PRA]ENJE PROMJENA KONCENTRACIJA TAS-a, URATA I TRANSFERINA U SERUMU BOLESNIKA NAKON UGRADNJE PREMOSNICA
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1 Lj. Mayer i sur. Pra}enje promjena koncentracija TAS-a, urata i transferina u serumu bolesnika... Stru~ni ~lanak Professional Paper PRA]ENJE PROMJENA KONCENTRACIJA TAS-a, URATA I TRANSFERINA U SERUMU BOLESNIKA NAKON UGRADNJE PREMOSNICA Ljiljana Romi}, Jasna Juri~ek, Lovorka \erek Zavod za laboratorijsku dijagnostiku KB Dubrava, Zagreb SA@ETAK U literaturi su prisutni rezultati ispitivanja usmjerenih na dokazivanje poja~anog stvaranja slobodnih radikala i nastanka oksidacijskog stresa u bolesnika podvrgnutih ugradnji premosnica. U ovo istra`ivanje uklju~eno je 29 pacijenata podvrgnutih spomenutom kirurškom zahvatu. Kod njih 11 je konvencionalna revaskularizacija miokarda na~injena uz potporu izvantjelesne cirkulacije, a u preostalih 18 nije korišten EKC. Odre ivane su promjene antioksidacijskih pokazatelja u serumu (TAS, urat, transferin). Koncentracija totalnog antioksidacijskog statusa smanjena je tijekom poslijeoperacijskog perioda. Koncentracija transferina je zna~ajno smanjena tijekom poslijeoperacijskog perioda, dok je koncentracija urata zna~ajno pove}ana. Rezultati dobiveni u ovom istra`ivanju još su jedna u nizu potvrda nastanka oksidacijskog stresa za vrijeme kirurškog zahvata ugradnje premosnica. Intenzitet promjena odre ivanih parametara ne razlikuje se obzirom na to je li korištena izvantjelesna cirkulacija ili nije. Klju~ne rije~i: oksidacijski stres, totalni antioksidacijski status, premosnica, urat, transferin UVOD Kirurški zahvat ugradnje sr~anih premosnica ~esto rezultira ošte}enjima drugih organa (plu}a, bubrezi, srce, pankreas), pri ~emu su poreme}aji funkcije plu}a i srca od vitalnog zna~aja. Mogu}i uzroci spomenutih ošte}enja, koja su ujedno odgovorna i za razvoj oksidacijskog stresa kod kardiokirurških bolesnika su: primjena visokog parcijalnog tlaka kisika tijekom operacije, primjena izvantjelesne (ekstrakorporalne) cirkulacije (EKC), ishemija miokarda pra}ena reperfuzijom visokooksigenirane krvi, smanjena perfuzija perifernih tkiva, trauma uslijed kirurškog zahvata i dr. (1,2). Dosadašnja istra`ivanja, u kojima je mjerena razina oksidacijskog stresa u kardiokirurških bolesnika, ukazuju na poja~ano stvaranje slobodnih radikala nakon operacije ugradnje premosnica. Adresa za dopisivanje: Mr. sci. Ljiljana Mayer, dipl. in`., Zavod za laboratorijsku dijagnostiku KB Dubrava, Av. Gojka Šuška 6, Zagreb, Hrvatska.Tel.: , fax.: , Lmayer@kbd.hr Pri tome se manji broj istra`ivanja temeljio na izravnom odre ivanju slobodnih radikala (3). Mnogo su ~eš}a istra`ivanja u kojima su odre ivani neizravni pokazatelji nastanka oksidacijskoga stresa (odre ivanje koncentracije lipidnih peroksida (4), konjugiranih diena (5), koncentracije vitamina E (6) i C, aktivnosti antioksidacijskih enzima te, manjim dijelom, odre ivanje koncentracija izvanstani~nih spojeva s antioksidacijskim svojstvima. Ovim radom pokušali smo definirati ovisnost promjena izvanstani~nih antioksidansa (TAS, urat i transferin) o primjeni EKC. Naime, u odre enom broju istra`ivanja upravo je izlaganje krvi nefiziološkoj površini tijekom EKC isticano kao temeljni ~imbenik razvoja oksidacijskog stresa kod bolesnika podvrgnutih operacijama ovakova tipa. Koncentracije odabranih parametara mjerene su u serumu prije operacije, te u razli~itim vremenima nakon završetka operacije (90 minuta, 6, 24 i 72 sata). BIOCHEMIA MEDICA god. 13, br. 1 2,
