,=20 to explore preventive and therapeutic effects. A = preliminary 20 % of rats receiving the honey = therapeutic=20 treatment in the selenite model presented delay = of=20 opacification. top=20
Rosedale, = R.	82. = CLINICAL=20 EXPERIENCE OF A DIET DESIGNED TO REDUCE=20 AGING R=20 Rosedale, E Westman Rosedale=20 Center, Denver CO - = ronrosedale@comcast.net The=20 neuroendocrine theory of aging is associated = with=20 elevated levels of glucose, insulin and = leptin. =20 The objective of this study is to describe the = metabolic=20 effects of a nutritional program designed to = reduce=20 these correlates of aging. A retrospective = chart=20 review was performed of patients attending an = outpatient=20 metabolic management program utilizing = instruction in a=20 high-fat, adequate-protein, low-carbohydrate = diet, the=20 use of nutritional supplements, and periodic = individual=20 visits. The general dietary recommendation = was=20 approximately 15% carbohydrate, 25% protein, and = 60%=20 fat. Recommended sources of fat included = raw nuts,=20 avocados, olives and olive oil, flax and cod = liver=20 oil. The intake of protein was limited to = 1.0 -=20 1.25 grams/kg lean body mass per day (increased = for=20 exercise to 1.25 grams/day). Recommended = sources=20 of protein included sardines, fish, eggs, tofu, = poultry,=20 wild meats, non-fat cheeses (cottage, ricotta, = cream),=20 and seafood. Only non-starchy, fibrous = vegetables=20 were acceptable. Nutritional supplements=20 recommended daily were: L-carnitine 2000mg, = alpha-lipoic=20 acid 400mg, coenzyme Q10 100 mg, 1 tbsp cod = liver oil,=20 magnesium 300mg, potassium 300mg, vitamin C = 1000mg,=20 vitamin E 800mg, and a multivitamin. Medications = were=20 adjusted if needed. The mean duration of = follow-up=20 was 91.5 days (range 36-211). Thirty-one = patients=20 were identified with baseline and follow-up body = weight,=20 and fasting laboratory tests. The mean age = of=20 patients was 57.6 years, 53% were female. = Over a=20 mean follow-up of 91.5 days, body weight = decreased 8.2%=20 (p<0.01), fasting serum glucose decreased = 8.3%=20 (p=3D0.001). There were approximate 50% = reductions=20 in insulin, leptin, fasting serum triglyceride, = and=20 triglyceride/HDL ratio (p<0.001). Free = T3=20 decreased 7% (p<0.001), while TSH did not = change=20 significantly. We conclude that a = high-fat,=20 adequate-protein, low-carbohydrate diet with = nutritional=20 supplementation led to improvements in serum = factors=20 related to the aging process top=20
Ross, Ian K.	83. =20 MITOCHONDRIA, SEX, AND MORTALITY=20 =20 I.=20 Ross=20 =20 Dept. of=20 Molecular, Cellular, and Developmental = Biology=20 University=20 of California, Santa Barbara, CA = 93106-9610=20 =20 Is a=20 universal cause of senescence in healthy = organisms the=20 expression of both alleles of nuclear-coded = proteins=20 that form part of the = oxidative-phosphorylation=20 complexes of the mitochondria? Could this = explain why=20 mitochondria are inherited from only one = parent? =20 And why clones (e.g. triplets) die of = different,=20 random, causes? Prior to the evolution = of sex,=20 the endosymbiotic relationship between = mitochondria=20 and nuclear genomes would have selected = mechanisms=20 that maintained the optimum interaction = between the=20 two genomes. Once sex evolved, mating would = introduce=20 different, competitive, mtDNA and /or = nDNA gene=20 products which could well upset the = balance.The=20 selection of mechanisms, such as the specific=20 degradation of one mitochondrial genome, would = have=20 prevented part of such competition. Unlike = most=20 protein complexes in the cell, the proteins of = the=20 multi-enzyme complexes of the ox-phos system = are=20 derived from both nuclear-genome-coded genes = and=20 mitochondrial-genome coded genes. Minor = mutations in=20 either mtDNA or nDNA coding for these proteins = are=20 known to lead to major and catastrophic = diseases of=20 humans, suggesting that very tight and precise = interactions are required. To maintain the=20 evolutionarily established balance after = mating,=20 monoallelic expression of the nuclear-coded = genes=20 would be advantageous and prevent subtly = different=20 competitive proteins from interacting with the = resident mitochondria. This would require = regulation=20 of the expression of those specific nuclear = genes,=20 possibly under the control of the resident=20 mitochondria. I have postulated that nuclear = genes=20 coding for these ox-phos complex proteins are = normally=20 expressed monoallelically and that biallelic=20 expression of these genes with age results in = adverse=20 effects in different tissues and organs = leading to=20 senescence and mortality. I will be = presenting a=20 fungal model to explore this hypothesis that = as=20 already produced evidence of monoallelic = expression of=20 at least one of these genes in diploids = top=20
Sagie, Doron	84. THE = RELATIONSHIP=20 BETWEEN THE ACTIVITY IN "AMCHA" AND THE HEALTH = STATUS=20 OF ELDERLY HOLOCAUST SURVIVORS = =20 AbstractAuthors:=20 D.Sagie(P),=20 A. Biderman=20 =20 Department = of=20 Gerontology and Department of Family=20 Medicine*,Ben-Gurion University of the Negev, = Beer-Sheva, Israel; = dsagi@bgumail.bgu.ac.il=20 =20 About 40%=20 of the total elderly population in Israel, are = Holocaust survivors. Previous studies have = shown that=20 this population of elderly survivors, is in = great risk=20 for different health problems. "Amcha" =96 The = Israeli=20 center for Psychosocial support of Holocaust=20 survivors, was established in 1987, in order = to=20 alleviate the emotional suffering of elderly = Holocaust=20 survivors. The treatment involves building a = framework=20 of mutual aid, memory processing and grief = resolution.=20 The aim of the current study was finding a=20 relationship between being an active member in = "Amcha", and the improvement of different = health=20 measures that are supposedly purely = biological.=20 We sampled=20 a study group that included "Amcha" members, = and a=20 control group composed of Holocaust survivors = who had=20 similar socio-demographic and health status=20 background, but were not "Amcha" = members.=20 The results=20 indicated that "Amcha" members evaluated their = health=20 status more positively)F=3D5.64, p<0.05). = This=20 measure of self-evaluation was proved in = previous=20 studies to be strongly related with future = morbidity=20 and mortality. "Amcha" members also tended to = utilize=20 less health services (F=3D.70, p<0.05). = This=20 research contributes not only to social = sciences, but=20 also to health and biological studies. It = shows that=20 an emotional treatment of the elderly can be = related=20 to an improvement in their health status, = which is=20 commonly associated with medical-biological = factors=20 and treatments. =20 =20 A person who used to live in Europe in the = years=20 1939-1945, in the time of the Nazi = regime.=20 top*=20
