DNA double strand breaks (DSBs), which may occur during DNA replication or due to the action of genotoxic agents, are extremely dangerous DNA lesions as they can cause chromosomal rearrangements and cell death. Therefore, accurate DSB repair is vital for genome stability and cell survival. Two main mechanisms serve to repair DNA DSBs: non-homologous end joining, which re-ligates DNA ends together, and homologous recombination (HR), which restores broken DNA using homologous sequence as a template for repair. One-ended DSBs are a subject for the specialised HR-dependent repair pathway known as break-induced replication (BIR). At low frequency, DNA breaks can also be healed by telomerase, which normally extends telomeres at natural chromosome ends, but may also add de novo telomeres to DSBs due to their similarity to chromosome ends. De novo telomere addition is a deleterious event, which is effectively inhibited by the nuclear Pif1 (nPif1) helicase phosphorylated at the TLSSAES motif in response to DNA damage. In this study, it is reported that the same regulatory motif of nPif1 is also required for DSB repair via BIR. The requirement of the nPif1 TLSSAES sequence in BIR is dependent on the functional DNA damage response (DDR). Thus, nPif1 phosphorylation by the DDR machinery might mediate the role of nPif1 in BIR. In contrast, the nPif1 regulatory motif is not essential for BIR at telomeres in cells lacking telomerase. These observations indicate that the mechanism of nPif1 function in DSB repair via BIR and in BIR at telomeres might be different. In this work, a protocol for nPif1 pull-down was optimized to reveal the mechanism of the phosphorylation-dependent nPif1 functions in cells undergoing DNA repair, i. e. the mechanism of nPif1-mediated inhibition of de novo telomere addition and promoting DSB repair via BIR. In future, this protocol can be used to dissect the role of nPif1 in DNA repair through the identification of its potential interacting partners. The Srs2 helicase negatively regulates HR via dismantling Rad51 filaments. According to preliminary data from the laboratory of Sveta Makovets, Srs2 also promotes de novo telomere addition at DSBs in a Rad51-dependent manner. The work presented here establishes that Srs2 is dispensable for telomerase-mediated addition of TG1-3 repeats to DSBs. Instead, Srs2 is required for the reconstitution of the complementary DNA strand after telomerase action, thus ensuring the completion of de novo telomere addition. Overall, this study demonstrates that recombination-dependent DSB repair and de novo telomere addition share common regulatory components, i. e. the nPif1 helicase phosphorylated in response to DNA damage and the Srs2 helicase. Phosphorylated nPif1 promotes DSB repair via BIR in addition to its known role in inhibition of telomerase at DSBs, whereas Srs2 uses its well established ability to remove Rad51 from ssDNA to promote the restoration of dsDNA and thus to complete de novo telomere addition.

