XXI srpski kongres medicinske i laboratorijske medicine

sa me|unarodnim u~e{}em

XXI Serbian Congress of Medical Biochemistry and Laboratory Medicine

with international participation

Posterske sekcije /Poster SessionsApstrakti /Abstracts

202

P001 ANALYSIS OF SHBG AND

INCREASED FREE BETA HCG IN THEFIRST TRIMESTER OF PREGNANCY

IN GLYCOREGULATION

Aleksandra \or|evi}1, Bosa Mirjani}-Azari}1,Dragica Draganovi}2, Aleksandra Rodi}2,

Dragana Vasi} Pavlovi}3, @eljka Kizi}3

1Institute of Laboratory Diagnostic, UniversityClinical Centre of the Republic of Srpska, Banja Luka

2Clinic of Gynecology and Obstetrics, UniversityClinical Centre of the Republic of Srpska, Banja Luka3Health Center Banja Luka, the Republic of Srpska

Gestational diabetes mellitus (GMD) is definedas any degree of glucose intolerance with the firstrecognition during pregnancy. The importance ofearly detection of GDM in pregnant women is a pre-vention of the complications that this disease carriesto the health of pregnant women and fetus. SHBG(sex hormone binding globulin) is a glycoprotein whorepresent a transport protein for sex hormones andits low values are associated with increased insulinresistance and hyperinsulinemia. The aim of thisstudy was to examine biochemical markers of gly-coregulation as well as SHBG level in pregnantwomen with increased free beta bhCG in the comm-bined screening test in the firs trimester of pregnancy.The prospective study was performed betweenNovembar 2015. and Februar 2018. in the Instituteof Laboratory Diagnostic, University Clinical Centreof the Republic of Srpska. The study include 89 preg-nant women divided into the two groups: the exam-ined group of pregnant women (n = 43) who hadfree bhCG ≥ 2.0 MoM in the commbined screeningtest in the first trimester of pregnancy and controlgroup (n = 46). Hormons were determined in serumelectrochemiluminescence immunoassay (ECLIA,Cobas, Roche Diagnostics, Mannheim, Germany).Statistical analysis results is performed using SPSSprogram version 22.0. The mean values obtained inthe examined group vs. the control group were:SHBG (nmol/L) 297.41±109.02 vs. 314.64±78.74(p = 0.277); glucose (mmol/L) 4.80±0.57 vs.4.79±0.45 (p = 0.902); insulin (mU/mL) 15.23±13.50 vs. 10.33±5.43 (p=0.081) i C-peptid (ng/mL)2.29±1.22 vs. 1.95±0.68 (p=0.549). Likewise, therewas not correlation of free bhCG with the examined

P001ANALIZA SHBG I POVEĆANE SLOBODNE bHCG U PRVOMTROMESE^JU TRUDNOĆE

U GLIKOREGULACIJI

Aleksandra \or|evi}1, Bosa Mirjani}-Azari}1,Dragica Draganovi}2, Aleksandra Rodi}2,

Dragana Vasi} Pavlovi}3, @eljka Kizi}3

1Zavod za laboratorijsku dijagnostiku, Univerzitetskiklini~ki centar Republike Srpske, Banja Luka

2Klinika za ginekologiju i aku{erstvo, Univerzitetskiklini~ki centar Republike Srpske, Banja Luka

3JZU Dom Zdravlja Banja Luka, Republika Srpska

Gestacijski diabetes mellitus (GDM) se defini{ekao bilo koji stepen glukozne intolerancije koji se jav -lja prvi put u trudno}i. Zna~aj ranog otkrivanja GDMkod trudnica predstavlja prevenciju od komplikacijakoje ova bolest nosi po zdravlje trudnica i ploda.SHBG (sex hormon binding globulin) je glikoproteinkoji je transportni protein za polne hormone, a nje-gove niske vrijednosti se povezuju sa pove}anominsulinskom rezistencijom i hiperinsulinemijom. Ciljrada je bio ispitati biohemijske markere glikoregu-lacije kao i nivo SHBG kod trudnica sa pove}anomslobodnom beta bhCG u kombinovanom skriningtestu prvog tromjese~ja trudno}e. Prospektivna studi-ja je izvedena u periodu od novembra 2015. do feb-ruara 2018. u Zavodu za laboratorijsku dijagnostikuUniverzitetskog klini~kog centra Republike Srpske. Uispitivanje je uklju~eno ukupno 89 trudnica, podjel-jene u dvije grupe: ispitivana grupa (n= 43) obuhva-ta trudnice koje su u kombinovanom skrining testu uprvom tromjese~ju trudno}e imale pove}anu vrijed-nost slobodne bhCG ≥ 2.0 MoM i kontrolna grupa(n=46). Hormoni su odre|ivani u serumu elektrohe -miluminiscentnom metodom (ECLIA, Cobas, RocheDiagnostics, Mannheim, Germany). Sta tis ti~ka anali -za rezultata je ra|ena pomo}u SPSS programa verzija22.0. U ispitivanoj grupi dobijene srednje vrijednostinaspram kontrolne grupe bile su: SHBG (nmol/L)297,41±109,02 vs. 314,64±78,74 (p=0.277);glu koza (mmol/L) 4,80±0,57 vs. 4,79±0,45 (p=0,902); insulin (mU/mL) 15,23±13,50 vs. 10,33±5,43 (p=0,081) i C-peptid (ng/mL) 2,29±1,22 vs.1,95±0,68 (p=0.549). Tako|e, nije postojala kore -lacija slobodne bhCG sa ispitivanim parametrimaglikoregulacije. Postojala je korelacija izme|u SHBG

UDK 577.1 : 61 ISSN 1452-8258

J Med Biochem 37: 202–217, 2018 Poster sessionsPosterske sekcije

J Med Biochem 2018; 37 (2) 203

i indeksa tjelesne mase (BMI) (r = - 0,486; p = 0,001vs. r = - 0,359; p = 0,018) kao i korelacija izme|uSHBG i PAPP-A (r = 0,296; p = 0,054 vs. r = 0,400;p = 0,006) u obe grupe trudnica. Izme|u BMI iPAPP-A je bila korelacija u obe grupe. Dvije trudnicesu imale spontani poba~aj u ispitivanoj grupi. Kodtrudnica sa slobodnom bhCG ≥ 2,0 MoM u prvomtromjese~ju trudno}e nije postojala zna~ajna razlika ubiohemijskim markerima glikoregulacije kao i SHBGproteina u odnosu na trudnice koje su imale normal-nu vrijednost slobodne bhCG.