Sagi, Orli	85. =20 EFFECT OF AGE AND LPS ON p69- SHCC INDUCTION = IN MOUSE=20 BRAIN =20 Sagi O.,=20 Wolfson M., Fraifeld V=20 =20 Department=20 of Microbiology and Immunology, CMRA, = Ben-Gurion=20 University of the Negev, Beer-Sheva, Israel;=20 fraifeld@bgumail.bgu.ac.il=20 =20 Recently,=20 Shc (Src homology 2 domain containing) = proteins were=20 implicated in the oxidative stress-induced = apoptosis=20 and longevity in mice. These proteins act as = adaptors=20 coupling activated receptors with tyrosine = kinase=20 activity to downstream signaling molecules, = including=20 the MAPK-associated pathway. The family of Shc = proteins includes ShcA, ShcB, and ShcC. ShcC = family=20 includes two isoforms of 55- and 69-kDa, and = their=20 expression appears limited to neural tissue. = The brain=20 is one of the targets for Lipopolysaccharide = (LPS), a=20 component of the cell wall of gram-negative = bacteria,=20 which is a classic trigger of inflammatory = reactions=20 accompanied by a generation of reactive oxygen = species=20 and activation of MAPKs. The response to LPS = is=20 altered with age. The present study was aimed = at=20 examining the expression of ShcC in the mouse = brain=20 with respect to animal=92s age and the effect = of LPS.=20 Male CD-1 mice aged 4 days to 12 months old, = were=20 injected intraperitoneally with 1 mg/kg LPS = and at=20 different times post-injection, their brains = were=20 removed for preparation of tissue extracts. = The latter=20 were processed through Western blot analysis = using=20 specific anti-ShcC antibodies. We found that = (i) the=20 basal level of ShcC in mouse brain increased = gradually=20 during the postnatal period, reaching the = maximum at 2=20 months. Middle-age mice had a lower level of = ShcC=20 compared to the young adult and young animals; = (ii)=20 both p55 kDa and p69 kDa isoforms of ShcC = displayed=20 similar age-related patterns; (iii) LPS = differentially=20 affects the expression of ShcC isoforms, = resulting in=20 a marked up-regulation of p69 and an = insignificant=20 effect on the level of p55; (iiii) the effect = of LPS=20 on ShcC was age-related, being more pronounced = in the=20 younger mice compared to the older animals. = =20 This study=20 was supported by a grant from the United = States-Israel=20 Binational Science Foundation (BSF), = Jerusalem,=20 Israel.=20 top*=20
Saretzki, Gabriele = C.	86. =20 A DNA DAMAGE CHECKPOINT-MEDIATED RESPONSE IN=20 TELOMERE-INITIATED SENESCENCE=20 =20 F. d=92Adda=20 di Fagagna, G. Saretzki, T. von Zglinicki, = S.P.=20 Jackson=20 =20 University=20 of Cambridge UK, The Wellcome Trust Sanger = Institute=20 Cambridge, UK =20 Replicative=20 senescence in human fibroblasts can be = triggered by=20 telomere uncapping, however, the signalling = pathways=20 leading to telomere-initiated activation of = cell cycle=20 checkpoints were not clear. = Specifically, it was=20 unknown whether uncapped telomeres could = signal arrest=20 directly or whether chromosomal breaks = following=20 telomeric fusions were main triggers of = senescence.=20 Here we=20 show that senescent human fibroblasts display=20 molecular markers characteristic of functional = DNA=20 damage foci including phosphorylated histone = H2AX in=20 colocalisation with DNA repair and DNA damage=20 checkpoint factors such as 53BP1, MDC1 and = NBS1.=20 Importantly, these senescence-associated DNA = damage=20 foci (SDF) associate preferentially with = telomeres=20 both after =91normal=92 telomere erosion and = after=20 telomere uncapping by induction of a = dominant-negative=20 TRF2. SDF remain present for many months in = senescent=20 cells. Presumably, they remain active because=20 inactivation of DNA damage checkpoint kinases = in=20 senescent cells can restore cell cycle = progression=20 into S phase. Thus, we=20 propose that telomere-initiated senescence = reflects a=20 DNA damage checkpoint that is activated with a = direct=20 contribution from uncapped telomeres. = Replicative=20 senescence appears to be a state that is = actively=20 maintained by sustained activation = of a=20 DNA damage response. Thus, DNA damage foci are = not=20 only indicators of acute DNA damage, but might = much=20 more resemble novel markers for the presence = of=20 senescent cells in vivo. =20 top*=20
Seluanov, Andrei	87. =20 DNA END JOINING BECOMES LESS EFFICIENT AND = MORE=20 ERROR-PRONE DURING CELLULAR = SENESCENCE=20 =20 A.=20 Seluanov, D. Mittelman, O. Pereira-Smith, J. = Wilson,=20 and V. Gorbunova=20 =20 Department=20 of Biochemistry and Molecular Biology, Baylor = College=20 of Medicine, Houston, Texas=20 =20 Accumulation=20 of somatic mutations is thought to contribute = to the=20 aging process. Genomic instability has = been=20 shown to increase during aging, suggesting an = aberrant=20 function of DNA double-strand break (DSB)=20 repair. Surprisingly, DSB repair has not = been=20 examined with respect to cellular = senescence. =20 Therefore, we have studied the ability of = young,=20 presenescent, and senescent normal human = fibroblasts=20 to repair DSBs in transfected DNA using a = fluorescent=20 reporter substrate. We have found that = the=20 efficiency of end joining is reduced up to 4.5 = fold in=20 presenescent and senescent relative to young=20 cells. Sequence analysis of end = junctions showed=20 that the frequency of precise ligation was = higher in=20 young cells, whereas end joining in old cells = was=20 associated with extended deletions. = These=20 results indicate that end joining becomes = inefficient=20 and more error-prone during cellular = senescence. =20 Furthermore, the ability to use = microhomologies for=20 end joining was compromised in senescent = cells,=20 suggesting that young and senescent cells may = use=20 different end joining pathways. We = hypothesize=20 that inefficient and aberrant end joining is a = likely=20 mechanism underlying the age-related genomic=20 instability and higher incidence of cancer in = the=20 elderly. top*=20
Shaikh, Aasef G.	88. =20 BLOCKADE OF=20 ION CONDUCTANCE IN AUDITORY NEURONS MIMIC = DISORDERED=20 CENTRAL PROCESSING IN AGED=20 A, Shaikh=20 (P); P, Finlayson Department=20 of Otolaryngology, Wayne State University, 550 = E=20 Canfield, Room # 327, Detroit, MI=20 48201 Presbycusis,=20 a senile hearing-loss, is an inevitable = sequelae of=20 aging. About one-third of Americans = older than=20 age 60 suffer from hearing disorders. In = addition, the elderly also have a diminished = ability=20 to understand time-compressed speech, which is = likely=20 due to a reduction in temporal resolving power = of=20 auditory neurons. Based on auditory brainstem=20 evoked-responses (ABR), affected auditory = temporal=20 processing in the aged has a central origin, = possibly=20 in Superior Olivary Complex (SOC). Yet the = precise=20 mechanism for its pathogenesis is not defined. = There=20 is mounting evidence that excitability in the = central=20 auditory neurons is affected in = presbycusis. A=20 range of ion conductances shape the auditory = neural=20 excitability.=20 = Hyperpolarization-activated-cationic-conductance (Ih)=20 is one of the predominant=20 conductances determining resting membrane = potential in=20 the SOC neurons and could regulate their = excitability.=20 In order to understand the pathophysiology of=20 disordered central processing in presbycusis, = it is=20 important to examine the influence of Ih on = the SOC=20 neural excitability. Auditory neurons also = exhibit an=20 important property, post-stimulatory = suppression, to=20 aid echo-suppression and thereby adequate = processing=20 of the time-compressed speech. As Ih is an = important=20 regulator determining the resting membrane = potential,=20 its role in the post-stimulatory suppression,=20 particularly in the SOC neurons needs to be=20 investigated. Here we document Ih has a = predominant=20 role in shaping the SOC neural excitability in = vivo.=20 We also report, selective blockade of Ih by = ZD7288=20 (100microM) significantly increases the = magnitude of=20 post-stimulatory suppression for shorter = inter-tone=20 intervals and raises the neural-response = threshold to=20 the second of the sequential-tones. As = Ih-blockade=20 decreases the neural excitability and enhances = the=20 post-stimulatory suppression for short = inter-tone=20 interval, in the SOC neurons; incompetent = function of=20 Ih could be a possible neurophysiological = correlate of=20 decreased amplitude of ABR = =93wave=94. top=20