2 Pra}enje promjena koncentracija TAS-a, urata i transferina u serumu bolesnika... Lj. Mayer i sur. MATERIJAL I METODE Bolesnici Nakon odobrenja Eti~kog povjerenstva KB Dubrava i uz pristanak bolesnika u klini~ku prospektivnu studiju uklju~eno je ukupno 29 bolesnika podvrgnutih operacijskom zahvatu aortokoronarnog premoštenja. Kriteriji uklju~enja bolesnika u istra`ivanje bili su trostruka koronarna bolest, te starosna dob izme u 60 i 70 godina. Kriteriji isklju~enja bolesnika iz istra`ivanja bili su: smanjena istisna frakcija lijevog ventrikula (<40%), kroni~ne upalne bolesti, še}erna bolest, te prethodni kardiokirurški operacijski zahvat. Bolesnici uklju~eni u ispitivanje nisu primali lijekove s antioksidacijskim u~inkom (alopurinol, manitol, kapropril i desferioksamin), niti protuupalne lijekove (kortikosteroidi, acetilsalicilna kiselina i njeni derivati). Bolesnici su bili podijeljeni u dvije skupine. Prva, skupina EKC+, brojila je 11 bolesnika kod kojih je konvencionalna revaskularizacija miokarda na~injena uz potporu EKC. Drugom, skupinom EKC- obuhva}eno je 18 bolesnika kod kojih je revaskularizacija miokarda na kucaju}em srcu na~injena bez potpore EKC. Anestezija, pra}enje i operacijska tehnika Svi bolesnici bili su premedicirani morfijem jedan sat prije operacijskog zahvata. Protokol op}e anestezije bio je isti za sve bolesnike. Tijekom indukcije primijenjen je midazolam, alfentanyl i atracurium. Nakon orotrahealne intubacije, bolesnici su bili priklju~eni na mehani~ku ventilaciju, a anestezija je odr`avana mješavinom dušika i kisika (60% : 40%) i isofluranom. Nakon pristupa srcu kroz medijalnu sternotomiju i perikardiektomiju, nakon prethodne pripreme presadaka, uspostavljena je EKC kaniliranjem aorte i desnog atrija dvofaznom kanilom (double-stage). Izazvana je artificijelna fibrilacija ventrikla, klema aorta i u bulbus aorte daje kardioplegijska otopina po Buckbergu. Nakon toga, tijekom provo enja anastomoza, primjenjuje se retrogradna kardioplegija kroz kanilu u koronarnom sinusu. Kod zahvata bez korištenja stroja za izvantjelesnu cirkulaciju pristup srcu je istovjetan prethodnom, a ekspozicija ciljnih `ila i stabilizacija miokardijalne stijenke posti`e se vakuum stabilizatorima. Stanje bolesnika pra}eno je s pomo}u EKG, pulsnog oksimetra i kapnografa. Uzorkovanje Uzorci krvi bolesnika su uzimani prijeoperacijski (dan prije operacije, na odjelu) i poslijeoperacijski: 90 minuta te 6, 24 i 72 sata nakon operacije. Sva poslijeoperacijska uzorkovanja na~injena su u jedinicama intenzivnog lije~enja u kojima svi takvi bolesnici borave prvih dana nakon operacije. Pri uzimanju uzorka krvi korišten je pribor za jednokratnu uporabu (BD Vacutainer-i) bez antikoagulansa. Nakon centrifugiranja 15 minuta / 3000 okretaja u minuti (Heraus Minifuga T), serum je alikvotiran, obilje`en i pohranjen do analize na temperaturi - 