Orientador: Valéria Cristina Sandrim / Resumo: Evidências apontam que a dieta vegetariana diminui a probabilidade de desenvolver doenças cardiovasculares (DCVs). Um dos principais mecanismos que levam às doenças cardiovasculares é a disfunção do endotélio, associada com a diminuição da biodisponibilidade de óxido nítrico (NO) e a produção excessiva de espécies reativas de oxigênio (ROS). Do ponto de vista de biomarcadores de estresse oxidativo/antioxidante e de envelhecimento biológico, enzimas com propriedades antioxidantes e o comprimento do telômero podem apresentar efeitos na modulação do sistema vascular. O presente estudo inclui dois manuscritos, sendo o primeiro relacionado a via de regulação NRF2/HO-1 e o segundo ao comprimento dos telômero em onívoros (ONI) e vegetarianos (VEG). No primeiro manuscrito, o objetivo foi verificar a concentração de HO-1 circulante, bem como investigar o efeito da incubação do plasma de ONI e VEG em células endoteliais sob a modulação da via NRF2/HO-1 e a produção de NO. Dos 745 indivíduos inicialmente recrutados, 44 ONI e 44 VEG do sexo masculino aparentemente saudáveis foram incluídos no estudo. A concentração de HO-1 circulante foi mensurado usando o ensaio de ELISA. As células endoteliais foram incubadas com amostras de plasma de ONI e VEG. Nós observamos que a concentração de HO-1 circulante foi maior nos ONI em relação aos VEG. A incubação das células endoteliais com o plasma de ONI induziu o aumento da expressão gênica/proteica do NRF2 e HO-1, bem como a atividade do ARE e a pr... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Several studies report that a vegetarian diet lowers the probability of developing cardiovascular diseases (CVDs). The endothelial dysfunction is one of the main mechanism that leads to CVDs, associated with decreased nitric oxide (NO) bioavailability and excessive production of reactive oxygen species (ROS). Regarding oxidative/antioxidant stress and biological aging biomarkers, enzymes with antioxidant properties and telomere length may have effects on vascular system modulation. The present study includes two manuscripts related to: 1) regulation of NRF2/HO-1 pathway and 2) telomere length in omnivorous (OMN) and vegetarians (VEG). In the first manuscript, our objectives were to verify circulating HO-1 levels and the effect of plasma incubation from omnivorous and vegetarians in endothelial cells on modulating of NRF2/HO-1 pathway and NO production. From 745 participants initially recruited, 44 omnivorous and 44 vegetarian men apparently healthy were included in this study and circulating HO-1 was measured using ELISA assay. Endothelial cells were incubated plasma samples from OMN and VEG. We found higher circulating HO-1 production in omnivorous compared to vegetarian. Moreover, the plasma collected from omnivorous was able to increase the gene/protein NRF2/HO-1 expression, ARE activity, and NO production in endothelial cells culture compared to vegetarian group. We suggest that HO-1 induction in omnivorous may indicate a pro-oxidative. Activation of the HO-1 / NRF2 pathw... (Complete abstract click electronic access below) / Doutor

Telomeres are the chromosome end structures consisting of telomere-associated proteins and short tandem repeat sequences, TTAGGG, in humans and mice. Telomeres prevent chromosome termini from being recognized as broken DNA ends. The structural integrity of DNA including telomeres is constantly threatened by a variety of DNA damaging agents on a daily basis. To counteract the constant threats from DNA damage, organisms have developed a number of DNA repair pathways to ensure that the integrity of genome remains intact. A number of DNA repair proteins localize to telomeres and contribute to telomere maintenance; however, it is still unclear as to what extent. Telomere shortening has been linked to rare human disorders that present with bone marrow failure including Fanconi anemia (FA). FANCC is one of the most commonly mutated FA genes in FA patients and the FANCC subtype tends to have a relatively early onset of bone marrow failure and hematologic malignancies. Here, we studied the role of Fancc in telomere length regulation in mice. We demonstrated that deletion of Fancc did not affect telomerase activity, telomere length or telomeric end-capping in mice with long telomeres. We also showed that Fancc deficiency accelerates telomere shortening during high turnover of hematopoietic cells and promotes telomere recombination initiated by short telomeres. Telomere shortening has also been linked to human aging and cancer development, with oxidative stress as a major contributing factor. 8-oxo-7, 8-dihydroguanine is among the most common oxidative DNA lesions, and is substrates for OGG1-initiated DNA base excision repair. Mammalian telomeres consist of triple guanine repeats and are subject to oxidative guanine damage. Here, we investigated the impact of oxidative guanine damage and its repair by OGG1 on telomere integrity. We demonstrated that oxidative guanine damage can arise in telomeres where it affects length homeostasis, recombination, DNA replication, and DNA breakage repair. We also examined if telomeric DNA is particularly susceptible to oxidative guanine damage and if telomere specific factors affect the incision of oxidized guanines by OGG1. We showed that the GGG sequence context of telomere repeats and certain telomere configurations may contribute to telomere vulnerability to oxidative DNA damage processing.