Shaikh, Aasef G.	89. =20 THE VESTIBULAR PROSTHESIS IN POSTURAL CONTROL = AND=20 BALANCE=20 =20 A. Shaikh,=20 G. Auner=20 =20 Wayne State=20 University, Detroit, MI 48202=20 =20 In older=20 age groups, spatial disorientation and falls = are one=20 of the most common reasons leading to = morbidity,=20 mortality and seeking for a physician-help. = Bilateral=20 vestibular end-organ failure is a major cause = of=20 spatial disorientation, which, in aged = patients, is=20 mainly due to Meniere=92s disease, ototoxicity = or age=20 induced vestibular hair-cell loss. Idiopathic = vertigo=20 is also a frequent malady in geriatric age = group.=20 Treatment of vestibular end-organ failure is = still=20 unknown and existing therapeutic principles = for=20 chronic vertigo, such as central vestibular=20 suppression, are just symptomatic. We have = pioneered=20 the state-of-art technology that will enable = us to=20 treat the vestibular end-organ failure and = chronic=20 vertigo by means of vestibular = electrical-stimulation.=20 The novel, prototype sensor is devised, which = encodes=20 head-angular-position in real time and thus = computes=20 head-velocity, a primary function of the = semicircular=20 canals. The sensor is implantable in the = mastoid=20 antrum and has three degrees of freedom. The = sensor is=20 linked to the decoder, processor, and sixteen = channels=20 stimulator chip; which are implantable = subcutaneously=20 on the squamous temporal-bone. The information = encoded=20 by the sensor is processed to compute = component of the=20 head velocity in a plane of each of the three=20 semicircular canals. Results of this = computation are=20 further utilized to modulate frequency of = biphasic=20 square wave electrical pulses, which are = delivered by=20 the sixteen channels stimulator chip to = nano-electric=20 arrays. These arrays are developed using the = novel=20 =93Self-Assembled Nano Array Platform = Approach=94 and=20 provide the best resolution for the=20 artificial-nerve-stimulation. The arrays are = attached=20 to self-retaining probe adapting the shape of = the=20 ampullae and providing the electrode array a = close=20 contact with the neuroepithelia for = specificity of the=20 individual three-canal-stimulation. = Integration of the=20 sensor feedback with multiple similar sensors=20 implanted at different body axes provides = basis of=20 functional-neural-stimulation to reinforce the = muscular control of posture. top=20
Shaikh, Aasef G.	88. =20 BLOCKADE OF ION CONDUCTANCE IN AUDITORY = NEURONS MIMIC=20 DISORDERED CENTRAL PROCESSING IN = AGED=20 =20 A, Shaikh;=20 P, Finlayson=20 =20 Department=20 of Otolaryngology, Wayne State University, 550 = E=20 Canfield, Room # 327, Detroit, MI 48201 = =20 Presbycusis,=20 a senile hearing-loss, is an inevitable sequel = of=20 aging. About one-third of Americans older than = age 60=20 suffer from hearing disorders. In = addition, the=20 elderly also have a diminished ability to = understand=20 time-compressed speech, which is likely due to = a=20 reduction in temporal resolving power of = auditory=20 neurons. Based on auditory brainstem = evoked-responses=20 (ABR), affected auditory temporal processing = in the=20 aged has a central origin, particularly in = Superior=20 Olivary Complex (SOC). Yet the precise = mechanism for=20 its pathogenesis is not defined. There is = mounting=20 evidence that excitability in the central = auditory=20 neurons is affected in presbycusis. A range of = ion=20 conductances shape the auditory neural = excitability.=20 = Hyperpolarization-activated-cationic-conductance (Ih)=20 is one of the predominant conductances = determining=20 resting membrane potential in the SOC neurons = and=20 could regulate their excitability. In order to = understand the pathophysiology of disordered = central=20 processing in presbycusis, it is important to = examine=20 the influence of Ih on the SOC neural = excitability.=20 Auditory neurons also exhibit an important = property,=20 post-stimulatory suppression, to aid = echo-suppression=20 and thereby adequate processing of the = time-compressed=20 speech. As Ih is an important regulator = determining=20 the resting membrane potential, its role in = the=20 post-stimulatory suppression, particularly in = the SOC=20 neurons needs to be investigated. Here we = document Ih=20 has a predominant role in shaping the SOC = neural=20 excitability in vivo. We also report, = selective=20 blockade of Ih by ZD7288 (100microM) = significantly=20 increases the magnitude of post-stimulatory=20 suppression for shorter inter-tone intervals = and=20 raises the neural-response threshold to the = second of=20 the sequential-tones. As Ih-blockade decreases = the=20 neural excitability and enhances the = post-stimulatory=20 suppression for short inter-tone interval, in = the SOC=20 neurons; incompetent function of Ih could be a = possible neurophysiological correlate of = decreased=20 amplitude of ABR =93wave V=94 (decreased SOC = neural=20 activity) in aged patients with speech = understanding=20 problems.=20 top=20
Sharman, Edward H.	90. MODULATION OF MRNA = EXPRESSION=20 PATTERNS IN MURINE CNS BY AGE AND DIETARY = MELATONIN:=20 FOCUS ON IMMUNITY =20 E.=20 Sharman, V. Perreau, K. Sharman, C. Cotman, = and S.=20 Bondy Center for Occupational and = Environmental=20 Health, Department of Community and = Environmental=20 Medicine, and Institute for Brain Aging and = Dementia,=20 University of California Irvine, CA = 92697-1825, U. S.=20 A. Age-dependent decline in brain = immune=20 response is a component of several = neurodegenerative=20 diseases such as Alzheimer=92s disease, = Parkinson=92s=20 disease and Huntington=92s disease. To assess = the impact=20 of age on CNS immune response, gene array = analysis was=20 used to identify changes in functional = patterns and=20 levels of mRNA expression induced by i.p. = injection of=20 lipopolysaccharide (LPS) in the cerebral = cortex of=20 both young (4-month) and old (27 month) CB6F1 = male=20 mice. Modulation of the LPS-induced CNS immune = response by the pineal secretory product = melatonin was=20 assessed by supplementing the diet of aged = animals=20 with 40 ppm melatonin for 8 weeks preceding = LPS=20 injection. A wide range of age-related changes = in=20 basal levels of genes within the brain was = found.=20 Dietary melatonin administered to aged mice = failed to=20 reverse most of these changes. LPS treatment = elevated=20 the expression of many genes in the younger = animal.=20 However, with senescence, the response to such = an=20 inflammatory stimulus was attenuated. = Administration=20 of melatonin was able to restore this reaction = to a=20 level more closely resembling that of the = younger=20 animal.=20 top*=20
