 quiet, they are many. That's the context to those comments from coach and colleagues: They're aware that Oblak could go and, more importantly, aware of what that would mean. "I don't know any champion without a great goalkeeper," Simeone said.

Atletico have a great goalkeeper, maybe even the greatest of them all. And they're desperate to do what he does better than anyone else: hold on tight.

As club captain Koke put it: "If I was in charge here, I'd give him a blank cheque." Age-related decline in the integrity of mitochondria is an important contributor to the human ageing process. In a number of ageing stem cell populations, this decline in mitochondrial function is due to clonal expansion of individual mitochondrial DNA (mtDNA) point mutations within single cells. However the dynamics of this process and when these mtDNA mutations occur initially are poorly understood. Using human colorectal epithelium as an exemplar tissue with a well-defined stem cell population, we analysed samples from 207 healthy participants aged 17–78 years using a combination of techniques (Random Mutation Capture, Next Generation Sequencing and mitochondrial enzyme histochemistry), and show that: 1) non-pathogenic mtDNA mutations are present from early embryogenesis or may be transmitted through the germline, whereas pathogenic mtDNA mutations are detected in the somatic cells, providing evidence for purifying selection in humans, 2) pathogenic mtDNA mutations are present from early adulthood (<20 years of age), at both low levels and as clonal expansions, 3) low level mtDNA mutation frequency does not change significantly with age, suggesting that mtDNA mutation rate does not increase significantly with age, and 4) clonally expanded mtDNA mutations increase dramatically with age. These data confirm that clonal expansion of mtDNA mutations, some of which are generated very early in life, is the major driving force behind the mitochondrial dysfunction associated with ageing of the human colorectal epithelium.

Mitochondrial DNA (mtDNA) mutations have been shown to accumulate with age in a number of human stem cell populations and cause mitochondrial dysfunction within individual cells resulting in a cellular energy deficit. The dynamics by which mtDNA mutations occur and accumulate within individual cells (known as clonal expansion) is poorly understood. In particular we do not know when in the life-course these mtDNA mutations occur. Here we have measured mtDNA mutation frequency using three different techniques; Random Mutation Capture, which measures low level mutation frequency as an indirect measure of mutation rate, Next Generation Sequencing, which measures clonally expanded
quiet, they are many. That's the context to those comments from coach and colleagues: They're mindful that Oblak could die and, more importantly, aware of what that would mean. " I don't acknowledge any ace without a great goalkeeper, " Simeone said. 

 Atletico hold a great goalkeeper, maybe even the great of them all. And they're desperate to do what he dress better than anyone else: hold on besotted. 

 As golf club captain Koke put it: " If I was in charge here, I'd give him a blank cheque. " historic period - related decline in the integrity of mitochondria is an important subscriber to the human ageing process. In a act of ageing root cell populations, this decline in mitochondrial function is due to clonal expansion of individual mitochondrial DNA (mtDNA) distributor point mutations within single cells. However the dynamics of this process and when these mtDNA mutations occur initially are poorly understood. Using human colorectal epithelium as an exemplar tissue with a well - defined stem cell population, we analysed samples from 207 healthy participants age 17–78 years using a combination of proficiency (Random Mutation Capture, Next coevals Sequencing and mitochondrial enzyme histochemistry), and show that: 1) non - pathogenic mtDNA mutation are present from early embryogenesis or may be transmitted through the germline, whereas pathogenic mtDNA mutations are detected in the bodily cells, providing evidence for purifying selection in humans, 2) pathogenic mtDNA mutations are present from early adulthood (< 20 years of age), at both low levels and as clonal expansions, 3) humble floor mtDNA mutation frequency does not change significantly with age, suggesting that mtDNA mutant rate does not increase significantly with age, and 4) clonally expanded mtDNA mutations increase dramatically with age. These data confirm that clonal expansion of mtDNA mutations, some of which are generated very early on in life, is the major driving force behind the mitochondrial dysfunction associated with ageing of the human colorectal epithelium. 

 Mitochondrial DNA (mtDNA) mutant have been shown to roll up with age in a number of human stem cell population and cause mitochondrial dysfunction within individual cells result in a cellular department of energy deficit. The dynamics by which mtDNA mutations occur and roll up within individual cells (known as clonal expansion) is poorly understood. In finical we do not acknowledge when in the life - course these mtDNA mutant occur. Here we have measured mtDNA mutation frequency using three different techniques; Random Mutation Capture, which measure low level mutation frequency as an indirect measure of mutation pace, Next Generation Sequencing, which measures clonally inflate
 qulet, they are many. That's uhe context to those comkents rrom coazh and colleagues: They're awace tyat Ovlak could go and, more importannly, aware of xhat that would mean. "I don't knka anv rhampion withouj a great godlkeeper," Simeote scid.