Telomeres are the outermost parts of linear chromosomes. They consist of tandemly repeated non-coding short nucleotide sequences (TTAGGG in all vertebrates), in humans spanning over the last 2 to 15 kilobase pairs of the chromosome. Due to the end-replication problem, telomeres shorten with each cellular division. A critically short telomere will trigger the cell to enter a state of cellular senescence or to apoptose. The rate of telomere shortening can be accelerated by factors such as oxidative stress and inflammation. Taken together, this contributed to making telomere length a candidate biomarker of health and aging. Studies have shown that leukocyte telomere length progressively shortens with age, and that it independent of age is associated with age-related morbidity, lifestyle factors, and mortality. This thesis was aimed at exploring the relationships of leukocyte telomere length with various functional and structural attributes of the brain. In Paper I, telomere length was shown to be longer among non-demented carriers of the apolipoprotein E (APOE) ε4 allele, a well-established risk factor for Alzheimer’s disease. However, the rate of telomere shortening was greater among the ε4 carriers, possibly due to the higher levels of oxidative stress and inflammation associated with this allele. Furthermore, performance on episodic memory tests was inversely related to telomere length among ε4 carriers. The results may contribute to a better understanding of the pathophysiology related to the APOE ε4 allele. The volume of the hippocampus, a structure in the brain critical for episodic memory function, was in Paper II found to be inversely related to telomere length among non-demented APOE ε3/ε3 carriers. No correlation between hippocampal volume and telomere length was discernible among ε4 carriers, but they fit the pattern exhibited by the ε3/ε3 carriers as they tended to have smaller hippocampi and longer telomere length compared with the ε3/ε3 carriers. The results are possibly explained by a low proliferative activity among subjects with smaller hippocampi, which might also explain the inverse association between telomere length and episodic memory performance in Paper I. In Paper III, we describe results corroborating earlier findings of shorter telomere length among individuals suffering from depression. Moreover, we found that the shorter telomere length among the patients to a large extent could be linked to a hypocortisolemic state; a state which has been associated with chronic stress. The findings corroborate the link between telomere length and stress, and underline the role of stress in depressive illness. Two prominent manifestations of the aging brain are atrophy and white matter hyperintensities. In Paper IV, we report that white matter hyperintensities and cerebral subcortical atrophy were associated with shorter telomere length in aged non-demented individuals. Cortical atrophy was not associated with telomere length. Inflammation may be the underlying cause of the associations, as it is linked to telomere attrition, subcortical atrophy, and white matter hyperintensities. Taken together, these results show that leukocyte telomere length has the potential of being used as a biomarker for structural and functional attributes of the brain. Furthermore, the findings can provide new insights into mechanisms of disease and aging of the brain

B-cell chronic lymphocytic leukemia (B-CLL) is the most common leukemia among the adult population in western countries and accounts for 30-40% of all leukemias. With survival time ranging from months to decades, the clinical course of individual CLL patients is highly variable. This heterogeneity and in the end the need for means to identify the patients with less favorable disease has encouraged the search for biomarkers that can predict the prognosis. Telomeres are repetitive structures protecting the chromosomal endings and shorten at each cell division. Telomere length (TL) has been indicated as a prognostic factor both in hematological malignancies and solid tumors. In B-CLL, TL is associated with mutation status of the immunoglobulin heavy chain variable (IGHV) gene and with clinical course. In the present thesis the main aim was to evaluate TL as a biomarker in B-CLL using a quantitative PCR-based method for TL determination. In paper I, TL was shown to be a prognostic factor for stage A and stage B/C patients, whereas IGHV mutation status predicted outcome only in stage A patients. Moreover, IGHV mutated CLL cases were subdivided by TL into two groups with different prognosis, a subdivision not seen for unmutated cases. Interestingly, the IGHV-mutated group with short telomeres had en overall survival close to that of the unmutated cases. Thus, a combination of IGHV mutation status and telomere length gave an improved subclassification of CLL identifying previously unrecognized patient groups with different outcomes. TL correlates with cellular origin of B-cell malignancies in relation to the germinal center (GC). In paper II different B-cell lymphoma/leukemia subtypes were