Shetty, Ritu A.	91. =20 SHORT-TERM COENZYME Q10 SUPPLEMENTATION = ENHANCES=20 COGNITIVE PERFORMANCE IN AGING = MICE.=20 =20 1) R.=20 A.Shetty=20 2)N.=20 Sumien.=20 3)R.Sohal.=20 4)M. J.=20 Forster.=20 =20 Dept of=20 Pharmacology and Neuroscience, UNTHSC, = FortWorth,=20 TX-76107. =20 Brain aging=20 is accompanied by an increase in oxidative = stress and=20 mitochondrial dysfunction. Coenzyme Q (CoQ) is = an=20 important component in the electron transport = chain=20 and is thought to serve as a potent = antioxidant. It is=20 also known that CoQ content declines with = aging and=20 the levels can be restored in older mice by=20 supplementation. However the effects of CoQ10=20 supplementation on behavior are not known. = Therefore=20 the purpose of this study was to determine the = short-term effects of CoQ10 supplementation on = psychomotor and cognitive performance in young = and old=20 mice. Separate groups of young (4 months) and = old mice=20 (18 months) were fed a control diet or a diet=20 supplemented with low or high concentrations = of CoQ10=20 for a period of 15 weeks. The low and high=20 supplemented diets yielded a daily CoQ10 = intake of=20 approximately 148- or 654-mg/kg, respectively. = After=20 6-weeks on the diets the mice were subjected = to a=20 battery of age-sensitive behavioral tests for=20 cognitive and psychomotor performance. These = tests=20 include locomotor activity, coordinated = running=20 performance, swim maze and startle response. = CoQ10=20 treated mice tended to exhibit faster learning = of the=20 swim maze task, an aging-sensitive measure of=20 cognitive performance. Diet supplementation = with CoQ10=20 also improved the ability of the old mice to = retain=20 information in swim maze task, an effect that = was not=20 evident in the younger mice. In tests for = auditory and=20 shock startle reflex and coordinated running = ability,=20 CoQ10 supplementation failed to improve = performance.=20 These results suggest that short-term CoQ10=20 supplementation improves impaired cognitive = function=20 in older mice, but fails to ameliorate = age-impaired=20 psychomotor function. top=20
Smith, Mark A.	94. =20 DIFFERENTIAL EXPRESSION OF METABOTROPHIC = GLUTAMATE=20 RECEPTORS (mGluRs) IN ALZHEIMER = DISEASE =20 =20 H-G Lee, X.=20 Zhu, M.J. O'Neill, G. Perry, M.A. = Smith=20 =20 Institute=20 of Pathology, Case Western Reserve University, = Cleveland, Ohio, USA=20 =20 Although=20 the mechanism underlying the loss and/or = dysfunction=20 of susceptible neurons in Alzheimer disease = (AD) is=20 unknown, it has been hypothesized that = excitotoxicity=20 may play a pivotal role. Since glutamate has = long been=20 thought to be important in the pathogenesis of = AD, we=20 hypothesized that select glutamatergic = receptor=20 populations (e.g., metabotropic glutamate = receptors=20 (mGluRs)) may correlate with those neurons = vulnerable=20 to AD. mGluRs are directly coupled to the=20 intracellular signaling system via GTP-binding = proteins and are thought to be related with = synaptic=20 transmission and neuroprotection. Since = synaptic loss=20 is a prominent feature of AD, in this study, = we=20 investigated the role of mGluR in AD. = Specifically, we=20 determined the expression of the family of = mGluR=20 receptors including group I, II and III in the = brains=20 of AD and age-matched control cases. We found = a=20 differential expression of mGluRs that related = directly to the pattern of neuronal = susceptibility in=20 AD. mGluR2 is specifically increased in = pyramidal=20 neurons in the hippocampus of AD and often=20 co-localizes with neurofibrillary pathology. = mGluR5=20 showed a similar expression pattern to mGluR2 = but was=20 also found in astrocytes surrounding amyloid = plaques.=20 The specificity of these findings is = demonstrated by=20 the fact that mGluR1 expression, which, like = mGluR5,=20 is a group I mGluR, is not different between = AD and=20 control cases. Our data suggest that specific = mGluRs=20 such as mGluR2 and mGluR5 are differentially = regulated=20 in AD and that this expression is directly = related to=20 neuronal populations vulnerable to = degeneration and to=20 amyloid-beta and tau protein lesions. = Therefore, the=20 differential regulation of specific mGluRs may = explain=20 the selective neuronal degeneration in AD and = provide=20 a unique therapeutic target for the treatment = of AD.=20 top*=20
Smith, Mark A.	92. =20 ACETYLATION: A NOVEL POSTTRANSLATIONAL = MODIFICATION IN=20 ALZHEIMER DISEASE=20 =20 A.K. Raina,=20 X. Zhu, A.D. Cash, L.M. Sayre, G. Perry, M.A.=20 Smith=20 =20 Institute=20 of Pathology, Case Western Reserve University, = Cleveland, Ohio, USA=20 =20 The=20 mechanism leading to the selective dysfunction = of=20 neurons in Alzheimer disease is incompletely=20 understood. Nonetheless, the formation of the=20 characteristic neurofibrillary pathology in = Alzheimer=20 disease is thought to play a pivotal role in = this=20 process. Associated with the formation of=20 neurofibrillary pathology, proteins are = subject to a=20 number of posttranslational modifications with = phosphorylation being the best studied. In = this study,=20 we report an entirely novel protein = modification,=20 namely acetylation of lysine, in vulnerable = neurons=20 that parallels the temporal pattern of = neuronal=20 vulnerability in cases of Alzheimer disease. = While the=20 exact role of protein acetylation in Alzheimer = disease=20 is unknown, it is likely, given the = physiological=20 roles of acetylation, that acetylation = promotes=20 neuronal survival by stabilizing the = cytoskeleton in=20 the face of occult, and otherwise = destabilizing, tau=20 phosphorylation events. This novel finding of = a=20 stabilizing modification in vulnerable neurons = in=20 Alzheimer disease will provide new insights = for=20 therapeutic interventions that could alter the = natural=20 history of this dementia.=20 top*=20
Smith, Mark A.	93. =20 A MECHANISTIC STUDY ON AMYLOID-BETA-INDUCED = JNK=20 ACTIVATION=20 =20 X. Zhu,=20 C.A. Rottkamp, Z. Kubat, A.K. Raina, S.L. = Siedlak,=20 C.S. Atwood, G. Perry, M.A. Smith=20 =20 Institute=20 of Pathology, Case Western Reserve University, = Cleveland, Ohio, USA=20 =20 Amyloid-β=20 protein (Aβ), the main constituent of = neuritic and diffuse plaques as well as=20 cerebrovascular amyloid deposits, = characterizes the=20 neuropathology of Alzheimer disease (AD). = While=20 Aβ is neurotoxic in vitro, the actual = biochemical mechanism remains to be = established. In=20 this study, we found that when Aβ is=20 pretreated with the iron chelator = deferoxamine,=20 neuronal toxicity is significantly attenuated = while=20 conversely, incubation of holo-Aβ = with excess=20 free iron restores toxicity to original = levels. These=20 data suggest that the toxicity of Aβ = is=20 mediated, at least in part, via redox-active = iron. The=20 in vivo significance of these findings is = amplified by=20 consideration of the enrichment of iron in AD=20 neuropil, and especially in association with=20 Aβ. To understand the mechanistic = consequence=20 of these findings, we also found that JNK/SAPK = is=20 activated in susceptible regions of the brain = in AD.=20 The co-localization of activated JNK/SAPK and=20 Aβ deposits, as well as concomitant = iron=20 accumulation, suggests that JNK/SAPK pathway = may play=20 an important role in metal-mediated = Aβ=20 toxicity. In this regard, we found that = although both=20 AβPP (K670N/M671L) transgenic mice = and=20 AβPP (V717I) transgenic mice develop = amyloid=20 senile plaques, iron accumulates only in the = former=20 and strikingly, activated JNK/SAPK is also = only found=20 in the former. Since such mice show cognitive=20 alterations, we hypothesize that neuronal = dysfunction=20 is, in part, mediated through alteration in = signal=20 transduction pathways such as JNK/SAPK = pathway.=20 Supported by the NIH (NS38648) to MAS and the=20 Alzheimer=92s Association (NIRG-02-3923) to = XZ.=20 top=20