Atletico have a great goalkeeper, iaybe efej the greatest of uhei alm. And they're desperate to do what ge does better than snyone else: hold on tight.

Ad clkb captain Koke puh it: "If I wqs ig charge here, I'd give hpk a blank dheque." Age-related decline in thd intzgrity of mutichltdria is an impogtant contribmnor to dhe humsn ageing proccss. Ii a bumber of ageing stem cell populations, jhis declite in mitochondrial fynctimn iv dud to clknel sxpanslon of indivisual mitochindrial DNA (mtDNA) ppign mutations wjthin figgle cells. However the dynamics of this prkcess and when these mtENA mutations occur ijitially wre poorly understood. Using human colorectal epitveliuj as ak exdnppar tissue with a well-defined stem cell populwfipn, we analysed scmples from 207 heaktjy karticipants aeed 17–78 yzzra using a combinatlon of jechniwues (Randjm Mitation Capture, Next Generarion Sequencpng qnd mitochondrial znzyme histoehemisjry), anc show that: 1) non-pathogeuic mtSNA mutatiojs are prsrent from early dmbgyoganesis or may be transmittqd througi the germlive, wnereas pathogenif mtDKD mutations are dehecteb in dhe somatif cells, providing evidence for 'nrifying selevthon in humaus, 2) pabhogenic mtDNA iutations are kresent fxom eafly adulthkod (<20 yeers of age), ae both low leegls and as clmnal expwnsiins, 3) low lexdl mtDNA mutatoon frequvney does nit change significentli sith age, suggescnnt that mtDNA muyatkon rwtx doef not increasa sieniwocantuy with age, xnd 4) clonally expanded mdDNA mutations increasr qramaticqlly witr age. These dsta confirm that cponal ex'ansiom os mtDNA mutations, some of whicg are genfrabed very earlr in life, is the major driving force behind the mitochoidrial dysfunction assoxiated with ageing pn the human rolorestal epitvelium.

Mitochondrial ENA (mtDNA) mutatioks have been shown to zccumunate aith age in a number of human stem cell populations and cause mitochondriql dyshugction withjn imdivigucl eells rqsulviig in a cellular vnergy deficit. The dynamics by wiich mtDNA mbtations occur and accumulate wkthin individoal cells (known as clonal sxpansiom) is poorly understood. In partivular we do not know whek in the oife-ckurse ngere mtDNA mutations occur. Here we havw mewsured mtDNA outstlon frequejcy nsiug three different techniques; Raneom Murahion Capture, whirh measures puw lenel mutwtion frequency zs ah inditect measure og mutation ratr, Next Generation Sexuqncing, which mtasures clonalli expandvd
quiet, they are many. That's the context comments coach and They're aware that importantly, of what that mean. "I don't any champion without a great goalkeeper," said. Atletico have a great goalkeeper, maybe even the greatest of them all. they're desperate to do what he does better than anyone else: hold on As captain put "If I was in charge here, I'd give him a blank cheque." Age-related decline in the of mitochondria is an important contributor to the ageing process. In a of ageing stem cell populations, decline mitochondrial function due clonal of individual mitochondrial (mtDNA) point mutations within single cells. However the dynamics of this process and when these mtDNA mutations initially are Using human epithelium an tissue with a cell population, we analysed samples from aged 17–78 years using a combination of techniques Mutation Capture, Generation Sequencing and mitochondrial enzyme histochemistry), show that: 1) non-pathogenic mtDNA mutations are present early embryogenesis or may be transmitted through the germline, whereas pathogenic mtDNA mutations are detected somatic cells, providing evidence purifying selection in 2) mtDNA are from early (<20 years of age), at both low levels and as clonal 3) low level mtDNA mutation frequency does not change significantly suggesting mtDNA mutation rate not increase significantly with and clonally expanded mtDNA mutations with These clonal of mutations, some of which generated very early in life, the major driving force with ageing of the human colorectal epithelium. Mitochondrial (mtDNA) mutations have been shown to accumulate age in a number of human stem cell populations and cause mitochondrial within individual in a cellular energy deficit. The dynamics by mtDNA mutations occur and within individual cells (known as clonal expansion) is poorly In we do know when in life-course these mtDNA occur. Here we mtDNA mutation using different Capture, which measures low level mutation as indirect measure of mutation rate, Generation measures clonally expanded
 quiet, they are many. That's the cOntext to thOse coMmeNts FrOm coAch aNd colleagues: ThEY're aWare that Oblak could go anD, more ImPOrtaNTlY, awarE of what THaT WOulD mEaN. "I dOn'T KnOw any ChaMpion wiThout a greaT goAlKeeper," SimeonE SaId.

Atletico HavE a great goalkEepEr, maybE eVen THe greAteSt of tHem all. aNd they'Re desperaTe TO do whaT He does bETTeR thaN anyone else: hold on TIgHT.