analyzed. Shortest telomeres were found in IGHV unmutated CLLs, differing significantly from IGHV mutated cases. Contrary to this, mantle cell lymphomas (MCL) demonstrated similar TL regardless of IGHV mutation status. TL differed significantly between GC-like and non-GC-like diffuse large B-cell lymphomas (DLBCL) and follicular lymphomas (FL) had shorter telomeres than GC-like DLBCL. Hairy cell leukemias, which display Ig gene intraclonal heterogeneity, had longer telomeres than FLs and non-GC-DLBCL, but shorter than GC-DLBCL. In conclusion, TL seemed not to simply correlate with GC origin. Paper III presents a B-CLL cohort assessed for TL, genomic aberrations, IGHV mutation status, CD38 and ZAP-70 expression. An inverse correlation existed between TL and IGHV homology, CD38 and ZAP-70 expression. The presence of genomic aberrations was similar among patients regardless of TL. In contrast, 13q deletion, a favorable biomarker, was more frequent in patients with long telomeres, while 11q and 17p deletions (markers of less favorable outcome) were more frequent in the subgroup with short telomeres. In paper IV a large group of mainly indolent CLL cases from a population based cohort was studied again showing an association between TL and prognosis, especially in “good” prognosis cases as defined by other biomarkers. Multivariate analysis indicated a strong connection between IGHV mutation status, lipoprotein lipase (LPL) expression and TL. A comparison of TL in diagnostic and follow up samples demonstrated a significant correlation, and also in the follow samples TL constituted a significant biomarker for survival.

G-quadruplex structures are found in important regions of the eukaryotic genome, such as telomeres and regulatory sequences of genes, and are likely to play important roles in regulation of biological events. The significant structural differences with duplex DNA make quadruplex DNA a very attractive target for anticancer drug design. The purpose of this study is to explore conformational space in a series of heterocyclic cations to discover novel structural motifs that can selectively bind and stabilize specific G-quadruplex arrangements. A variety of biophysical techniques such as thermal melting experiments, biosensor surface plasmon resonance, circular dichroism, fluorescence displacement assay and mass spectrometry were employed to evaluate the affinity of the compounds and their recognition properties. The screening of the molecules allowed the identification of not only selective G-quadruplex ligands but also potential quadruplex groove binders. These results can be useful for the development of new efficient telomerase inhibitors which are endowed with pharmacological activity.

G-quadruplex forming human telomere sequence (HTS) DNA, has been widely studied due to the telomere's implied role in biological processes, including cellular ageing and cancer physiology. The goal of these previous efforts has been to characterize the physiologically relevant structures and their stability and dynamics in order to develop therapeutic applications. Unfortunately, understanding the biologically relevant form of the human telomere DNA is complicated by the fact that HTS-derived sequences are highly polymorphic. To further complicate the issue, recent investigations have demonstrated the ability of "cell-like" co-solvents to alter the preferred G-quadruplex fold of HTS DNA. However, the origins of G-quadruplex structure selection, the relative contributions of crowding versus dehydration, and the possible effects of co-solvents on kinetically determined folding pathways remain unresolved. Towards answering these questions, I investigated HTS DNA G-quadruplex in extreme anhydrous and high viscosity conditions utilizing a deep eutectic solvent (DES) consisting of choline chloride and urea. Herein I report that the water-free DES supports an extremely stable parallel stranded structure, consistent with observations that diminished water activity is the main cause of structural transitions to the "parallel-propeller" form. Furthermore, my research shows that the highly viscous nature of the solvent enables significant diffusion based control over HTS g-quadruplex folding rates and topology, fully consistent with Kramers rate theory. To the best of my knowledge, this is the first example of the kinetic exploration of G-quadruplex folding utilizing high friction solvent; the results of which display a decreased intramolecular folding rate of HTS DNA to a never before encountered time scale on the order of days at physiological temperature. Moreover, I have demonstrated that the folding pathway of a G-quadruplex can be altered with increased solvent friction. These discoveries are important because they highlight the need to consider the viscosity when exploring the dynamics of human telomeres specifically drug binding and folding of G-quadruplexes in vivo where cellular viscosity has been reported to be as high as 140cP. Lastly, it appears that tuning solvent viscosity could prove useful to the continued study of G-quadruplex dynamics.