Tsui, Justine C. Y.	95. =20 TELOMERE LENGTH AS A POSSIBLE-AGE MARKER IN=20 FISH=20 =20 Justine,=20 C.Y. Tsui, Stephen, B. Pointing and Kenneth, = M.Y.=20 Leung=20 =20 The Swire=20 Institute of Marine Science, Department of = Ecology=20 & Biodiversity, The University of Hong = Kong,=20 Pokfulam, Hong Kong SAR, China=20 =20 Telomeres=20 are nucleoprotein structures at the ends of = linear=20 chromosomes consisting of DNA sequences = arranged in=20 tandem repeated units (TTAGGG). They act to = prevent=20 aberrant recombination and degradation of the=20 chromosomal ends, and thus maintain chromosome = integrity. Since DNA polymerases are unable to = copy=20 chromosomal DNA completely, telomere sequences = typically shorten with the number of cell = divisions=20 and hence with age. Previous research has = demonstrated=20 a negative relationship between telomere = length and=20 age for various vertebrate species. In this = study, we=20 describe for the first time an assessment of = telomere=20 length as a function of age for the mangrove = snapper,=20 Lutjanus argentimaculatus, a commercially = important=20 teleost species in Asia and the Middle East.=20 Individuals with known chronological ages = (2.5-36=20 months) were obtained from a fish farm in = Shenzhen,=20 China. Genomic DNA was extracted from whole = blood and=20 dorsal muscle tissues. Telomere length was = quantified=20 using the Telo TAGGG Telomere Length Assay = (Roche=20 Molecular Biochemicals), which is based on = Southern=20 blot-hybridization with terminal restriction = fragments=20 of the telomere. Telomere length in blood was=20 significantly reduced with age of individuals = (r2 =3D=20 0.2484, F1,22 =3D 6.942, n =3D 23, p < = 0.05),=20 suggesting a telomere shortening rate of 0.39 = kbp/yr.=20 Telomere length in muscle tissues also = decreased=20 significantly in relation to fish age (r2 =3D = 0.1513,=20 F1,26 =3D 4.459, n =3D 27, p<0.05), = resulting in a=20 slightly faster shortening rate of 0.44 = kbp/yr. The=20 results suggest that patterns in fish telomere = shortening are similar to that in other = vertebrates=20 such as mammals and birds. Measurement of = telomere=20 length in blood may have potential as a=20 non-destructive method for estimating age in=20 fish.=20 top*=20
van = Steeg,=20 Harry	96. =20 LONGEVITY AND CROSS SECTIONAL STUDIES IN = DNA-REPAIR=20 DEFICIENT MOUSE MODELS Harry van = Steeg1, Susan=20 Wijnhoven1, Rudolf Beems1, Martijn Doll=E91, Jan = Vijg2,=20 Paul Lohman1, Jan Hoeijmakers3 and Bert van der = Horst3=20 1Natl Inst of Public Health and the = Environment,=20 Department of Toxicology, Pathology and = Genetics,=20 Bilthoven, The Netherlands; 2University of Texas = Health=20 Science Center, Department of Physiology, San = Antonio,=20 Texas, USA; 3MGC department of Cell Biology and=20 Genetics, Erasmus University, Rotterdam, The=20 Netherlands The accumulation of somatic = DNA=20 damage is considered to be a major cause of the = aging=20 process in various species including mice and = humans.=20 Among the sources of DNA damage, reactive oxygen = species=20 (ROS) are often thought to be the ultimate cause = of=20 aging. However, the mechanisms involved remain = obscure.=20 To counteract the effects of DNA damage, an = intricate=20 network of DNA repair pathways has evolved. One = major=20 pathway is nucleotide excision repair (NER), = which=20 removes a broad range of bulky lesions including = some=20 forms of oxidative damage. Patients with a = defect in NER=20 proteins like CSB and XPD, both involved in = repair as=20 well as transcription of DNA, appeared to have a = decreased life span. In order to = investigate=20 whether defects in genome maintenance are = correlated to=20 accelerated aging, we have successfully = conducted=20 several longevity and cross sectional studies = with mice=20 having defect in DNA repair and/or RNA = transcription=20 (i.e. Xpa-, Csb-, XpdTTD- deficient mice as well = as=20 C57Bl/6 wild type controls). The mean survival = of female=20 XpdTTD as well as Xpa mice appeared to be much = shorter=20 (appr. 90 weeks) than those found for Csb and = wild type=20 littermate controls (104-110 weeks). Full = histopathology=20 has been performed in aged female mice, and = gross=20 examination at autopsy revealed small posture, = kyphosis,=20 large spleen, small thymus and abnormal skin and = hair=20 especially in XpdTTD mice. The terminal body = weights in=20 XpdTTD and Csb females were decreased, with an = increase=20 in the relative weights of several organs of = XpdTTD=20 mice, especially in the kidney, spleen and the = heart.=20 Furthermore, lipofuscin pigmentation (an aging = feature,=20 correlated to oxidative damage) was found to be=20 accumulated in the liver of XpdTTD mice as = compared to=20 the other phenotypes. Finally, a variety of = immune=20 parameters were determined in mice of the = various=20 genotypes and ages. top=20
von Zglinicki, = Thomas	98. =20 VARIATION IN TELOMERE SHORTENING RATE CAUSES=20 HETEROGENEITY IN REPLICATIVE LIFESPAN = =20 T.von=20 Zglinicki, C. Martin-Ruiz, J. Petrie, G.=20 Saretzki=20 =20 Biogerontology=20 Laboratory, Newcastle University, UK =20 The=20 replicative lifespan of human fibroblasts is=20 heterogeneous, with a fraction of cells = senescing at=20 every population doubling. To find out whether = this=20 heterogeneity is due to premature senescence, = i.e.=20 driven by a non-telomeric mechanism, = fibroblasts with=20 a senescent phenotype were isolated from = growing=20 clones by flow cytometry. These senescent = cells had=20 shorter average telomere length than their = cycling=20 counterparts at all population doubling levels = both in=20 mass culture and in individual sub-clones, = indicating=20 heterogeneity in the rate of telomere = shortening.=20 Ectopic expression of telomerase stabilised = telomere=20 length in the majority of cells and rescued = these from=20 early senescence, suggesting a causal role of = telomere=20 shortening. Under standard cell culture = conditions,=20 there was a minor fraction of cells which = showed a=20 senescent phenotype and short telomeres = despite active=20 telomerase. This fraction increased under = chronic mild=20 oxidative stress, which is known to accelerate = telomere shortening. The data show that = heterogeneity=20 of human fibroblast replicative lifespan can = be caused=20 by significant stochastic cell-to-cell = variation in=20 telomere shortening. top=20