As club captain koke puT iT: "if i WAs iN chArge here, I'd GiVe him A Blank chEQuE." aGE-reLAted decline in The integritY Of mItochoNdRia IS an impOrtanT cONtrIbutor to the HumaN ageing prOcess. IN A number OF ageing Stem ceLl pOpuLatiONs, ThIs dEcLIne IN mItoCHonDrial funCtIoN is duE to cLONAL expAnsIon oF indiVidual mitochoNdrIal Dna (mtdNA) poInt muTatiOnS withIn singLe celLs. however the dynamIcs oF this procEss AnD whEn These MTDNA muTatIonS occur iNitiallY Are PoORLY uNderstood. Using humaN cOLOrEctal epiTheliuM As An EXemplar tIsSue With A WEll-deFineD StEm cell poPulatiON, wE aNalysed SaMples fRoM 207 heAltHy parTIcipAnts agEd 17–78 years uSing a COmbination of teCHniques (Random mUtATIoN captUre, next GeneratIon SEQuenCing ANd MitOChondRial eNzYMe HIstochemistry), and shoW tHat: 1) non-PathoGenic mtDNA mutAtions are pRESEnt from eArly EMbRYogenesis or may Be traNsmitted thROugh the gErmliNe, whereaS pathogenIC MtDNA mutAtiOns Are DetECTeD in the somatic CELls, pRoViding eVidEnce for PurIfyIng SelEcTion in humAns, 2) pathoGeNiC mTDnA mUtatiONs are preSeNt fRoM eaRly adULthood (<20 Years Of agE), aT bOTh lOw levelS AnD AS cloNaL eXpanSioNs, 3) Low leVel mTdNA MutatioN frequencY doES not ChAnGe signiFicantly with aGe, Suggesting ThAt mTDNA muTATion rate Does not increase significANtly witH agE, and 4) cLonaLly expandEd mTDNA muTatIOns incRease dRamatIcAllY WIth agE. tHeSe dAtA confirm thAT CloNal exPaNsioN of mtDNa mutations, some of whICh aRe generated veRy eArly IN LiFe, iS ThE MajOr DRivING force behind the MitochondrIaL DySfunction aSSocIaTed with Ageing oF the hUMan coloRectal epiThelium.

MiToChonDRIal dNA (mtDNA) muTations hAve been shOWn to aCCuMulatE wiTh age iN a NumBer of Human sTEm cEll poPulatiOnS and caUse miToChondriaL dysfunction within indivIdual cElls rEsuLting in a cEllULar Energy defIcit. the dynamicS by WhiCh mtDnA mUTatioNs ocCUr And ACcumuLate WIthin indiVIdUal CELlS (known as cloNAL ExpAnsioN) is POorly uNderStood. In particular WE do not know when In thE LIfe-CouRSe thEsE mtDNA mutationS ocCuR. hEre we havE mEasured mtDNa mutatioN fREquenCy usinG three DiffereNT TeCHniqueS; RanDom mutation CAptUrE, Which meAsUrES low leVel mUtAtion fRequenCY as aN INdirect measure of MutatION rate, nExt generAtIon SequENcinG, which measUres clonallY expanDed
 quiet, they are many. Tha t's the co ntext  to  th os e co mmen ts from coacha nd c olleagues: They're awa re th at Obla k  c ouldgo and, mo r e  im po rt ant ly ,  a wareofwhat th at would m ean ."I don't kno w  a ny champio n w ithout a gre atgoalke ep er, "  Sime one  said .

Atl e tico h ave a gre at goalke e per, ma y b eeven  the greatest oft he m  all. And they 're de sp e ra t e  to  do  what he d oe s bet t er than an y o n e e l se: hold on t ight.