Telomeres are fundamental structures found at the end of all eukaryotic chromosomes that function to protect the end of chromosomes from end-to-end fusion, erosion and subsequent telomere dysfunction. Telomerase and alternative lengthening of telomere (ALT) mechanisms maintain the telomeres by compensating natural telomeric loss. ALT is found to be present in 15% of human tumours lines and it may be expressed at low levels in the normal mouse tissues. However, the exact mechanism behind ALT depression and/or activation in the mammalian cells is not fully understood. Previous studies have highlighted the role of BRCA1 in telomere dysfunction. Also, it has recently been shown that BRCA1 co-localises at telomeres in the ALT + human cells through BLM and Rad50. However, it is still unclear whether BRCA1 plays a direct role on telomere length maintenance and integrity. The aim of this project was to examine the role of BRCA1 in telomere maintenance associate with ALT in BRCA1 defective mammalian cells. Therefore to achieve this, we have set up series of experiments to look at, (a) hallmarks of ALT activity at the cytological level, (b) measuring of ALT activity using biochemical and immunocytochemistry techniques and (c) understanding the role of BRCA1 in DNA damage response mechanism and telomere dysfunction. Firstly, we found elevated levels of recombination at telomeres in the two human BRCA1 carrier cell lines and mouse embryonic stem cell with deficiency in Brca1-/-. Secondly, our data showed that human and mouse BRCA1 defective cells are significantly more sensitive to ionizing radiation in line with the DNA repair function of BRCA1. Moreover, we found persistent DNA damage at telomeres in the BRCA1 defective environment when after exposure of cells to ionizing radiation. Thirdly, we found evidence of ALT activity in some mouse cell lines, and elevated ALT in mouse cells defective in Brca1. Finally, we examined some other ALT markers using immunofluorescence. Our data indicate differences between human and mouse cells in regulating ALT. Taken together data presented in this thesis revealed that (i) BRCA1 plays a major role in telomere maintenance and defective BRCA1 mammalian cells show evidence of telomere dysfunction and telomere length shortening in line with previous publish data, (ii) BRCA1 defective mouse cells have elevated levels of ALT, (iii) the mouse lymphoblastoid LY-S cells have complete absence of ALT.