von Zglinicki, = Thomas	97. =20 TELOMERE SHORTENING IN HUMAN FIBROBLASTS IS = NOT=20 DEPENDENT ON THE SIZE OF THE TELOMERIC = 3=92-OVERHANG=20 =20 T. von=20 Zglinicki, B. Keys, V. Serra, G. = Saretzki=20 =20 Biogerontology=20 Laboratory, Newcastle University, UK=20 =20 Telomeres=20 shorten in human somatic cells with each round = of DNA=20 replication, and this shortening is thought to = ultimately trigger replicative senescence. = Telomere=20 shortening is caused partly by the inability = of=20 semi-conservative DNA replication to copy a = linear=20 strand of DNA to its very end. = Post-replicative=20 processing of telomeric ends, producing=20 single-stranded G-rich 3=92 overhangs, has = also been=20 suggested to contribute to telomere = shortening. This=20 suggestion implies that a positive correlation = should=20 exist between the length of 3=92 overhangs and = the rate=20 of telomere shortening. Measuring overhang = length by=20 in-gel hybridisation, we could confirm = shortening of=20 overhangs as human MRC5 and BJ fibroblasts = approach=20 senescence. However, a large study comprising = of=20 fibroblast strains from 21 donors under = conditions=20 leading to two orders of magnitude of = variation in=20 telomere shortening rate ruled out any = correlation=20 between telomere overhang length and = shortening rate,=20 suggesting that overhang length is neither = causal for=20 nor indicative of telomere shortening.=20 =20 top=20
Wang, = Zhihui	99. =20 EXPRESSION OF GENES RELATED TO INSULIN = SENSITIVITY AND=20 LIPID METABOLISM IN AMES DWARF AND CALORIC = RESTRICTED=20 MICE Zhihui Wang, = K.A.=20 Al-Regaiey, M.M.Masternak, = A.Bartke Southern=20 Illinois University, School of Medicine, = Geriatrics=20 Research, 801 N. Rutledge Springfield, IL=20 62794 Ames dwarf (df) mice display = primary=20 deficiency of growth hormone (GH), prolactin = (PRL), and=20 thyroid simulating hormone (TSH), improved = insulin=20 sensitivity, diminutive body size, and prolonged = longevity. Young adult df mice have higher % = body fat=20 compared with their normal littermates (Wild = type, WT).=20 In order to study the mechanism of improved = insulin=20 sensitivity in these mice, we have used real = time PCR to=20 examine gene expression of adipocytokines = related to=20 insulin sensitivity including adiponectin, = leptin,=20 resistin, TNF-, and interleukin = (IL)-6, and=20 genes related to fat metabolism including fatty = acid=20 synthase (FAS), hormone-sensitive lipase (HSL),=20 uncoupling protein (UCP)-2, and = 3-adrenergic=20 receptors (3AR) in white adipose = tissue=20 (WAT). We also analyzed expression of the same = genes in=20 df and WT animals subjected to 30% caloric = restriction=20 (CR). In WAT from df mice fed ad libitum (AL), = the=20 expression of adiponectin, TNF- , = FAS, HSL,=20 and UCP2 were significantly diminished, and the=20 expression of IL-6 was dramatically suppressed, = whereas=20 the expression of leptin, resistin, and = 3AR=20 were not changed. CR increased gene expression = of=20 resistin, FAS, and 3AR, and = diminished gene=20 expression of UCP2 and IL-6 in WT mice compared = with=20 their AL littermates. CR also diminished = expression of=20 leptin in df mice compared with AL df mice. In = both WT=20 and df mice, CR did not alter the expression of=20 adiponectin, HSL, although their expression and = the=20 expression of resistin, FAS, UCP2, = 3AR in CR=20 df mice were much lower than in CR WT mice. = These=20 results suggest that decreased expression of=20 TNF-, FAS, and IL-6 might play a = role in=20 improving insulin sensitivity in df mice. CR = might=20 down-regulate the gene expression of leptin via=20 increasing expression of 3AR, and=20 down-regulate the expression of UCP2, thus = maintaining=20 the energy balance of the body. (Supported by = NIA and=20 Ellison Medical Foundation) top=20
Wilson, Mark A.	100. =20 BLUEBERRY-DERIVED PHYTONUTRIENTS CAN AFFECT = AGING AND=20 STRESS TOLERANCE IN THE NEMATODE, = CAENORHABDITIS=20 ELEGANS=20 =20 M. Wilson,=20 B. Shukitt-Hale, G. Lee, D. Ingram, J. Joseph, = C.=20 Wolkow=20 =20 Laboratory=20 of Neuroscience, National Institute on Aging, = IRP,=20 5600 Nathan Shock Drive, Baltimore, MD = 21224=20 =20 Plant-derived=20 antioxidants have been shown to provide = beneficial=20 health effects. In order to explore the = mechanisms=20 behind these observations, we investigated the = effects=20 of blueberries on lifespan and stress = resistance in=20 the nematode, C. elegans. Either crude = extracts or=20 partially purified polyphenol fractions of = blueberries=20 increased mean lifespan and slowed age-related = declines in pharyngeal pumping rates in = wild-type=20 animals. Resistance to thermal stress was also = increased significantly. One possibility is = that=20 blueberry extracts inhibited bacterial growth, = a=20 treatment known to extend mean C. elegans = lifespan. We=20 examined this possibility by comparing the = effects of=20 antibiotics and blueberry. While treatment = with either=20 ampicillin or blueberry extract extended C. = elegans=20 lifespan, there were no additive effects on = longevity=20 when these treatments were combined. = However,=20 while blueberry significantly increased = thermal=20 tolerance, ampicillin had no effect. The = differential=20 effect on stress tolerance suggests that = blueberry can=20 directly affect C. elegans physiology, = independent of=20 effects on bacterial growth. Through what = genetic=20 pathways can blueberries alter stress = tolerance? =20 Reductions in insulin/IGF-like signaling are=20 correlated with increased stress resistance = and=20 lifespan in a wide range of organisms. = Longevity=20 and stress resistance in C. elegans = insulin-like=20 pathway mutants requires the DAF-16/FOXO = transcription=20 factor. To determine whether blueberry affects = insulin-like signaling, we examined whether = the=20 beneficial effects of blueberry extracts = required=20 daf-16 activity. Fractional increases in = lifespan, and absolute increases in = thermotolerance,=20 were similar in wild-type and daf-16(mgDf50)=20 animals. In untreated daf-16(mgDf50)=20 animals, pharyngeal pumping declined = more=20 rapidly with age than in wildtype animals. = This=20 decline was slowed to the wild-type rate with=20 blueberry extract. These experiments = indicate=20 that blueberry's longevity-promoting effects = do not=20 require DAF-16/FOXO activity, and probably = occur=20 independently of insulin-like signaling. We=20 hypothesize that treatment with blueberry = extract=20 increases C. elegans longevity by increasing = intrinsic=20 resistance to environmental and/or metabolic = stresses=20 that limit mean lifespan.=20 top=20