Klju~ne rije~i: gestacionalni, dijabetes, trud -no}a.

glycoregulation parameters. There was a correlationbetween SHBG and body mass index (BMI) (r=- 0.486;p = 0.001 vs. r = - 0.359; p = 0.018), as well as thecorrelation between SHBG and PAPP-A (r = 0.296;p = 0.054 vs. r = 0.400; p = 0.006) in both groupsof pregnant women. Between BMI and PAPP-A wascorrelation in both groups. Two pregnant women hadspontaneous abortion in the examined group.Pregnant women with free bhCG ≥ 2.0 MoM in thefirst trimester of pregnancy there was not significantdifference in the biochemical markers of glycoregula-tion and SHBG proteins compared to pregnantwomen who had a normal value of free bhCG.

Key words: gestational, diabetes, pregnancy.

P002 REFERENTNI INTERVAL

ZA TIROKSIN NA ADVIA CENTAUR ANALIZATORU

Marijana Dajak1, Ivana Draga{evi}1, Svetlana Ignjatovi}1,2

1Centar za medicinsku biohemiju, Klini~ki centar Srbije, Beograd, Srbija,

1Centar za medicinsku biohemiju, Klini~ki centarSrbije i Katedra za medicinsku biohemiju,

Farmaceutski fakultet, Univerzitet u Beogradu,Beograd, Srbija

Laboratorijsko merenje tiroidnih hormona je po`eljnosredstvo za dijagnozu i pra}enje pacijenta sa dis-funkcijom {titaste `lezde. Referentni intervali suneophodni u ocenjivanju rezultata ispitivanja. Ve}inadru{tava za klini~ku hemiju preporu~uje da svaka la -boratorija treba da odredi svoje lokalne referentnevred nosti. U na{oj laboratoriji koristimo referentneintervale koje daju proizvo|a~i. Tiroidni hormoni semere na dva analizatora: Siemens Advia Centaur XPi Roche Cobas e601. Uo~eno je, pregledom labora-torijskih podataka, u slu~ajevima kada su svi tiroidnitestovi bili u referentnom intervalu, u pribli`no 7%slu ~ajeva, odre|ivanje tiroksina (T4) pomo}u Cen -taur metode dalo je visoke vrednosti. Dakle, cilj ovestudije bio je upore|ivanje vrednosti T4 sa dva ana -lizatora i verifikacija referentnog intervala za T4 zaCentaur metodu. Pore|enje vrednosti T4 izme renihCentaur i Cobas metodama pokazalo je da nije po -stojala zna~ajna razlika izme|u vrednosti za merniopseg od 63,1–208,5 nmol/L. Me|utim, vrednostiT4 iz Centaur metode bile su ve}e od vrednosti izCobas metode u rasponu od 124,7–172,0 nmol/L(t-test, p < 0,05). 95% referentni interval (normalnadistribucija) za vrednosti Centaur T4, dobijenih od160 o~igledno zdravih osoba (73 mu{karaca i 87 `ena,od 21 do 79 godina starosti), iznosio je 73,5–149,3

P002 THE REFERENCE INTERVAL

FOR THYROXINE ON ADVIA CENTAUR ANALYSER

Marijana Dajak1, Ivana Draga{evi}1, Svetlana Ignjatovi}1,2

1Center of Medical Biochemistry, Clinical Center of Serbia, Belgrade, Serbia,

2Center of Medical Biochemistry, Clinical Center ofSerbia and Department of Medical Biochemistry,

Faculty of Pharmacy, University of Belgrade,Belgrade, Serbia

The laboratory measurement of thyroid hor-mones is the preferred means to diagnose and mon-itor patient with thyroid dysfunction. The referenceintervals are indispensable in evaluating test results.Most of the societies for clinical chemistry have rec-ommended that each laboratory should determine itsown local reference values. In our laboratory we usethe reference intervals provided by the manufac -turers. Thyroid hormones are measured on two ana-lyzers: Siemens Advia Centaur XP and Roche Cobase601. It was observed, by reviewing laboratory data,in the occasions when all thyroid tests were withinreference interval, in about 7% of cases, the determi-nation of thyroxine (T4) using Centaur method gavehigh values. So, the aim of this study was to comparethe T4 values from two analyzers and to verifyCentaur reference interval for T4. The comparison ofT4 values measured by Centaur and Cobas methodsshowed that there was no significant differencebetween values for measuring range of 63.1–208.5nmol/L. However, T4 values from Centaur methodwere higher than values from Cobas method in rangeof 124.7–172.0 nmol/L (t-test, p < 0.05). The 95%reference interval (normal distribution) for CentaurT4 values, obtained from 160 apparently healthysubjects (73 men and 87 women, aged 21–79 years)

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nmol/L, sa srednjom vredno{}u od 108,3 nmol/L.Ovaj interval nije sasvim u skladu sa referen tnimintervalom koji je obezbedio proizvo|a~ Centaur(58,1–140,6 nmol/L), a srednja vrednost je zna~ajnoveca od vrednosti koju je dao proizvo|a~ (90,3nmol/L). Dobijena srednja vrednost se nije statisti~kirazlikovala od medijane date za Cobas metodu(105,0 nmol/L), {to ukazuje da su blago povi{enevrednosti odre|ene Centaur metodom, vi{e u saglas-nosti sa referentnim intervalom koji je dao Cobasproizvo|a~ (66–181 nmol/L). Rezultati studije pred-la`u referentni interval od 73,5–149,3 nmol/L za od -re|ivanje T4 kori{}enjem Advia Centaur analizatora.

was 73.5–149.3 nmol/L, with mean value of 108.3nmol/L. This interval is not completely in line withthe reference interval provided by the Centaur man-ufacturer (58.1–140.6 nmol/L), and the mean wassignificantly higher than the manufacturer’s mean(90.3 nmol/L). The obtained mean didn`t statisticallydiffer from median given in Cobas method (105.0nmol/L), which indicates that slightly elevated valuesdetermined by Centaur method are more in accor-dance with the reference interval provided by Cobasmanufacturer (66–181 nmol/L). The results of studysuggested the reference interval of 73.5–149.3nmol/L for determination of T4 using Advia Centauranalyzer.

