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  • 1
    Keywords: brain ; COMBINATION ; Germany ; GENERATION ; SYSTEM ; liver ; PROTEIN ; PROTEINS ; HEART ; COMPLEX ; COMPLEXES ; MECHANISM ; RAT ; PHOSPHORYLATION ; TARGET ; RAT-LIVER ; SUBUNIT ; MEMBRANE ; STRESS ; MODULATION ; MITOCHONDRIA ; OXYGEN ; antioxidants ; PROTEOMICS ; reactive oxygen species ; glutathione-S-transferase ; GENE-EXPRESSION PROFILE ; ageing ; GEL-ELECTROPHORESIS ; assembly ; proteome ; REACTIVE OXYGEN ; ROS ; CALORIE RESTRICTION ; SHORT-TERM ; COMPLEX-I ; MEMBRANE-PROTEINS ; CYTOCHROME-C-OXIDASE ; DIGE ; Species ; ROS PRODUCTION ; SHIFT ; RESPIRATORY-CHAIN
    Abstract: Mitochondria being the major source and target of reactive oxygen species (ROS) play a crucial role during ageing. We analyzed ageing and calorie restriction (CR)-induced changes in abundance of rat liver mitochondrial proteins to understand key aspects behind the age-retarding mechanism of CR. The combination of blue-native (BN) gel system with fluorescence Difference Gel Electrophoresis (DIGE) facilitated an efficient analysis of soluble and membrane proteins, existing as monomers or multi-protein assemblies. Changes in abundance of specific key subunits of respiratory chain complexes I, IV and V, critical for activity and/or assembly of the complexes were identified. CR lowered complex I assembly and complex IV activity, which is discussed as a molecular mechanism to minimize ROS production at mitochondria. Notably, the antioxidant system was found to be least affected. The GSH:GSSG couple could be depicted as a rapid mean to handle the fluctuations in ROS levels led by reversible metabolic shifts. We evaluated the relative significance of ROS generation against quenching. We also observed parallel and unidirectional changes as effect of ageing and CR, in subunits of ATP synthase, cytochrome P450 and glutathione S-transferase. This is the first report on such 'putatively hormetic' ageing-analogous effects of CR, besides the age-retarding ones
    Type of Publication: Journal article published
    PubMed ID: 19894137
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  • 2
    Keywords: CANCER ; DISEASE ; POLYMORPHISMS ; AGE ; LIFE-SPAN ; LENGTH ; METAANALYSIS ; GENOME-WIDE ASSOCIATION ; OLDEST-OLD ; NORMAL HUMAN-CELLS
    Abstract: Leukocyte telomere length (LTL) has been observed to be hereditable and correlated with longevity. However, contrasting results have been reported in different populations on the value of LTL heritability and on how biology of telomeres influences longevity. We investigated whether the variability of genes correlated to telomere maintenance is associated with telomere length and affects longevity in a population from Southern Italy (20-106 years). For this purpose we analyzed thirty-one polymorphisms in eight telomerase-associated genes of which twelve in the genes coding for the core enzyme (TERT and TERC) and the remaining in genes coding for components of the telomerase complex (TERF1, TERF2, TERF2IP, TNKS, TNKS2 and TEP1). We did not observe (after correcting for multiple testing) statistically significant associations between SNPs and LTL, possibly suggesting a low genetic influence of the variability of these genes on LTL in the elderly. On the other hand, we found that the variability of genes encoding for TERF1 and TNKS2, not directly involved in LTL, but important for keeping the integrity of the structure, shows a significant association with longevity. This suggests that the maintenance of these chromosomal structures may be critically important for preventing, or delaying, senescence and aging. Such a correlation was not observed in a population from northern Italy that we used as an independent replication set. This discrepancy is in line with previous reports regarding both the population specificity of results on telomere biology and the differences of aging in northern and southern Italy.