40 o C. Metode Koncentracija totalnog antioksidacijskog statusa (TAS) odre ena je u serumu s pomo}u Randox-ovog test-paketa (Randox, Crumlin, UK) (7). Preciznost je kontrolirana s pomo}u Randox Total Antioxidant Control (Randox, Crumlin, UK) Koncentracije transferina i urata odre ene su gotovim test-paketima tvrtke Olympus na analizatoru Olympus AU 600. Mjerenjem koncentracije odre ivanih parametara u pool serumu odre ivana je nepreciznost iz dana u dan. Statisti~ka analiza Zna~ajnost promjena koncentracija mjerenih parametara izme u dviju skupina u svim to~kama va enja ispitana je s pomo}u Mann-Whitney-og U testa. Zna~ajnost razlikovanja pojedinog parametra u odnosu na prijeoperacijske vrijednosti na~injena je primjenom Wilcoxonovog parnog testa. Primjenom statisti~ke analize ANOVA/MANOVA ispitana je sveobuhvatna razlika izme u dviju skupina u kretanju pojedinog parametra tijekom cijeloga razdoblja mjerenja. Vrijednost p < 0,05 pokazala se kao statisti~ki zna~ajna. REZULTATI Koncentracija TAS-a u serumu op}enito je smanjena tijekom poslijeoperacijskog razdoblja u odnosu na prijeoperacijske vrijednosti (1,41±0,15 mmol/l). Spomenuto smanjenje koncentracije TAS-a najizra`enije je u prvoj poslijeoperacijskoj to~ci mjerenja (1,31±0,15 mmol/l), u kojoj su odre ene vrijednosti smanjene u prosjeku za 14% od prijeoperacijskih. Šest i 24 sata nakon operacije koncentracija TAS-a u serumu se zadr`ava na sli~nim vrijednostima (1,33 ± 0,13 i 1,36 ± 0,15 mmol/l), a 72 sata nakon operacije TAS se pribli`ava prijeoperacijskim vrijednostima, ali je smanjenje još uvijek zna~ajno. Koncentracija TAS-a se u svim poslijeoperacijskim to~kama odre ivanja statisti~ki zna~ajno razlikuje (Wil- 34 BIOCHEMIA MEDICA god. 13, br. 1 2, 2003.
3 Lj. Mayer i sur. Pra}enje promjena koncentracija TAS-a, urata i transferina u serumu bolesnika... coxon-ov parni test) u odnosu na koncentracije izmjerene prije operacije u obje skupine bolesnika (slika 1.). Slika 1. Koncentracija TAS-a u serumu bolesnika skupina EKC+ ( ) i EKC ( ) (statisti~ki zna~ajno u odnosu na prijeoperacijske vrijednosti; p < 0,05, s pomo}u Wilcoxonovog testa) Figure 1. TAS concentration in the sera of EKC+ ( ) and EK ( ) groups of patients (statistically significant vs. preoperative values; p < 0.05, by Wilcoxon test) Primjenom Mann-Whitney-eva testa uspore- ivane su koncentracije TAS-a u serumu bolesnika iz skupina EKC+ i EKC dobivenog istovremenim odre ivanjem. U niti jednoj od pet to~aka va enja nije prona ena statisti~ki zna- ~ajna razlika u koncentraciji TAS-a u serumu dviju skupina bolesnika. Primjenom statisti~ke analize ANOVA/MA- NOVA nije utvr ena zna~ajna razlika izme u kretanja koncentracije TAS-a u serumu skupina EKC+ i EKC tijekom cijeloga mjernog perioda (p=0,8602). Ispitivanje zna~ajnosti razlikovanja koncentracija urata i transferina u pojedinoj to~ci va- enja izme u dviju skupina bolesnika na~injeno je tako er s pomo}u Mann-Whitney-eva testa. Dobivene vrijednosti (p>0.05) ukazuju na nepostojanje statisti~ki zna~ajne razlike u koncentraciji ovih izvanstani~nih molekula s antioksidacijskim svojstvima izme u skupina bolesnika razdvojenih s obzirom na to je li revaskularizacija na~injena uz EKC