P003ZNA^AJ ODRE\IVANJA

GLIKOZILIRANOG HEMOGLOBINA U PROCENI RIZIKA OD AMPUTACIJAKOD PACIJENATA SA SINDROMOM

DIJABETESNOG STOPALA

Tatjana Stefanovi}1, Sre}ko Bosi}2, Jelena Veki}3, Vesna Spasojevi}-Kalimanovska3,

Zorana Jeli}-Ivanovi}3

1Klini~ko-biohemijska laboratorija, Op{ta bolnica, Po`arevac

2Hirur{ka slu`ba, Op{ta bolnica, Po`arevac3Katedra za medicinsku biohemiju,

Univerzitet u Beogradu – Farmaceutski fakultet

Glikozilirani hemoglobin (HbA1c) je va`an para-metar za pra}enje dugoro~ne metaboli~ke kontrole iprocenu rizika za razvoj hroni~nih komplikacija kodosoba sa dijabetes melitusom (DM). U ovoj studijiispi tivali smo povezanost postignute metaboli~kekon trole u DM sa rizikom za razvoj i primenjenimmodalitetom le~enja sindroma dijabetesnog stopala(SDS). U istra`ivanju je u~estvovalo 111 pacijenata(80 mu{karaca i 31 `ena) sa tipom 2 DM, starosti65,5 ± 9,8 godina. Razvoj SDS nastupio je 13,9 ±8,8 godina nakon postavljene dijagnoze dijabetesa.Kod 41 pacijenta SDS je le~en neoperativnim (kon -zervativnim) pristupom. Hirur{ka intervencija manjegobima (amputacija u predelu stopala) je izvr{ena kod41, a opse`na hirur{ka intervencija (amputacija upre delu potkolenice) kod 29 pacijenata. Koncen tra -cije biohemijskih parametara i HbA1c) su odre|enestandardnim laboratorijskim metodama. U ispitivanojgrupi naju~estaliji faktor rizika za nastanak SDS (82%pacijenata) bila je lo{e regulisana glikemija (HbA1c)>8%). Utvrdili smo da su pacijenti koji su le~enihirur{kim putem imali statisti~ki zna~ajno vi{e vred-nosti HbA1c) u pore|enju sa pacijentima koji su le ~e -ni neoperativnim pristupom (P<0,05). Ukupan broj

P003SIGNIFICANCE OF GLYCOSILATED

HEMOGLOBIN DETERMINATIONFOR THE ASSESSMENT OF LOWER-EXTREMITY AMPUTATION RISK INPATIENTS WITH DIABETIC FOOT

Tatjana Stefanovi}1, Sre}ko Bosi}2, Jelena Veki}3, Vesna Spasojevi}-Kalimanovska3,

Zorana Jeli}-Ivanovi}3

1Clinical Chemistry Laboratory, General Hospital, Po`arevac

2Department of Surgery, General Hospital, Po`arevac3Department of Medical Biochemistry,

Faculty of Pharmacy, University of Belgrade

Glycosylated hemoglobin (HbA1c) is importantparameter for the assessment of long term metaboliccontrol and risk for development of chronic compli-cations in patients with diabetes mellitus (DM). Thisstudy investigates associations between achievedmetabolic control in DM with the risk for the develop-ment of diabetic foot (DF) and its treatment modali-ties. The study included 111 patients (80 men and31 women) with type 2 DM, aged 65.5 ± 9.8 years.DF lesions occurred 13.9 ± 8.8 years after diagnosisof DM. In 41 patients DF was treated by conservativeapproach, 41 patients underwent a minor limbamputation (foot amputation) and 29 patients under-went a major limb amputation (below-knee amputa-tion). The levels of HbA1c and serum biochemicalparameters were measured by standard laboratorymethods. The most frequent risk factor for DF in theexamined group (82% patients) was poor glycaemiccontrol (HbA1c >8%). The level of HbA1c value wassignificantly higher in surgically treated patients thanin conservative treated group (P<0.05). The numberof patients who underwent surgical treatment weresignificantly higher in group with poor glycaemic con-trol (62.8% patients with poor glycaemic control vs.

J Med Biochem 2018; 37 (2) 205

pacijenata koji je podvrgnut hirur{kom le~enju bio jezna~ajno ve}i u grupi sa lo{om glikoregula cijom(68,2% pacijenata sa lo{e regulisanom vs. 40% paci-jenata sa dobro regulisanom glikemijom; P<0,05).Utvrdili smo da je verovatno}a za amputaciju 3,2puta ve}a ukoliko pacijent ima lo{e regulisanuglikemiju (OR=3,21; 95%CI:1,18–8,69; P<0,05).Tako|e, rezultati su pokazali da rizik za amputacijuraste za 54% ukoliko se koncentracija HbA1c pove}aza 1% (OR=1,54; 95% CI:1,02–2,31; P<0,05).Na{i rezultati su pokazali da se odr`avanjem dobremetaboli~ke kontrole u tipu 2 DM smanjuje rizik zarazvoj SDS, ali i stepen invazivnosti postupka le~enjaSDS.

40% patients with optimal glycaemic control;P<0.05). We found that the patients with poor gly-caemic control had 3.2 times higher risk for amputa-tion (OR=3.21; 95%CI: 1.18–8.69; P<0.05). Also,our results have shown that an increase of HbA1clevel by 1% was associated with 54% higher risk foramputation (OR=1.54; 95%CI: 1.02–2.31; P<0.05).In conclusion, our results demonstrated that optimalmetabolic control in type 2 DM reduces the risk forDF development, as well as the invasiveness of DFtreatment.

P004IZVE[TAJ O SLU^AJU IGG NETOLERANCIJE HRANE

Despina Efremova Veskovska, V. Soleva, B. Jaglikovski, P. Desoski

PZU Avicena laboratorija, Skopje, Makedonija

IgG netolerancija hrane (ili alergija hrane tipaIII) je kada imunolo{ki sistem proizvodi specifi~na IgGantitela nakon konzumacije triger hrane. Ova IgGantitela mogu dovesti do inflamatornih procesa kojiizazivaju specifi~ne (npr. migrena, sindrom iritabilnogcreva, psorijaza, ekzem) ili nespecifi~ne (npr. umor,zadr`avanje vode, variranje telesne te`ine) simptome.IgG test hrane poma`e u ograni~avanju sumnje natipu hrane. Posle ovog testiranja preporuka je da seizbegava hrana sa pove}anim titrom IgG 10–12ne delja. Devetogodi{nje dete `alilo se na gubitakkose i nelagodnost u stomaku. Redovno se vakcini{e,u dobrom je op{tom stanju bez ikakve bolesti ili aler -gije. Dermatolo{ki nije bilo parazita ili mikrospora naskalpu. Napravio je sve biohemijske parametre zaautoimune bolesti (ANA, ANCA, anti dsDNA, RF...),enzime (AST, ALT) i total IgE. Rezultati su bili nega-tivni. Zatim, pedijatar predla`e da se testira IgG ne to -le rancija hrane. Na{a IgG netolerancija hrane sadr`i90 namirnica. Titar IgG bio je veoma visok za le{nike,p{enice i gluten; visok za kravlje mleko, krav lje kiselomleko, ov~ji sir, jaja i kikiriki, i pove}ani titar IgG bilisu za kravljeg sira, kozjeg sira, semena suncokreta,bibera, vanile i narand`e. Posle ovih rezultata, majkadeteta rekla je da pacijent u du`em vremenskomperiodu konzumira proizvode koji sadr`e le{nike(~okolade sa le{nicima), a istovremeno se poklapa saintenzivnijim gubicima kose. Mi smo mu dali pre-poruke za izbegavanje hrane sa visokim ti trom IgG utoku 3 meseca, a hrana sa IgG vredno stima u porastuda se uklju~uje u 4 dnevan ciklus rotacije. Posle 6meseci, dete je do{lo na kontrolu. Anamnesti~ki po -daci, koje je dala njegova majka, su bili da se pacijent

