Clinical Clin Inveslia (19941 72.920-936 Investigator Molecular Medicine Sprineet- Velaf Determination of DNA single-strand breaks in lymphocytes of smokers and nonsmokers exposed to environmental tobacco smoke using the nick translation assay Scherer: . AR. Tricker:. Held' , Krause Warnck: Holz Meibner . M. Einhaus Adlkofer , H W Rudiger' Mcdizin [V Cnivetsilae Wien Winrineer Guri: 18-J0 A-1090 Ver Austnin Abicilun? Arbzitsmedizin Klnik Ff Jnnene Oermznt 20. D-8ojjo Munenen Adikolet. Loclme5l255. Fors-Junzsizoo: Prol D: Analyuisch-blolozischEs 19 June Aczeple : Pecelle - and cotinine Ievels cells COHb. Plasma nicoting DNA   single-strand The detection Abstraci; considerabl; increased in both smokers Jnd mononuciczted #hite blood Wen belskMSSBCT Usummat modificucversion of {he icc ETS on  days anc nonsmokers exposed cellslaioB Cssu5in3 predtecd DNA-SSB were detected in all volunieers #th This allows 35571 translauon 3552} interindividual daj t0 0z, an morning and sensitive examination of SSB using only and 5apid After smoking: SSB increased eightfold determina eering Vatiations: heparinized blood for an smokers; in nonsmokers standardized by incubation of on da; Jnc on d; Tne 3552! ton exposed {0 ETS no exposure-rellled variation in ~methyl-A" nitro-N-nitro MBWC in vitro with We concluce tnut th: SSB levels was (ound Modi- known genotoxic agent; soguanidine (MNNGL sensitite enouzh [0 In`vitro ircubation of MWBC with MNIG Iied nick Ianslation 4552} Dy' exposure detect   SSB induced "Fo dependen: increase DNA-SSB 2t duced dosc genotoxic agcn: and could therefore b: uscd In DiO- and S00uM MNNG. The delec doses berwcen monitoring J1 th? workpiace. LM MNNG_ logica tior Iimit for the as52 "a5 SSB in sess Ihe sumtabili:y of this t0 detect 74552# DNA [- DNA single-strand breaks which Key controlled stud; was performed vords: Uvo Environmeil- Smoking Nick: trinsiation voluntee: smokers (=31 nonsmokers PWIT Biological monitoring tobacco smoke (ETSI and tohacco smoke I0 environmenta posed The nonsmoker controls ("= compared Wete and experimental days conitol stuc; lasted and control days (days exposure days functions DNA in living Owing t0 ils physiological unventilzted and nonsmokers sal smokers highly reucuive 3nd unstable molecui: cells dars (days room ior On Ine exposute 4m DNA is modilied at a rate cstimated t0 be 10` bases smoked cach of   the Mive smorers and Per cell per da}. cven without exogcnous damage h while the fiva nonsmokers Verc ciposete{O the ETSheneraed &, noz smoking voi: [26]: For the deieclion ol single strand breaks ISSB exposcc sevcral techniques havc becn developed Fucn t0 tobucco smoke Ws con unteets; High exposure 114] sedi- conventional alkaline sucrose Erdicoi ICO- carboxyhemoglobin lrmed by' dosimetry alkalinc elution [I- mentation anulysis [19] Hbl; plasma nicoting and cotininc levels Blood #as Jnc 13]. The use of thc nick Iransiation (NT) [0 deicct drawn bcfore and alcr cach cxposure SSB in DNA was lirst described in /983 in permc d4ys for delermination o DNA-SSB in perimenia abilizcd humun (ihroblasis alcr cxposuirc t0 ivmphocytes immedialely afcr isolation of blood (MxxGi N-nitrosogunnidine methyl-] niro h; > hecn demonsirled [ult DMEM Dul. (20]. In tnc mcantimc curhorhemoglohin: cohb AhnerCinUS high snecilicity and sensitivity modi-ution Exupte < thc NT nus Cni Mcumn ; bcecu > Minimm Iah gcnolaie Canounus Untfuinninij Tuthacc SS8 ETS widc rangc of chcmicl sinti- einc oreak YYYG L-micina #hel(481) _IL;4I1ILIIIL . ~mula ~nint 16. W|ad to herut /// Morcovcr; Ihis :ssJ} I0 inonaiuic|e;ted u #le hhonl eeik17 Miac" t ~ ilso bc used I0 sludy thc kinctics o DNA rcplir 1jlof [67. 28. 291 Rudiear Crer '4nu unzilt 'c 4 7
