ACCEPTED ABSTRACTS
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O R A L |
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Brewer, Gregory J. |
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AGING NEURON MITOCHONDRIAL FUNCTION AND REJUVENATION WITH ESTROGEN OR
MITOSIS
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G.
Brewer, J. Nash, T. Jones, J. Reichensperger, M. Parihar
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PO
Box 19626, Springfield, IL 62794-9626
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The mitochondrial
oxyradical theory of aging is gaining increased support from studies of
brain, heart and muscle tissue. Is this aging caused by systemic
hormones or intrinsic to aging mitochondria in neurons, for example.
The intrinsic hypothesis is supported by our findings with isolated neurons
of increased susceptibility with age to glutamate and A-beta toxicity, two
endogenous toxins important to age-related neurodegenerative disease
(Brewer, 1998). For neuron-specific function, we have established
culture techniques that allow us to compare neurons isolated from embryonic,
middle-age (12 month) and old (24 month) rat hippocampus (Brewer, 1997).
For use with these cultured neurons, we monitored cytochrome oxidase
activity (COX), cardiolipin specific staining of mitochondria with
nonyl-acrydine orange (NAO), as well as labeling with mitotracker-red and
immunostain for cytochrome C, fluorescent NADH, glutathione levels and
respiration. Cytochrome C immunostain and mitotracker red stained equal
numbers of mitochondria per cell for middle-age and old neurons, compared to
higher numbers per embryonic neuron. In contrast to mitochondrial
numbers, COX activity/cell declined sharply with age of the neurons: levels
of old neurons were reduced by 30% of those of middle-age neurons.
Similarly, NAO stain for mitochondria was 40% lower for old neurons compared
to middle-age neurons. By addition of FGF2 and low density passage,
old neurons divide in culture (Brewer, 1999). COX activity in these
dividing old neurons was equivalent to that of middle-age neurons.
Age-related reductions in glutathione levels were exacerbated by exposure to
glutamate. NADH levels and respiration in old neurons were normal, but
showed dramatic deficits in response to glutamate. In addition to
restoring youthful calcium dynamics, treatment of old neurons with
17-beta-estradiol restored NAO staining levels/cell to those of middle-age
neurons and provided full neuroprotection from glutamate and A-beta
toxicity. These results demonstrate that mitochondrial function is
impaired in old neurons, which correlates with increased susceptibility to
toxic stress, but function can be restored by treatment with FGF2 or
estradiol.
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Carter, Christy S. |
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ACE inhibition intervention: Implications for improving age-related
declines in physical performance and longevity.
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CS
Carter, M Cesari, M Pahor
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Wake Forest Univeristy School of Medicine, J. Paul Sticht Center on Aging, 1
Medical Center Blvd. Winston-Salem NC, 27157
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Age-related disregulation
of the renin-angiotensin system (RAS) leads to variety of pathologies
including end organ damage to kidney, heart, blood vessels, and more
recently has been suggested to exacerbate declining cognitive and physical
performance. Reversing or attenuating these effects could have a significant
impact on maintaining independence and extending longevity in the elderly.
In fact, both pharmacological and genetic studies have demonstrated that
lower circulating levels of angiotensin converting enzyme (ACE) results in
enhanced responsiveness to exercise, increased muscle strength, and lower
adiposity. The relationship between low physical performance in healthy
older adults, shifting body composition (increases in fat and decreases in
fat-free mass) and longevity has been documented and more recently this same
assessment has been made in aging rodents. Therefore, interventions
that retard declining performance such as ACE inhibition (ACEi) may also
have validity for studies of longevity. We have conducted studies to
assess the effects of ACEi on body composition and physical performance in
aged rats. Animals were randomized to daily injections of 40 or 80 mg/kg of
enalapril or saline at baseline (24 months of age) and were followed for 6
months. ACEi attenuated declining physical performance as measured by
both grip strength and inclined plane tasks, and was most likely explained
by a significant reduction in total fat mass, as no differences in lean mass
were observed between groups (as measured using DXA). A possible
interpretation is that beyond its powerful hemodynamic effects, ACEi
regulates many aspects of metabolic functioning, decreases oxidative stress
in many tissues, and may act ubiquitously to reduce age and disease related
chronic inflammatory states. Therefore, our understanding of these
results could be enhanced by a more thorough and mechanistic
characterization of the effects of long-term ACEi on lifespan and declining
performance.
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Chang, Sandy |
- ESSENTIAL ROLE OF LIMITING TELOMERES IN THE PATHOGENESIS OF
WERNER SYNDROME
AbstractAuthors: S. Chang*, A. Multani, N. Cabrera, M. Naylor, P. Laud, D.
Lombard, S. Pathak, R. de Pinho
MD Anderson Cancer Center, Houston, TX and Dana Farber
Cancer Center, Boston, MA
Mutational inactivation of the Wrn helicase gene causes Werner Syndrome (WS), an
autosomal recessive disease characterized by premature aging, elevated genomic
instability and increased cancer incidence. The capacity of enforced telomerase
expression to rescue premature senescence of cultured human WS cells and the
lack of a phenotype in Wrn deficient mice with long telomeres have implicated
telomere attrition in WS pathogenesis. Here, we show that the varied and complex
in vivo and cellular phenotypes of WS are precipitated upon exhaustion of
telomere reserves in mice. In mice doubly null for Wrn and the telomerase RNA
component (Terc), telomere dysfunction elicits a classical WS premature aging
syndrome typified by premature death, hair graying, alopecia, osteoporosis, type
II diabetes, and cataracts. This model also exhibited accelerated replicative
senescence and accumulation of DNA-damage foci in cultured cells as well as
increased chromosomal instability and cancer, particularly non-epithelial
malignancies typical of WS. This faithful mouse model of WS establishes that the
complex and pleiotropic in vivo sequelae of Wrn deficiency are provoked largely
by critical telomere shortening.
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Heilbronn, Leonie K. |
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THE EFFECTS OF 6-MONTHS OF CALORIE RESTRICTION ON BIOMARKERS OF AGING IN
NON-OBESE HUMANS.
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Leonie K Heilbronn*, Eric Ravussin and the Pennington CALERIE group.
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Pennington Biomdeical Research Center, 6400 Perkins Rd, Baton Rouge, LA
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Caloric restriction (CR)
extends life span and retards age-related diseases in rats, mice, flies,
worms and yeast. Several biomarkers of aging (fasting glucose, insulin,
DHEAS and core temperature) have been identified in human and monkey studies
of longevity. Whether prolonged CR improves biomarkers of aging in humans is
unknown. Forty-eight healthy, nonsmoking, male (25-50y) and female (25-45y),
overweight participants (25 < BMI < 30) were randomized into one of four
groups; group 1 = 25% CR, group 2 = 12.5% CR + 12.5% increase in physical
activity, group 3 = low calorie diet until 15% weight reduction, group 4 =
Control. Subjects were fed by the metabolic kitchen from Weeks 1-12 whilst
they were taught a calorie counting system to allow them to prepare their
own meals (Weeks 12-22). Subjects returned to infeeding from Weeks 22-24.
Fasting glucose, insulin, DHEAS and 24h core temperature were assessed at
baseline, weeks 12 and 24. At the time of writing, only 30 subjects had
completed the study and so CR groups (Groups 1-3) were combined for
analysis. Average weight loss was 10% in the CR groups. Weight was not
changed significantly in the controls. Fasting insulin was reduced 24% in CR
groups as compared to a 5% increase in the controls (p<0.05). Glucose was
reduced 1% in CR groups as compared to a 4% increase in controls (p<0.03).
DHEAS and daytime core temperature were not significantly changed after 6
months. However, nighttime temperature (recorded from 10pm to 6am) was
reduced by 0.2ºC in CR groups as compared to a 0.3ºC increase in controls
(p=0.06). This study suggests that 6-mo. of calorie restriction in non-obese
humans was sufficient to observe improvements in 3 out of 4 known biomarkers
of aging. Whether these changes are sustained over longer periods of time is
unknown.
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Kitani, Kenichi |
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THE PRESENCE OF AN OPTIMAL DOSE RANEGE MAY EXPLAIN DISCREPANCIES IN THE
EFFECTS OF (-)DEPRENYL (D) ON SURVIVALS OF ANIMALS IN PAST REPORTS
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K.Kitani*1, S.Kanai2, K.Miyasaka2, M.C.Carrillo3 ,GO Ivy4
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Tokyo Metropolitan Institute of Gerontology
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THE PRESENCE OF AN OPTIMAL
DOSE RANGE MAY EXPLAIN DISCREPANCIES IN THE EFFECTS OF (-)DEPRENYL (D) ON
SURVIVALS OF ANIMALS IN PAST REPORTS
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K.Kitani*1, S.Kanai2,
K.Miyasaka2, M.C.Carrillo3 ,GO Ivy4
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*1National Institute for
Longevity Sciences, 36-3, Gengo, Moriokacho, Obu-shi; 2Tokyo Metropolitan
Institute of Gerontology, Tokyo, Japan; 3Univ.Rosario,Rosario, Argentina;
4Univ.Toronto, Scarborough,Canada
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To
explain discrepancies in the effect of D on life spans of animals in past
reports, we examined the effect of D at a dose of 0.25 mg/kg/inj s.c. 3x a
week starting from 18 months of age on life spans of F344Du/Crj rats of both
sexes. Average life spans (days) were significantly increased in
both male (895.2±109.7,mean ±S.D., n=30; 967.8±88.6, n=30, control vs. D
treated,+8.1%, P<0.01) and female (924.7±132.2, n=38; 987.1±133.4,
n=39,+6.7%, P<0.05) rats that received D injections compared to control
animals given saline. Increases in life expectancy from 24 months of
age were 44% in males and 32% in females. We previously reported that
a dose of 0.5mg/kg/inj significantly increased the life span of male F344
rats (1). However, a dose of 1.0 mg/kg/inj somewhat shortened the life span
of animals, although not statistically significantly (2). Together,our data
indicate that a proper dose range of D can significantly increase the life
spans of rats of both sexes, however, a greater dose becomes less effective
and may adversely affect the life span of animals. The presence of
this effective dose range of D may explain discrepancies in the effect of D
on life spans of animals previously reported. Furthermore, the fact
that the effective dose range observed for the life prolonging effect of D
parallels that for increasing activities of SOD and catalase strengthens our
contention that these two effects are causally related. Our new
observation that the same dosage of D (0.25mg/kg) increases life spans of
both male and female rats further supports our contention, since we
previously reported that the optimally effective dose of D to increase
antioxidant enzyme activities becomes closer for male and female rats as
they get older, while at young ages, the optimal dose is 10 times greater in
male rats (3). Ref. 1) Life Sci. 52:281-288, 1993. 2) Life Sci.
67:577-585, 2000.3)Life Sci. 52:1925-1934,1993
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Ladiges, Warren C. |
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AGE-ASSOCIATED CELLULAR STRESS IN HSP-40 MOLECULAR CHAPERONE MUTANT MICE
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S
Knoblaugh, J Morton, G Moore-Sanders, A MacAuley, W Ladiges*
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Department of Comparative Medicine, University of Washington, Seattle,
WA 98195
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Cellular stress associated with a malfunctioning endoplasmic reticulum (ER)
is causally related to the inability of molecular chaperones to maintain
structural integrity of proteins. Proteins which are structurally damaged
under conditions of oxidative stress or aggregation during the aging process
place an increased burden on the processing capacity of the ER. The result
is an exaggerated and extended ER stress response, which triggers
transcriptional and translational pathways of cell death and decreased
protein synthesis. ER dysfunction has been implicated in a number of
age-related diseases including Parkinsons, Alzheimers, cardiovascular
conditions, and diabetes. The HSP-40 molecular chaperone family member,
p58ipk, functions in the later stages of the ER stress response as an off
switch to prevent excessive cell death and promote protein synthesis.
Mutant mice deficient in p58ipk have evidence of an exaggerated ER stress
response in several cell types including pancreatic beta cells, hepatocytes,
and plasma cells. Gene expression, Western immunoblots and
immunohistochemistry in cells from p58ipk mutant mice confirm the
upregulation of ER stress response genes and an enhanced cell death pattern
reflective of an age-dependent onset of diabetes and immune dysfunction, and
an average life span of 18 months. We conclude that ER resident
proteins, including HSP-40/p58ipk, are targets for investigating specific
aging and age-related disease questions. The p58ipk mutant mouse model will
be of interest in intervention studies to determine capabilities of
decreased disease and increased life span mediated by a functional ER.
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Nikolich-Zugich, Janko |
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PHENOTYPIC, FUNCTIONAL AND GENETIC PROFILES OF THE AGING T- LYMPHOCYTES
IN PRIMATES AND THEIR ALTERATION BY CALORIC RESTRICTION
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Janko Nikolich-Zugich, Jessica Warner, Bree Fisher, Dragana Nikolich-Zugich
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and Ilhem Messaoudi
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Vaccine and Gene Therapy Institute and the Oregon National Primate Research
Center, Oregon Health & Science University, Beaverton, OR, USA
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Numerous T-cell functions
are diminished or dysregulated in old age, including T-cell population
dysbalance (altered na ve:memory and CD4/CD8 T-cell ratios), diminished
T-cell responsiveness to a variety of signals and altered cytokine networks.
Yet, despite intense research, numerous gaps in the understanding of T-cell
senescence still exist. Moreover, much of our current knowledge on T-cell
biology and aging comes from inbred, specific pathogen-free rodents, and it
is unclear which observations translate to human immune aging.
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We begun to develop a
primate model of human immune senescence using Rhesus macaques (RM). We
evaluated T-cell surface phenotype, in vivo and in vitro turnover, the
complexity of T-cell receptor (TCR) repertoire and correlated them to
functional studies of isolated cell subsets and gene expression profile
analysis. We also evaluated the ability of an experimental manipulation -
caloric restriction, the only intervention that consistently and
reproducibly increases longevity and quality of life in a variety of animal
models - to impact upon these parameters.
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We showed that sharp
differences exist in the phenotypic and functional T-cell aging between the
CD8 and CD4 T-cell subsets in: (i) cell cycle programs (as assessed by both
in vitro proliferation and in vivo turnover measurement); (ii) CD28
regulation upon cell cycle entry; (iii) accumulation of immediate effector
cells amongst the CD28- cells, believed to be close to or at replicative
senescence. These results underscore poor reliability of CD28 as a marker
for senescence, and further suggest that some of the T-cell aging
phenomenology in RM can be ascribed to accentuation over time of the
inherent differences in activation programs in CD8 and CD4 T-cells. We also
found age-related differences in T-cell turnover, and modest, but
consistent, gene expression changes in certain resting T-cell subsets.
Caloric restriction was able to modulate most, if not all, of these
parameters, resulting in young-like characteristics of examined
T-cells. We conclude that the results so far are consistent with resource
conservation induced by caloric restriction.
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Shaikh, Aasef G. |
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cAMP
forms a neurochemical
correlate of tinnitus
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A, Shaikh; P, Finlayson
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Department of Otolaryngology, Wayne State
University, Detroit, MI 48201
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Cochlear damage that commonly occurs in
elderly is either due to ototoxicity of anticancer drugs (cisplatin) or a
process of aging. Such hearing loss is often accompanied by tinnitus. It was
suggested that increased spontaneous neural activity (SA) may be the
underlying mechanism for tinnitus. Neurochemical consequences of cochlear
ablation (CA) have also been widely explored. A significant synaptic
plasticity has been reported following cochlear insults.
Yet a direct relationship of increased SA with plastic neurochemical
alterations remains to be recognized. In central auditory neurons, the
signals emerging after CA are transduced by range of mechanisms including
extracellular signal-regulated kinase (ERK) pathway. One of the functional
roles of this pathway is to enhance the inhibition of phosphodisterase E 4
(PDE4), thereby elevating intracellular cAMP concentrations ([cAMP]i). We
sought to determine if increasing [cAMP]i affects SA, and if so what
possible mechanisms may be involved and the possible
physiological/pathological ramifications. We also investigated such
elevations in SA could mimic the neural code for the pure-tone. Forskolin
(an agent that systematically increases [cAMP]i) specifically increases
tone-evoked responses and SA of the SOC neurons in dose-dependent manner.
Interestingly, the increased SA following application of 50 µM
forskolin mimics the neural code that is normally generated by the tonal
stimulus. We also report that effects of forskolin are predominantly due
to PKA independent pathway that involves hyperpolarization activated inward
conductances. These results provide for the first time, direct evidence that
systematic increase in the [cAMP]i, in the auditory brainstem neurons, by
application of forskolin elevates the SA, which mimics the neural code for
the pure-tone. Here we suggest, increased levels of [cAMP]i that could
follow cochlear insults, forms the basis of increased SA and therefore could
be the “neurochemical correlate” of tinnitus.
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Sun, Liou Y. |
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HIPPOCAMPAL GH AND IGF-1 EXPRESSION IN GH-DEFICIENT MICE
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Liou
Y. Sun*, Khalid Al-Regaiey, Michal M. Masternak, Jian Wang and Andrzej
Bartke
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Geriatrics Research, Department of Medicine and Department of Physiology
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Southern Illinois
University, Springfield, IL 62794, USA
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Liou Y. Sun*, Khalid Al-Regaiey,
Michal M. Masternak, Jian Wang and Andrzej Bartke
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Geriatrics Research,
Department of Medicine and Department of Physiology
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Southern Illinois
University, Springfield, IL 62794, USA
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Beneficial effects of GH and IGF-1 on the development and function of the
central nervous system are well documented. In spite of primary deficiency
of GH and secondary IGF-1 deficiency, Ames dwarf mice live considerably
longer than normal animals, exhibit apparently normal cognitive functions
and maintain them into advanced age. In an attempt to reconcile these
findings, we have examined local expression of GH and IGF-1 in the
hippocampus, brain region involved in learning and memory, of these mice.
RNA and protein was extracted from the hippocampus of dwarf and normal mice
and assayed for the GH and IGF-1 transcripts and their encoded proteins.
With the real-time RT-PCR analyses, hippocampus of Ames dwarf mice was found
to express normal levels of GH and IGF-1 messenger RNA indicating ectopic GH
expression and the integrity of transcription capability in dwarf mice. The
identities of the PCR products were confirmed by sequencing. Hippocampal
levels of GH and IGF-1 were evaluated by western blotting using the
antibodies specific for the respective proteins. Both GH and IGF-1 protein
levels are significantly increased in the hippocampus of Ames dwarf compared
with normal mice suggesting a compensatory mechanism of peripheral hormonal
deficiency. In contrast, IGF-1 expression in the liver of Ames dwarf mice is
profoundly suppressed, consistent with congenital GH deficiency and lack of
detectable GH and IGF-1 in peripheral circulation. In addition, Increased
phosphorylation of Akt and CREB were also detected in the hippocampus of
Ames dwarf mice. Our results suggest that increase in hippocampal GH and
IGF-1 protein expression and subsequent activation of PI3K/Akt-CREB signal
transduction cascade might contribute to the maintenance of cognitive
function and is likely to be responsible for the integrity of neuronal
structure, and maintenance of youthful levels of cognitive function in these
long-lived mice during aging.