P004CASE REPORT OF IGG FOOD INTOLERANCE

Despina Efremova Veskovska, V. Soleva, B. Jaglikovski, P. Desoski

PHI Avicena Laboratory, Skopje, Macedonia

IgG food intolerance (type III food allergy) iswhen the immune system produces specific IgG anti-bodies after the consumption of a trigger food. TheseIgG antibodies can lead to inflammatory processescausing specific (e. g. migraine, Irritable Bowel Syn -drome, psoriasis, eczema) or unspecific (e.g. fatigue,retention of water, variation of body weight) symp-toms. An IgG food screen test helps to limit the sus-pecting food. After this testing the recommendationis to avoid foods with elevated IgG titer 10–12 weeks.9 years old twin child is complaining on hair loss anddiscomfort in the abdomen. He is regularly vaccinat-ed. He is in good general condition without any ill-ness or allergy. Dermatologically, there were no scalpparasites or microspores. He made all biochemicalparameters for autoimmune diseases (ANA, ANCA,anti dsDNA, RF...), the enzymes (AST, ALT) and totalIgE. The results were negative. Next, the pediatriciansuggests him for IgG food intolerance testing. TheIgG food screen plate contains 90 foods. The IgGtiter were very high for hazelnuts, wheat and gluten;high for cow’s milk, cow’s sour milk, sheep’s cheese,eggs and peanuts, and increased IgG titer were forcow’s cheese, goat’s cheese, sunflower seeds, pep-per, vanilla and orange. After this results child’smother said that the patient for a long period con-sume products that contains hazelnuts (chocolateswith hazelnuts), and at the same time it coincideswith a more intense hair loss. We gave him re -commendations for avoiding foods with high IgG titerfor 3 months and including foods with increased IgGvalues in 4 day rotating cycle. After 6 months, childcame into control. Anamnestic data, given by his

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pridr`avao preporuka i promene u rastu kose su bilevidljive. Posle 6 meseci, titar IgG hrane sa vi sokimvrednostima IgG I IgG u porastu zna~ajno je sma -njen. U skladu sa dobijenim rezultatima nakon IgGnetolerancija hrane, kod deteta vi dljive su kli ni~kepromjene.

mother, was that child have complied with the recom-mendations and there were visible changes in thescalp and hair growth. After 6 months, the IgG titerof foods with high and elevated IgG values were sig-nificantly decreased. In accordance with the obtainedcontrol results in the child, a visible clinical changesare observed.

P005BIOHEMIJSKA LABORATORIJA

TREBA DA OBJAVLJA VERIFICIRANE REFERENTNE

INTERVALE

Irena Kostovska, Gordana Bosilkova, Svetlana Cekovska, Verica Ristevska,

Katerina Tosheska-Trajkovska

Institut za medicinsku i eksperimentalnu biohemiju,Medicinski Fakultet, Skopje, Makedonija

Kvalitet referentnih intervala u klini~koj hemijije veoma va`an za tuma~e rezultata. ISO 15189ka`e da referentne rangove treba pregledati perio-di~no. Kako je u praksi nemogu}e izvesti ekstenzivnestudije za sve svoje testove, {iroko je prihva}eno dase ovi podaci preuzmu iz literature, proizvo|a~a, ilidrugih laboratorija. Cilj ove studije je da se uradi ve -rifikacija referentnih intervala naj~e{}ih laboratorij skihtestova (glukoze, lipida, urea, kreatinina, mo kra} neki seline, ukupnih proteina, albumina, kalcijuma, na -trijuma, kalijuma, neorganskog fosfata, i nekoliko en -zima) utvr|enih od strane proizvo|a~a testova za invitro dijagnozu (IVD), a koji se obavljaju u na{oj la -boratoriji. Za verifikaciju je primenjen pristup koji supreporu~ili IFCC and CLSI. Skupili smo 20 uzorakaod zdravih osoba koje smo analizirali sa standardnimmetodama na Gesan Chem 200 analizatoru, pri~emu smo prihvatili da 20 osoba prestavljaju lokalnupopulaciju, utvrdili da su pre-analiti~ki uslovi kao iranije, a da analiti~ki rad na dan testiranja pretstavljadugoro~ni analiti~ki performans. Skoro za sve testo -ve, 18 od 20 rezultata su bili unutar referentnih inter-vala koji su ranije bili prihva}eni i uklju~eni u izve{tajuo rezultatima. Samo za obe transaminaze, 3 rezulta-ta su bili van dometa, tako da smo po istom postup-ku analizirali novi izbor od 20 biolo{kih uzoraka.Samo je jedan uzorak bio van referentnog ranga,~ime su referentni intervali proizvo|a~a bili verifici-rani. Zaklju~ak je izveden iz IFCC preporuke: … pre-gled ili verifikacija objavljenih referentnih granicaomogu}ava prenos podataka proizvedenih od drugela boratorije ili IVD proizvo|a~a, koriste}i pri tomveoma jednostavan process validacije {to mo`e dabude od velike pomo}i.