Wolkow, Catherine A.	101. =20 AGING ALTERS THE COORDINATION OF SENSORY = INPUTS WITH=20 MOTOR RESPONSES IN THE NEMATODE, = CAENORHABDITIS=20 ELEGANS=20 =20 C. Wolkow,=20 D. Chow, C. Glenn, L. David, I. = Goldberg=20 =20 Laboratory=20 of Neuroscience, National Institute on Aging, = IRP,=20 5600 Nathan Shock Drive, Baltimore, MD = 21224=20 =20 Many=20 behavioral responses require the coordination = of=20 sensory inputs with motor outputs. Aging = is=20 associated with progressive declines in both = motor=20 function and muscle structure. However, = the=20 consequences of age-related motor deficits = upon=20 behavior have not been clearly defined. = To=20 investigate this question, we have examined = the=20 effects of aging upon behavior in the = nematode,=20 Caenorhabditis elegans. We first = analyzed the=20 effect of age on responses to sensory = stimuli. =20 When touched gently with a hair, both young = and aged=20 animals responded appropriately by backing = away from=20 the stimulus. When presented with an = aversive=20 odor, octanol, both young and aged animals = stopped=20 forward movement similarly. However, = aged=20 animals exhibited defects in backing away from = the=20 odorant source and these defects were = exacerbated when=20 octanol was diluted 100-fold. Aged = animals=20 displayed significant deficits in chemotaxis = behavior,=20 as older animals failed to progress fully to = the=20 source of an attractive stimulus. = Together,=20 these results indicate that age was not = associated=20 with significant declines in sensory = ability. =20 However, locomotory responses to stimuli were = severely=20 compromised.To determine the basis for these=20 behavioral deficits, we analyzed the patterns = of=20 movement in aged animals. Older animals = moved=20 50% slower than young animals and failed to = maintain=20 forward movement for sustained = intervals. =20 Examination of muscle structure at these ages = showed=20 that reduced muscle tissue integrity was = correlated=20 with age-related declines in locomotory = behavior,=20 although widespread muscle deterioration was = rare at=20 the ages examined. Finally, treatment = with a=20 muscarinic agonist improved locomotory = behavior in=20 aged animals, indicating that improved = neuromuscular=20 signaling may be one strategy for reducing the = severity of age-related behavioral = impairments.=20 top=20
Xu, Dongsheng	102. =20 GENETIC SELECTION AND INITIAL CHARACTERIZATION = OF=20 MOUSE LINES FOR HIGH AND LOW STRESS=20 SUSCEPTIBILITY=20 =20 DS.=20 Xu,1,3, T. Lambert 1, D. Paul 2, Laura = Meyerle=20 1 , C. Nekl 1 , J. Potts 2, M. Nielsen 2, Y. = Zhou=20 1=20 =20 1=20 Department of Veterinary & Biomedical = Sciences, 2=20 Department of Animal Sciences, University of = Nebraska=20 - Lincoln; 3 Center for Neurovirology = and=20 Neurodegenerative Disorders, University of = Nebraska=20 Medical Center, Omaha =20 Two mouse=20 lines were generated by genetic selection for = high and=20 low stress susceptibility (SH and SL lines,=20 respectively), from baseline populations of = mice=20 selected for high (base for SH) and low (base = for SL)=20 rates of metabolism, which also showed = differences in=20 locomotor activity and stress responses.=20 Corticosterone concentrations in sera = collected at=20 various time points from the SH and SL mice = with or=20 without exposure to acute or repeated = restraint stress=20 were determined by radioimmunoassay and used = as one of=20 the key criteria for selection. The SH mice = showed=20 significantly (P < 0.05) higher levels of = serum=20 corticosterone, at all tested time points, = than the SL=20 mice. Initial reverse transcription polymerase = chain=20 reaction (RT-PCR) analysis using brain RNA = revealed=20 differences in expression of three major=20 stress-response regulatory genes:=20 corticotrophin-releasing hormone (CRH), = glucocorticoid=20 receptor (GR) and mineralocorticoid receptor = (MR).=20 Expression of CRH was higher while GR and MR=20 expressions were down-regulated in the SH mice = as=20 compared to SL mice. Microarray assays were = used to=20 compare gene expression in the brains of the = two lines=20 of mice under stressed and non-stressed = conditions,=20 and revealed significant differential = expression=20 patterns of many genes (up or down regulated) = between=20 the two lines as well as within the same line = of=20 mice. Subsequent RT-PCR experiments = using=20 specific primers for 25 genes of interest, = selected=20 based on the microarray data, confirmed the = genetic=20 and stress-induced alterations in gene = expression=20 between the SH and SL mice, such as=20 microtubule-associated protein-2, = P21-activated=20 kinase-3, calcium/calmodulin-dependent protein = kinase=20 IIa, and calcium channel proteins. = Further=20 establishment of the SH and SL mouse lines, = divergent=20 through selection for altered stressed = susceptibility,=20 may become a useful tool to study = stress-induced early=20 aging and neurodegeneration.=20 top*=20
Yin, Zhirong	103. =20 AGE-RELATED DIFFERENCES OF GENE EXPRESSIONS = AND DNA=20 DAMAGE IN HUMAN FIBROBLASTS AFTER EXPOSURE TO = COBALT=20 CHROME PARTICLES=20 =20 Yin Z.R.,=20 Papageorgiou I., Clerkin J.S., Learmonth I.D., = Case=20 C.P.=20 =20 Bristol=20 Implant Research Centre, University of = Bristol, BS10=20 5NB, U.K.=20 =20 Wear debris=20 from worn cobalt chrome orthopaedic joint = replacements=20 causes an increase in chromosomal = translocations and=20 aneuploidy. In this study the levels of DNA = damage and=20 certain gene expressions (TGF-=DF2, p38 MAPK,=20 Integrinβ1, SOD1, Caspase 10, PURA, = FRA-1 and=20 VNR) have been compared in human fibroblasts = of=20 different age (10 and 35 population doublings, = PD)=20 after exposure to different doses of cobalt = chrome=20 particles and at different times of exposure = (6 and 24=20 hours). The 35 PD fibroblasts showed = significantly=20 more immunostaining of the = senescence-associated=20 β-galactosidase (57.4%) than the 10 = PD=20 fibroblasts (1.4%). The level of DNA = damage, as=20 detected with alkaline comet assay, was = greater at=20 higher doses, at longer exposures (up to 24 = hours) and=20 in younger (10 PD) fibroblasts. No significant = change=20 in cell viability was noted using MTT assay or = LDH=20 assay. The expression of all the genes listed = above=20 was generally lower after exposure to cobalt = chrome=20 particles using semi-quantitative reverse=20 transcription-polymerase chain reaction = (RT-PCR). The=20 reduction in gene expressions, like the = increase in=20 DNA damage was greater at higher dose and at = longer=20 time of exposure. Interestingly these changes = in DNA=20 damage and gene expression were more = pronounced in=20 younger than in older fibroblasts but not = until after=20 24 hours of exposure. These results show that = levels=20 of gene expression of TGF-=DF2, p38 MAPK,=20 Integrinβ1, SOD1, Caspase10, PURA, = FRA-1 and=20 VNR may be correlated with the level of DNA = damage and=20 that this depends on the dose and length of = exposure=20 and the age of the cells. This highlights the=20 potential importance of these genes in the=20 mutagenicity of cobalt chrome particles in = human=20 fibroblasts. top=20