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Wilcock, Donna M. |
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PASSIVE AMYLOID IMMUNOTHERAPY CLEARS AMYLOID AND TRANSIENTLY ACTIVATES
MICROGLIA IN A TRANSGENIC MOUSE MODEL OF AMYLOID DEPOSITION
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Donna M. Wilcock1, Amyn Rojiani2, Arnon Rosenthal3, Jennifer Alamed1, David
Wilson1, Nedda Wilson1, Melissa J. Freeman1, Marcia N. Gordon1, Dave
Morgan1.
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AbstractLocation: 1,2:
Alzheimer’s Research Laboratory, 1Department of Pharmacology, 2Departments
of Interdisciplinary Oncology and Pathology, University of South Florida,
12901 Bruce B Downs Blvd, Tampa, FL 33612, USA.
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3: Rinat Neuroscience Corp.
3155 Porter Drive, Palo Alto, California, 94304, USA.
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The role of microglia in
the removal of amyloid deposits following systemically administered anti-Aß
antibodies remains unclear. In the current study we injected Tg2576 APP
transgenic mice weekly with anti-Aß antibody for a period of one, two or
three months such that all mice were 22 months at the end of the study. In
mice immunized for three months we found an improvement in alternation
performance in the Y maze. Histologically, we were able to detect mouse IgG
bound to congophilic amyloid deposits in those mice treated with anti-Aß
antibody but not in those treated with control antibody. We found that
Fcgamma receptor expression on microglia was increased following one month
of treatment while CD45 was increased following two months of treatment.
Associated with these microglial changes was a reduction in both diffuse and
compact amyloid deposits following two months of treatment.
Interestingly, the microglia markers were reduced to control levels
following three months of treatment while amyloid levels remained reduced.
Serum Aß levels and anti Aß antibody levels were elevated to similar levels
at all three survival times in mice given anti-Aß injections rather than
control antibody injections. These data show that antibody is able to enter
the brain and bind to the amyloid deposits, likely opsonizing the Aß and
resulting in Fcgamma receptor mediated phagocytosis. Together with our
earlier work, our data argue that all proposed mechanisms of anti-Aß
antibody mediated amyloid removal can be simultaneously active.
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P O S T E R |
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Al-Regaiey, Khalid |
43. EFFECTS OF LONG-TERM CALORIC
RESTRICTION ON WILD-TYPE AND GROWTH HORMONE RECEPTOR KNOCKOUT MICE
Khalid Al-Regaiey,
Michal M. Masternak, Michael Bonkowski, Liou Sun, Andrzej Bartke
Southern Illinois University School of Medicine, Springfield, IL 62794
Reduced IGF-1/insulin signaling and caloric restriction (CR) are known to extend
life span and delay age related diseases. To address the interaction of these
two interventions, we subjected normal (N) and growth hormone receptor knockout
(KO) mice to CR starting at weaning for 20 months. Molecules involved in
glucose homeostasis were investigated. CR resulted in decreased plasma
glucose levels in both phenotypes (N-CR and KO-CR). Circulating IGF-1 was
reduced in N-CR and was undetectable in KO animals. Insulin was also
reduced in N-CR to levels comparable to those in KO animals. Corticosterone and
adiponectin levels were higher in KO than in N animals while leptin was reduced
by CR in both phenotypes. We also analyzed hepatic gene and protein expressions
of molecules involved in insulin signaling using real-time PCR and western
blotting, respectively. Diet and phenotype did not affect Akt gene and total
protein expression while KO mice exhibited reduced Akt activation with no diet
effect. Phosphorylation of protein kinase C γ/ζ was not affected by
either diet or phenotype. KO mice exhibited increased CREB activation. Gene
expression and protein levels of Foxo1 and PGC-1 were increased in KO mice with
little diet effects. Genes that are known to be regulated by CREB, Foxo1, and/or
PGC-1including pepck, g6pase, igfbp-1, and MnSOD had increased expression in KO
mice. Genes encoding proteins that are involved in lipolysis, including
hormone-sensitive lipase (HSL) and lipo-protein lipase (LPL) were upregulated in
KO mice. We conclude that CR and growth hormone resistance probably affect
longevity of mice by different mechanisms and that gluconeogenesis pathway as
well as fatty acids mobilization are more active in KO mice. These data also
suggest that liver of KO mice is more protected against oxidative stress as
indicated by increased Foxo proteins and MnSOD mRNA.
Supported by NIA.
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Anderson, Rozalyn M. |
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44. AGING RETARDATION BY CALORIE RESTRICTION: ROLE OF REGULATORS OF ENERGY
METABOLISM.
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R.
Anderson (1), J.Barger (1), T. Pugh (1), S. Park (2), T. Prolla (2) and R.
Weindruch (1).
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(1)Department of Medicine, University of Wisconsin and GRECC, VA Hospital,
Madison WI 53705 USA. (2) Department of Genetics and Medical Genetics,
University of Wisconsin, Madison WI 53706 USA.
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University of Wisconsin
Madison, GRECC, VA Hospital, Madison WI 53705, USA.
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Caloric restriction (CR) extends lifespan in a broad spectrum of organisms
and retards the progression of a wide range of age-associated biological
changes; however, the underlying mechanisms are unclear. In mice,
there is an inverse linear relationship between calorie intake and lifespan
extension suggesting that factors central to energy metabolism may underlie
aging retardation by CR. One hypothesis is that CR triggers an active
response involving a reprogramming of energy metabolism that reduces the
rate of aging and the development of several age-related diseases.
Transcription profiling of tissues from control and CR animals points to
mitochondrial function as a central feature in this process. We have
used a combination of techniques to identify factors that may be involved in
the mechanism of lifespan extension by CR. We then investigated how
these potential effectors of CR are regulated and examined their
relationship to elements of known longevity pathways, including members of
the FOXO and Sirtuin families.
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Armbrecht, Harvey J. |
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45. REGULATION OF THE RENAL VITAMIN D 1ALPHA-HYDROXYLASE CYTOCHROME
P450(CYP27B1) BY DIETARY PHOSPHORUS CHANGES WITH AGE
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H.J. Armbrecht*, M.A. Boltz
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Geriatric Center, St. Louis VA Medical Center, St. Louis, MO 63125
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Young rats adapt to a low phosphorus diet by increasing plasma levels of
1,25-dihydroxyvitamin D (1,25D), the biologically active metabolite of
vitamin D, which then increases intestinal phosphate absorption.
Previous studies have shown that the capacity of rats to adapt to low
dietary phosphorus declines with age. The purpose of this study was to
determine whether this decreased adaptation was due to decreased expression
of the renal 1alpha-hydroxylase (1-OHase), which makes 1,25D. The
1-OHase consists of ferredoxin reductase, ferredoxin, and a terminal
cytochrome P450 – CYP27B1. Young (2 months) and adult (12 months) F344
rats were placed on a low phosphorus (0.1%) or high phosphorus (1.0%) diet
for 2 weeks. Plasma 1,25D was markedly increased by the low phosphorus
diet in young animals but not in adults. To determine whether this
difference was due to decreased 1-OHase expression, mRNA levels of CYP27B1
were measured by ribonuclease protection assay. In young animals, the
low phosphorus diet increased renal CYP27B1 mRNA levels 8-fold compared to
the high phosphorus diet, but there was no significant effect in adults.
The effect of low dietary phosphorus on plasma calcium, phosphate, and
parathyroid hormone levels was similar in both age groups. These
results suggest that the decreased CYP27B1 expression in the adult in
response to dietary phosphorus depravation is not due to changes in the
major regulators of 1-OHase activity.
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Ball, Sheldon S. |
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46. SENEX: INTEGRATING GERIATRICS AND MOLECULAR GERONTOLOGY
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S
Ball, A Nhalil and V Mah
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Department of Veterans Affairs, Greater Los Angeles Healthcare, 11301
Wilshire Boulevard, GRECC 11G, Los Angeles CA 90073
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Senex is a computer application 15 years in development integrating
geriatrics and molecular gerontology (from bench to bedside). It includes
information (and tools for interpretation of data) that span internal
medicine, molecular pathology (biochemistry, molecular biology, pathology),
laboratory medicine, radiology, pharmacology, anatomy, and statistics.
Clinical and molecular information is needed from laboratory bench to
patient bedside, during contact between physician and patient, educator and
student, scientist and technician. The information needs to be
structured so that specific and real-time retrieval is achievable. The
information needs to be presented in a manner that facilitates a conceptual
understanding of the details presented. Additionally, an information
system should display a certain degree of intelligence, including
flexibility in accepting input from the user, the capacity to reason with
structured information, and the esthetic display of context-specific
information. Implementation of such an information system should allow use
to become better physicians, scientists, educators, students and/or informed
citizens. Senex functions independently as a stand-alone application (i.e.
not dependent upon internet access). Thus it is fast, reliable, and mobile.
Senex also provides direct links into the world-wide-web, to molecular and
clinical databases and to the original literature when access to the
internet is available. Senex is a large application with: 1514
organisms, 2417 anatomic structures, 222 cells, 131 cellular compartments,
13,138 molecules, 9607 proteins, 628 genes, 795 motifs, 67 molecular
pathways, 1580 diseases, 540 clinical laboratory tests, 18,680
database links (6140 Swiss Prot, 6280 OMIM, 870 Prosite, 2000 PIR,
3390 Locus Link). Senex allows a user to add proprietary information
on top of the core Senex knowledge base, thus customizing the application
for a user’s own information needs. Senex runs on Macintosh and
Windows computers. The Macintosh version has a microarray data analysis and
data mining module. Reference: www.senex-medical-software.com
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Bergamini, Ettore |
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47. THE AGE-RELATED ACCUMULATION OF DOLICHOL IN TISSUES SATISFIES ALL
CRITERIA TO BE QUALIFIED A BIOMARKER OF AGING
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E. Bergamini, G. Cavallini, A. Donati, Z. Gori, A. Manfrini, I. Parentini
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-
Centro di Ricerca di Biologia e Patologia dell’Invecchiamento
dell’Universita’ di Pisa, Dipartimento di Patologia sperimentale, Via Roma
55, 56126 PISA (Italy); Tel 0039-050-2218584 Fax 0039-050-2218581
e-mail ebergami@med.unipi.it
-
-
It was stated that the
identification of specific biomarkers of aging would be an important
milestone in gerontological research, and criteria for defining biomarkers
were suggested (Mooradian, 1990). In this communication evidence on male and
female Sprague Dawley and Lewis rats fed ad-libitum or on regimens of food
restriction and on human tissues are reviewed, showing that accumulation of
dolichol in tissues of older animals meet these criteria: levels of dolichol
exhibit a quantitative correlation with age in all tissues; the biological
parameter is not altered by several age-dependent diseases in the same
direction as that of aging; data with transplanted organs show that the
age-related accumulation is not secondary to metabolic changes of aging;
factors that modulate the aging rate like caloric restriction and physical
exercise appropriately alter the accumulation of dolichol; this putative
biomarker of aging appears to be applicable to different tissues across
mammalian species including human; biomarker is applicable to the
hypothalamic digoxin-mediated model for trisomy 21 (Kurup & Kurup, 2003).
Reliable changes in tissue dolichol levels are seen in relatively short
intervals of time compared to life-time, and levels can be tested on a small
amount of tissue without causing death of the animal, which are desirable
features of a biomarker of aging. The age-associated alteration in dolichol
levels is likely to reflect an age-dependent derangement of free radical
metabolism in membranes. Useful applications are shown, which include a
study of the effect of the donor-recipient age-mismatch on the biological
age of the graft tissues, and the detection of gender-related differences in
aging.
-
Research was supported in
part by a grant of MIUR (Ministero Istruzione, Universita’ e Ricerca)
-
Mooradian, A.D. (1990) J
Gerontol 45(8) B183-186.
-
Kurup R.K., Kurup, P.A.
(2004) Pediatr Pathol Mol Med 22(5) 411-422.
|
|
Bowen,
Richard |
107. LIVING AND DYING FOR SEX: A THEORY OF AGING
BASED ON THE MODULATION OF CELL CYCLE SIGNALING OF REPRODUCTIVE HORMONES
R. Bowen, C. Atwood*
Voyager Pharmaceutical Corporation, Raleigh, North Carolina, USA
* Department of Pathology, Case Western Reserve University, Cleveland, Ohio and
School of Medicine, Section of Geriatrics and Gerontology, Department of
Medicine, University of Wisconsin, Madison, Wisconsin, USA
We put forth a new theory based on a novel definition of aging - any change in
an organism over time. This definition includes not only the changes associated
with the loss of function (i.e. senescence), but also the changes associated
with the gain of function (growth and development). Using this definition, the
rate of aging would be synonymous with the rate of change. The rate of
change/aging is most rapid during the fetal period when organisms develop from a
single cell at conception to a multicellular organism at birth. Therefore,
“fetal aging” would be determined by factors regulating the rate of mitogenesis,
differentiation, and cell death. We suggest these factors also are responsible
for regulating aging throughout life. Thus, whatever controls mitogenesis,
differentiation and cell death must also control aging. Since life-extending
modalities consistently affect reproduction, and reproductive hormones are known
to regulate mitogenesis and differentiation, we propose that aging is primarily
regulated by the hormones that control reproduction (hence, the
Reproductive-Cell Cycle Theory of Aging). In mammals, reproduction is controlled
by the hypothalamic-pituitary-gonadal (HPG) axis hormones. Longevity inducing
interventions, including caloric restriction, decrease fertility by suppressing
HPG axis hormones and HPG hormones are known to affect signaling through the
well-documented longevity regulating GH/IGF-1/PI3K/Akt/Forkhead pathway. This is
exemplified by genetic alterations in C. elegans where homologues of the HPG
axis pathways, as well as the daf-2 and daf-9 pathways, all converge on daf-16,
the homologue of human Forkhead that functions in the regulation of cell cycle
events. In summary, we propose that the hormones that regulate reproduction act
in an antagonistic pleiotrophic manner to control aging via cell cycle
signaling; promoting growth and development early in life in order to achieve
reproduction, but later in life, in a futile attempt to maintain reproduction,
become dysregulated
|
|
Chacon, M.A. |
108. UP-REGULATION OF FREE
FATTY ACIDS - A POTENTIAL MASTER SWITCH OF CALORIC RESTRICTION PROTECTION
P. Wills1, H.
Brown-Borg2,and M.A. Chacon1(P)
1Irazu
Biodiscovery, LLC, Baltimore, MD 21224; 2Department of Pharmacology,
Physiology and Therapeutics, University of North Dakota School of Medicine and
Health Sciences, Grand Forks, ND 58203.
Seventy years of scientific
investigation has demonstrated that caloric restriction (CR) without
malnutrition is the only experimental intervention that can extend life span,
and decrease the incidence and delay the onset of pathologies associated with
aging. Nevertheless, the precise mechanism by which caloric restriction induces
its life-extending effect remains poorly understood. One consequence of caloric
restriction observed in experimental rats, which has not been previously
investigated, is an elevation in serum levels of free fatty acids (FFA). In our
studies, we found that calorie restricted rats, mice, and dwarf mice all have
elevated free fatty acids. In addition to their well-characterized role as an
energy substrate, recent scientific studies have demonstrated that these
molecules possess signaling properties. Therefore, we tested the hypothesis
that free fatty acids may induce (mimic) some of the protective effects seen in
calorie restricted animals. Using an in vitro, cell-based ROS assay, we
demonstrated that supplementation of serum from ad lib animals with free fatty
acids to levels comparable to those found in the serum of caloric restricted
animals conferred protection from the cytotoxic effects of H2O2. We contend
that the up-regulation of FFA and their CoA derivatives, through their effects
on ion channels, oxygen utilization, and gene expression, offer a plausible
mechanism for the protective phenotype associated with CR.
|
|
Chai,
Weihang |
48. SENESCENT NORMAL HUMAN FIBROBLASTS MAINTAIN
THEIR 3' TELOMERIC OVERHANG LENGTH
W. Chai, J.W. Shay, and W.E. Wright
Department of Cell Biology, University of Texas
Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75390-9039
Replicative senescence involves progressive telomere shortening but the exact
molecular mechanisms triggering the growth arrest are poorly understood.
Recently, using a telomere-oligonucleotide ligation assay (T-OLA), it was
reported1 that the 3’ G-rich overhang was eroded at senescence, leading to the
hypothesis that loss of the 3’ G-rich overhang is the molecular signal that
triggers senescence. To better understand the molecular mechanisms triggering
replicative senescence, we have developed a quantitative assay to measure the
length of the G-rich 3’ telomeric overhangs from cultured cells. By using this
assay and the conventional non-denaturing hybridization assay for measuring
G-rich overhangs, we show that normal human fibroblasts maintain their 3’ G-rich
overhangs at senescence. Furthermore, cells maintain their G-rich overhangs when
they bypass senescence after the inactivation of p53 and retinoblastoma proteins
by human papillomavirus type 16 (HPV) oncoproteins E6 and E7 (E6/E7).
Interestingly, cells expressing just HPV E7 exhibit longer overhangs when they
growth arrest. We also found that expression of simian virus 40 large T-antigen
induces some telomeric overhang shortening. These results demonstrate that a
significant reduction in overhang length is not the molecular signal that
triggers senescence.
Reference:
1. Stewart, S.A., I. Ben-Porath, V.J. Carey, B.F. O'Connor, W.C. Hahn, and R.A.
Weinberg. 2003. Erosion of the telomeric single-strand overhang at replicative
senescence. Nat Genet 33: 492-6.
|
|
Chow, Vivian W.H. |
-
49. APOPTOTIC DNA FRAGMENTATION IN THE BRAINS OF YOUNG AND AGED eNOS-, iNOS-
and nNOS-KNOCKOUT MICE: MEASUREMENT BY NEW ULTRASENSITIVE CE-LIF TECHNIQUE
-
-
VW
Chow*, RR Fiscus, SB Chew
-
-
The
Department of Physiology, The Chinese University of Hong Kong, Shatin, N.T.
Hong Kong.
-
-
Previously, we showed that
elevation of cGMP levels in PC12 cells exposed to atrial and brain
natriuretic peptides (ANP & BNP) inhibits onset of apoptosis and promotes
survival [1]. We further showed that 24h-preincubation of NG108-15 cells
with ANP protects against pro-apoptotic effects of nitric oxide (NO) donor
(high concentrations) [2]. At lower physiological concentrations, NO, via
cGMP elevations, inhibits apoptosis and improves neural survival [3].
Depending on the source of NO, whether from nNOS, iNOS or eNOS, and
depending on age, NO may promote or protect against apoptosis. The present
study determines if aging-induced-apoptosis in brain is affected by genetic
deletion of nNOS, iNOS and eNOS using nNOS-, iNOS-, eNOS-knockout mice.