    Type of Publication: Journal article published
    PubMed ID: 25631672
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  • 3
    ISSN: 1573-6768
    Keywords: aging ; antibody ; functional genomics ; phage display
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Filamentous bacteriophage offers the possibility oflinking genotype with phenotype in one geneticpackage. By creating a library of different proteinsas fusion to one of the coat proteins of filamentousbacteriophage it becomes possible to isolate proteinsbased on their binding characteristics. Phagedisplayed libraries of varying formats, ranging fromsmall peptides to antibody fragments, have beenselected successfully for ligands toward a large panelof targets. These peptides and antibody fragments areapplicable in analysis of the behaviour of certainproteins with respect to changes in biologicalprocesses such as aging, thus providing valuable toolsfor a general understanding of the aging process.Alternatively, display of antibody fragments orpeptides on filamentous bacteriophage can beinstrumental in the discovery of novel antigens asexemplified by selections on cell surfaces or oncomplex protein mixtures such as sera from agingindividuals. Although phage display has been appliedsuccessfully in a large number of studies relating tocancer, viral infections and other biologicalprocesses, its application in the field of agingresearch is yet to be realised.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-6768
    Keywords: gene deletion ; Podospora anserina ; readthrough ; senescence ; translation elongation factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract eEF1A is encoded by a unique gene in the filamentousfungus Podospora anserina. We show here that (1)this gene is essential for vegetative growth, (2)readthrough at UGA stop codon level is positivelycorrelated with eEF1A level, (3) eEF1A level isregulated in P. anserina. (4) Increasing eEF1Agene dosage does not modify P. anserina lifecycle parameters, especially longevity is not changed.These data confirm and extend those previouslyobtained in yeast and Drosophila.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-6768
    Keywords: diphtheria ; elderly ; hemodialysis ; immunology ; vaccination ; zinc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Zinc deficiency causes abnormalities of the immuneresponse. In chronic hemodialysis therapyabnormalities in zinc metabolism as well as animpaired immune response to vaccination have beenreported. Therefore we performed a vaccination studyagainst diphtheria and hypothesized that the responseto diphtheria vaccination is related to serum zincdeficiency in hemodialysis patients. Serum zincconcentrations were assayed in 16 chronic hemodialysispatients (10 male, 6 female; mean age 65 years)without a documented vaccination history againstdiphtheria. Nine of these patients were tripleimmunized against diphtheria while seven received asingle vaccination. The response to diphtheriavaccination was measured by ELISA detecting specificantibodies to diphtheria-toxoid. Seroconversion 6 and12 months after vaccination was defined as thedoubling of antibody titers in patients ≥ 0.1IU/ml prior to vaccination or as titers %gt; 0.1 IU/mlin all other patients. Only 6/16 hemodialysispatients responded to immunization against diphtheriaby specific antibody production (〉 0.1 IU/ml).Twelve months after the single injection 3/7 patientsseroconverted while six months after the triplevaccination 3/9 patients responded to immunization.This was not age-dependent, whereas in non-responderswe detected significantly decreased serum zinc levels.In contrast, responders showed similar serum zinclevels as age-matched controls. Furthermore, wemeasured a decreased α2-macroglobulinconcentration only in the responders amongst thehemodialysis patients. Protection against diphtheriaand the immune response to diphtheria vaccination inhemodialysis patients is poor. The failure to respondto active diphtheria vaccination is related to asignificantly decreased serum zinc concentration inhemodialysis patients.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Biogerontology 1 (2000), S. 1-1 
    ISSN: 1573-6768
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-6768
    Keywords: dipeptide ; life span ; rat ; thymus ; tumorigenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Immunomodulatory molecule L-Glu-L-Trp wasisolated from natural calf thymic peptide complexThymalin by reverse-phase high performance liquidchromatography. On the basis of the synthesizeddipeptide a pharmaceutical was designed containg this compound, which later receives the brand nameThymogen®. The agent activated T-celldifferentiation, T-cell recognition of peptide-MHCcomplexes, induced changes in intracellularcomposition of cyclic nucleotides, and activatedneutrophilic chemotaxis and phagocytosis. The effectof dipeptide on survival, life span and spontaneoustumor development was studied in female rats. Seventy-six, five-month-old outbred