ili ne (i u posljednjoj to~ci mjerenja koncentracija urata izmjerena u dvije skupine se razlikuje; p=0,0532). Primjenom Wilcoxon-ova parnoga testa na- ena je statisti~ki zna~ajna promjena koncentracije urata u odnosu na prijeoperacijske vrijednosti 1,5 i 6, 24 i 72 sata nakon operacije (p<0,05) u skupini EKC, kao i 6 (p=0,008) i 24 (p=0,028) sati nakon operacije u skupini EKC+ (slika 2). Slika 2. Koncentracija urata u serumu bolesnika skupina EKC+ ( ) i EKC ( ) (statisti~ki zna~ajno u odnosu na prijeoperacijske vrijednosti; p < 0,05, s pomo}u Wilcoxonovog testa) Figure 2. Urate concentration in the sera of EKC+ ( ) and EKC ( ) groups of patients (statistically significant vs. preoperative values; p < 0.05, by Wilcoxon test) Primjenom statisti~ke analize ANOVA/MA- NOVA ispitana je zna~ajnost razlike kretanja koncentracije urata u serumu skupina EKC+ i EKC tijekom cijeloga mjernog perioda. Dobivena vrijednost (p=0,0598), iako grani~na, ipak ukazuje da se promjene koncentracije urata tijekom razdoblja mjerenja ne razlikuju zna~ajno u dvije ispitivane skupine bolesnika. Koncentracija transferina zna~ajno se smanjuje tijekom cijelog poslijeoperacijskog razdoblja u odnosu na prijeoperacijske vrijednosti (2,3±0,7 g/l). Spomenuto smanjenje tijekom poslijeoperacijskog razdoblja se u odnosu na prijeoperacijske vrijednosti kre}e u rasponu od 26-36% prijeoperacijskih koncentracija transferina (slike 3). Slika 3. Koncentracija transferina u serumu bolesnika skupina EKC+ ( ) i EKC ( ) (statisti~ki zna~ajno u odnosu na prijeoperacijske vrijednosti; p < 0,05, s pomo}u Wilcoxonovog testa) Figure 3. Concentration of transferine in the sera of EKC+ ( ) and EKC ( ) groups of patients (statistically significant vs. preoperative values; p < 0.05, by Wilcoxon test) Sli~no kao kod prethodna dva parametra, s pomo}u statisti~ke ANOVA/MANOVA analize is- BIOCHEMIA MEDICA god. 13, br. 1 2,
4 Pra}enje promjena koncentracija TAS-a, urata i transferina u serumu bolesnika... Lj. Mayer i sur. pitana je zna~ajnost razlike u kretanju koncentracije transferina u serumu skupina EKC+ i EKC tijekom cijeloga mjernog perioda. Dobivena vrijednost (p=0,7376) ukazuje da ne postoji statisti~ki zna~ajna razlika u kretanju ovog parametra s obzirom na primjenu EKC. RASPRAVA Postojanje oksidacijskog stresa nakon kirurškog zahvata ugradnje premosnica ispitivano je u dosadašnjim istra`ivanjima odre ivanjem klasi~nih neizravnih pokazatelja (mjerenje koncentracije produkata lipidne peroksidacije, koncentracije ili aktivnosti endogenih i egzogenih antioksidanasa), ili izravnim odre ivanjem slobodnih radikala metodom elektronske spinske rezonancije. Rezultati tih istra`ivanja uglavnom ukazuju da oksidacijski stres nastaje vrlo brzo, mjerljiv je ~ak i tijekom operacije te, što je vrlo zna~ajno, nije isklju~ivo lokalnog karaktera, ve} je sistemskog podrijetla (6). S obzirom na navedeno, u ovom ispitivanju pra}ene su koncentracije izvanstani~nih pokazatelja oksidacijskog stresa (TAS, urat, transferin) u serumu bolesnika podvrgnutih ugradnji premosnica prije operacije i u prvim danima nakon operacije. Tako er je ispitivana ovisnost promjene koncentracije navedenih parametara o primjeni EKC. Za