Young (2-4-month-old) and aged (12-22-month-old) nNOS-/-, iNOS-/-, eNOS-/-
knockout and control mice (C57BL/6J & B6129SF2/J) were used to determine
levels of apoptosis in half-brain and different brain regions by a new
ultrasensitive technique using capillary electrophoresis with laser-induced
fluorescent detector (CE-LIF) to accurately quantify apoptotic-DNA
fragmentation. Using half-brain, amounts of apoptotic-DNA-fragments were
11-fold higher in aged than in young B6129SF2/J (nNOS-/-control) mice
(p<0.0001). Apoptotic-DNA-fragments in young nNOS-/- were 23-fold higher
than in young B6129SF2/J (p<0.0001). Apoptotic levels in aged B6129SF2/J
were 2.5-fold higher than in aged nNOS-/- (p<0.001). These data suggest that
nNOS contributes to aging-dependent-increase in brain apoptosis of
B6129SF2/J, but provide neuroprotective effect in young adults.
Neuroprotective effects of nNOS were especially large in young hippocampus.
Apoptotic-DNA-fragments in young iNOS-/- were 30-fold higher than in young
C57 controls (p<0.001). Aged iNOS-/- showed 6.5-fold lower level of
apoptosis compared to young adult (p<0.001). iNOS had anti-apoptotic effect
in young C57 but not aged C57. eNOS showed significant neuroprotective
effect in medulla oblongata of both young and aged C57. All NOS isoforms
provided neuroprotective function in young adult brain, but this
neuroprotection appeared change during aging.
-
1. Fiscus
RR, Tu AW, Chew SB. Neuroreport 2001;12:185-189.
-
2. Cheng
Chew SB, Leung PY, Fiscus RR. Histochem Cell Biol. 2003;120:163-171.
-
3. Fiscus
RR. Neurosignals 2002;11:175-190.
-
Support: Competitive
Earmarked Grant from RGC of Hong Kong to RRF. (#CUHK 4169/02M)
-
|
|
Citron, Bruce |
- 50. GENE EXPRESSION CHANGES IN NEURODEGENERATIVE MICE AND CULTURED
CELLS
- B.A. Citron*1,2, A. Surguchov3,4, I. Surgucheva3,4, and B. W.
Festoff3,4,5
1Lab of Molecular Biology, VA Medical Center, Bay Pines, FL, 2Dept. of
Biochemistry & Molecular Biology, USF College of Medicine, Tampa, FL,
3Neurobiology Research Lab, VA Medical Center, Kansas City, MO, Depts. of
4Neurology and 5Pharmacology, University of Kansas Medical School, Kansas City,
KS
A variety of genes have been implicated in late onset neurodegenerative
disorders. Several reports linked synucleins to motor neuron diseases including
ALS. Present in aggregates, α-synuclein is important for the long term survival
of neurons. The role of γ-synuclein is less clear but recent studies indicate
that it regulates other neurodegenerative factors. Synucleins modulate certain
proteases, however their own proteolytic degradation may be essential in the
process of formation of inclusion bodies. We have been studying cultured neurons
and wobbler mice, which undergo an age dependent loss of motor neurons in the
cervical region of the spinal cord and model sporadic ALS. We have measured
activation of transcription factors (with electrophoretic mobility shift assays)
in wobbler spinal cords and cultured neurons exposed to insults. Specifically,
we found that activated transcription factors SP1 and NF-kB, but not AP1 are
increased in wobbler mice and in motor neurons exposed to apoptosis inducing insults (thrombin or mechanical injury). We
have also examined mRNAs levels with Affymetrix gene arrays. Genes responsible
for CNS maintenance (e.g. myelin-associated oligodendrocytic basic protein) are
downregulated in the wobbler spinal cord. Ubiquitin C-terminal hydrolase is also
expressed at lower levels in the wobbler cord and this could affect the
ubiquitination of proteins that aggregate in the degenerating CNS. α-synuclein
is upregulated in the wobbler compared to the wild-type littermate spinal cord.
Directed elevation of α-synuclein, γ-synuclein or both, in transfected neurons
resulted in a marked dysregulation of gene expression in several functional
categories. For example, of the genes that are significantly expressed in these
cells, more genes involved in cell adhesion are upregulated compared to genes in
several other categories. The over and underrepresented functional classes in
gene groups upregulated and downregulated by α- and/or γ-synuclein introduction have been determined to obtain
a picture of synuclein regulatory influences.
|
|
Cohen, Haim Y. |
- 51.
The SIRT1 NAD+-dependent
deacetylase regulates Bax-mediated apoptosis
Haim Y. Cohen*, Christine Miller, Kevin Bitterman and David A. Sinclair.
Department of Pathology, Harvard Medical School. Boston MA USA 02115.
Decreased apoptosis and increased cell survival is observed in organisms whose
lifespan has been extended by calorie restriction or genetic manipulation. A
crucial but poorly understood step in the stress-induced apoptotic pathway is
activation of the pro-apoptotic factor Bax. In yeast, lifespan extension by
calorie restriction requires Sir2, a stress-responsive, NAD+-dependent
deacetylase. Here we show that the human Sir2 homolog, SIRT1, controls cell
survival in response to stress by inhibiting Bax-mediated apoptosis. SIRT1
deacetylates specific lysines in the Ku70 C-terminus, which increases the
Ku70-Bax interaction and prevents Bax from relocalizing to mitochondria. We show
that this pathway can be manipulated using small molecules and discuss the
implications of this finding to the general decline in physiological function
with age.
|
|
Coumoul, Xavier |
- 52. Modulation of IGF members expression in Brca1-knockout Mice
X Coumoul*, L Cao and C Deng
National Institute of Health, National Institute of
Diabetes and Digestive and Kidney Diseases, 9000 Rockville Pike, Bethesda MD
20892, USA
Breast cancer affects 200.000 women in the United States which results in 40.000
deaths every year. Heterozygous carriers of Breast Cancer Associated Protein 1
(Brca1) mutations are at great risk to develop breast or ovarian cancers through
their lifetime. Our mice models of Brca1 deletions have shown that indeed lack
of this gene plays a role in mammary tumorigenesis but also interestingly in
aging process. Here, we shown that deletion of Brca1 modulates expression of
several Insulin-like Growth Factor (IGF) members in mice liver. IGF members
expression has previously been linked to tumorigenesis and aging process.
Interestinlgy, IGF1 serum levels but also mRNA liver levels are increased as
shown by microarrays and RT-PCR analysis. Other members like IGF1-Receptor,
Insulin Receptor Substrate 1 or IGF-Binding Protein 1 and 2 also appears to be
modulated. We also use a human cell culture system UBR60, allowing modification
of Brca1 expression, to shown that those results are reproducible with a
different model. Those results allow us to hypothesize that modulation of IGF
members expression by Brca1 could be one of the mechanism that links lack of
this protein to aging and tumorigenesis process.
|
|
de Grey, Aubrey D. |
-
53.
THE UNFORTUNATE
INFLUENCE OF THE WEATHER ON THE RATE OF AGING
-
-
A. de Grey
-
-
Department of Genetics,
University of Cambridge, Downing Street, Cambridge CB2 3EH, UK
-
-
Much research interest, and
recently even commercial interest, has been predicated on the assumption
that reasonably closely-related species -- humans and mice, for example --
should in principle respond to aging-retarding interventions with an
increase in life expectancy roughly proportional to their control lifespan
(life expectancy without the intervention). Here it is argued that the
best-studied life-extending manipulations of mice are examples of a category
that is highly unlikely to follow this rule, and more likely to exhibit only
a similar absolute increase in lifespan from one species to the next,
independent of the species' control lifespan. That category -- deprivation
of dietary calories or of the organism's ability to metabolise or sense them
-- is widely recognised to extend lifespan as an evolutionary adaptation to
transient starvation in the wild, a situation which alters the organism's
optimal partitioning of resources between maintenance and reproduction. What
has been generally overlooked is that the extent of the evolutionary
pressure to maintain adaptability to a given duration of starvation varies
with the frequency of that duration, something which is -- certainly for
terrestrial animals, and less directly for others -- determined principally
by the weather. The pattern of starvation that the weather imposes is
suggested here to be of a sort that will tend to cause all terrestrial
animals, even those as far apart phylogenetically as nematodes and mice, to
possess the ability to live a similar maximum absolute (rather than
proportional) amount longer when food is short than when it is plentiful.
This generalisation seems to be strikingly in line with available data,
leading to the biomedically and commercially sobering conclusion that human
interventions which manipulate caloric intake or its sensing are unlikely
ever to confer more than two or three years' increase in lifespan at the
most.
-
|
|
Ding, Shi-Ying |
-
54. CRP LEVELS ARE STRONGLY ASSOCIATED WITH AGING
-
-
SY.
Ding, X.T. Tigno, and B.C. Hansen
-
-
Obesity, Diabetes, and Aging Animal Resource Center, Department of
Physiology, School of Medicine, University of Maryland at Baltimore, 10 S.
Pine St, Baltimore MD 21201
-
-
Epidemiological studies
have consistently shown that the plasma level of C-Reactive Protein (CRP), a
marker of inflammation, is a strong, independent predictor of future
cardiovascular events, both in patients with a history of coronary heart
disease (CHD) and in apparently healthy subjects. Inflammatory markers have
also lately been implicated in aging and in type 2 diabetes. The plasma
concentrations of CRP were determined by ELISA in a cohort of 33 diabetic
and non-diabetic rhesus monkeys: 13 young normal (Y) with a mean age of
9.1±2.9 yr, 11 old normal (O, 25.9±1.5 yr), 8 old type 2 diabetic monkeys
(DM, 26.2±2.3 yr) and 4 calorie-restricted (for more than 20 years) monkeys
(CR, 30.1± 2.8 yr). Results show that CRP levels were positively correlated
to age (r=0.46, p<0.01) and body weight (r=0.33, p<0.02). The mean levels of
CRP were found to be highest among the O (246±16µg/L), a value significantly
greater than that among the Y (184±14µg/L). However, mean CRP values did not
differ significantly between O, DM (221.4±18µg/L) and CR (214±26µg/L), all
of whom were elderly. The data suggests that the proportion of the variation
in CRP that can be accounted for by variation in age is 0.21, and that age
is a stronger predictor of CRP than weight (R2= 0.18) or metabolic status,
as reflected by circulating levels of CRP in the non-human primate.
|
|
Dolle, Martijn |
55. AGE-RELATED GENOME INSTABILITY IN DNA
REPAIR-DEFICIENT MICE
M. Dollé1, R. Busuttil2, L. Niedernhofer3, G. van der Horst3, J. Hoeijmakers3,
S. Wijnhoven1, P. Lohman1, H. van Steeg1, J. Vijg2
1National Institute of Public Health and the Environment, Department of
Toxicology, Pathology and Genetics, Bilthoven, the Netherlands; 2University of
Texas Health Science Center, Department of Physiology, San Antonio, TX, USA;
3Erasmus University, Department of Cell Biology and Genetics, Rotterdam, the
Netherlands
Genomic instability has been implicated as a major cause of both cancer and
aging. Previously we have shown that mutations accumulate with age in an
organ-specific manner, using a transgenic mouse model with a chromosomally
integrated lacZ mutational reporter gene. To examine the correlation between age
and mutation load, we are investigating genomic instability in aging DNA
repair-deficient mice. Here we present our preliminary findings on different
mouse mutants with deficiencies in nucleotide excision repair (NER) and DNA
interstrand crosslink (ICL) repair. NER removes a broad range of lesions, such
as UV-induced damage, other bulky adducts and some forms of oxidative damage.
Life span studies and mutant frequency determinations in liver and kidney were
performed on male mice with defects in global and/or transcription-coupled
repair (Xpa, Csb or Xpd(ttd)). We observed a reduced life span for Xpa and
Xpd(ttd), as compared to Csb and wild type control animals. Conform to our
initial studies with just wild type mice, all NER-deficient models showed an
age-related increase of mutant frequencies in liver and kidney. However, only
the Xpa-mice showed elevated mutant frequencies over control animals in both
organs. In addition, we examined mutant frequencies in liver of the very
short-lived Ercc1(*292) mouse. This mouse model lacks the last 7 amino acids of
Ercc1 and is deficient in both NER and DNA ICL repair. We observed a two-fold
increase in mutant frequency over control mice at 23 weeks, which is close to
their maximum life span. A comparison of mutation spectra in liver revealed that
the general NER-deficiency of XPA-mice resulted in point mutations,
predominantly consisting of one base pair deletions. The increased mutant
frequency in the Ercc1(*292) mice was due to both point mutations and
translocations, the latter presumably reflecting the additional cross-link
repair deficiency compared to Xpa-mice.
|
|
Falcon, Alaric A. |
-
56. DNA EPISOMES AND REPLICATIVE SENESCENCE IN
S. CEREVISIAE
-
- Alaric A.
Falcón*, Diego M. Ayo, Natalie Rios, and John P. Aris
-
- Department
of Anatomy and Cell Biology, Health Science Center, University of Florida,
Gainesville, FL 32610-0235
-
- We have shown that
plasmids containing an autonomously replicating sequence (ARS; yeast DNA
replication origin) reduce yeast replicative life span, most likely due to
their accumulation during replicative aging (Plasmid accumulation reduces
life span in Saccharomyces cerevisiae(Falcón and Aris, 2003). This raises
the question: how is plasmid accumulation mechanistically linked to
replicative senescence in yeast? One possibility is that accumulated DNA
exerts an effect on other cellular DNA, perhaps by influencing DNA
replication and/or inheritance. To explore this, we have studied
“interactions” between different types of episomes in yeast: recombinant
plasmids, extrachromosomal rDNA circle (ERCs), and the naturally-occurring 2
micron plasmid. A short-lived sir2∆ strain that contains elevated
ERC levels transmits recombinant ARS1-plasmids to daughter cells more
often than a SIR2 control strain. However, an ARS1-CEN4-plasmid
is transmitted to daughter cells equally well in sir2∆ and SIR2
strains. Levels of ARS1-plasmids are reduced in old sir2∆
cells, compared to old SIR2 cells. Unexpectedly, an ARS1-plasmid,
but not an ARS1-CEN4-plasmid, significantly increased the
frequency of loss of the 2 micron plasmid (i.e., curing of a cir+ strain
to cir0).
Consistent with this, levels of the 2 micron plasmid are reduced in old
cells compared to young cells. Thus, accumulation of different types of
nuclear episomal DNAs can have significant and unexpected effects on other
nuclear DNAs. These findings support a relationship between episomal DNA
accumulation, effects on cellular DNA metabolism, and cell senescence in S. cerevisiae.
|
|
Fiscus, Ronald R. |
-
57.
INHIBITOR OF APOPTOSIS PROTEIN-2 (IAP-2) EXPRESSION LEVELS ARE
DOWN-REGULATED BY CYCLIC GMP DEPLETION AND UP-REGULATED BY CYCLIC GMP
ELEVATIONS INDUCED BY NATRIURETIC PEPTIDES OR ANALOGS OF CYCLIC GMP IN
NG108-15 CELLS: MOLECLULAR MECHANISM OF CYCLIC GMP/PROTEIN KINASE G (PKG)-MEDIATED
NEUROPROTECTION
-
-
R.R.
Fiscus* and J.P. Yuen
-
-
Dept. Physiology (Fac. Medicine), Epithelial Cell Biology Res. Ctr. and Ctr.
for Gerontology & Geriatrics, The Chinese University of Hong Kong, Shatin,
NT, Hong Kong
-
-
Previously, we showed that cGMP elevations protect hippocampal neurons
against glutamate toxicity [1], inhibit apoptosis and prolong cell survival
in stressed PC12 cells [2] and protect NG108-15 cells against the
pro-apoptotic effects of nitric oxide (NO, used at high/neurotoxic
concentrations mimicking pathogenesis of Alzheimer’s and Parkinson’s
disease) [3]. Activation of cGMP/PKG signaling pathway in neurons may
represent an important physiological mechanism protecting against
neurotoxicity and neurodegeneration (reviewed [4]). Even basal levels
of cGMP appear sufficient (and necessary) to prevent spontaneous development
of apoptosis in some neural cells (e.g. NG108-15 cells), because depletion
of cGMP by ODQ triggers apoptosis [4]. Recently, cAMP was shown to
inhibit apoptosis in intestinal epithelial cells via protein kinase A (PKA)-mediated
phosphorylation of CREB and increased IAP-2 expression [5]. Because
PKG phosphorylates CREB at the same site as PKA [4], we hypothesized that
cGMP/PKG-mediated neuroprotection may involve IAP-2 expression. The
present study found that ODQ (10-100 micromolar, 24 h), which lowered basal
cGMP levels, greatly reduced protein content of IAP-2 (Western blot) in a
concentration-dependent manner in NG108-15 cells. Elevation of cGMP
levels with either atrial or brain natriuretic peptides (ANP or BNP, 10-100
nM) or with 8-pCPT-cGMP (1-100 micromolar, a cell-permeable direct activator
of PKG), all of which by-pass the ODQ block, completely reversed the ODQ-induced
down-regulation of IAP-2. Indeed, ANP and BNP up-regulated IAP-2
levels above normal control. The results suggest that up-regulation of
IAP-2 expression in neural cells may represent one of the molecular
mechanisms that mediates cGMP/PKG-induced protection against apoptosis.
1. Barger et al (1995) J Neurochem 64:2087-2096, 2. Fiscus et al
(2001) NeuroReport 12:185-189, 3. Chew et al (2003), Histochem Cell
Biol 120:163-171, 4. Fiscus (2002) NeuroSignals 11:175-190, 5.
Nishihara et al (2003) PNAS 100:8921-8926. (Support: Grant #
CUHK4169/02M from Research Grants Council of Hong Kong to RRF)
|
|
Gemma,
C |
58. nINHIBITION OF CASPASE 1 IMPROVES CONTEXTUAL FEAR CONDITIONING IN AGED RATS
C. Gemma (P)*, M. Fister, C.
Hudson, P.C. Bickford
James A Haley VAH, Tampa
Florida and Center for Aging and Brain Repair, USF, 12901 Bruce B. Downs BLVD,
Tampa, FL 33612
An interplay between the immune
system and the central nervous system may underlie the neuropsychological
changes occurring with aging. It has been long accepted that brain levels of
certain cytokines increase as a function of age, even in absence of a pathologic
stimulus. In aged rats there is an increase in IL1b that has been implicated in
declines of synaptic plasticity in the hippocampus and performance on cognitive
tasks. IL-1b receptor expression is high in the hippocampus, an area of the
brain, which plays a pivotal role in memory and learning, suggesting that the
effects of IL-1b may be specific to hippocampal-dependent memory processes.