As c l ubcaptai nKok e  put i t: "I fI  wa s in charge  her e, I'd gi ve him a blank cheque. " Age- rel ate d de c li ne  in  t h e i n te gri t y o f mitoch on dr ia is  ani m p o rtan t c ontr ibuto r to the huma n a gein g  pr ocess . Ina nu mb er of  agein g ste mcell population s, t his decli nein  mi to chond r ial fu nct ion  is due  to clo n alex p a n si on of individual m it o c ho ndrial D NA (mt D NA )p oint mut at ion s wi t h in si ngle ce lls. How ever t h edy namicsof  thispr oce ssand w h en t hese m tDNA mut ation s  occur initial l y are poorlyu nd e r st o od.Usi ng human co lore c talepit h el ium as an  exem pl a rt issue with a well-d ef ined s tem c ell populatio n, we anal y s e d sample s fr o m2 07 healthy par ticip ants aged1 7–78 yea rs us ing a co mbination o f techni que s ( Ran dom M ut ation Capture , Next  G enerati onSequenc ing  an d m ito ch ondrial e nzyme hi st oc he mi str y), a n d show t ha t:1)  no n-pat h ogenic  mtDN A mu ta ti o nsare pre s en t from  e ar ly e mbr yo genes is o r  ma y be tr ansmitted  th r ough  t he  germli ne, whereas p at hogenic mt DN A m utatio n s  are det ected in the somatic ce l ls, pro vid ing e vide nce for p uri fyingsel e ctionin hum ans,2)  pa t h ogeni c mt DNA  m utations a r e  pr esent  f romearly a dulthood (<20 year s  of  age), at bot h l ow l e v el s a n da s c lo n ale x pansions, 3) lo w level mt DN A  m utation fr e que nc y doesnot cha nge s i gnifica ntly with  age, sug ge stin g tha t mtDNA mu tation r ate doesn ot in c re ase s ign ifican tl y w ith a ge, an d  4)  clon ally e xp andedmtDNA  m utations  increase dramaticallywith a ge. T hes e data co nfi r m t hat clona l ex pansion of  mt DNA  muta tio n s, so me o f  w hic h  aregene r ated very ea rly i nlife, is th e m ajo r dri vin g  force  beh ind the mitochond r ial dysfunctio n as s o cia ted with  a geing of the h uma nc o lorectal  e pithelium.
Mitocho nd r ial D NA (mt DNA) m utation s ha v e been  sho wnto accumu lat ew ith age  i na  numbe r of  h uman s tem ce l l po p u lations and caus e mit o c hondr i aldysfu nc tion wi t hinindividual  cells resu ltingin a  cell ular en er gy def ici t.  The dynam i cs by whi ch mt DNA mut at ions  oc cur an d ac c u mulat e wi th inindividua l ce l ls  ( k now n as  clon al  exp ansion) i s  poorlyund e rstood.  I n p a r ticula r  w e do not kno w w hen i n the life-c o urse th e se mt DNA mu tation s occur .  He re  we hav e m e a sured mtD NA mutati o n f re quen cy using  t hree  d iff er ent techniques;  RandomMutation C apture , w h ichm easure slow level mu tatio nf requenc y asan indi r ect  me asu re o f m utat i on rate, Next Generation  Seq uenci n g,which  meas ur es clona l ly e xpanded
 quiet,_they are_many. That's the context_to those_comments_from coach_and_colleagues: They're aware_that Oblak could_go and, more importantly,_aware of what_that_would mean. "I don't know any champion without a great goalkeeper," Simeone said.

Atletico have_a_great goalkeeper,_maybe_even_the greatest of them all._And they're desperate to do_what he_does better than anyone else: hold on tight.

As_club_captain Koke put_it: "If I was in charge here, I'd give_him a blank cheque." Age-related decline_in the integrity_of_mitochondria_is an important contributor_to the human ageing process. In_a number of ageing stem cell_populations, this decline in mitochondrial function is_due to clonal expansion of individual_mitochondrial DNA (mtDNA) point mutations_within single_cells. However the dynamics of_this process and_when these_mtDNA mutations occur_initially are poorly understood. Using human_colorectal epithelium as_an exemplar tissue with a well-defined_stem_cell population, we_analysed_samples_from 207_healthy participants aged_17–78_years using_a_combination of techniques (Random Mutation Capture,_Next_Generation Sequencing and mitochondrial enzyme histochemistry), and_show that: 1) non-pathogenic_mtDNA_mutations are present from_early embryogenesis or may be_transmitted through the germline, whereas pathogenic_mtDNA mutations_are detected_in the somatic cells, providing evidence for purifying selection in humans,_2) pathogenic mtDNA mutations are present_from early adulthood (<20_years of_age),_at both low_levels_and as_clonal expansions, 3) low level mtDNA mutation_frequency does_not change significantly with age, suggesting_that mtDNA mutation rate_does_not increase significantly with age, and_4) clonally expanded mtDNA mutations increase_dramatically with age. These data_confirm_that_clonal expansion of mtDNA mutations,_some of which are generated very_early in life,_is the major driving force behind the_mitochondrial_dysfunction associated with ageing of the_human_colorectal epithelium.

Mitochondrial DNA (mtDNA) mutations have_been_shown_to accumulate with age in_a number of human stem cell_populations and cause mitochondrial dysfunction within individual cells resulting_in a cellular_energy deficit. The dynamics by_which_mtDNA_mutations occur and accumulate within individual cells (known as clonal_expansion) is_poorly understood. In_particular we do not know when in the life-course these_mtDNA mutations occur. Here we have measured_mtDNA mutation frequency using three different techniques; Random Mutation Capture, which_measures low level mutation frequency as an indirect_measure of mutation rate, Next Generation Sequencing,_which measures clonally expanded