koncentraciju TAS-a kao parametar izbora odlu~ili smo se stoga što neki autori kretanju njegove koncentracije pripisuju prognosti~ko zna~enje, u smislu definiranja dugotrajnijeg ishoda kirurškog zahvata ugradnje premosnica (8). Pretrage za odre ivanje ukupnog antioksidacijskog kapaciteta na~injene su zbog mjerenja svih antioksidansa u biološkim sustavima. Smatra se tako da je ukupni antioksidacijski kapacitet rezultat slo`enog sinergisti~kog djelovanja neenzimskih ~imbenika (npr. tokoferola, karotenoida, vitamina C, selena), enzimskih ~imbenika (npr. katalaza, SOD, GPX), odnosno stani~nih i izvanstani~nih ~imbenika, kao i još nepoznatih ili rijetko odre ivanih ~imbenika antioksidacijskog statusa u organizmu. Doprinos pojedine komponente ukupnom antioksidacijskom statusu razli- ~it je u stanicama i izvanstani~noj teku}ini (9). U literaturi se nalaze uglavnom ispitivanja odre ivanja koncentracije TAS-a u plazmi ili serumu, ~emu najve}im dijelom doprinose sljede}e komponente: albumin (kao glavni izvor tiolnih skupina), urat i vitamin C, dok na ostale poznate i nepoznate komponente otpada 40% ukupnog antioksidacijskog kapaciteta krvne plazme (9). Tijekom cijelog poslijeoperacijskog razdoblja, poglavito 1,5 sati poslije operacije, zna~ajno su manje koncentracije TAS-a u odnosu na prijeoperacijske vrijednosti. Do sli~noga rezultata došli su i drugi autori, tuma~e ga prije svega primjenom EKC tijekom operacije (8). U istra`ivanju Gu i sur. (2) izmjerene su u kardiokirurških bolesnika zna~ajno sni`ene vrijednosti pojedina~nih komponenti antioksidacijskog statusa izvanstani~nog odjeljka krvi. Tako je u plazmi smanjena koncentracija askorbata i glutationa, a u serumu beta karotena i vitamina E, zna~ajnih komponenti TAS-a. Takav rezultat ukazuje na pove}ano stvaranje slobodnih radikala na sistemskoj razini, a ne lokalizirano, odnosno da oni ne potje~u samo i isklju~ivo od ošte- }enih miocita. Od endogenih izvanstani~nih antioksidanasa u serumu mjerili smo koncentraciju transferina kao preventivnog antioksidansa, odnosno biomolekule koja vezanjem `eljeza onemogu}uje jedan od na~ina stvaranja visokoreaktivnog hidroksil- radikala, te koncentraciju urata. Prijeoperacijske vrijednosti navedenih antioksidanasa nisu se zna~ajno razlikovale od vrijednosti izmjerenih u zdravoj populaciji. Tijekom poslijeoperacijskog perioda izmjerili smo zna~ajno smanjenu koncentraciju transferina u svim, a pove}anu koncentraciju urata u nekim to~kama mjerenja. Promjene koncentracije urata kod operacija srca naj~eš}e su, prema literaturnim podatcima, odre ivane s dva aspekta. S jedne strane, urat kao derivat purina (hipoksantina i ksantina) mo- `e biti neizravni biljeg procjene razine ishemije (10), a s druge pak strane upotrebljiv je pokazatelj za pra}enje primjene imunosupresiva, poput alopurinola, nakon transplantacija (11). Koncentracije TAS-a i urata se tijekom perioda mjerenja ponašaju razli~ito. Naime, koncentracija TAS-a se smanjuje, a urata pove}ava tijekom mjerenja. Kako je i urat obuhva}en odre ivanjem koncentracije TAS-a u serumu, ovaj rezultat ukazuje da se nu`no ne moraju svi raspolo`ivi izvanstani~ni antioksidansi mijenjati istovjetnim smjerom i dinamikom. Dobiveni rezultat ukazuje da je porast urata u ukupnom antioksidacijskom kapacitetu prekrio još zna- ~ajniji pad jedne ili više komponenti obuhva}enih mjerenjem ovoga analita. Neki autori isti~u da je dinami~ka ravnote`a izme u prooksidansa i antioksidansa u odre enoj okolini vjerodostojnije prikazana odre ivanjem ukupnog antioksidacijskog kapaciteta, nego odre ivanjem svake od komponeneta zasebno. Pri tome treba obratiti 36 BIOCHEMIA MEDICA god. 13, br. 1 2, 2003.