P005BIOCHEMICAL LABORATORY

SHOULD REPORT VERIFICATED

REFERENCE INTERVALS

Irena Kostovska, Gordana Bosilkova, Svetlana Cekovska, Verica Ristevska,

Katerina Tosheska-Trajkovska

Department of Medical and ExperimentalBiochemistry, Medical Faculty, Skopje, Macedonia

The quality of the reference intervals in clinicalchemistry is very much important in result interpreta-tion. ISO 15189 states that reference ranges shouldbe reviewed periodically. As in practice it is impossi-ble for any single laboratory to perform their ownextensive studies for all of the tests it is widely accept-ed that this data may come from the literature, man-ufacturers, or other laboratories. The aim of thisstudy was to review the reference intervals of themost common laboratory tests (glucose, lipids, urea,creatinine, uric acid, total protein, albumin, calcium,sodium, potassium, inorganic phosphate, and severalenzymes) established by a manufacturer of in vitrodiagnostic devices (IVD) performed in our laboratory.Verification was done from a sample of apparentlyhealthy subjects using the approach proposed in therecommendations of the IFCC and CLSI. 20 sampleswere collect and analyzed with standard methods onGesan Chem 200 analyzer. The assumptions consid-ered were that the 20 individuals represented thelocal population, the pre-analytics conditions asbefore and analytical performance on the day of test-ing represented the long term analytical perform-ance. For almost all tests, 18 to 20 results were with-in the range that had been previously accepted to beincluded in the results report. Only for both transam-inases 3 results were out of range so a new selectionof 20 biological samples had been analyzed with thesame procedure, after which the proposed referencelimits were accepted with only one of 20 out ofrange. Conclusion is derived from IFCC recommen-dation: …it is proposed that only a »review or verifi-cation« of the published reference limits shall bemade transfer of data produced by other laboratoriesor IVD manufacturers, combined with a simple vali-dation process, could be a great help.

J Med Biochem 2018; 37 (2) 207

P006STATUS OKSIDATIVNOG STRESA

I GENSKA EKSPRESIJA IZOENZIMASUPEROKSID-DISMUTAZE KODPACIJENATA U TERMINALNOJ

FAZI BUBREŽNE BOLESTI

Ana Nini}1, Miron Sopi}1, Jelena Munjas1,Jelena Kotur-Stevuljevi}1, Nata{a Bogavac-Stanojevi}1, Sanja Simi}-Ogrizovi}2,3, MilicaKravlja~a2, Vesna Spasojevi}-Kalimanovska1

1Katedra za medicinsku biohemiju, Univerzitet u Beogradu – Farmaceutski fakultet, Srbija

2Klinika za nefrologiju, Klini~ki centar Srbije,Beograd, Srbija

3Medicinski fakultet, Univerzitet u Beogradu, Srbija

Brojne studije su ukazuju na to da su mnogifaktori uklju~eni u nastanak oksidativnog stresa kodpacijenata sa hroni~nom bubre`nom bole{}u (HBB)na hemodijalizi. Osim metaboli~kih i funkcionalnihporeme}aja mnogih organa koji se javljaju u HBB,sama procedura hemodijalize dodatno doprinosipove }anom oksidativnom stresu, koji se odnosi napove}anu produkciju reaktivnih jedinjenja kiseonika ioslabljenu enzimsku i ne-enzimsku antioksidativnuza{titu. Cilj ove studije je bio da se ispita povezanostgenske ekspresije izoenzima superoksid-dismutaze(Cu/Zn SOD i Mn SOD) u mononuklearnim }elijamaperiferne krvi (M PK) sa markerima oksidativnogstresa i antioksidativne za{tite kod pacijenata nahemodijalizi. Nivoi informacione ribonukleinskekiseline (iRNK) izoenzima SOD su odre|eni Real timePCR metodom. Totalni antioksidantni status (TAS),koncentracija tiobarbituratna kiselina-reaguju}ihsupstanci (TBKRS) i nivoi superoksid-anjon radikala(O2

.-) su, tako|e, odre|eni. Nivoi Cu/Zn SOD i MnSOD iRNK su bili zna~ajno ve}i kod pacijenata nahemodijalizi nego u kontrolnoj grupi (p<0,001 ip=0,011). Zna~ajno ve}e koncentracije TBKRS(p<0,001), nivoi O2

.- (p<0,001) i ni`i TAS(p<0,001) su dobijeni kod pacijenata na hemo -dijalizi u odnosu na kontrolnu grupu. Multivarijantnalinearna regresiona analiza je pokazala da je TASnezavisno povezan sa smanjenjem nivoa iRNK Cu/ZnSOD (b=-0,317, p=0,036) kao i sa smanjenjemnivoa iRNK Mn SOD (b=-0,331, p = 0,041). TASpred stavlja nezavistan prediktor za gensku ekspresijuizoenzima SOD u M PK.

P006OXIDATIVE STRESS STATUS

AND SUPEROXIDE DISMUTASEISOENZYMES’ GENE EXPRESSIONS

IN PATIENTS WITH END-STAGERENAL DISEASE

Ana Nini}1, Miron Sopi}1, Jelena Munjas1,Jelena Kotur-Stevuljevi}1, Nata{a Bogavac-Stanojevi}1, Sanja Simi}-Ogrizovi}2,3, MilicaKravlja~a2, Vesna Spasojevi}-Kalimanovska1

1Department of Medical Biochemistry, University of Belgrade – Faculty of Pharmacy, Serbia

2Clinic of Nephrology, Clinical Center of Serbia,Belgrade, Serbia

3School of Medicine, University of Belgrade, Serbia

Strong evidences indicate that many factors areinvolved in the pathogenesis of oxidative stress inpatients with chronic kidney disease (CKD) onhemodialysis. Besides metabolic and functional disor-ders of many organs which occur in CKD, thehaemodialysis procedure itself additionally con-tributes to enhancing oxidative stress, which involvesan increased production of reactive oxygen speciesand weakened enzymatic and non-enzymatic antiox-idant protection. The aim of this study was to exam-ine possible associations of superoxide dismutaseisoenzymes’ (Cu/Zn SOD and Mn SOD) geneexpressions in peripheral blood mononuclear cells(PBMC) with oxidative stress and antioxidative pro-tection parameters in patients on hemodialysis. SODisoenzymes messenger ribonucleic acid (mRNA) lev-els were determined by Real-time PCR method. Totalantioxidant status (TAS), thiobarbituric acid-reactivesubstances (TBARS) and superoxide anion radical(O2

.-) were also determined. Cu/Zn SOD and MnSOD mRNA levels were significantly higher inpatients on hemodialysis than in the control group(p<0.001 and p=0.011, respectively). The signifi-cantly higher TBARS (p<0.001), O2

.- (p<0.001)levels and significantly lower TAS levels (p<0.001)were found in hemodialysis patients than in controlgroup. Multivariate linear regression analysis demon-strated that TAS was independently associated with adecrease in Cu/Zn SOD mRNA (b=-0.317, p=0.036)and, as well as, with a decrease in Mn SOD mRNAlevels (b=-0.331, p=0.041). Only TAS could serveas an independent predictor for both SOD isoen-zymes gene expression levels in PBMC.