A injúria renal aguda representa um estado de senescência precoce induzida por estresse, e as células-tronco mesenquimais podem ser uma alternativa para seu tratamento. Células-tronco jovens reduzem o fenótipo de envelhecimento em rins quando comparadas a células idosas. O objetivo deste estudo foi avaliar se o tratamento com jovens células-tronco mesenquimais derivadas de cordão umbilical humano podem interferir na senescência renal induzida por lesão de isquemia-reperfusão em ratos. Ratos machos foram submetidos ao modelo de isquemia de artérias renais bilateralmente por 45 minutos, com reperfusão após, e alguns animais receberam 1 X 106 células por via intraperitoneal após 6 horas da indução da lesão. Os animais foram eutanasiados no segundo ou no sétimo dia pós-isquêmico. No segundo dia após a lesão de isquemia-reperfusão, o tratamento com as células melhorou a filtração glomerular e a função tubular, melhorou a expressão renal de aquaporina-2 e reduziu a infiltração de macrófagos nos rins. Proteínas relacionadas à senescência (-galactosidase, p21, p16 e fator de transformação do crescimento ) e microRNAs (mir-29a e miR-34a) estiveram com a expressão aumentada após a isquemia-reperfusão, e houve redução nesses parâmetros com o tratamento. A redução na expressão de Klotho e o estado pró-oxidativo gerados pela isquemia-reperfusão também foram revertidos pelo tratamento. A senescência induzida pela injúria renal aguda é um processo independente de telômeros. Ao sétimo dia pós-lesão, os ratos isquêmicos mantinham defeito de concentração urinária, que foi revertido nos animais tratados. Além disso, o tratamento reduziu o índice de necrose tubular aguda em tecido renal e reduziu o infiltrado macrofágico túbulo-intersticial. O marcador pró-senescência p16 foi completamente restabelecido nos animais tratados. Nossos dados demonstram que o tratamento com jovens células-tronco mesenquimais derivadas de cordão umbilical humano atenua a resposta inflamatória e de estresse oxidativo que ocorre na injúria renal aguda, e reduz a expressão de proteínas e microRNAs relacionados à senescência. Nossos achados expandem as perspecivas para o tratamento da injúria renal aguda / Acute kidney injury represents a status of premature stress-induced senescence, and mesenchymal stem cells are an alternative for treatment. Young stem cells reduce aging phenotype in kidneys when compared to old cells. The objective of this study was to evaluate if treatment with young human umbilical cord mesenchymal stem cells could interfere in kidney senescence induced by renal ischemia-reperfusion in rats. Male rats were induced to ischemia-reperfusion injury by 45-minutes clamping of both renal arteries; some rats received 1X106 cells intraperitonally six hours later. Rats were euthanatized on post-renal ischemia reperfusion days two and seven. At day 2 after ischemia-reperfusion injury, treatment with cells improved glomerular filtration, tubular function, improved renal expression of aquaporin 2 and decreased macrophage kidney infiltration. Senescence-related proteins (?-galactosidase, p21, p16 and transforming growth factor ?) and microRNAs (miR-29a and miRNA-34a) were overexpressed after ischemia-reperfusion, and reversed by the treatment. The Klotho reduced expression and the pro-oxidative status induced by ischemia-reperfusion were reversed by the treatment. Senescence induced acute kidney injury is a telomere-independent process. At day 7, ischemic rats maintained urinary concentrating defect, which is reversed in treated animals. Moreover, treatment decreased the index of acute tubular necrosis in kidney tissue and decreased macrophage kidney infiltration. Senescence marker p16 was completely restored in treated animals. Our data demonstrate that young human umbilical mesenchymal stem cells treatment attenuates the inflammatory and oxidative stress response occurring in acute kidney injury, and reduces the protein and microRNA expression related to senescence. Our findings broaden the perspectives for the treatment of AKI