5 Lj. Mayer i sur. Pra}enje promjena koncentracija TAS-a, urata i transferina u serumu bolesnika... pozornost na sam postupak odre ivanja TAS-a, budu}i da je doprinos individualnih antioksidanasa pri mjerenju ukupnog antioksidacijskog kapaciteta razli~it, ovisan o primijenjenom postupku odre ivanja (12, 13). U literaturi je opisano da operacije ovakvoga tipa mogu biti uzrokom pove}anog zasi}enja transferinom koje je odraz suviška `eljeza. Na taj na~in je umanjena njegova djelotvornost vezanja u oksidacijskom smislu reaktivnog oblika `eljeza, što je uostalom i mehanizam njegovog antioksidacijskog djelovanja (14). Stoga, pad koncentracije transferina (kakav smo mi izmjerili nakon operacije) mo`e doista biti neizravan uzro~nik pove}ane razine oksidacijskog stresa. Rezultati našeg ispitivanja su još jedna u nizu potvrda nastanka oksidacijskog stresa za vrijeme kirurškog zahvata ugradnje sr~anih premosnica. Pokazuju, nadalje, da do oksidacijskog stresa dolazi neovisno o tome je li tijekom operacije primijenjena EKC ili ne, da organizam vrlo brzo reagira promjenom statusa antioksidansa u serumu. Zaklju~ujemo da rezultati upu}uju na opravdanost preventivnog antioksidacijskog tretmana bolesnika prije ili tijekom kirurškog zahvata kako bi se smanjile mogu}e poslijeoperacijske komplikacije, koje su posljedica nastalog sistemskog oksidacijskog stresa. Daljnja dobro planirana i kontrolirana ispitivanja s razli~itim vrstama antioksidanasa ili kombinacijama antioksidanasa mogla bi ukazati na naju~inkovitiji antioksidans, njegov najbolji oblik i na~in primjene. MONITORING CONCENTRATION CHANGES IN TAS, URATE AND TRANSFERRIN IN THE SERA OF PATIENTS AFTER BYPASS PROCEDURE ABSTRACT Results of numerous studies aimed at demonstrating enhanced production of free radicals and occurrence of oxidative stress in patients undergoing bypass procedures. Twenty-nine patients who underwent surgical bypass were included in our research. In eleven patients conventional myocardial revascularization was performed with CPB support which was not used in the remaining eighteen patients. Samples were collected before surgery and an hour and half, six, twenty-four and seventy two hours after the operation. Changes in the concentration of antioxidative parameters were monitored in the sera (TAS, urate, transferrin). Total antioxidant status (TAS) was decreased during the entire postoperative period. Transferrin concentration was significantly reduced during the entire postoperative period, while the concentration of urate was significantly increased. The results of our study provided an additional confirmation of the occurrence of oxidative stress during the operative bypass procedure. Intensity of changes in all determined