P007POVEZANOST

ANTROPOMETRIJSKIH PARAMETARA I LIPIDA U SERUMU KOD DECE

Ana Nini}1, Jelena Mladenovi}2, Vesna Spasojevi}-Kalimanovska1, Nata{a

Bogavac-Stanojevi}1, Jelena Kotur-Stevuljevi}1,Danijela Kornic-Ristovski3

1Katedra za medicinsku biohemiju, Univerzitet u Beogradu – Farmaceutski fakultet, Srbija

2Medicinska {kola, Beograd, Srbija3Dom zdravlja Pan~evo, Srbija

Aterosklerotski proces, hroni~na i inflamatornabolest, po~inje u detinjstvu i prethodi klini~kim ma -nifestacijama kao {to su kardiovaskularne bolesti kas-nije u `ivotu. Nepovoljan lipidni status, definisan kaopovi{eni nivoi holesterola u lipoproteinima niske gus-tine (LDL-c), triglicerida (TG) i smanjeni nivoi ho -lesterola u lipoproteinima visoke gustine (HDL-c), jeva`an faktor rizika za ubrzanu aterosklerozu u ranom`ivotu i obi~no je povezan sa gojazno{}u. Stoga, ciljove studije je bio da se utvrdi povezanost izme|u an -tro pometrijskih parametara i lipidnog statusa koddece. Studija je obuhvatila 186 zdrave dece (95de~aka i 91 devoj~icu) od 7 do 15 godina `ivota.Devoj ~ice i de~aci su podeljeni u podgrupe premagodinama starosti i indeksu telesne mase (ITM). Kon -centracije HDL-c i TG su odre|ene rutinskim enzim-skim metodama, a koncentracija LDL-c je izra~unataFriedewald-ovom formulom. ITM i odnos obima stru-ka i kuka (S/K) su izra~unati pomo}u formula. Poka -zane su zna~ajne razlike u koncentracijama HDL-c iLDL-c izme|u starosnih i ITM podgrupa devoj~ica.Kod devoj~ica, LDL-c je bio u pozitivnoj korelaciji saITM i obimom kukova. Kod de~aka, postojale su zna -~ajne razlike samo u koncentraciji HDL-c izme|urazli~itih starosnih podgrupa. Kod de~aka, HDL-c jebio u negativnoj korelaciji sa svim antropometrijskimparametrima osim sa S/K. Ordinalna regresiona ana -liza je pokazala nezavisnu negativnu prediktivnu ulogustarosti na koncentracije HDL-c kod oba pola. ITM jebio zna~ajan nezavisni prediktor za koncentracijeHDL-c i LDL-c samo kod devoj~ica.

P007RELATIONSHIPS

BETWEEN ANTHROPOMETRICPARAMETERS AND SERUM

LIPIDS IN CHILDREN

Ana Nini}1, Jelena Mladenovi}2, Vesna Spasojevi}-Kalimanovska1, Nata{a

Bogavac-Stanojevi}1, Jelena Kotur-Stevuljevi}1,Danijela Kornic-Ristovski3

1Department of Medical Biochemistry, University of Belgrade – Faculty of Pharmacy, Serbia

2Medical School, Belgrade, Serbia3Health Centre Pan~evo, Serbia

The atherosclerotic process, as chronic andinflammatory disease, begins at childhood and pre-cedes its clinical manifestations, such as cardiovascu-lar diseases later in life. Unfavourable lipid status,defined as elevated levels of low density lipoprotein-cholesterol (LDL-c), triglycerides (TG) and decreasedlevels of high density lipoprotein-cholesterol (HDL-c),have been well established as important risk factor foraccelerated atherosclerosis in early life and it is usu-ally associated with a high percentage of body fat.Therefore, the aim of this study was to determineassociations between anthropometric and lipid statusparameters in children. The study included 186apparently healthy children (95 boys and 91 girls)from 7 to 15 years of age. Girls and boys were divid-ed in age and in body mass index (BMI) subgroups.HDL-c and TG concentrations were assayed by rou-tine enzymatic methods and LDL-c concentration wascalculated by Friedewald formula. BMI and waist-to-hips ratio (WHR) were calculated. There were signif-icant differences in HDL-c and LDL-c in girls’ ageand BMI subgroups. In girls, LDL-c positively correlat-ed with BMI and hip circumference. In boys, therewere significant differences only in HDL-c concen -trations between different age subgroups. In boys,HDL-c negatively correlated with all anthropometricparameters except WHR. Ordinal regression analysisshowed independent negative predictive role of ageson HDL-c concentrations in both genders. BMI wassignificant independent predictor for HDL-c andLDL-c concentrations only in girls.

208

J Med Biochem 2018; 37 (2) 209

P008GENSKA EKSPRESIJA

RECEPTORA ZA KRAJNJE PRODUKTE GLIKACIJE I

TRANSFORMI[UĆI FAKTOR RASTA B1 KOD DECE SA TIPOM 1

DIJABETES MELITUSA

Dragana Bojanin1, Ana Nini}2, Marija Mihajlovi}2, Tatjana Milenkovi}3,

Ljubica Zatezalo1, Miron Sopi}2, Jelena Munjas2, Aleksandra Stefanovi}2,

Jelena Veki}2, Vesna Spasojevi}-Kalimanovska2

1Biohemijska laboratorija, Institut za zdravstvenuza{titu majke i deteta »Dr Vukan ^upi}«, Beograd2Katedra za medicinsku biohemiju, Univerzitet u

Beogradu – Farmaceutski fakultet, Beograd3Odeljenje za ispitivanje i le~enje `lezda sa

unutra{njim lu~enjem, Institut za zdravstvenu za{titumajke i deteta »Dr Vukan ^upi}«, Beograd