However, although at pathophysiological levels IL-1b produces detrimental
effects on learning and memory processes, under physiological circumstances
IL-1b seems to be required for normal learning and memory processes. IL-1b is a
proinflammatory cytokine initially synthesized in an inactive precursor form
that is cleaved to generate the biological mature 17kDa form by a protease named
caspase-1. In the present study we chronically inhibit the cleavage of IL-1b
using a specific inhibitor of Caspase-1(Ac-YVAD-CMK, 10 pmol), both in old (22
months) and young (4 months) rats, and observe the effect on contextual fear
conditioning paradigm. Ac-YVAD-CMK was delivered for 28 days icv through a
brain infusion cannula connected to an osmotic minipump (Alzet, Model 2004
pumping rate, 0.25ml/h; total volume 200ml) implanted subcutaneously. On day 20
the animals were exposed on contextual fear conditioning and the memory for
context was tested on day 22. Chronic infusion of a specific Caspase-1
inhibitor in aged rats resulted in an improvement in the memory for context.
|
|
Gorbunova, Vera |
59.
EXPRESSION OF HUMAN TELOMERASE (hTERT) DOES NOT PREVENT STRESS-INDUCED
SENESCENCE IN NORMAL HUMAN FIBROBLASTS BUT PROTECTS THE CELLS FROM
STRESS-INDUCED APOPTOSIS AND NECROSIS
Vera Gorbunova1 (P),
Andrei Seluanov1, and Olivia M. Pereira-Smith2
1Verna
and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor
College of Medicine, Houston, TX 77030
2Department
of Cellular and Structural Biology, Sam and Ann Barshop Center for Longevity and
Aging Studies, University of Texas Health Science Center, San Antonio, TX
78229-3900
Cells subjected to sub-lethal
doses of stress such as irradiation or oxidative damage enter a state that
closely resembles replicative senescence. What triggers stress-induced
premature senescence (SIPS) and how similar this mechanism is to replicative
senescence are not well understood. It has been suggested that stress-induced
senescence is caused by rapid telomere shortening resulting from DNA damage. In
order to test this hypothesis directly, we examined whether overexpression of
the catalytic subunit of human telomerase (hTERT) can protect cells from SIPS.
We therefore analyzed the response of four different lines of normal human
fibroblasts with and without hTERT to stress induced by UV, -irradiation, and
H2O2. SIPS was induced with the same efficiency in normal and hTERT-immortalized
cells. This suggests that SIPS is not triggered by telomere shortening and that
nonspecific DNA damage serves as a signal for induction of SIPS. Although
telomerase did not protect cells from SIPS, fibroblasts expressing hTERT were
more resistant to stress-induced apoptosis and necrosis. We hypothesize that
healing of DNA breaks by telomerase inhibits the induction of cell death, but
because healing does not provide legitimate DNA repair, it does not protect
cells from SIPS.
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|
Guerin, John C. |
-
60. ROCKFISH: RETARDING AGING WITH "NEGLIGIBLE SENESCENCE"
-
-
J.
Guerin
-
-
Portland, OR
-
-
Field observations have suggested for quite some time that certain fish,
turtles and invertebrates have extremely long maximum lifespan potential.
Age validation techniques have since confirmed these observations.
Negligible senescence is defined in part as no observable age-related
increase in mortality rate or decrease in reproduction rate after maturity,
and no observable age-related decline in physiological capacity or disease
resistance. Recent data compiled on rockfish (genus Sebastes) have
documented ages exceeding 200 years. However, many rockfish species
are reported under 30 years maximum observed lifespan, raising the
intriguing possibility of intra-species lifespan comparison. The
Centenarian Species and Rockfish Project has 14 total pilot studies.
The three studies reported in this poster are: “Rockfish liver microarrays
using existing 16,000+ zebrafish gene chips”, a collaboration between Glenn
S. Gerhard, Dartmouth Medical Center, and Renee Malek, TIGR (The Institute
for Genomic Research); “Heat Shock Protein comparison between younger and
older rockfish”, Marcelle Morrison-Bogorad, Associate Director, Neuroscience
and Neuropsychology of Aging Program, NIA; and “Electron transport
abnormalities and mitochondrial DNA mutations in rockfish heart tissue”,
Judd Aiken, University of Wisconsin-Madison.
|
|
Halaschek-Wiener, Julius |
- 61. ANALYSIS OF LONG LIVED C. ELEGANS DAF-2 MUTANTS USING SERIAL
ANALYSIS OF GENE EXPRESSION
J Halaschek-Wiener*, S McKay, S Jones, M Marra, D Riddle, A
Brooks-Wilson
Genome Sciences Centre, BC Cancer Agency, 600 West 10th
Avenue, Vancouver, BC V5Z 4E6, Canada
Serial Analysis of Gene Expression (SAGE) was used to identify
longevity-associated genes in a long-lived daf-2 C. elegans mutant. The daf-2
gene encodes an insulin/IGF-1 receptor-like protein and mutations therein lead
to a 100% increase in mean life span. SAGE is a method to efficiently count
large numbers of mRNA transcripts by sequencing short tags. We prepared adult C.
elegans SAGE libraries of days 1, 6 and 10 for daf-2 and days 1 and 6 for
control worms. Analyses of gene expression profiles within daf-2 libraries and
between daf-2 versus control SAGE libraries (day 6 vs. 6 and day 10 vs. 6)
identified not only single genes but also whole gene families that were
differentially regulated. Furthermore, daf-2 mutants at day 6 show a strong
hypometabolic phenotype when compared to equally aged control worms that
diminishes at advanced age. Identified gene families regulate important
metabolic processes including stress response, lipid-, DNA/RNA-, protein, and
energy metabolisms as well as intracellular signaling and cell structure.
Identical expression patterns of various members of several gene families
emphasize the importance of these types of genes in longevity-related processes.
Our results suggest that long-lived daf-2 mutants are severely hypometabolic in
mid-life and we present evidence that specific gene families are involved in
these metabolic changes.
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|
Harper, James M. |
-
62. SERUM IGF-I AND LIFESPAN IN CROSSES BETWEEN WILD AND LABORATORY STOCKS OF
MICE
J.M. Harper
(P)1, S.N. Austad1, R.A. Miller1, R.C.
Dysko2
1Department
of Pathology and Geriatrics Center, University of Michigan School of
Medicine, 2Unit for Laboratory Animal Medicine, University of
Michigan-
-
Using F2 hybrid stocks
constructed as a cross between two stocks of wild-derived mice (Id and Ma)
and the C57BL/6J inbred strain, we have assessed the ability of a suite of
life-history and physiological traits to act as predictors of lifespan, as a
prelude to mapping genes that may regulate lifespan in mice. The Id
parental strain, the Ma parental strain, and the (Id x B6)F2 hybrid stock
are all significantly longer-lived than a laboratory-derived control stock
(DC), generated as the offspring of (BALB/cJ x C57BL/6J)F1 females and (C3H/HeJ
x DBA/2J)F1 males (log-rank test p £ 0.02). The lifespan of the (Ma x B6)F2
hybrid stock is indistinguishable from the control (log-rank, p = 0.84).
Because serum IGF-I levels measured in 6-month old mice are lower in both
the Id and Ma stocks than in the DC control (p < 0.001), we tested to see if
IGF-I levels could predict life expectancy in mice of the two segregating F2
stocks. We found in each case that low IGF-I levels at 6 months of age
were associated with longer life span (r2 £ 0.19, p = 0.02). IGF-I
levels did not predict life span in either of the wild-derived parental
stocks Ma and Id, nor in the laboratory control DC mice (p > 0.2 for all).
These data suggest that endogenous levels of early life IGF-I may play a key
role in the regulation of lifespan, and that there are polymorphisms between
at least the Id and C57BL/6J stocks of mice that influence lifespan.
The QTL mapping of the loci responsible for these effects will provide
valuable insight into the hormonal and genetic control of mammalian aging.
|
|
Kirchman, Paul A. |
-
63. ANALYSIS OF THE INFLUENCE OF MITOCHONDRIAL DNA POINT MUTATIONS ON
LONGEVITY IN SACCHAROMYCES CEREVISIAE
-
-
P.
Kirchman
-
-
Harriet L. Wilkes Honors College, Florida Atlantic University, 5353 Parkside
Dr., Jupiter, FL 33458
-
-
Evidence from several species points to mitochondrial DNA (mtDNA) as one
determinant of longevity. Interpretation of these studies is hampered
by nuclear DNA heterogeneity of the test subjects and, in some cases, by the
variable environments in which subjects live(d). To eliminate
variability in both the nuclear genome and the environment, the budding
yeast, Saccharomyces cerevisiae, was used to examine the influence of mtDNA
variation on longevity. Mitochondrial DNA’s containing
point mutations in the COX1, COX3, or CYTB gene were transferred to a yeast
strain that lacks mtDNA (rho0). The created strains vary only in the
point mutations on the mtDNA. These strains were analyzed for
variation in longevity relative to a control strain containing the
non-mutated mtDNA. No statistically significant variation in longevity
has been found between any of the strains tested. To influence
longevity, variation in mitochondrial DNA may need to be more extensive than
the single nucleotide mutations tested. Analysis of additional
mitochondrial mutant strains is ongoing and additional results will be
reported.
|
|
Lee, Yongwoo |
-
64. MICROARRAY ANALYSIS OF GENE EXPRESSION CHANGES IN
INTERLEUKIN-4-STIMULATED HUMAN VASCULAR ENDOTHELIAL CELLS
-
-
YW
Lee*, M Toborek
-
-
Department of Surgery/Division of Neurosurgery, University of Kentucky
College of Medicine, Lexington, Kentucky 40536
-
-
Oxidative stress-mediated
inflammatory reactions within vascular endothelium have been implicated in
the development of age-related human diseases including atherosclerosis. We
have reported that interleukin-4 (IL-4) can induce the pro-oxidative and
pro-inflammatory pathways in human vascular endothelial cells. The cellular
and molecular regulatory mechanisms underlying this process, however, are
not fully understood. In the present study, we performed GeneChip microarray
analysis to investigate global gene expression patterns in human vascular
endothelial cells after treatment with IL-4 using the Affymetrix GeneChip®
Human Genome U133A Arrays, which contain more than 22,000 human genes. Our
results showed that mRNA levels of a total of 106 genes were significantly
up-regulated and 41 genes significantly down-regulated with more than a
twofold change. Majority of these genes are critically involved in the
regulation of inflammatory responses, apoptosis, signal transduction,
transcription factors, metabolism; functions of the remaining genes are
unknown. The changes in gene expression of selected genes related to
inflammatory reactions such as VCAM-1, E-selectin, MCP-1 and IL-6 were
verified by quantitative real-time RT-PCR and ELISA, respectively. IL-4
treatment also significantly increased the adherence of inflammatory cells
to endothelial cell monolayers in a dose-dependent manner. These results may
help determine the molecular mechanisms of action of IL-4 in human vascular
endothelium. In addition, a better understanding of IL-4-induced vascular
injury at the level of gene expression could lead to the identification of
new therapeutic strategies for atherosclerosis. (This work was supported by
the American Heart Association, Ohio Valley Affiliate and University of
Kentucky Microarray Facility Program)
|
|
Li, Hong |
-
65. GENDER DIFFERENCES IN NEURONAL CELL DEATH AFTER OGD INJURY AND NMDA-MEDIATED
EXCITOTOXICITY IN ORGANOTYPIC HIPPOCAMPAL CULTURES
-
-
Hong
Li*, MD, Katrin Andreasson, MD, Louise McCullough, MD, PhD
-
-
Department of Neurology, Johns Hopkins University, Baltimore, Maryland,
21287
-
-
Background and Objective:
Increasing evidence has demonstrated striking sex differences in outcomes
after acute neurological injury. Females are less vulnerable to acute
insults associated with experimental cerebral ischemia. It is believed that
the greater neuroprotection seen in females is due to circulating estrogens.
However, this endogenous female neuroprotection may not be due solely to
hormonal influences. The objective of this study was to determine if there
are gender differences in neuronal cell death in organotypic hippocampal
cultures after oxygen glucose deprivation (OGD) injury and
M-Methyl-d-Aspartic Acid (NMDA)-mediated excitotoxicity. Methods:
Organotypic hippocampal slice cultures were prepared from individual
postnatal day 8 Sprague–Dawley rat pups. Gender genotyping was accomplished
by PCR analysis from pup tail DNA. The cultures were exposed to 10 mM NMDA
for 1 h or subjected to OGD for 45 min, in presence or absence of 30 mM 7NI
(7 Nitroindazole; a selective nNOS inhibitor), or 10nm 17b-estradiol (E2)
for 7 days prior to OGD. Neuronal death was quantified with propidium iodide
(PI). Data were analyzed by one-way ANOVA followed by Newman–Keuls post hoc
test. Results: Baseline neuronal cell death in control, 7NI, and E2 treated
groups was equivalent in males and females. Cell death markedly increased
after stimulation with NMDA or OGD (P < 0.001) in both groups, but there was
a significant increase in cell death in male cultures (P < 0.001 ~ 0.05)
after both OGD and NMDA treatment. Treatment with7NI and E2 reduced neuronal
damage induced by NMDA and OGD (P < 0.001 ~ 0.05). Interestingly, this
effect was more notable in males than in females after OGD injury (P <0.01).
Conclusions: There is a consistent gender difference after OGD and NMDA
toxicity, demonstrating neuroprotection in female cells. Dimorphisms in cell
survival may underlie enhanced neuronal survival (or decreased apoptosis) in
female brain.
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|
Lopez-Cruzan, Marisa |
-
66. ROLE OF MITOCHONDRIAL CASPASE-2 ACTIVITY IN OXIDATIVE STRESS-INDUCED
APOPTOSIS
-
-
Marisa Lopez-Cruzan, Victoria Centonze, and Brian Herman
-
-
Cellular and Structural Biology Department, University of Texas Health
Science Center at San San Antonio. 7703 Floyd Curl Drive, San Antonio, Texas
78229-3900
-
-
The role of mitochondrial
caspase-2 in oxidative stress-induced apoptosis was examined by first
determining if mitochondria contain oxidative stress-inducible capase-2
activity and second, what the effect of loss of mitochondrial caspase-2
would be on mitochondrial-specific oxidative stress-induced apoptosis.
Oxidative stress-induced mitochondrial-specific caspase-2 activation and
apoptosis were assayed in murine NIH-3T3 fibroblasts or fibroblasts obtained
from caspase-2 knockout mice using Florescence Resonance Energy Transfer
(FRET)/optical microscopy (mitochondrial caspase-2 activity) and uptake of
propidium iodide (apoptosis). Mitochondrially targeted FRET caspase-2 fusion
protein containing the preferential caspase-2 substrate flanked by CFP and
YFP, as donor and acceptor fluorophores respectively, were used as an
in-situ sensor of mitochondrial caspase-2 activity. FRET efficiency,
measured using acceptor photobleaching, demonstrated a time-dependent loss
of FRET after exposure of cells to tert-butyl hydroperoxide, indicative of
mitochondrial specific caspase-2 activation. Inhibition of complex I or III
of the mitochondrial electron transport chain is known to induce reactive
oxygen species (ROS) within the mitochondria. Treatment with the complex I
inhibitor rotenone, the complex III antimycin A or staurosporine (STS), a
non-ROS inducer of apoptosis, produced a significant amount of cell death in
fibroblasts isolated from wild type mice. However, cells isolated from
caspase-2 deficient mice were highly resistant to rotenone and antimycin A,
but not STS-induced apoptosis. These results indicate that mitochondria
contain oxidative stress-inducible caspase-2 and that caspase-2 activation
is required for mitochondrial oxidative stress-induced apoptosis.
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|
Mamczarz, Jacek A. |
-
67.
PATTERN OF GENE EXPRESSION IN LIVER OF YOUNG AND AGED RATS AFTER SHORT-TERM
DIET RESTRICTION OR 2-DEOXY-D-GLUCOSE INJECTION: cDNA
-
-
Jacek Mamczarz*, Min Zhu, Jonna Bowker, Kara Duffy, Donald Ingram
-
-
Lab. Exp. Gerontology, National Institute on Aging, NIH, 5600 Nathan Shock
Drive, Baltimore, MD 21224
-
-
Diet restriction (DR) in
rodents increases lifespan, reduces age-related disease and pathology,
increases stress responses, and maintains better function later into life
compared to conventional ad libitum (AL) feeding. We have been investigating
DR regimen and also DR mimetics that stimulate stress response pathways that
are activated by DR. By inhibiting glycolysis, feeding or injection of
2-deoxy-D-glucose (2DG) has been proposed as a DR mimetic. In the current
study we investigated the pattern of gene expression using cDNA microarray
in liver of young (4 mo) and aged (26 mo) male Fischer-344 rats subjected to
3 weeks of 40% DR or 2DG injections once a day. 40% DR reduced initial body
weight (BW) around 15% in young and 14% in old rats, respectively. 2DG in
the dose of 125 mg/kg attenuated food intake and developmental gains in BW
in young rats, while in aged rats 2DG decreased BW and food intake below
initial levels. 2DG at a dose of 250 mg/kg also decreased BW and food intake
in both young and aged rats; however, in aged rats the decrease was at the
level of DR. In general, DR and 2DG treatment up-regulated more genes
in aged rats than recorded in young rats (around twice in DR, 6 times in
2DG-125, and 3.5 times more in 2DG-250). Down-regulation of gene expression
was similar in young and old DR rats, twice higher in young 2DG-125, and
twice lower in young 2DG-250, respectively, compared to aged rats. DR and
2DG-125 shared much more common patterns of gene expression in aged rats
than in young, but for 2DG-250 there was a generally common pattern shared
in both age groups. Analysis of gene expression patterns can be used to
assess potential effectiveness of DR mimetics.
|
|
Martin, Rolf J. |
-
68. BLUEBERRY STUDY WEB SITES FOR COGNITIVE PERFORMANCE MEASUREMENT APPEAR
SUFFICIENTLY PRECISE TO DETECT A 0.1% CHANGE IN PERFORMANCE DURING AGING
-
-
R. Martin*, MMT Corp.,
Sherman, CT 06784, R.J. Coppings, Lane College, Jackson, TN 38301, K.E.
Gerstmann, NY, NY 10014, William Holme, Bethel, CT 06801, K. Hull, New
Fairfield Senior Center, New Fairfield, CT 06812, J.A. Joseph, Human
Nutrition Research Center on Aging at Tufts University, Boston, MA 02111,
A.C. Kokesh, Charleston, WV 25301, B. Kristal, Weill Medical College-Cornell
University Medical Center, NY, NY 10021 and Burke Medical Research
Institute, White Plains, NY 10605, M. Luzi and T. Millard, Sherman IGA
Supermarket and B. Sachs, HR Herbs, Sherman, CT 06784, H.A. Raphaelson,
Mansfield Senior Center, Mansfield, CT 06268, A. Pruchnicki, Mount Sinai
Medical Center, NY, NY 10029, R. Schnoll, Brooklyn College, Brooklyn, NY
11210, and A. Wetherell, Defence Science & Technology Laboratory, UK.