parameters did not differ regardless of whether CPB support was used or not. Key words: oxidative stress, total antioxidant status, bypass, urate, transferrine LITERATURA 11. Kirklin JK. The postperfusion syndrome: inflammation and the damaging effects of cardiopulmonary bypass. In: Tinker JH, editor. Cardiopulmonary bypass: current concepts and controversis. Philadelphia: WB Saunders, 1989: (4): Gu M, Love H, Schofield D, Turkie W, Odom N and Braganza JM. A pilot study of blood antioxidant and free radical marker profiles in patients awaiting coronary artery bypass grafting. Clin Chim Acta 1996; 252: Clermont G, Vergely C, Jazayeri S, Lahet JJ, Goudeau JJ, Lecour S, David M, Rochette L and Girard C. Systemic free radical activation is a major event involved in myocardial oxidative stress ralated to cardiopulmonary bypass. Anesthesiology 2002; 96: Davies SW, Duffy JP, Wickens DG, Underwood SW, Hill A, Alladine F, Feneck RO, Dormandy TL and Walesby RK. Time-course of free radical activity during coronary artery operations with cardiopulmonary bypass. J Thorac Cardiovasc Surg 1993; 105: Tovoinen HJ, Ahotupa M. Free radical reaction products and antioxidant capacity in arterial plasma during coronary artery bypass grafting. J Thorac Cardiovasc Surg 1994; 108: Coghlan JG, Flitter WD, Cluton SM, Ilsley CDJ, Rees A and Slater TF. Lipid peroxidation and vitamin E levels during coronary artery bypass grafting. J Thorac Cardiovasc Surg 1993; 106: Miller NJ, Rice-Evans C, Davies MJ, Gopinthan V, Milner A. A novel method for measuring antioxidant capacity and its application to monitoring antioxidant status in premature neonates. Clin Sci 1993; 84: McColl AJ, Keeble T, Hadjinikolaou L, Cohen A, Aitkenhead H, Glenville B, Richmond W. Plasma antioxidants: evidence for a protective role against reactive oxygen species following cardiac surgery. Ann Clin Biochem 1998; 35: Abuja PM, Albertitni R. Methods for monitoring oxidative stress, lipid peroxidation and oxidation resistance of lipoproteins. Clin Chim Acta 2001; 306: Gerritsen WB, Aarts LP, Morshuis WJ, Haas FJ. Indices of oxidative stress in urine of patients undergoing coronary artery bypass grafting. Eur J Clin Chem Clin Biochem 1997; 35: BIOCHEMIA MEDICA god. 13, br. 1 2,
6 Pra}enje promjena koncentracija TAS-a, urata i transferina u serumu bolesnika... Lj. Mayer i sur. 11. Tarkka MR, Kaukinen S, Holm P, Kaukinen U, Sisto T, Kataja J, Huang WQ. Allopurinol does not increase free radical scavenging capacity during reperfusion in coronary artery bypass graft patients. Scand Cardiovasc J 2000; 34: Ghiselli A, Serafini M, Natella F, Scaccini C. Total antioxidant capacity as a tool to assess redox status: critical view and experimental data. Free Rad Biol Med 2000; 29: Janaszewska A, Bartosz G (2002) Assay of total antioxidant capacity and comparison of four methods as applied to human blood plasma. Scand J Clin Lab Invest 2002; 62: Mumby S, Chaturvedi RR, Brierley J Lincoln C, Petros A, Redington AN, Gutteridge JM. Iron overload in paediatrics undergoing cardiopulmonary bypass. Bioch Biophys Acta 2000; 1500: BIOCHEMIA MEDICA god. 13, br. 1 2, 2003.
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