Clin Inveslia (19941 72.920-936 Clinical Investigator Molecular Medicine Sprineet- Velaf Determination of DNA single-strand breaks in lymphocytes of smokers and nonsmokers exposed to environmental tobacco smoke using the nick translation assay Meibner . Krause Warnck: Held' , Scherer: . AR. Tricker:. M. Einhaus Holz Adlkofer , H W Rudiger' Mcdizin [V Cnivetsilae Wien Winrineer Guri: 18-J0 A-1090 Ver Austnin Abicilun? Arbzitsmedizin Klnik Ff Jnnene Adikolet. Loclme5l255. 20. D-8ojjo Munenen Oermznt Analyuisch-blolozischEs Fors-Junzsizoo: Prol D: Aczeple : 19 June Pecelle - cells COHb. Plasma nicoting and cotinine Ievels Wen considerabl; increased in both smokers Jnd nonsmokers exposed ETS on  days anc DNA-SSB were detected in all volunieers #th and interindividual daj t0 0z, an morning eering Vatiations: After smoking: SSB increased on da; Jnc on d; smokers; in nonsmokers exposed {0 ETS no exposure-rellled variation in SSB levels was (ound We concluce tnut th: Modi- Iied nick Ianslation 4552} sensitite enouzh [0 detect   SSB induced "Fo Dy' exposure genotoxic agcn: and could therefore b: uscd In DiO- logica monitoring J1 th? workpiace. Abstraci; The detection DNA   single-strand mononuciczted #hite blood belskMSSBCT Usummat modificucversion of {he icc cellslaioB Cssu5in3 predtecd translauon 3552} This 35571 allows and sensitive examination of SSB using only 5apid heparinized blood for an eightfold determina ton Tne 3552! standardized by incubation of MBWC in vitro with ~methyl-A" nitro-N-nitro soguanidine (MNNGL known genotoxic agent; In`vitro ircubation of MWBC with MNIG duced dosc  dependen: increase DNA-SSB 2t doses berwcen and S00uM MNNG. The delec tior Iimit for the as52 "a5 LM MNNG_ sess Ihe sumtabili:y of this 74552# t0 detect SSB in Uvo controlled stud; was performed which voluntee: smokers (=31 nonsmokers posed I0 environmenta tobacco smoke (ETSI and nonsmoker controls ("= Wete compared The stuc; lasted experimental days conitol and exposure days control days (days and smokers and nonsmokers sal unventilzted 4m room ior On Ine exposute dars (days and cach of   the Mive smorers smoked h while the fiva nonsmokers Verc ciposete{O the ETSheneraed &, noz smoking voi: exposcc unteets; High exposure t0 tobucco smoke Ws con lrmed by' dosimetry carboxyhemoglobin ICO- Hbl; plasma nicoting and cotininc levels Blood #as drawn bcfore and alcr cach cxposure perimenia d4ys for delermination o DNA-SSB in ivmphocytes immedialely afcr isolation of blood Key vords: DNA single-strand breaks DNA [- PWIT Nick: trinsiation Smoking Environmeil- tohacco smoke Biological monitoring Owing t0 ils physiological functions DNA in living cells highly reucuive 3nd unstable molecui: DNA is modilied at a rate cstimated t0 be 10` bases Per cell per da}. cven without exogcnous damage [26]: For the deieclion ol single strand breaks ISSB sevcral techniques havc becn developed Fucn conventional alkaline sucrose Erdicoi 114] sedi- mentation anulysis [19] Jnc alkalinc elution [I- 13]. The use of thc nick Iransiation (NT) [0 deicct SSB in DNA was lirst described in /983 in permc abilizcd humun (ihroblasis alcr cxposuirc t0 methyl-] niro N-nitrosogunnidine (MxxGi (20]. In tnc mcantimc h; > hecn demonsirled [ult thc NT Iah nus high snecilicity and sensitivity widc rangc of chcmicl gcnolaie Canounus I0 16. W|ad to herut /// Morcovcr; Ihis :ssJ} ilso bc used I0 sludy thc kinctics o DNA rcplir [67. 28. 291 AhnerCinUS cohb curhorhemoglohin: DMEM Dul. bcecu > modi-ution Exupte < Minimm  Cni Mcumn ; sinti- einc oreak ETS Untfuinninij Tuthacc SS8 ~mula YYYG L-micina ~nint #hel(481) _IL;4I1ILIIIL . Miac" inonaiuic|e;ted u #le hhonl eeik17 t ~ 1jlof Crer '4nu unzilt 'c 4 Rudiear 7