female rats were randomlysubdivided into two groups and were subcutaneouslyinjected with 0.2 ml of normal saline (controls, 32 rats) or with 5 µg/rat of the dipeptideL-Glu-L-Trp, dissolved in 0.2 ml of saline (44 rats),5 times per week for 12 months. Animals were monitoredup to their natural death and all the tumorsdiscovered were studied microscopically. Mean lifespan of rats in both groups was similar but that of10% maximum survived control rats constituted949 ± 16.1 days, whereas in the dipeptide-treatedrats this value was 1048 ± 21.1 days (P 〈 0.001).Six out of 44 rats treated with the drug survived overthe maximum life span of control rats (965 days). Theaging rate indicated as α in the Gompertz equation,was 0.0071 days−1 in controls and 0.0041 days−1 in rats exposed to L-Glu-L-Trp. Totaltumor incidence was 1.5 times lower (P 〈 0.01),malignant tumor incidence 1.7 times lower (P 〈 0.01),and hematopoietic malignancies (leukemias andlymphomas) 3.4 times lower (P 〈 0.02) in rats exposedto the dipeptide in comparison with controls. Thus,treatment with L-Glu-L-Trp delayed aging rate anddecreased spontaneous tumor incidence in rats.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Biogerontology 1 (2000), S. 157-162 
    ISSN: 1573-6768
    Keywords: ageing ; circadian activity patterns ; clock ; Drosophila ; fusion ; green fluorescent protein ; per
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A growing body of evidence indicates that one of theage-associated changes in the central nervous systemthat affect most old people is the loss of functionof the circadian clock system. This loss results inimpaired timing and quality of sleep, with consequentcognitive and other behavior problems. Failure of theclock contributes to the difficulties encountered withAlzheimer's disease. It also results in adversechanges in the hormonal regulation of intermediarymetabolism, stress resistance and sexual function.Drosophila melanogaster is proposed as a modelorganism where this age-related change may be studiedmore readily. Circadian patterns are disrupted inDrosophila, with considerable differences betweenstrains. In addition a fusion gene product of a keygene involved in the clock (per), and GreenFluorescent Protein, shows a 50% fall with age.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1573-6768
    Keywords: Drosophila melanogaster ; hypergravity ; longevity ; mild stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Drosophila melanogaster flies were exposed to hypergravity starting at two days of age, the range of gravity levels used being 2.58–7.38 g. No longevity change was observed for exposures of less than 14 days. The longevity of males increased if they were submitted to hypergravity for durations ranging from 14 to 24 days. This increase in longevity was never observed in females. The positive effect of exposure to hypergravity has been replicated in two laboratories using two wild-type strains and different rearing conditions. A short hypergravity exposure seems to be a mild stress, yielding positive effects on longevity. This is in accordance with two previous studies showing a slight longevity increase after heat shock in the nematode Caenorhabditis elegans and in Drosophila melanogaster.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1573-6768
    Keywords: alternative methods ; drug testing ; in vitro toxicology ; irreversible effects ; premature senescence ; stress-induced ; sublethal stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract No alternative in vitro method exists fordetecting the potential long-term genotoxic effects ofmolecules at subcytotoxic concentrations, in terms ofdays and weeks after exposure(s) to the moleculetested. A theoretical model of cellular senescence ledto the concept that subcytotoxic stresses under anymolecules at subcytotoxic doses, such as moleculesunder development in the pharmaceutical, cosmetics andfood industry, might lead human fibroblasts into a stateclosely related to in vitro senescence. Thisconcept was then experimentally confirmed invitro: many biomarkers of replicative senescence ofhuman fibroblasts were found 72 h after theirexposure to various kinds of stressors used at non-cytotoxic concentrations. This phenomenon has beentermed stress-induced premature senescence (SIPS).Moreover, proteomics studies have revealed that,besides their effects on the appearance of thebiomarkers of senescence, sublethal stresses under avariety of stressors also lead to long-term specificchanges in the expression level of proteins which arestress-specific. These changes have been coined themolecular scars of stress. The proteins correspondingto these molecular scars may be identified using thelatest developments in mass spectrometry. This modelof stress-induced premature senescence may be appliedto the toxicological sciences when testing for thepotential irreversible long-term effects of moleculeson the cell fate.
    Type of Medium: Electronic Resource
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