Dijabetes melitus tipa 1 (T1DM) je jedna odnaj~e{}ih autoimunih bolesti mladih. Hroni~nevaskularne komplikacije su glavni uzrok morbiditeta imortaliteta u T1DM. Pored toga {to su receptor zakrajnje produkte glikacije (RAGE) i transformi{u}ifaktor rasta (TGF-b1) uklju~eni u razvoj i progresijumikro- i makrovaskularnih komplikacija u dijabetesu,oni imaju va`nu ulogu i u regulaciji funkcija imun-skog sistema. Interakcija RAGE-AGE ima ulogu u re -gulaciji inflamatornih citokina, dok je TGF-b1 klju~niregulator u osiguranju imunolo{ke tolerancije. Ciljove studije je bio da se ispita ekspresija gena za RAGEi TGF-b1 u mononuklearnim celijama periferne krvikod 74 zdrava i 203 deteta sa T1DM. Nivoi informa-cione ribonukleinske kiseline (iRNA) RAGE i TGF-b1su odre|eni Real time PCR metodom. Tako|e,odre|eni su i osnovni biohemijski parametri i C-reak-tivni protein visoke osetljivosti. Nivoi iRNK RAGE iTGF-b1 bili su zna~ajno veci kod zdrave dece negokod dece sa T1DM. Univarijatna logisti~ka regre-siona analiza je pokazala zna~ajnu povezanost nivoaiRNK TGF-b1 sa prisustvom T1DM; OR=0,312,95% CI (0,215–0,453), p<0,001. Multivarijantnalogisti~ka regresiona analiza pokazala je da su nivoiiRNK TGF-b1 nezavisno povezani sa prisustvom T1DM;OR = 0,319, 95% CI (0,162–0,628), p<0,001 kadasu testirani u modelu sa drugim klini~kim markerima.Primenom logisti~ke regresione analize nije poka -zana zna~ajna povezanost izme|u nivoa iRNK RAGEi prisustva T1DM (p=0,699). Na{i rezultati bi moglida uka`u na poreme}aj regulacije ekspresije gena zaRAGE i TGF- b1 kod dece sa T1DM.

P008RECEPTOR FOR ADVANCED

GLYCATED ENDPRODUCTS ANDTRANSFORMING GROWTH FACTOR

B1 GENE EXPRESSIONS IN CHILDREN WITH TYPE 1

DIABETES MELLITUS

Dragana Bojanin1, Ana Nini}2, Marija Mihajlovi}2, Tatjana Milenkovi}3,

Ljubica Zatezalo1, Miron Sopi}2, Jelena Munjas2, Aleksandra Stefanovi}2,

Jelena Veki}2, Vesna Spasojevi}-Kalimanovska2

1Biochemical laboratory, Mother and Child HealthCare Institute »Dr Vukan ^upi}«, Belgrade

2Department of Medical Biochemistry, University of Belgrade –

Faculty of Pharmacy, Belgrade3Department of Endocrinology, Mother and ChildHealth Care Institute »Dr Vukan ^upi}«, Belgrade

Diabetes mellitus type 1 (T1DM) is one of themost frequent autoimmune diseases in youth.Chronic vascular complications are responsible formajor morbidity and mortality in T1DM. Although,rreceptor for glycated end products (RAGE) andtransforming growth factor-b1 (TGF-b1) are implicat-ed in developement and progression of diabeticmicro- and macrovascular complications, they alsohave important roles in immune system regulation.RAGE-AGE interaction is involved in regulation ofinflammatory cytokines and TGF-b1 is a key regula-tor of ensuring immune tolerance. The aim of thisstudy was to determine RAGE and TGF-b1 geneexpressions in peripheral blood mononuclear cells in74 healthy children and 203 children with T1DM.RAGE and TGF-b1 messenger ribonucleic acid(mRNA) levels were determined by Real-time PCRmethod. Basic biochemistry parameters and highsensitivity C-reactive protein were also determined.RAGE and TGF-b1 mRNA levels were significantlyhigher in control group than in children with T1DM.Univariate logistic regression analysis revealed signif-icant association of TGF-b1 mRNA levels with thepresence of T1DM, OR=0.312; 95%CI (0.215–0.453), p<0.001. Multivariate logistic regressionanalysis demonstrated that TGF-b1 was independ-ently associated with the presence of T1DM;OR=0.319, 95%CI (0.162–0.628), p<0.001 whenused in model with other clinical markers as con-founders. No significant association between RAGEmRNA levels and presence of T1DM (p=0.699)were determined in tested population. Our resultscould indicate gene expression dysregulation ofRAGE and TGF-b1 in children with T1DM.

P009 IgG – MARKER ZA

TOLERANCIJU HRANE

Violeta Soleva, D. E. Veskovska, B. Jaglikovski, P. Desoski

Avicena Laboratorija, Skopje, Macedonia

Postoje razli~iti razlozi zbog kojih hrana mo`eprouzrokovati probleme, me|u njima klasi~na IgEalergija na hranu i odlo`ena IgG alergija na hranu.Alergija hrane tipa III je kada imuni sistem proizvodispecifi~na IgG antitela protiv uobi~ajeno bezopasnihnamirnica. Imunolo{ki sistem la`no smatra da su to{tetne supstance, jer se crevni zid mo`e o{tetiti ilipostati propustljiv kao rezultat stresa, infekcije ililekova, a velike koli~ine neprilago|ene hrane mogupro}i kroz krvotok. IgG antitela i antigena formirajuimuni kompleksi koji se talo`e u organima i tkivima iaktiviraju sistem komplementa. Kada su imuni kom-pleksi razgra|eni, okolna tkiva su tako|e o{te}ena,{to dovodi do niskog stepena inflamatornog stanja.Re dovnim unosom hrane koja je u pitanju mo`e iza-zvati hroni~nu upalu. Pribli`no 45% ljudi ima netoler-anciju za hranu, mnogi od njih pate od hroni~nihsimptoma (gastrointestinalne, ko`ne, glavobolje,migrene, pre komerne te`ine itd.) sa odlo`enompojavom (nakon nekoliko sati ili ~ak i dana), {to bimoglo biti sma njeno ili eliminisano samo ako se utvr-di uzrok problema kroz testiranje netolerancije hrane.Na{ interes je da saznamo koja vrsta hrane naj~estodaje porast ili visok titar antitela slu~ajnim pacijenti-ma iz na{e zemlje. Analiza se radi sa ELISA meto -dom. IgG titar se mo`e kvantifikovati u nekolikogrupa (negativno, nisko, pove}ano i visoko). Na{imtestom se ana lizira 90 vrsta hrane. U na{oj laboratori-ji, analiza je uve|ena u januaru 2014. Od tada jeizvr{eno 740 analiza. Analizirali smo 160 randompacijenata na uzrast od 18 do 65 godina. Od svih 90vrsta hrane, na{i pacijenti su obi~no netolerantni nakravlje mle ko (p(pove}an)-10,6%, v(visok titar)-44,4%), kravlje kiselo mleko (p-11,9%, v-41,9%),kozji sir (p-16,3%, v-23,1), ov~i sir (p-15,0%, v-25,6%), jaja (p-18,1%, v-42,5%), p{enicu (p-26,3%,v-36,3%), gluten (p-22,5%, v-48,8%), badem (p-15,0%, v-21,9%), crni biber (p-37,5%, v-29,4%) ivanilu (p-11,3%, v-32,5%). Gluten je jedan od naj -~e{}ih uzroka intolerancije hrane. Pacijent mo`e daizbegava da jede problemati~nu hranu navedenu unalazima i zameni je hranom koju organizam toleri{e.