Contact email: BlueberryStudy@aol.com
-
-
1
Barlow Farm Road, Sherman, Connecticut 06784
-
-
To
evaluate methods for reducing age-associated cognitive decline during 3-year
study periods, measurement precision should detect with 95% confidence
differences of 0.1% or more between treatment and control groups having ~1%
average annual decline. To develop such high precision for our 2000, 2002
and 2004 blueberry studies, self-calibrating web sites were developed for
online measurement of 4-choice and 2-choice decision speed and immediate and
delayed word recall (patent no. 6,712,615). These sites enable participants
to measure their performance repeatedly and thereby obtain very precise
average measures of these aspects of cognition. Annual test-retest
reliability values for decision speed during 2000-2004 ranged from 0.95 to
over 0.99. Results collected from these sites were used to develop computer
simulations which indicate that both our blueberry study design, with two
major types of performance measurement (decision speed and recall), and Alex
Comfort's (1970) approach to measuring human aging, with 60 different types
of physiological and performance measurement, are capable of identifying
agents that can reduce a 1% rate of annual decline to 0.95% for studies
involving 10,000 or more participants. The major problem of accurately
monitoring food and supplement intake over long study periods can be solved
in part with supermarket purchase logs and weekly food sharing and spoilage
reports. Purchase logs received to date indicate that 18 participants
purchased 2,845 items at the Sherman IGA Supermarket with electronic
discount cards that provide automatic "health discounts" of ~40% on
blueberries and 35 other health-related foods. Automatic discount cards can
significantly aid recruitment and retention and provide a high-density data
stream to help examine whether local ad campaigns or health news reports
cause unbalanced diet or health supplement shifts in either treatment or
control groups. Blueberrystudy.com measurement and diet-tracking software
are expected to be available to other research groups later this year.
-
|
|
Masternak, Michal M. |
-
69. THE
EFFECT OF GROWTH HORMONE RECEPTOR KNOCKOUT AND CALORIC RESTRICTION ON
EXPRESSION OF GENES RELATED TO INSULIN SIGNALING IN MICE
-
-
Michal M Masternak, Khalid A. Al-Regaiey, Michael S. Bonkowski, Andrzej
Bartke
-
-
Departments of Internal Medicine and Physiology, Geriatrics Research,
Southern Illinois University School of Medicine, 801 N. Rutledge,
Springfield IL 62794
-
-
Growth hormone receptor
knockout (GHR-KO) mice are long-lived, hypoinsulinemic and hypoglycemic and
exhibit enhanced sensitivity to injected insulin (Ins). Using Real-time PCR
(RT-PCR) we analyzed hepatic and muscle levels of insulin receptor (IR),
insulin receptor substrate 1 (IRS1), IRS2, peroxisome
proliferators-activated receptor gamma (PPARg) glucose transporter 4
(GLUT4), insulin-like growth factor 1 (IGF1) and IGF1 receptor (IGF1R) mRNA
expression in GHR-KO and in normal (N) animals from the same strain. We have
also analyzed gene expression in GHR-KO (KO) and normal mice subjected to 30
% caloric restriction (CR). In the liver, gene expression was affected by
genotype but not diet. The expression of IR, IRS1 and IRS2 mRNA was
increased (P<0.0044, P<0.024, P<0.0003, respectively) in GHR-KO animals.
PPARg mRNA expression was also increased in KO mice (P<0.0042). However,
IGF1 mRNA expression in the liver was decreased in GHR-KO mice as expected
(P<0.0001). Different effects were detected in the skeletal muscles of these
animals. The mRNA expression was not affected by genotype but not by CR. The
expression of IR, IRS1 and IRS2 was decreased by CR in N (P<0.02, P<0.0034,
P<0.025, respectively) and in KO mice (P<0.0005, P<0.0003 and P<0002,
respectively). PPARg mRNA was decreased by CR in normal and KO animals
(P<0.0096 and P<0.0009, respectively). The expression of GLUT4 mRNA in
muscle showed significant down regulated by CR in both N and KO mice
(P<0.0044 and P<0.0003 respectively). The expression of IGF1 mRNA was
decreased by CR in normal and KO animals (P<0.0053 and P<0.0001). Moreover,
the expression of IGF1 mRNA was reduced in KO vs. normal mice (P<0.0001). In
contrast, the expression of IGF1R mRNA was not affected by CR in N mice, but
was up regulated in KO in comparison to normal mice (P<0.014) and reduced by
CR in KO mice (P<0.0005).
-
Supported by NIA
|
|
Mattison, Julie A. |
- 70. THE INFLUENCE OF SODIUM INTAKE ON CARDIOVASCULAR RESPONSES IN
RHESUS MONKEYS
J. A. Mattison, A. Bagrov, H. Spurgeon, P. Pullen, M. A.
Lane, G. S. Roth, D. K. Ingram, E. G. Lakatta
NIA, NIH Animal Center, Poolesville, MD 20837
Although there is considerable evidence linking salt intake to hypertension, how
this dietary variable is involved in remodeling of the vascular wall to affect
arterial stiffness is still unknown. The effect of an incrementally increased
salt load on vascular stiffness and modulation of this response by production of
an endogenous ligand was studied in nine old normotensive male rhesus macaques
(mean age at start = 19.1 +/- 0.67 years). Serial measurements of aortic pulse
wave velocity (PWV), blood pressure (BP), urine and sodium output, and
marinobufagenin (MBG) were made before and following each stepwise increase in
dietary sodium chloride (NaCl) from 0.8% to 6.0% of daily intake. Sodium
excretion significantly increased with each incremental increase in dietary
intake. PWV significantly increased as a direct nonlinear function of Na intake
independent of changes in BP.
Although there was considerable variability among monkeys in BP response, there
was an immediate pressure increase that was attenuated at higher dietary Na
loads. MBG excretion was positively correlated to Na excretion at baseline and
with a 1.7% and 2.6% NaCl diet. This direct relationship was not evident at
intakes of 3.5% and 6% NaCl. These results suggest that old monkeys are
susceptible to NaCl dependent vascular changes that can be partially compensated
by increased MBG to enhance Na excretion.
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|
Mounkes, Leslie C. |
-
71. ABNORMAL SPLICING OF LMNA CAUSES PROLIFERATIVE DEFECTS LEADING TO A
HUTCHINSON GILFORD PROGERIA SYNDROME PHENOTYPE IN MICE
-
-
Leslie C. Mounkes*, Lidia Hernandez, Serguei Kozlov, Colin L. Stewart
-
-
Cancer & Developmental Biology Laboratory, National Cancer
Institute—Frederick, MD
-
-
Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare genetic disorder
resulting in the acceleration of age-related phenotypes, including shortened
stature, craniofacial disproportion, parchment thin skin, alopecia, and
osteoporosis, with death predominantly due to premature atherosclerosis in
the early teens1. We derived mice carrying a splicing defect in the
Lmna gene and found that mice homozygous for the mutation display defects
remarkably similar to progeria1. Lmna encodes the A-type lamins, major
components of the nuclear lamina with putative functions in nuclear
structure, chromatin organization, gene regulation, and intracellular
signaling. Mutant mice (LmnaHGPS) showed marked reduction in growth
rate and died by 4 weeks of age. Pathology of the heart, bone, muscle
and skin are consistent with phenotypes associated with aging of these
tissues. The Lmna mutation resulted in nuclear morphology defects and
decreased lifespan of homozygous fibroblasts from postnatal, but not
embryonic tissues, suggesting a developmentally regulated mechanism of
premature cell death. Furthermore, in a background deficient for
p16/p19ARF, the premature death of the mutant progeric cells is rescued in
vitro, suggesting the nuclear lamina plays a role in the control of cell
growth, possibly through pathways involving pRB and/or p53. Muscle
myoblast cultures from progeric mice displayed a slower response to
differentiation cues and made myotubes, which were shorter with disorganized
nuclei compared to myotube differentiation of wild type myoblasts.
Defects in proliferation and differentiation could contribute to the
incomplete development of muscle and other mesenchymal tissues in progeria.
-
-
1LC Mounkes, S Kozlov, L
Hernandez, T Sullivan & CL Stewart. A progeroid syndrome in mice
is caused by defects in A-type lamins. Nature 425: 298-301 (2003).
|
|
Overton, James M. |
-
72. PHYSIOLOGICAL RESPONSES TO VERY MILD AND STANDARD LEVELS OF CALORIC
RESTRICTION IN RATS
-
-
J.
M. Overton, A.D. Parsons, S.A. Evans
-
-
Department of Nutrition, Food and Exercise Sci., Florida State Univ.,
Tallahassee, FL 32306-4340
-
-
Very mild caloric restriction (CR) is commonly used as a control
intervention to assess the effects of more substantive CR on lifespan.
The purpose of this study was to compare the physiological effects of
short-term, very mild CR to standard levels of CR in rats. Male FBNF1
rats weighing about 360-380 g were instrumented with telemetry devices for
measurement of mean arterial pressure (MAP) and heart rate (HR) and
continuously housed in room calorimeters for assessment of oxygen
consumption (VO2) and respiratory quotient (RQ). Rats were acclimated
to thermoneutral (TMN; 30°C) conditions prior to imposing very mild (5-10%)
or standard levels of caloric restriction (CR; 40% restriction of ad lib)
for two weeks. Controls continued to consume food ad libitum (5-6
rats/group). Over the two-week period, controls gained 15 grams, while
mild CR reduced weight by about 3 grams and 40% CR reduced weight by 30
grams. As expected, 40% CR significantly reduced body weight, VO2, HR
and RQ; while increasing HR variability. Unexpectedly, mild CR, which
had no effect on body weight, RQ and HR variability, produced significant
reductions in HR and VO2. Weight maintenance produced 50% of the
physiologic effect associated with 40%CR. The results suggest that in
rats housed at thermonuetrality, the cardiovascular and metabolic responses
to CR are engaged by very mild stimulus that does not require weight loss.
|
|
Paik, David C |
73. DETECTION OF NOVEL CROSS-LINK COMPOUNDS
FROM THE NITRITE/COLLAGEN REACTION
DC Paik
Department of Ophthalmology, Columbia University, New York, New York and
Department of Surgery, North General Hospital. New York, New York
Introduction: Increased collagen cross-linking is one of the hallmarks
of human aging and can contribute to the functional decline of aging
organ systems. Nitrite ion, whose sources include inflammation and
smoking, can mediate damaging nitration, nitrosation, and hydroxylation
reactions. The in vitro neutral pH reaction of nitrite with collagen
produces changes that mimic collagen aging and include increased
cross-linking, UV absorbance, and fluorescence. The current study was
undertaken in order to identify covalent collagen cross-links produced
specifically by reaction with nitrite. Methods: Model studies were
performed using gelatin. Samples (10mg/ml) were incubated with 0 to
200mM sodium nitrite at neutral pH for up to 10 days. The reaction was
stopped with ammonium sulphamate. UV/vis and fluorescence spectra were
obtained on the protein mixtures. Cross-link enrichment was performed on
acid hydrolysates using Skinner’s (1982) cellulose mini-column method.
Isodesmosine was added as an internal standard (0.5ug/mg)
pre-hydrolysis. The water fraction was collected and analyzed by HPLC
(C18) with diode array and fluorescence detection. Gradient conditions
using 20mM heptafluorobutyric acid and 2-20% acetonitrile over 60 min
were employed. Pump speed was 1ml/min. Results: Nitrite modification of
gelatin produced spectroscopic changes similar to those observed
previously for collagen types I and IV and include increased UV
absorbance ca. 350nm and increased fluorescence with excitation peaks
ca. 290 and 340, and an emission peak ca. 430. Cross-link analysis
revealed 6 unique products of the reaction as detected by UV at 280nm.
Five of the compounds also showed fluorescence (ex290/em410). The
unknowns elute prior to pyridinoline and isodesmosine. Conclusions: This
is the first identification of cross-link compounds specific to the
nitrite/collagen reaction. The detected compounds may serve as
biomarkers of the reaction in future studies.
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|
Paredes, Daniel |
-
74. NOREPINEPHRINE RELEASE IN CEREBELLUM DURING DELAY
CLASSICAL EYEBLINK CONDITIONING DECLINES IN AGED RATS
D. A. Paredes, A.N. Samec, M. Fister and P.C.
Bickford
Center for Aging and Brain Repair, USF, 12901
Bruce B Downs Blvd, Tampa, Fl 33612
Delay Classical Eyeblink Conditioning is an important model of
associative, cerebellar dependent learning. Aged animals and humans show
deficits in learning on this paradigm. Our laboratory has been
investigating the role of norepinephrine (NE) in this paradigm. We have
demonstrated that blocking either the β-noradrenergic receptor or
protein kinase A with local infusions into cerebellar lobule HVI and
interpositus nucleus can interfere with learning. The goal of this study
was to determine whether NE release is observed in HVI during delay
eyeblink conditioning and if this is altered in aged rats. In vivo
microdialysis coupled to eyeblink conditioning was performed on three
month old or twenty month old F344 rats. Dialysate samples were
collected every 10 minutes. After 5 baseline samples were collected rats
received 50 training trials (45 paired with a 12 psi airpuff and 5 tone
only [3kHz 85 dB] trials), or Rescorla type pseudo conditioning. Rats
showed a significant release of NE during and after training, for young
rats with 47.3 ± 5.0 nM of NE released as a max value reached 30 min
after training started; while aged rats showed a max of NE release of
23.2 nM at 80 min after training started. This data clearly shows a
decline on NE release in aged rats compared with young, this could
explain some of the deficiencies of aged animals to learn new motor
tasks which could be linked with the NE reuptake system deficiencies.
Support Contributed By: NSF 019674
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Pendergrass, Bill R. |
- 75. Mitochondrial function, organelle degradation, and cataract in
lenses from aging: normal mice, GPX-1 KO mice, and Ghr-KO mice.
B Pendergrass,P Penn, A Bartke, H Van Remmen, and N Wolf
1Department of Pathology, University of Washington
School of Medicine, Department of Pathology, University of Washington School of
Medicine, Seattle, WA. 98195. USA
Cataract incidence was increased dramatically in old mice of each genotype
relative to genotype matched young mice. Old Glutathione Peroxidase-1 knock out
mice (GPX-1 KO), had an increased cataract incidence relative to age-matched
controls. Older long lived growth hormone receptor knock out mice (GHr-KO mice)
had significantly less cataract than age-matched controls. Mitochondrial
oxidative cycling (a measure of O2 use) in lens epithelial cells (LECs) on the
surface of mouse lenses was determined from the rate of oxidation of reduced
Mitotracker red (H2MTR) to Mitotracker Red (MTR). This was significantly reduced
in old compared to young control mice lenses. It was even more reduced in old
GPX-1 KO mice lenses compared to age-matched controls. The reduction in
oxidation was apparent whether measured relative to cell surface area, or to DNA
in the cells. We also found a parallel decrease in total mitochondrial mass in
old mouse LECs. The relative mitochondrial membrane potential (MMP) in the LECs, was determined from the fluorescence ratio of
the vital stains Mitotracker Red (sensitive to MMP) to Mitotracker Green
(sensitive to mitochondrial mass). This MMP ratio was slightly higher in older
LECs than young LECs. Young GHr-KO mice had significantly reduced MMP relative
to young controls, possibly indicating mild uncoupling of oxidative
phosphorylation. We also monitored the degradation of DNA and mitochondrial
proteins using specific fluorescent stains in internalized LEC fiber cells. The
lens fiber cells in the interior of the lens from old mice were deficient in the
ability to degrade nuclear and mitochondrial organelles. Cataractous areas of
the lenses were especially heavily stained for DNA and mitochondrial debris.
Dichlorofluorescein (DCF) staining of ROS was also significantly increased in
lens fibers from old mice and this was frequently associated with cataractous
foci in old lenses.
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Pepper, Evan |
76. SOS-INDUCED DNA POLYMERASES
ENHANCE LONG-TERM SURVIVAL AND EVOLUTIONARY FITNESS
E. Pepper, B. Yeiser, MF Goodman, and SE Finkel
Department of Biological Sciences, SHS 172, University of Southern California,
Los Angeles, CA. 90089-1340
Escherichia coli encodes three SOS-induced DNA polymerases: pol II, pol IV, and
pol V. We show here that each of these polymerases confers a competitive fitness
advantage during the stationary phase of the bacterial life cycle, in the
absence of external DNA-damaging agents known to induce the SOS response. When
grown individually, wild-type and SOS pol mutants exhibit indistinguishable
temporal growth and death patterns. In contrast, when grown in competition with
wild-type E. coli, mutants lacking one or more SOS polymerase suffer a severe
reduction in fitness. These mutants also fail to express the “growth advantage
in stationary phase” (GASP) phenotype as do wild-type strains, instead
expressing two additional new classes of GASP. These polymerases contribute to
survival by providing essential functions to ensure replication of the
chromosome and by generating genetic diversity.
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Quanjian, Yan |
77. NANOG KEEP EARLY EMBRYO AND
TROPHOBLASTIC INVINCIBILITY FROM AGING
Quanjian Yan (P), Xiangmei
Chen, Yumei Zhang, Daqiang Li, Quan Hong, Bo Fu.
State Key Laboratory of Nephrology, Kidney Department of 301 Hospital Fuxing
Road 28. Haidian, 100853, Beijing. P.R.China.
In our co-culture system, we
found that early embryo and trophoblastic cell of KM mouse can invade malignant
tumor cells and keep youth. Nanog is a newly identified homeodomain gene that
functions to sustain the pluripotency of embryonic stem cells. So we investigate
whether Nanog is necessary for early embryo and trophoblastic cell keep
invincibility invasion and youth. Morphological analysis showed that mouse
blastocysts have the invincibility of adhesion, migration and invasion after
implant in many normal cells or tumor cells. The normal cell strains include
L929 and NIH3T3 mouse fibroblasts, 9HTE human tracheal epithelial cells and the
ECV-304 human umbilical vein endothelial cells. The malignant tumor cell strains
include differently histological origins, the HNE1 human nasopharyngeal
carcinoma cells include BGC-823 and SGC-7901, human kidney carcinoma cells
786-0, human bladder carcinoma cells BIU-87 and BADM-60, human osteoblastoma
cells Ros17/2.8 and Saos-2, human endometrial carcinoma cells RL95-2, rat breast
carcinoma cells SHZ-88, human hepatocarcinoma cell strains HepG2, SMMC-7721,
QGY-7703 and MHCC97-H; human breast carcinoma cell strains T47D, MCF-7, ZR75-30,
Bcap-37, MDA-MB-231 and MDA-MB-435s. After 72 h of co-culture, integrin alpha V,
FAK, MMP-9, Cyclin D1, CD44 protein and Ki-67 mRNA were lower expressed in the
trophoblastic cells in the group without the inner cell cluster or Nanog
expression embryonic stem (ES) cell cluster with immunocytochemical and in situ
hybridization than that with Nanog expression ES cell cluster. And the outgrowth
area in the group without the inner cell cluster or Nanog null ES cell cluster
is smaller than that with the inner cell or Nanog expression ES cell cluster
(P>0.05).
These
results strongly suggest that Nanog activation keep the inner cell or ES cell
cluster from aging, promote trophoblastic adhesion, migration and invasion
(etc).