Young, J.	104. =20 Assessment of motor performance in rhesus = monkeys on=20 long-term calorie restriction J.=20 Young1, Zhiming Zhang2, D. Gash2, G. Gerhardt2, = G.=20 Roth3, M. Lane,3 J. Mattison,3 D. = Ingram3 1Sobran,=20 Inc, 4401 Dayton-Xenia Road, Dayton, OH = 45432;=20 2Dept. of Anatomy and Neurobiology, University = of=20 Kentucky Medical Center, Lexington, KY 40536;=20 3Laboratory of Experimental Gerontology, = National=20 Institute on Aging, NIH, Gerontology Research = Center,=20 5600 Nathan Shock Drive, Baltimore, MD 21224;=20 In=20 numerous rodent studies, calorie restriction = (CR) has=20 been demonstrated consistently to increase = lifespan,=20 reduce the incidence and onset of age-related = chronic=20 disease, and attenuate age-related decline in = many=20 physiological functions. Emerging evidence = from=20 studies in rhesus monkeys suggests that CR can = also=20 retard aging processes in a species closely = related to=20 humans. An important question regarding = the=20 effects of long-term CR in monkeys is whether = behavioral=20 function will be impacted, either negatively or=20 positively. We have initiated studies to = evaluate=20 motor performance in male and female rhesus = monkeys=20 involved in a long-term study of CR. = Control=20 monkeys were provided a highly nutritious diet = while=20 monkeys on CR have had their intake of this diet = reduced=20 30 percent from the level of comparable aged=20 controls. Monkeys were introduced to the = study at=20 different stages in the lifespan, including = juvenile,=20 adult, and aged groups of monkeys. Fine = motor and=20 coarse motor performance were measured in an = automated=20 movement assessment panel attached to the home = cage of=20 the monkey. During trials of 45 sec = duration,=20 monkeys are given access to the apparatus during = which=20 they must reach with their hand through an = opening and=20 then must maneuver their hand at a 90o angle = from entry=20 into an elevated center chamber to retrieve a = visible=20 food treat (soft lifesaver candy). During=20 different sessions, the treat is positioned = either on a=20 flat platform, a straight rod, or a hook in the = center=20 chamber. Performance on each task is = measured as=20 the time required to retrieve the treat once the = hand=20 has entered the apparatus. Age-related = decline in=20 performance (longer retrieval times) of rhesus = monkeys=20 on all three tasks has been previously = documented (Zhang=20 et al. J.Gerontol. Biol. Sci. 55:B473, 2000), = and we=20 have replicated these findings in the current=20 study. To assess effects of CR on = performance,=20 comparisons of fine motor retrieval time were = made for=20 male and female monkeys between the ages of 12.1 = - 24.3=20 yr that had been on CR for 12 -18 yr. For = males,=20 we found that retrieval time was equivalent = between diet=20 groups on the easiest tasks (platform and = straight rod),=20 but times on the more difficult hook task were=20 significantly faster for CR group. For = females, CR=20 monkeys had significantly faster retrieval times = on both=20 the platform and the hook tests, but had = significantly=20 slower times on the straight rod task. = Considering=20 performance on only the most difficult task, we = can=20 conclude that CR improves motor performance. = top=20
Zachariah, Sally B.	105. =20 BOTULINUM TOXIN TYPE A (BoNT/A) FOR CHRONIC = HEADACHES=20 IN ELDERLY=20 =20 S.Zachariah,M.D,=20 T.Ranjan,M.D and Aveen Zachariah.=20 =20 Background:=20 Headache is reported to be the tenth and = fourteenth=20 most common symptom in elderly women and men=20 respectively. Five to ten percent of patients = over 65=20 develop chronic daily headaches. BoNT/A has=20 dramatically improved the treatment of a = variety of=20 neurological disorders, including headache. = =20 Methods: Data reviewed of 30 patients (average = age-60.8year) involved in an open label trial = showed=20 21 had typical vascular headaches, 7 had = chronic daily=20 headaches [>20 headaches in a month] and 2 = had=20 tension headaches. They received BoNT/A for = headaches=20 (average duration 10 yrs) between 1995 and = 2003. =20 All patients met the following criteria: = headache duration of >2 yrs; absence of = known=20 allergy to BoNT/A; severity of headache > 5 = [on=20 0-10 scale]; failure to Triptans, Beta = blockers,=20 Calcium Channel blockers, Anticonvulsant and=20 Antidepressants. Forty to seventy-five = unit=20 intra muscular injections were given in = corrugators=20 and bilateral upper & lower frontalis and=20 temporalis at the depth of 1-3 mm using number = 30=20 needle. Patients with tension headache = were=20 given semispinalis and splenius capitis = injections,=20 following the path of pain. Injection = was=20 repeated after 3 months. Evaluation entailed: = headache=20 scale, satisfaction survey, medication use, = life style=20 questionnaire and neurological exam.=20 =20 Results: Most of the patients had effect in = 3-5 days=20 with optimum benefit in 2 weeks. The average = pain=20 intensity reduced from 8.66 to 1. First = injection=20 effects lasted from 45 to100 days with later=20 injections lasting longer. Twenty-four = patients out of=20 30 (80%) had clinical response, of which 20 = patients=20 were very satisfied, 4 patients were satisfied = and 6=20 patients [20%] had no response to therapy. = Respondents=20 had long-term benefits like pain relief, = stoppage or=20 reduction of medications, better life style, = and=20 improvement in productivity.=20 =20 Conclusions: Older patients with chronic = headache with polypharmacy failures tolerate = and=20 benefit from BoNT/A therapy with minimal acute = side=20 effects. top=20
Zienko, Sarah C.	106. =20 UNDERSTANDING THE GENETICS OF AGING: A CANINE=20 MODEL=20 =20 S. Zienko,=20 K.Greer, M. Breen, K. Murphy=20 =20 Dept. of=20 Veterinary Pathobiology; Texas A&M Univ. = College=20 of Veterinary Medicine; College Station, = Texas,=20 U.S.A.; Dept. of Molecular Biomedical = Sciences, North=20 Carolina State Univ. College of Veterinary = Medicine;=20 Raleigh, North Carolina, U.S.A. =20 The biology=20 of aging is extremely complex, and is, = therefore,=20 poorly understood. Research pertaining = to aging=20 has many potential benefits, the most = important of=20 which are extension of life spans and = improvement of=20 the quality of life in aged populations. = Select=20 human populations have significantly longer = life=20 expectancies than others, suggesting an = underlying=20 genetic component associated with the aging=20 process. The genetic factors governing = longevity=20 are numerous, difficult to define, and may = possess=20 regulatory functions. The hereditary = component=20 of longevity applies to animal = populations. Our=20 interest in the dog as a model stems from the = fact=20 that the dog serves as excellent model animals = for=20 many human hereditary conditions, and that = average=20 life expectancy of purebred dogs decreases as = both=20 height and weight increase, as shown by = preliminary=20 regression analysis of 117 breeds. As a = first=20 step to understanding the potential genetic = factors=20 associated with longevity in dogs, comparative = gene=20 mapping data is being generated by cataloging = and=20 mapping 56 candidate genes. Variation in = 23 of=20 these genes is associated with either extended = or=20 truncated life spans in the human or = mouse. The=20 additional 23 genes, that give approximately = 0.5Mb=20 coverage of a 22Mb region of human chromosome = 4, were=20 selected because a microsatellite marker, = D4S1564,=20 shows strong association with excessive = longevity in=20 an Italian population. Following mapping = of=20 these genes, detailed analysis will be = conducted to=20 obtain polymorphic markers in close vicinity = of the=20 mapped genes. These markers will be = analyzed=20 across a range of breeds with contrasting = degrees of=20 longevity. Additionally, comprehensive = analysis=20 will be carried out to identify variations = within=20 selected genes (from the group of 56) across = breeds=20 with diverse longevity. These pilot = studies will=20 serve as a platform to initiate broader = research aimed=20 at understanding genetic factors associated = with=20 longevity in dogs and other animals. top=20 =

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R=20 Rosedale, E Westman

Rosedale=20 Center, Denver CO - = ronrosedale@comcast.net