Abstract Physical inactivity is an increasing health problem worldwide and one of the most important risk factors for global premature mortality. Along with gained years, it is highly essential for both individuals and society that elderly persons perceive their health as good and are able to cope well with everyday life and enjoy it. Hence, our aim was to investigate the long-term associations of midlife physical activity (PA) with health-related quality of life (HRQL), frailty stage, leukocyte telomere length (LTL), a potential indicator of biological ageing, and mortality. The Helsinki Businessmen Study cohort has been followed since the 1960s and it has been studied from several perspectives over the years. In the baseline examinations in 1974, 782 healthy men (mean age 48) completed a questionnaire about their PA pattern. These men form the population of this study. According to their global description of their PA pattern, the men were categorized into low, moderate and high PA groups. After a 26-year follow-up in 2000, the HRQL and disease prevalences were appraised in 552 (mean age 73) men using a postal questionnaire including the RAND-36 instrument. PA was significantly associated with better physical function, one of the eight domains of HRQL. From the same questionnaire, frailty stage was appraised for 514 men using modified Fried’s criteria: weight loss, physical inactivity, low vitality and physical weakness. After adjusting for CVD risk factors at baseline, the risk for frailty was 80% lower in the high PA group compared with the low PA group. After a 29-year follow-up in 2003, 204 randomly selected survivors were invited to laboratory tests. The mean LTL was longer in the moderate PA group than in the low PA group and, contrary to our hypothesis, in the high PA group. We found that midlife PA was related to mortality during a 34-year follow-up and the protective effect of PA was independent of the cardiovascular disease (CVD) risk factors, including body mass index, age, cholesterol, glucose, systolic blood pressure and smoking at baseline. In conclusion, the overall results of this study lend support to the view that those who have adopted a physically active lifestyle in middle age have better physical function and lower risk of frailty in old age and lower mortality. / Tiivistelmä Riittämätön fyysinen aktiivisuus on kasvava terveysongelma ja ennenaikaisen kuoleman riskitekijä kaikkialla maailmassa. Elinvuosien lisääntymisen ohella on tärkeää, että ikääntyvä ihminen kokee terveytensä hyväksi ja että hän nauttii jokapäiväisestä elämästään. Tarkoituksenamme oli tutkia keski-iän liikunnan yhteyksiä terveyteen liittyvään elämänlaatuun, hauraus-raihnaus -oireyhtymään, telomeerien pituuteen sekä kuolleisuuteen 1960-luvulta lähtien seuratussa Helsingin Johtajat -kohortissa. Vuonna 1974, Helsingin Johtajat -tutkimuksen alkuvaiheessa, 782 tervettä miestä (keski-ikä 48 vuotta) täytti liikuntatottumuksiaan koskevan kyselylomakkeen. He muodostavat tämän tutkimuksen aineiston. Liikuntaharrastuksestaan antamansa yleiskuvauksen mukaan miehet jaettiin matalan, keskitason ja korkean liikuntatason ryhmiin. Vuonna 2000 terveyteen liittyvä elämänlaatu ja sairauksien esiintyvyys määritettiin 26 vuoden seuranta-ajan jälkeen 552 mieheltä (keski-ikä 73 vuotta) käyttäen postitettua kyselylomaketta, joka sisälsi RAND-36 -instrumentin. Liikunta oli tilastollisesti merkitsevästi yhteydessä parempaan fyysiseen toimintakykyyn, yhteen kahdeksasta terveyteen liittyvän elämänlaadun ulottuvuudesta. Saman kyselylomakkeen vastauksista selvitettiin hauraus-raihnaus -oireyhtymän esiintyvyys käyttäen seuraavia, Friedin kriteereistä muunneltuja kriteerejä: tahaton painon lasku, vähäinen liikunta, tarmokkuuden puute ja fyysinen heikkous. Riski hauraus-raihnaus -oireyhtymään oli 80 % pienempi korkean liikuntatason ryhmässä kuin matalan liikuntatason ryhmässä. Vuonna 2003, 29 vuoden seurannan jälkeen, 204 satunnaisesti valittua elossa olevaa miestä pyydettiin laboratoriotesteihin. Keskimääräinen leukosyyttien telomeerien pituus oli suurempi keskitasoisen liikunnan ryhmässä matalan ja korkean liikuntatason ryhmiin verrattuna. Tutkimuksessamme keski-iän liikuntaharrastus oli yhteydessä kuolleisuuteen 34 vuoden seurannan aikana, ja liikunnan suojaava vaikutus oli havaittavissa lähtövaiheen sydän- ja verisuonitautien riskitekijöistä (painoindeksi, ikä, kolesteroli, glukoosi, systolinen verenpaine ja tupakointi) riippumattomasti. Tämän tutkimuksen tulokset tukevat ajatusta, että fyysisesti aktiivinen elämäntapa keski-iässä on yhteydessä parempaan fyysiseen toimintakykyyn ja vähäisempään hauraus-raihnaus -oireyhtymään vanhalla iällä ja vähentää kuolleisuutta.