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Rikke, Brad A. |
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78.
GENETIC
DISSECTION OF DIETARY RESTRICTION
-
-
Brad A. Rikke*, Thomas E.
Johnson
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-
University of Colorado at
Boulder
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-
Dietary restriction (DR) is
the most validated method of extending health and longevity n mammals,
extending the life span of rodents by as much as 50%. Many
physiological responses to DR have been put forth as causal factors
underlying DR’s benefits. As a first step towards testing some of
these factors, we’ve examined 83 strains of mice for significant differences
in early responses to DR. These responses include lowered body
temperature, lowered body weight, reduced growth rates (tail and hair), and
reduced motor activity (runwheel and home cage). We’ve also examined
the extension of female fertility after extended DR. For each
response, we find that there is a phenomenal amount of strain variation,
with surprisingly little covariation among responses. These findings
could be used to critically test whether any of these responses covary with
DR-induced longevity. Responses validated to covary with life
extension could be used as a tool to screen for mutations that are likely to
affect DR-induced longevity. In addition, we are mapping quantitative
trait loci (QTLs) specifying each response by studying strains from the
largest murine panel of recombinant inbred strains ever developed, the LXS
panel of 77 strains. This mapping makes it possible to subsequently
identify the genes underlying these QTLs by positional cloning, ultimately
leading to a better understanding of the molecular mechanism by which DR
extends mammalian life span. (Supported by the Ellison Medical
Foundation and NIH R01 AA11984)
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Perez-Perez, E. M. |
79. ANTIOXIDANT CAPACITY OF
HONEY TYPES ACCORDING TO BOTANICAL AND ENTOMOLOGICAL ORIGIN
E. M. Pérez-Pérez1,
A. J. Rodriguez-Malaver1(P), P. Vit2
1Lab. de
Bioquímica Adaptativa, Dep. de Bioquímica, Fac. de Medicina; 2Dep.
Ciencia de los Alimentos, Fac. de Farmacia y Bioanálisis, Universidad de Los
Andes, Mérida 5101, Venezuela.
At the
present, there is overwhelming evidence that free radicals cause oxidative
damage to lipids, proteins, carbohydrates and nucleic acids. Therefore,
reactive oxygen species (ROS) such as O2-●, OH●, or lipid peroxyl
radical (LOO●) might lead to many biological complications including
carcinogenesis, mutagenesis, aging,
atherosclerosis and
neurodegenerative diseases. Honey has been known to exert a significant in vitro
antioxidant activity, in part due to its phenolic content. In this work, we
studied the antioxidant capacity of 12 types of honeys from different places and
floral origins, from Apinae and
Meliponinae bee subfamilies, on superoxide anion
and hydroxyl radical. We found that honey samples inhibited the formation of
superoxide anion and hydroxyl radical, under a range from 86 to 25% depending on
the sample. This inhibition capacity was, for some samples, higher than
quercetin and melatonin. On the other hand, we measured the antioxidant activity
(AOA) values of honey samples in comparison to uric acid at 1 mM. In this case,
honey samples had AOA values between 0.32 and 0.84 Mm which were considerably
high as compared to melatonin and quercetin.
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Perez-Perez, E. M. |
80. ANTIOXIDANT CAPACITY OF
VENEZUELAN BLACKBERRY (RUBUS BOGOTENSIS KUNTH) WINE
Pérez-Pérez EM1, Rodríguez-Malaver AJ1(P),
Vit P2
1Laboratorio
de Bioquímica Adaptativa, Departamento de Bioquímica, Facultad de Medicina,
2Departamento Ciencia de los Alimentos, Facultad de Farmacia y Bioanálisis;
Universidad de Los Andes, Mérida 5101, Venezuela
The
reported health benefits of moderated wine drinking, especially in theprevention
of chronic diseases associated with oxidative stress, may be related to the
antioxidant activity of flavonols; quercetin and myricetin, which can readily
reduce free radical by donating an unpaired electron. Rubus Bogotensis Kunth (Rosaceae)
is the blackberry cultivar in the Venezuelan Andes, where a fermented product of
the berries is marketed as blackberry wine. We studied the antioxidant capacity
of 12 samples of blackberry wine produced in Mérida state, on the formation of
superoxide anion and hydroxyl radical. Wine samples inhibited the formation of
superoxide anion and hydroxyl radical under a range from 17% to 53%. For some
wine samples, this inhibition capacity was close to those of quercetin (71%, for
superoxide anion, 53% for hydroxyl radical) and melatonin (79%, for superoxide
anion, 54%, for hydroxyl radical). We also measured the antioxidant activity (AOA)
value of wine samples in comparison to 1 mM uric acid. Wine samples had AOA
values between 0.95 and 0.80 mM, which were, for some samples, actually high if
were compared to those of melatonin (0.83 mM) and quercetin (0.86 mM), used as
reference in this study. This antioxidant activity can be due to the phenolic
content of wine samples, which are very efficient scavengers of free radicals
because its molecular structure includes an aromatic ring with hydroxyl groups
containing mobile hydrogens, and they have the capacity to reduce or chelate
divalent ions which catalyze redox reactions.
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Rodriguez-Malaver, A. |
81. HONEY AND LUTEOLIN
EFFECT ON STREPTOZOTOCIN, SELENITE AND OSMOTIC INDUCED CATARACTS
P. Vit1,5,
R. De Jesus2, M. Gudiño3, E. Perez4, A.
Rodriguez-Malaver4(P), A. Melendez5
1Dep.
Food Science, Fac. Pharmacy, 2 Dep. Biology, Fac. Sciences, 3BIOULA
Animal House, 4Dep. Biochemistry, Fac. Medicine, Universidad de Los
Andes, Mérida, Venezuela, 5Dep. Physiology, Fac. Medicine, National
University of Singapore, Singapore
Ocular
cataracts are of epidemiological interest in human populations. The intraocular
lens implantation is a successful technique to improve vision by removing the
damaged ocular lens. However, a medical therapy to prevent, retard or reverse
lens opacity is not available. In the Neotropics, stingless bee (Meliponinae)
honey eyedrops are instilled to treat cataracts. Trigona Tetragonisca angustula
angustula honey is currently used for this purpose, therefore this honey type
was harvested from a hive and refrigerated until use. Antioxidant capacity,
luteolin and luteolin derivatives were detected in previous studies with honey
extracts. From a screening with commercial flavonoids, luteolin tetra-methyl
ether, luteolin 4’-glucosyde, luteolin 3’-7-diglucosyde reduced osmotic induced
opacification in an ovine lens culture model assessed by digital image analysis.
We further investigated two in vivo rat models of cataracts induced by
streptozotocin and selenite, with slit lamp evaluations. Luteolin tetra-methyl
ether and honey eyedrops were applied before and after cataract onset, to
explore preventive and therapeutic effects. A preliminary 20 % of rats receiving
the honey therapeutic treatment in the selenite model presented delay of
opacification.
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Rosedale,
R. |
82. CLINICAL EXPERIENCE OF A
DIET DESIGNED TO REDUCE AGING
R
Rosedale, E Westman
The neuroendocrine theory of
aging is associated with elevated levels of glucose, insulin and leptin. The
objective of this study is to describe the metabolic effects of a nutritional
program designed to reduce these correlates of aging. A retrospective chart
review was performed of patients attending an outpatient metabolic management
program utilizing instruction in a high-fat, adequate-protein, low-carbohydrate
diet, the use of nutritional supplements, and periodic individual visits. The
general dietary recommendation was approximately 15% carbohydrate, 25% protein,
and 60% fat. Recommended sources of fat included raw nuts, avocados, olives and
olive oil, flax and cod liver oil. The intake of protein was limited to 1.0 -
1.25 grams/kg lean body mass per day (increased for exercise to 1.25
grams/day). Recommended sources of protein included sardines, fish, eggs, tofu,
poultry, wild meats, non-fat cheeses (cottage, ricotta, cream), and seafood.
Only non-starchy, fibrous vegetables were acceptable. Nutritional supplements
recommended daily were: L-carnitine 2000mg, alpha-lipoic acid 400mg, coenzyme
Q10 100 mg, 1 tbsp cod liver oil, magnesium 300mg, potassium 300mg, vitamin C
1000mg, vitamin E 800mg, and a multivitamin. Medications were adjusted if
needed. The mean duration of follow-up was 91.5 days (range 36-211).
Thirty-one patients were identified with baseline and follow-up body weight, and
fasting laboratory tests. The mean age of patients was 57.6 years, 53% were
female. Over a mean follow-up of 91.5 days, body weight decreased 8.2%
(p<0.01), fasting serum glucose decreased 8.3% (p=0.001). There were
approximate 50% reductions in insulin, leptin, fasting serum triglyceride, and
triglyceride/HDL ratio (p<0.001). Free T3 decreased 7% (p<0.001), while TSH did
not change significantly. We conclude that a high-fat, adequate-protein,
low-carbohydrate diet with nutritional supplementation led to improvements in
serum factors related to the aging process
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Ross, Ian K. |
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83. MITOCHONDRIA, SEX, AND MORTALITY
-
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I. Ross
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Dept. of Molecular,
Cellular, and Developmental Biology
-
University of California,
Santa Barbara, CA 93106-9610
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Is
a universal cause of senescence in healthy organisms the expression of both
alleles of nuclear-coded proteins that form part of the oxidative-phosphorylation
complexes of the mitochondria? Could this explain why mitochondria are
inherited from only one parent? And why clones (e.g.
triplets) die of different, random, causes? Prior to the evolution of
sex, the endosymbiotic relationship between mitochondria and nuclear genomes
would have selected mechanisms that maintained the optimum interaction
between the two genomes. Once sex evolved, mating would introduce different,
competitive, mtDNA and /or nDNA gene products which could well upset
the balance.The selection of mechanisms, such as the specific degradation of
one mitochondrial genome, would have prevented part of such competition.
Unlike most protein complexes in the cell, the proteins of the multi-enzyme
complexes of the ox-phos system are derived from both nuclear-genome-coded
genes and mitochondrial-genome coded genes. Minor mutations in either mtDNA
or nDNA coding for these proteins are known to lead to major and
catastrophic diseases of humans, suggesting that very tight and precise
interactions are required. To maintain the evolutionarily established
balance after mating, monoallelic expression of the nuclear-coded genes
would be advantageous and prevent subtly different competitive proteins from
interacting with the resident mitochondria. This would require regulation of
the expression of those specific nuclear genes, possibly under the control
of the resident mitochondria. I have postulated that nuclear genes coding
for these ox-phos complex proteins are normally expressed monoallelically
and that biallelic expression of these genes with age results in adverse
effects in different tissues and organs leading to senescence and mortality.
I will be presenting a fungal model to explore this hypothesis that as
already produced evidence of monoallelic expression of at least one of these
genes in diploids
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Sagie, Doron |
-
84.
THE RELATIONSHIP BETWEEN THE ACTIVITY IN "AMCHA" AND THE HEALTH STATUS OF
ELDERLY HOLOCAUST SURVIVORS
-
-
AbstractAuthors:
D.Sagie(P)*, A. Biderman**
-
-
Department of Gerontology* and Department of
Family Medicine**,Ben-Gurion University of the Negev, Beer-Sheva, Israel;
dsagi@bgumail.bgu.ac.il
-
-
About 40% of the total elderly population in Israel, are Holocaust
survivors*. Previous studies have shown that this population of elderly
survivors, is in great risk for different health problems. "Amcha" – The
Israeli center for Psychosocial support of Holocaust survivors, was
established in 1987, in order to alleviate the emotional suffering of
elderly Holocaust survivors. The treatment involves building a framework of
mutual aid, memory processing and grief resolution. The aim of the current
study was finding a relationship between being an active member in "Amcha",
and the improvement of different health measures that are supposedly purely
biological.
-
We sampled a study group
that included "Amcha" members, and a control group composed of Holocaust
survivors who had similar socio-demographic and health status background,
but were not "Amcha" members.
-
The results indicated that
"Amcha" members evaluated their health status more positively)F=5.64,
p<0.05). This measure of self-evaluation was proved in previous studies to
be strongly related with future morbidity and mortality. "Amcha" members
also tended to utilize less health services (F=.70, p<0.05).
-
This research contributes
not only to social sciences, but also to health and biological studies. It
shows that an emotional treatment of the elderly can be related to an
improvement in their health status, which is commonly associated with
medical-biological factors and treatments.
-
-
* A person who used to live
in Europe in the years 1939-1945, in the time of the Nazi regime.
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Sagi, Orli |
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85.
EFFECT OF AGE AND LPS ON p69- SHCC INDUCTION IN MOUSE BRAIN
-
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Sagi O*.,
Wolfson M., Fraifeld V
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Department of Microbiology and Immunology, CMRA, Ben-Gurion University of
the Negev, Beer-Sheva, Israel; fraifeld@bgumail.bgu.ac.il
-
-
Recently, Shc (Src homology 2 domain containing) proteins were implicated in
the oxidative stress-induced apoptosis and longevity in mice. These proteins
act as adaptors coupling activated receptors with tyrosine kinase activity
to downstream signaling molecules, including the MAPK-associated pathway.
The family of Shc proteins includes ShcA, ShcB, and ShcC. ShcC family
includes two isoforms of 55- and 69-kDa, and their expression appears
limited to neural tissue. The brain is one of the targets for
Lipopolysaccharide (LPS), a component of the cell wall of gram-negative
bacteria, which is a classic trigger of inflammatory reactions accompanied
by a generation of reactive oxygen species and activation of MAPKs. The
response to LPS is altered with age. The present study was aimed at
examining the expression of ShcC in the mouse brain with respect to animal’s
age and the effect of LPS. Male CD-1 mice aged 4 days to 12 months old, were
injected intraperitoneally with 1 mg/kg LPS and at different times
post-injection, their brains were removed for preparation of tissue
extracts. The latter were processed through Western blot analysis using
specific anti-ShcC antibodies. We found that (i) the basal level of ShcC in
mouse brain increased gradually during the postnatal period, reaching the
maximum at 2 months. Middle-age mice had a lower level of ShcC compared to
the young adult and young animals; (ii) both p55 kDa and p69 kDa isoforms of
ShcC displayed similar age-related patterns; (iii) LPS differentially
affects the expression of ShcC isoforms, resulting in a marked up-regulation
of p69 and an insignificant effect on the level of p55; (iiii) the effect of
LPS on ShcC was age-related, being more pronounced in the younger mice
compared to the older animals.
-
-
This study was supported by
a grant from the United States-Israel Binational Science Foundation (BSF),
Jerusalem, Israel.
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Saretzki, Gabriele C. |
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86. A
DNA DAMAGE CHECKPOINT-MEDIATED RESPONSE IN TELOMERE-INITIATED SENESCENCE
-
-
F.
d’Adda di Fagagna, G. Saretzki*, T. von Zglinicki, S.P. Jackson
-
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University of Cambridge UK, The Wellcome Trust Sanger Institute Cambridge,
UK
-
-
Replicative senescence in human fibroblasts can be triggered by telomere
uncapping, however, the signalling pathways leading to telomere-initiated
activation of cell cycle checkpoints were not clear. Specifically, it
was unknown whether uncapped telomeres could signal arrest directly or
whether chromosomal breaks following telomeric fusions were main triggers of
senescence.
-
Here we show that senescent
human fibroblasts display molecular markers characteristic of functional DNA
damage foci including phosphorylated histone H2AX in colocalisation with DNA
repair and DNA damage checkpoint factors such as 53BP1, MDC1 and NBS1.
Importantly, these senescence-associated DNA damage foci (SDF) associate
preferentially with telomeres both after ‘normal’ telomere erosion and after
telomere uncapping by induction of a dominant-negative TRF2. SDF remain
present for many months in senescent cells. Presumably, they remain active
because inactivation of DNA damage checkpoint kinases in senescent cells can
restore cell cycle progression into S phase.
-
Thus, we propose that
telomere-initiated senescence reflects a DNA damage checkpoint that is
activated with a direct contribution from uncapped telomeres. Replicative
senescence appears to be a state that is actively maintained by
sustained activation of a DNA damage response. Thus, DNA damage foci
are not only indicators of acute DNA damage, but might much more resemble
novel markers for the presence of senescent cells in vivo.
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Seluanov, Andrei |
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87. DNA END JOINING BECOMES LESS EFFICIENT AND MORE ERROR-PRONE DURING
CELLULAR SENESCENCE
-
-
A.
Seluanov*, D. Mittelman, O. Pereira-Smith, J. Wilson, and V. Gorbunova
-
-
Department of Biochemistry and Molecular Biology, Baylor College of
Medicine, Houston, Texas
-
-
Accumulation of somatic
mutations is thought to contribute to the aging process. Genomic
instability has been shown to increase during aging, suggesting an aberrant
function of DNA double-strand break (DSB) repair. Surprisingly, DSB
repair has not been examined with respect to cellular senescence.
Therefore, we have studied the ability of young, presenescent, and senescent
normal human fibroblasts to repair DSBs in transfected DNA using a
fluorescent reporter substrate. We have found that the efficiency of
end joining is reduced up to 4.5 fold in presenescent and senescent relative
to young cells. Sequence analysis of end junctions showed that the
frequency of precise ligation was higher in young cells, whereas end joining
in old cells was associated with extended deletions. These results
indicate that end joining becomes inefficient and more error-prone during
cellular senescence. Furthermore, the ability to use microhomologies
for end joining was compromised in senescent cells, suggesting that young
and senescent cells may use different end joining pathways. We
hypothesize that inefficient and aberrant end joining is a likely mechanism
underlying the age-related genomic instability and higher incidence of
cancer in the elderly.
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Shaikh, Aasef G. |
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88.
BLOCKADE OF ION
CONDUCTANCE IN AUDITORY NEURONS MIMIC DISORDERED CENTRAL PROCESSING IN AGED
A, Shaikh (P); P,
Finlayson
Department of
Otolaryngology, Wayne State University, 550 E Canfield, Room # 327, Detroit,
MI 48201
Presbycusis, a senile
hearing-loss, is an inevitable sequelae of aging. About one-third of
Americans older than age 60 suffer from hearing disorders. In addition, the
elderly also have a diminished ability to understand time-compressed speech,
which is likely due to a reduction in temporal resolving power of auditory
neurons. Based on auditory brainstem evoked-responses (ABR), affected
auditory temporal processing in the aged has a central origin, possibly in
Superior Olivary Complex (SOC). Yet the precise mechanism for its
pathogenesis is not defined. There is mounting evidence that excitability in
the central auditory neurons is affected in presbycusis. A range of ion
conductances shape the auditory neural excitability. Hyperpolarization-activated-cationic-conductance
(Ih) is one of the
predominant conductances
determining resting membrane potential in the SOC neurons and could regulate
their excitability. In order to understand the pathophysiology of disordered
central processing in presbycusis, it is important to examine the influence
of Ih on the SOC neural excitability. Auditory neurons also exhibit an
important property, post-stimulatory suppression, to aid echo-suppression
and thereby adequate processing of the time-compressed speech. As Ih is an
important regulator determining the resting membrane potential, its role in
the post-stimulatory suppression, particularly in the SOC neurons needs to
be investigated. Here we document Ih has a predominant role in shaping the
SOC neural excitability in vivo. We also report, selective blockade of Ih by
ZD7288 (100microM) significantly increases the magnitude of post-stimulatory
suppression for shorter inter-tone intervals and raises the neural-response
threshold to the second of the sequential-tones. As Ih-blockade decreases
the neural excitability and enhances the post-stimulatory suppression for
short inter-tone interval, in the SOC neurons; incompetent function of Ih
could be a possible neurophysiological correlate of decreased amplitude of
ABR “wave”.
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Shaikh, Aasef G. |
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89. THE
VESTIBULAR PROSTHESIS IN POSTURAL CONTROL AND BALANCE
-
-
A. Shaikh, G. Auner
-
-
Wayne State University,
Detroit, MI 48202
-
-
In older age groups,
spatial disorientation and falls are one of the most common reasons leading
to morbidity, mortality and seeking for a physician-help. Bilateral
vestibular end-organ failure is a major cause of spatial disorientation,
which, in aged patients, is mainly due to Meniere’s disease, ototoxicity or
age induced vestibular hair-cell loss. Idiopathic vertigo is also a frequent
malady in geriatric age group. Treatment of vestibular end-organ failure is
still unknown and existing therapeutic principles for chronic vertigo, such
as central vestibular suppression, are just symptomatic. We have pioneered
the state-of-art technology that will enable us to treat the vestibular
end-organ failure and chronic vertigo by means of vestibular
electrical-stimulation. The novel, prototype sensor is devised, which
encodes head-angular-position in real time and thus computes head-velocity,
a primary function of the semicircular canals. The sensor is implantable in
the mastoid antrum and has three degrees of freedom. The sensor is linked to
the decoder, processor, and sixteen channels stimulator chip; which are
implantable subcutaneously on the squamous temporal-bone. The information
encoded by the sensor is processed to compute component of the head velocity
in a plane of each of the three semicircular canals. Results of this
computation are further utilized to modulate frequency of biphasic square
wave electrical pulses, which are delivered by the sixteen channels
stimulator chip to nano-electric arrays. These arrays are developed using
the novel “Self-Assembled Nano Array Platform Approach” and provide the best
resolution for the artificial-nerve-stimulation. The arrays are attached to
self-retaining probe adapting the shape of the ampullae and providing the
electrode array a close contact with the neuroepithelia for specificity of
the individual three-canal-stimulation. Integration of the sensor feedback
with multiple similar sensors implanted at different body axes provides
basis of functional-neural-stimulation to reinforce the muscular control of
posture.
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Shaikh, Aasef G. |
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88. BLOCKADE OF ION CONDUCTANCE IN AUDITORY NEURONS MIMIC DISORDERED CENTRAL
PROCESSING IN AGED
-
-
A,
Shaikh; P, Finlayson
-
-
Department of Otolaryngology, Wayne State University, 550 E Canfield, Room #
327, Detroit, MI 48201
-
-
Presbycusis, a senile hearing-loss, is an inevitable sequel of aging. About
one-third of Americans older than age 60 suffer from hearing disorders.
In addition, the elderly also have a diminished ability to understand
time-compressed speech, which is likely due to a reduction in temporal
resolving power of auditory neurons. Based on auditory brainstem
evoked-responses (ABR), affected auditory temporal processing in the aged
has a central origin, particularly in Superior Olivary Complex (SOC). Yet
the precise mechanism for its pathogenesis is not defined. There is mounting
evidence that excitability in the central auditory neurons is affected in
presbycusis. A range of ion conductances shape the auditory neural
excitability. Hyperpolarization-activated-cationic-conductance (Ih) is one
of the predominant conductances determining resting membrane potential in
the SOC neurons and could regulate their excitability. In order to
understand the pathophysiology of disordered central processing in
presbycusis, it is important to examine the influence of Ih on the SOC
neural excitability. Auditory neurons also exhibit an important property,
post-stimulatory suppression, to aid echo-suppression and thereby adequate
processing of the time-compressed speech. As Ih is an important regulator
determining the resting membrane potential, its role in the post-stimulatory
suppression, particularly in the SOC neurons needs to be investigated. Here
we document Ih has a predominant role in shaping the SOC neural excitability
in vivo. We also report, selective blockade of Ih by ZD7288 (100microM)
significantly increases the magnitude of post-stimulatory suppression for
shorter inter-tone intervals and raises the neural-response threshold to the
second of the sequential-tones. As Ih-blockade decreases the neural
excitability and enhances the post-stimulatory suppression for short
inter-tone interval, in the SOC neurons; incompetent function of Ih could be
a possible neurophysiological correlate of decreased amplitude of ABR “wave
V” (decreased SOC neural activity) in aged patients with speech
understanding problems.
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|
Sharman, Edward H. |
- 90. MODULATION OF MRNA EXPRESSION PATTERNS IN MURINE CNS BY AGE
AND DIETARY MELATONIN: FOCUS ON IMMUNITY
- E. Sharman*, V. Perreau, K. Sharman, C. Cotman, and S. Bondy
Center for Occupational and Environmental Health, Department
of Community and Environmental Medicine, and Institute for Brain Aging and
Dementia, University of California Irvine, CA 92697-1825, U. S. A.
Age-dependent decline in brain immune response is a component of several
neurodegenerative diseases such as Alzheimer’s disease, Parkinson’s disease and
Huntington’s disease. To assess the impact of age on CNS immune response, gene
array analysis was used to identify changes in functional patterns and levels of
mRNA expression induced by i.p. injection of lipopolysaccharide (LPS) in the
cerebral cortex of both young (4-month) and old (27 month) CB6F1 male mice.
Modulation of the LPS-induced CNS immune response by the pineal secretory
product melatonin was assessed by supplementing the diet of aged animals with 40
ppm melatonin for 8 weeks preceding LPS injection. A wide range of age-related
changes in basal levels of genes within the brain was found. Dietary melatonin
administered to aged mice failed to reverse most of these changes. LPS treatment
elevated the expression of many genes in the younger animal. However, with
senescence, the response to such an inflammatory stimulus was attenuated.
Administration of melatonin was able to restore this reaction to a level more
closely resembling that of the younger animal.
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Shetty, Ritu A. |
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91. SHORT-TERM COENZYME Q10 SUPPLEMENTATION ENHANCES COGNITIVE PERFORMANCE IN
AGING MICE.
-
-
1) R. A.Shetty
-
2)N. Sumien.
-
3)R.Sohal.
-
4)M. J. Forster.
-
-
Dept of Pharmacology and Neuroscience, UNTHSC, FortWorth, TX-76107.
-
-
Brain aging is accompanied
by an increase in oxidative stress and mitochondrial dysfunction. Coenzyme Q
(CoQ) is an important component in the electron transport chain and is
thought to serve as a potent antioxidant. It is also known that CoQ content
declines with aging and the levels can be restored in older mice by
supplementation. However the effects of CoQ10 supplementation on behavior
are not known. Therefore the purpose of this study was to determine the
short-term effects of CoQ10 supplementation on psychomotor and cognitive
performance in young and old mice. Separate groups of young (4 months) and
old mice (18 months) were fed a control diet or a diet supplemented with low
or high concentrations of CoQ10 for a period of 15 weeks. The low and high
supplemented diets yielded a daily CoQ10 intake of approximately 148- or
654-mg/kg, respectively. After 6-weeks on the diets the mice were subjected
to a battery of age-sensitive behavioral tests for cognitive and psychomotor
performance. These tests include locomotor activity, coordinated running
performance, swim maze and startle response. CoQ10 treated mice tended to
exhibit faster learning of the swim maze task, an aging-sensitive measure of
cognitive performance. Diet supplementation with CoQ10 also improved the
ability of the old mice to retain information in swim maze task, an effect
that was not evident in the younger mice. In tests for auditory and shock
startle reflex and coordinated running ability, CoQ10 supplementation failed
to improve performance. These results suggest that short-term CoQ10
supplementation improves impaired cognitive function in older mice, but
fails to ameliorate age-impaired psychomotor function.
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Smith, Mark A. |
-
94.
DIFFERENTIAL EXPRESSION OF METABOTROPHIC GLUTAMATE RECEPTORS (mGluRs) IN
ALZHEIMER DISEASE
-
-
H-G
Lee, X. Zhu, M.J. O'Neill, G. Perry, M.A. Smith*
-
-
Institute of Pathology, Case Western Reserve University, Cleveland, Ohio,
USA
-
-
Although the mechanism
underlying the loss and/or dysfunction of susceptible neurons in Alzheimer
disease (AD) is unknown, it has been hypothesized that excitotoxicity may
play a pivotal role. Since glutamate has long been thought to be important
in the pathogenesis of AD, we hypothesized that select glutamatergic
receptor populations (e.g., metabotropic glutamate receptors (mGluRs)) may
correlate with those neurons vulnerable to AD. mGluRs are directly coupled
to the intracellular signaling system via GTP-binding proteins and are
thought to be related with synaptic transmission and neuroprotection. Since
synaptic loss is a prominent feature of AD, in this study, we investigated
the role of mGluR in AD. Specifically, we determined the expression of the
family of mGluR receptors including group I, II and III in the brains of AD
and age-matched control cases. We found a differential expression of mGluRs
that related directly to the pattern of neuronal susceptibility in AD.
mGluR2 is specifically increased in pyramidal neurons in the hippocampus of
AD and often co-localizes with neurofibrillary pathology. mGluR5 showed a
similar expression pattern to mGluR2 but was also found in astrocytes
surrounding amyloid plaques. The specificity of these findings is
demonstrated by the fact that mGluR1 expression, which, like mGluR5, is a
group I mGluR, is not different between AD and control cases. Our data
suggest that specific mGluRs such as mGluR2 and mGluR5 are differentially
regulated in AD and that this expression is directly related to neuronal
populations vulnerable to degeneration and to amyloid-beta and tau protein
lesions. Therefore, the differential regulation of specific mGluRs may
explain the selective neuronal degeneration in AD and provide a unique
therapeutic target for the treatment of AD.
|
|
Smith, Mark A. |
-
92. ACETYLATION: A NOVEL POSTTRANSLATIONAL MODIFICATION IN ALZHEIMER DISEASE
-
-
A.K.
Raina, X. Zhu, A.D. Cash, L.M. Sayre, G. Perry, M.A. Smith*
-
-
Institute of Pathology, Case Western Reserve University, Cleveland, Ohio,
USA
-
-
The mechanism leading to
the selective dysfunction of neurons in Alzheimer disease is incompletely
understood. Nonetheless, the formation of the characteristic neurofibrillary
pathology in Alzheimer disease is thought to play a pivotal role in this
process. Associated with the formation of neurofibrillary pathology,
proteins are subject to a number of posttranslational modifications with
phosphorylation being the best studied. In this study, we report an entirely
novel protein modification, namely acetylation of lysine, in vulnerable
neurons that parallels the temporal pattern of neuronal vulnerability in
cases of Alzheimer disease. While the exact role of protein acetylation in
Alzheimer disease is unknown, it is likely, given the physiological roles of
acetylation, that acetylation promotes neuronal survival by stabilizing the
cytoskeleton in the face of occult, and otherwise destabilizing, tau
phosphorylation events. This novel finding of a stabilizing modification in
vulnerable neurons in Alzheimer disease will provide new insights for
therapeutic interventions that could alter the natural history of this
dementia.
|
|
Smith, Mark A. |
-
93. A MECHANISTIC STUDY ON AMYLOID-BETA-INDUCED JNK ACTIVATION
-
-
X.
Zhu, C.A. Rottkamp, Z. Kubat, A.K. Raina, S.L. Siedlak, C.S. Atwood, G.
Perry, M.A. Smith
-
-
Institute of Pathology, Case Western Reserve University, Cleveland, Ohio,
USA
-
-
Amyloid-β protein
(Aβ), the main constituent of neuritic and diffuse plaques as well as
cerebrovascular amyloid deposits, characterizes the neuropathology of
Alzheimer disease (AD). While Aβ is neurotoxic in vitro, the actual
biochemical mechanism remains to be established. In this study, we found
that when Aβ is pretreated with the iron chelator deferoxamine,
neuronal toxicity is significantly attenuated while conversely, incubation
of holo-Aβ with excess free iron restores toxicity to original levels.
These data suggest that the toxicity of Aβ is mediated, at least in
part, via redox-active iron. The in vivo significance of these findings is
amplified by consideration of the enrichment of iron in AD neuropil, and
especially in association with Aβ. To understand the mechanistic
consequence of these findings, we also found that JNK/SAPK is activated in
susceptible regions of the brain in AD. The co-localization of activated JNK/SAPK
and Aβ deposits, as well as concomitant iron accumulation, suggests
that JNK/SAPK pathway may play an important role in metal-mediated Aβ
toxicity. In this regard, we found that although both AβPP
(K670N/M671L) transgenic mice and AβPP (V717I) transgenic mice develop
amyloid senile plaques, iron accumulates only in the former and strikingly,
activated JNK/SAPK is also only found in the former. Since such mice show
cognitive alterations, we hypothesize that neuronal dysfunction is, in part,
mediated through alteration in signal transduction pathways such as JNK/SAPK
pathway. Supported by the NIH (NS38648) to MAS and the Alzheimer’s
Association (NIRG-02-3923) to XZ.
|
|
Tsui, Justine C. Y. |
-
95. TELOMERE LENGTH AS A POSSIBLE-AGE MARKER IN FISH
-
-
*Justine, C.Y. Tsui, Stephen, B. Pointing and Kenneth, M.Y. Leung
-
-
The
Swire Institute of Marine Science, Department of Ecology & Biodiversity, The
University of Hong Kong, Pokfulam, Hong Kong SAR, China
-
-
Telomeres are nucleoprotein
structures at the ends of linear chromosomes consisting of DNA sequences
arranged in tandem repeated units (TTAGGG). They act to prevent aberrant
recombination and degradation of the chromosomal ends, and thus maintain
chromosome integrity. Since DNA polymerases are unable to copy chromosomal
DNA completely, telomere sequences typically shorten with the number of cell
divisions and hence with age. Previous research has demonstrated a negative
relationship between telomere length and age for various vertebrate species.
In this study, we describe for the first time an assessment of telomere
length as a function of age for the mangrove snapper, Lutjanus
argentimaculatus, a commercially important teleost species in Asia and the
Middle East. Individuals with known chronological ages (2.5-36 months) were
obtained from a fish farm in Shenzhen, China. Genomic DNA was extracted from
whole blood and dorsal muscle tissues. Telomere length was quantified using
the Telo TAGGG Telomere Length Assay (Roche Molecular Biochemicals), which
is based on Southern blot-hybridization with terminal restriction fragments
of the telomere. Telomere length in blood was significantly reduced with age
of individuals (r2 = 0.2484, F1,22 = 6.942, n = 23, p < 0.05), suggesting a
telomere shortening rate of 0.39 kbp/yr. Telomere length in muscle tissues
also decreased significantly in relation to fish age (r2 = 0.1513, F1,26 =
4.459, n = 27, p<0.05), resulting in a slightly faster shortening rate of
0.44 kbp/yr. The results suggest that patterns in fish telomere shortening
are similar to that in other vertebrates such as mammals and birds.
Measurement of telomere length in blood may have potential as a
non-destructive method for estimating age in fish.
|
|
van Steeg,
Harry |
96. LONGEVITY AND CROSS SECTIONAL STUDIES IN
DNA-REPAIR DEFICIENT MOUSE MODELS
Harry van Steeg1, Susan Wijnhoven1, Rudolf Beems1, Martijn Dollé1, Jan Vijg2,
Paul Lohman1, Jan Hoeijmakers3 and Bert van der Horst3
1Natl Inst of Public Health and the Environment, Department of Toxicology,
Pathology and Genetics, Bilthoven, The Netherlands; 2University of Texas Health
Science Center, Department of Physiology, San Antonio, Texas, USA; 3MGC
department of Cell Biology and Genetics, Erasmus University, Rotterdam, The
Netherlands
The accumulation of somatic DNA damage is considered to be a major cause of the
aging process in various species including mice and humans. Among the sources of
DNA damage, reactive oxygen species (ROS) are often thought to be the ultimate
cause of aging. However, the mechanisms involved remain obscure. To counteract
the effects of DNA damage, an intricate network of DNA repair pathways has
evolved. One major pathway is nucleotide excision repair (NER), which removes a
broad range of bulky lesions including some forms of oxidative damage. Patients
with a defect in NER proteins like CSB and XPD, both involved in repair as well
as transcription of DNA, appeared to have a decreased life span.
In order to investigate whether defects in genome maintenance are correlated to
accelerated aging, we have successfully conducted several longevity and cross
sectional studies with mice having defect in DNA repair and/or RNA transcription
(i.e. Xpa-, Csb-, XpdTTD- deficient mice as well as C57Bl/6 wild type controls).
The mean survival of female XpdTTD as well as Xpa mice appeared to be much
shorter (appr. 90 weeks) than those found for Csb and wild type littermate
controls (104-110 weeks). Full histopathology has been performed in aged female
mice, and gross examination at autopsy revealed small posture, kyphosis, large
spleen, small thymus and abnormal skin and hair especially in XpdTTD mice. The
terminal body weights in XpdTTD and Csb females were decreased, with an increase
in the relative weights of several organs of XpdTTD mice, especially in the
kidney, spleen and the heart. Furthermore, lipofuscin pigmentation (an aging
feature, correlated to oxidative damage) was found to be accumulated in the
liver of XpdTTD mice as compared to the other phenotypes. Finally, a variety of
immune parameters were determined in mice of the various genotypes and ages.
|
|
von Zglinicki, Thomas |
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98. VARIATION IN TELOMERE SHORTENING RATE CAUSES HETEROGENEITY IN REPLICATIVE
LIFESPAN
-
-
T.von Zglinicki, C. Martin-Ruiz, J. Petrie, G. Saretzki
-
-
Biogerontology Laboratory, Newcastle University, UK
-
-
The replicative lifespan of
human fibroblasts is heterogeneous, with a fraction of cells senescing at
every population doubling. To find out whether this heterogeneity is due to
premature senescence, i.e. driven by a non-telomeric mechanism, fibroblasts
with a senescent phenotype were isolated from growing clones by flow
cytometry. These senescent cells had shorter average telomere length than
their cycling counterparts at all population doubling levels both in mass
culture and in individual sub-clones, indicating heterogeneity in the rate
of telomere shortening. Ectopic expression of telomerase stabilised telomere
length in the majority of cells and rescued these from early senescence,
suggesting a causal role of telomere shortening. Under standard cell culture
conditions, there was a minor fraction of cells which showed a senescent
phenotype and short telomeres despite active telomerase. This fraction
increased under chronic mild oxidative stress, which is known to accelerate
telomere shortening. The data show that heterogeneity of human fibroblast
replicative lifespan can be caused by significant stochastic cell-to-cell
variation in telomere shortening.
|
|
von Zglinicki, Thomas |
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97. TELOMERE SHORTENING IN HUMAN FIBROBLASTS IS NOT DEPENDENT ON THE SIZE OF
THE TELOMERIC 3’-OVERHANG
-
-
T. von
Zglinicki, B. Keys, V. Serra, G. Saretzki
-
-
Biogerontology Laboratory, Newcastle University, UK
-
-
Telomeres shorten in human
somatic cells with each round of DNA replication, and this shortening is
thought to ultimately trigger replicative senescence. Telomere shortening is
caused partly by the inability of semi-conservative DNA replication to copy
a linear strand of DNA to its very end. Post-replicative processing of
telomeric ends, producing single-stranded G-rich 3’ overhangs, has also been
suggested to contribute to telomere shortening. This suggestion implies that
a positive correlation should exist between the length of 3’ overhangs and
the rate of telomere shortening. Measuring overhang length by in-gel
hybridisation, we could confirm shortening of overhangs as human MRC5 and BJ
fibroblasts approach senescence. However, a large study comprising of
fibroblast strains from 21 donors under conditions leading to two orders of
magnitude of variation in telomere shortening rate ruled out any correlation
between telomere overhang length and shortening rate, suggesting that
overhang length is neither causal for nor indicative of telomere shortening.
-
|
|
Wang, Zhihui |
99. EXPRESSION OF GENES RELATED TO
INSULIN SENSITIVITY AND LIPID METABOLISM IN AMES DWARF AND CALORIC RESTRICTED
MICE
Zhihui Wang, K.A. Al-Regaiey,
M.M.Masternak, A.Bartke
Southern Illinois University, School of Medicine, Geriatrics Research, 801 N.
Rutledge Springfield, IL 62794
Ames dwarf (df) mice display primary deficiency of growth hormone (GH),
prolactin (PRL), and thyroid simulating hormone (TSH), improved insulin
sensitivity, diminutive body size, and prolonged longevity. Young adult df mice
have higher % body fat compared with their normal littermates (Wild type, WT).
In order to study the mechanism of improved insulin sensitivity in these mice,
we have used real time PCR to examine gene expression of adipocytokines related
to insulin sensitivity including adiponectin, leptin, resistin, TNF-,
and interleukin (IL)-6, and genes related to fat metabolism including fatty acid
synthase (FAS), hormone-sensitive lipase (HSL), uncoupling protein (UCP)-2, and
3-adrenergic receptors (3AR) in white adipose tissue (WAT). We
also analyzed expression of the same genes in df and WT animals subjected to 30%
caloric restriction (CR). In WAT from df mice fed ad libitum (AL), the
expression of adiponectin, TNF- , FAS, HSL, and UCP2 were significantly
diminished, and the expression of IL-6 was dramatically suppressed, whereas the
expression of leptin, resistin, and 3AR were not changed. CR increased
gene expression of resistin, FAS, and 3AR, and diminished gene
expression of UCP2 and IL-6 in WT mice compared with their AL littermates. CR
also diminished expression of leptin in df mice compared with AL df mice. In
both WT and df mice, CR did not alter the expression of adiponectin, HSL,
although their expression and the expression of resistin, FAS, UCP2, 3AR
in CR df mice were much lower than in CR WT mice. These results suggest that
decreased expression of TNF-, FAS, and IL-6 might play a role in
improving insulin sensitivity in df mice. CR might down-regulate the gene
expression of leptin via increasing expression of 3AR, and down-regulate
the expression of UCP2, thus maintaining the energy balance of the body.
(Supported by NIA and Ellison Medical Foundation)
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|
Wilson, Mark A. |
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100. BLUEBERRY-DERIVED PHYTONUTRIENTS CAN AFFECT AGING AND STRESS TOLERANCE IN
THE NEMATODE, CAENORHABDITIS ELEGANS
-
-
M.
Wilson, B. Shukitt-Hale, G. Lee, D. Ingram, J. Joseph, C. Wolkow
-
-
Laboratory of Neuroscience, National Institute on Aging, IRP, 5600 Nathan
Shock Drive, Baltimore, MD 21224
-
-
Plant-derived antioxidants have been shown to provide beneficial health
effects. In order to explore the mechanisms behind these observations, we
investigated the effects of blueberries on lifespan and stress resistance in
the nematode, C. elegans. Either crude extracts or partially purified
polyphenol fractions of blueberries increased mean lifespan and slowed
age-related declines in pharyngeal pumping rates in wild-type animals.
Resistance to thermal stress was also increased significantly. One
possibility is that blueberry extracts inhibited bacterial growth, a
treatment known to extend mean C. elegans lifespan. We examined this
possibility by comparing the effects of antibiotics and blueberry. While
treatment with either ampicillin or blueberry extract extended C. elegans
lifespan, there were no additive effects on longevity when these treatments
were combined. However, while blueberry significantly increased
thermal tolerance, ampicillin had no effect. The differential effect on
stress tolerance suggests that blueberry can directly affect C. elegans
physiology, independent of effects on bacterial growth. Through what genetic
pathways can blueberries alter stress tolerance? Reductions in
insulin/IGF-like signaling are correlated with increased stress resistance
and lifespan in a wide range of organisms. Longevity and stress
resistance in C. elegans insulin-like pathway mutants requires the DAF-16/FOXO
transcription factor. To determine whether blueberry affects insulin-like
signaling, we examined whether the beneficial effects of blueberry extracts
required daf-16 activity. Fractional increases in lifespan, and
absolute increases in thermotolerance, were similar in wild-type and
daf-16(mgDf50) animals. In untreated daf-16(mgDf50) animals,
pharyngeal pumping declined more rapidly with age than in wildtype animals.
This decline was slowed to the wild-type rate with blueberry extract.
These experiments indicate that blueberry's longevity-promoting effects do
not require DAF-16/FOXO activity, and probably occur independently of
insulin-like signaling. We hypothesize that treatment with blueberry extract
increases C. elegans longevity by increasing intrinsic resistance to
environmental and/or metabolic stresses that limit mean lifespan.
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|
Wolkow, Catherine A. |
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101. AGING ALTERS THE COORDINATION OF SENSORY INPUTS WITH MOTOR RESPONSES IN
THE NEMATODE, CAENORHABDITIS ELEGANS
-
-
C.
Wolkow, D. Chow, C. Glenn, L. David, I. Goldberg
-
-
Laboratory of Neuroscience, National Institute on Aging, IRP, 5600 Nathan
Shock Drive, Baltimore, MD 21224
-
-
Many behavioral responses require the coordination of sensory inputs with
motor outputs. Aging is associated with progressive declines in both
motor function and muscle structure. However, the consequences of
age-related motor deficits upon behavior have not been clearly defined.
To investigate this question, we have examined the effects of aging upon
behavior in the nematode, Caenorhabditis elegans. We first analyzed
the effect of age on responses to sensory stimuli. When touched gently
with a hair, both young and aged animals responded appropriately by backing
away from the stimulus. When presented with an aversive odor, octanol,
both young and aged animals stopped forward movement similarly.
However, aged animals exhibited defects in backing away from the odorant
source and these defects were exacerbated when octanol was diluted 100-fold.
Aged animals displayed significant deficits in chemotaxis behavior, as older
animals failed to progress fully to the source of an attractive stimulus.
Together, these results indicate that age was not associated with
significant declines in sensory ability. However, locomotory responses
to stimuli were severely compromised.To determine the basis for these
behavioral deficits, we analyzed the patterns of movement in aged animals.
Older animals moved 50% slower than young animals and failed to maintain
forward movement for sustained intervals. Examination of muscle
structure at these ages showed that reduced muscle tissue integrity was
correlated with age-related declines in locomotory behavior, although
widespread muscle deterioration was rare at the ages examined.
Finally, treatment with a muscarinic agonist improved locomotory behavior in
aged animals, indicating that improved neuromuscular signaling may be one
strategy for reducing the severity of age-related behavioral impairments.
|
|
Xu, Dongsheng |
-
102. GENETIC SELECTION AND INITIAL CHARACTERIZATION OF MOUSE LINES FOR HIGH
AND LOW STRESS SUSCEPTIBILITY
-
-
DS.
Xu*,1,3, T. Lambert 1, D. Paul 2, Laura Meyerle 1 , C. Nekl 1 , J.
Potts 2, M. Nielsen 2, Y. Zhou 1
-
-
1
Department of Veterinary & Biomedical Sciences, 2 Department of Animal
Sciences, University of Nebraska - Lincoln; 3 Center for Neurovirology
and Neurodegenerative Disorders, University of Nebraska Medical Center,
Omaha
-
-
Two mouse lines were generated by genetic selection for high and low stress
susceptibility (SH and SL lines, respectively), from baseline populations of
mice selected for high (base for SH) and low (base for SL) rates of
metabolism, which also showed differences in locomotor activity and stress
responses. Corticosterone concentrations in sera collected at various time
points from the SH and SL mice with or without exposure to acute or repeated
restraint stress were determined by radioimmunoassay and used as one of the
key criteria for selection. The SH mice showed significantly (P < 0.05)
higher levels of serum corticosterone, at all tested time points, than the
SL mice. Initial reverse transcription polymerase chain reaction (RT-PCR)
analysis using brain RNA revealed differences in expression of three major
stress-response regulatory genes: corticotrophin-releasing hormone (CRH),
glucocorticoid receptor (GR) and mineralocorticoid receptor (MR). Expression
of CRH was higher while GR and MR expressions were down-regulated in the SH
mice as compared to SL mice. Microarray assays were used to compare gene
expression in the brains of the two lines of mice under stressed and
non-stressed conditions, and revealed significant differential expression
patterns of many genes (up or down regulated) between the two lines as well
as within the same line of mice. Subsequent RT-PCR experiments using
specific primers for 25 genes of interest, selected based on the microarray
data, confirmed the genetic and stress-induced alterations in gene
expression between the SH and SL mice, such as microtubule-associated
protein-2, P21-activated kinase-3, calcium/calmodulin-dependent protein
kinase IIa, and calcium channel proteins. Further establishment of the
SH and SL mouse lines, divergent through selection for altered stressed
susceptibility, may become a useful tool to study stress-induced early aging
and neurodegeneration.
|
|
Yin, Zhirong |
-
103. AGE-RELATED DIFFERENCES OF GENE EXPRESSIONS AND DNA DAMAGE IN HUMAN
FIBROBLASTS AFTER EXPOSURE TO COBALT CHROME PARTICLES
-
-
Yin
Z.R., Papageorgiou I., Clerkin J.S., Learmonth I.D., Case C.P.
-
-
Bristol Implant Research Centre, University of Bristol, BS10 5NB, U.K.
-
-
Wear debris from worn
cobalt chrome orthopaedic joint replacements causes an increase in
chromosomal translocations and aneuploidy. In this study the levels of DNA
damage and certain gene expressions (TGF-ß2, p38 MAPK, Integrinβ1,
SOD1, Caspase 10, PURA, FRA-1 and VNR) have been compared in human
fibroblasts of different age (10 and 35 population doublings, PD) after
exposure to different doses of cobalt chrome particles and at different
times of exposure (6 and 24 hours). The 35 PD fibroblasts showed
significantly more immunostaining of the senescence-associated
β-galactosidase (57.4%) than the 10 PD fibroblasts (1.4%). The
level of DNA damage, as detected with alkaline comet assay, was greater at
higher doses, at longer exposures (up to 24 hours) and in younger (10 PD)
fibroblasts. No significant change in cell viability was noted using MTT
assay or LDH assay. The expression of all the genes listed above was
generally lower after exposure to cobalt chrome particles using
semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR).
The reduction in gene expressions, like the increase in DNA damage was
greater at higher dose and at longer time of exposure. Interestingly these
changes in DNA damage and gene expression were more pronounced in younger
than in older fibroblasts but not until after 24 hours of exposure. These
results show that levels of gene expression of TGF-ß2, p38 MAPK, Integrinβ1,
SOD1, Caspase10, PURA, FRA-1 and VNR may be correlated with the level of DNA
damage and that this depends on the dose and length of exposure and the age
of the cells. This highlights the potential importance of these genes in the
mutagenicity of cobalt chrome particles in human fibroblasts.
|
|
Young, J. |
104. Assessment of motor
performance in rhesus monkeys on long-term calorie restriction
J. Young1, Zhiming Zhang2, D.
Gash2, G. Gerhardt2, G. Roth3, M. Lane,3 J. Mattison,3 D. Ingram3
1Sobran, Inc, 4401
Dayton-Xenia Road, Dayton, OH 45432; 2Dept. of Anatomy and Neurobiology,
University of Kentucky Medical Center, Lexington, KY 40536; 3Laboratory of
Experimental Gerontology, National Institute on Aging, NIH, Gerontology Research
Center, 5600 Nathan Shock Drive, Baltimore, MD 21224;
In numerous rodent studies,
calorie restriction (CR) has been demonstrated consistently to increase
lifespan, reduce the incidence and onset of age-related chronic disease, and
attenuate age-related decline in many physiological functions. Emerging
evidence from studies in rhesus monkeys suggests that CR can also retard aging
processes in a species closely related to humans. An important question
regarding the effects of long-term CR in monkeys is whether behavioral function
will be impacted, either negatively or positively. We have initiated studies to
evaluate motor performance in male and female rhesus monkeys involved in a
long-term study of CR. Control monkeys were provided a highly nutritious diet
while monkeys on CR have had their intake of this diet reduced 30 percent from
the level of comparable aged controls. Monkeys were introduced to the study at
different stages in the lifespan, including juvenile, adult, and aged groups of
monkeys. Fine motor and coarse motor performance were measured in an automated
movement assessment panel attached to the home cage of the monkey. During
trials of 45 sec duration, monkeys are given access to the apparatus during
which they must reach with their hand through an opening and then must maneuver
their hand at a 90o angle from entry into an elevated center chamber to retrieve
a visible food treat (soft lifesaver candy). During different sessions, the
treat is positioned either on a flat platform, a straight rod, or a hook in the
center chamber. Performance on each task is measured as the time required to
retrieve the treat once the hand has entered the apparatus. Age-related decline
in performance (longer retrieval times) of rhesus monkeys on all three tasks has
been previously documented (Zhang et al. J.Gerontol. Biol. Sci. 55:B473, 2000),
and we have replicated these findings in the current study. To assess effects
of CR on performance, comparisons of fine motor retrieval time were made for
male and female monkeys between the ages of 12.1 - 24.3 yr that had been on CR
for 12 -18 yr. For males, we found that retrieval time was equivalent between
diet groups on the easiest tasks (platform and straight rod), but times on the
more difficult hook task were significantly faster for CR group. For females,
CR monkeys had significantly faster retrieval times on both the platform and the
hook tests, but had significantly slower times on the straight rod task.
Considering performance on only the most difficult task, we can conclude that CR
improves motor performance.
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Zachariah, Sally B. |
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105.
BOTULINUM TOXIN TYPE A (BoNT/A) FOR CHRONIC HEADACHES IN ELDERLY
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S.Zachariah,M.D, T.Ranjan,M.D and Aveen Zachariah.
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Background: Headache is reported to be the tenth and fourteenth most common
symptom in elderly women and men respectively. Five to ten percent of
patients over 65 develop chronic daily headaches. BoNT/A has dramatically
improved the treatment of a variety of neurological disorders, including
headache.
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Methods: Data reviewed of 30 patients (average age-60.8year) involved in an
open label trial showed 21 had typical vascular headaches, 7 had chronic
daily headaches [>20 headaches in a month] and 2 had tension headaches. They
received BoNT/A for headaches (average duration 10 yrs) between 1995 and
2003. All patients met the following criteria: headache duration
of >2 yrs; absence of known allergy to BoNT/A; severity of headache > 5 [on
0-10 scale]; failure to Triptans, Beta blockers, Calcium Channel blockers,
Anticonvulsant and Antidepressants. Forty to seventy-five unit intra
muscular injections were given in corrugators and bilateral upper & lower
frontalis and temporalis at the depth of 1-3 mm using number 30 needle.
Patients with tension headache were given semispinalis and splenius capitis
injections, following the path of pain. Injection was repeated after 3
months. Evaluation entailed: headache scale, satisfaction survey, medication
use, life style questionnaire and neurological exam.
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Results: Most of the patients had effect in 3-5 days with optimum benefit in
2 weeks. The average pain intensity reduced from 8.66 to 1. First injection
effects lasted from 45 to100 days with later injections lasting longer.
Twenty-four patients out of 30 (80%) had clinical response, of which 20
patients were very satisfied, 4 patients were satisfied and 6 patients [20%]
had no response to therapy. Respondents had long-term benefits like pain
relief, stoppage or reduction of medications, better life style, and
improvement in productivity.
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Conclusions: Older patients with chronic headache with polypharmacy
failures tolerate and benefit from BoNT/A therapy with minimal acute side
effects.
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Zienko, Sarah C. |
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106. UNDERSTANDING THE GENETICS OF AGING: A CANINE MODEL
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S.
Zienko, K.Greer, M. Breen, K. Murphy
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Dept. of Veterinary Pathobiology; Texas A&M Univ. College of Veterinary
Medicine; College Station, Texas, U.S.A.; Dept. of Molecular Biomedical
Sciences, North Carolina State Univ. College of Veterinary Medicine;
Raleigh, North Carolina, U.S.A.
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The biology of aging is
extremely complex, and is, therefore, poorly understood. Research
pertaining to aging has many potential benefits, the most important of which
are extension of life spans and improvement of the quality of life in aged
populations. Select human populations have significantly longer life
expectancies than others, suggesting an underlying genetic component
associated with the aging process. The genetic factors governing
longevity are numerous, difficult to define, and may possess regulatory
functions. The hereditary component of longevity applies to animal
populations. Our interest in the dog as a model stems from the fact
that the dog serves as excellent model animals for many human hereditary
conditions, and that average life expectancy of purebred dogs decreases as
both height and weight increase, as shown by preliminary regression analysis
of 117 breeds. As a first step to understanding the potential genetic
factors associated with longevity in dogs, comparative gene mapping data is
being generated by cataloging and mapping 56 candidate genes.
Variation in 23 of these genes is associated with either extended or
truncated life spans in the human or mouse. The additional 23 genes,
that give approximately 0.5Mb coverage of a 22Mb region of human chromosome
4, were selected because a microsatellite marker, D4S1564, shows strong
association with excessive longevity in an Italian population.
Following mapping of these genes, detailed analysis will be conducted to
obtain polymorphic markers in close vicinity of the mapped genes.
These markers will be analyzed across a range of breeds with contrasting
degrees of longevity. Additionally, comprehensive analysis will be
carried out to identify variations within selected genes (from the group of
56) across breeds with diverse longevity. These pilot studies will
serve as a platform to initiate broader research aimed at understanding
genetic factors associated with longevity in dogs and other animals.
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