P009 IgG – MARKER FOR FOOD TOLERANCE

Violeta Soleva, D. E. Veskovska, B. Jaglikovski, P. Desoski

Avicena Laboratory, Skopje, Macedonia

There are various reasons why a food can causeproblems, among them a classic IgE food allergy(type I) and a delayed IgG food allergy (type III).Atype III food allergy is when the immune system (Bcells and plasma cells) produces specific IgG antibod-ies against usually harmless foodstuffs. The immunesystem falsely considers these to be harmful sub-stances, because the intestinal wall can be damagedor become permeable as a result of stress, infectionor medication and large quantities of undigestedfoods can make their way into the bloodstream. IgGantibodies and the antigens agglomerate to formimmune complexes, which adhere to organs and tis-sues and activate the complement system. When theimmune complexes are destroyed, the surroundingtissues are also damaged, which leads to low-gradeinflammatory condition. A regular intake of the con-cerned food may then cause a chronic inflammation.Approximately 45% of people have an intolerance toa food, many of these people suffer with chronicsymptoms (gastrointestinal, skin, headache, mig raine,overweight etc.) with delayed appearance (after afew hours or even days), that could be reduced oreliminated if only they could pinpoint the cause ofthe problem through food intolerance testing. Ourinterest is to find out which type of food often givesan increase or high titer of antibodies to randompatients from our country. Food screen analyses isworks whit ELISA method. IgG titer can be quantifiedin several groups (negative, low, increased and high).Our food screen analyses IgG antibody reactions to90 possible type of foods. In our laboratory, theanalysis was introduced in January 2014. Since then,740 analysis have been made. We analyzed 160randomized patients aged 18 to 65 years. Of all 90type of foods, our patients are usually in tolerant ofcow’s milk (increased-i 10.6% high-h 44.4%), cowsour milk (i-11.9% h-41.9%), goat cheese (i-16.3%h-23.1%), sheep cheese (i-15.0% h-25.6%), eggs(i-18.1% h-42.5%), wheat (i-26.3% h-36.3%), glu ten(i-22.5% h-48,8%), almond (i-15.0% h-21,9%), blackpepper (i-37.5% h-29.4%) and vanilla (i-11.3%h-32.5%). Gluten is one of the most common causesof food intolerance. The patient can avoid eating theproblematic foods listed in the findings and replacethem with foods he tolerates.

210

J Med Biochem 2018; 37 (2) 211

P010 PRAĆENJE PRE-ANALITI^KIH

GRE[AKA U KLINI^KOJ LABORATORIJI

Ana Marinkovi}, Mirjana Stanojkovi}, Dragana Miti}

Centar za medicinsku biohemiju, Klini~ki centar Ni{, Srbija

Procenjeno je da pre i post-analiti~ke gre{ke~ine 93% od svih gre{aka u laboratorijskom radu.Gre{ke u bilo kojoj fazi procesa prikupljanja, testira -nja i interpretiranja rezultata mogu da dovedu doozbiljne gre{ke u postavljanju dijagnoze pacijenta.Gre{ke tokom procesa prikupljanja nisu neizbe`ne,ali mogu se spre~iti primenom kontrole kvaliteta,kon ti nuiranog obrazovanja i efikasnosti sistema zaprikupljanje podataka. U ovom radu vr{ena je per-spektivna analiza dobijenih rezultata iz Centra za me -dicinsku biohemiju, Klini~kog centra u Ni{u, Srbija ogre{kama u preanalitickoj fazi i izvr{eno je sumiranjei obrada dobijenih podataka. Osoblje u laboratoriji jeevidentiralo gre{ke i sam prijem uzoraka koji su prih-vatani ili odbijani za dalju laboratorijsku obradu. Od48 328 pristiglih epruveta u laboratoriju za skrining uperiodu od 8 meseci, pre analiti~ke gre{ke su prime -} ene kod oko 4,9% od ukupnog broja primljenih uzo-raka. Zatim su izra~unati procenti razli~itih tipovapre-analiti~kih gre{aka. Ve}ina odbijenih uzoraka pri -stiglih na analizu u laboratoriju je usledila usled po -stojanja hemolize uzoraka, {to ~ini 1,1% od ukupnogbroja uzoraka pristiglih tokom ovog perioda. Koli~inakrvi koje nije bilo dovoljno za potpunu analizu je ~i -nila 0,08% svih evidentiranih gre{aka. Ukupno 0,4%od svih primljenih uzoraka sa odeljenja dono{eno jeu laboratoriju u pogre{noj epruveti. Ljudska uloga usakupljanu uzoraka krvi i drugih telesnih te~nostimo`e imati za cilj potpunu eliminaciju gre{aka u vezisa laboratorijskim ispitivanjima. Dobra praksa i uskla -|enost sa novim strategijama za spre~avanje gre{akamo`e dovesti do zna~ajnog smanjenja pre-analiti~kihgre{aka. Praksa vo|enja evidencije o gre{kama u svimfazama analize i korektivne strategije za njihovu pre-venciju mo`e postepeno spre~iti laboratorije od po -sto janja takvih gre{aka a samim tim i unaprediti kva -litet rada u biomedicinskim laboratorijama.

P010FOLLOWING THE PRE-ANALYTICAL

ERRORS IN THE CLINICAL LABORATORY

Ana Marinkovi}, Mirjana Stanojkovi}, Dragana Miti}

Centre for Medical Biochemistry, Clinical Centre of Nis, Serbia

Pre- and post-analytical errors are estimated toconstitute 93% of errors in the biomedical laboratory.Errors at any stage of the collection, testing andreporting process can potentially lead to a seriouspatient misdiagnosis. Errors during the collectionprocess are not inevitable but can be prevented witha diligent application of quality control, continuingeducation and effective collection systems. A per-spective analysis of the results obtained from the bio-medical laboratory of Clinical centre of Nis, Serbiafor errors of the preanalytical phase has been carriedout to summarize data. Laboratory personnel wereasked to register rejections, and causes for rejectionof wards. Out of the 48328 blood collection tubesscreened over a period of 8 months, pre-analyticalerrors were observed in approximately 4.9% of thetotal number of samples received. The distribution ofthe different types of errors was then calculated. Themajority of the rejected samples were haemolysed,which accounts for 1.1% of the total number of sam-ples received during this period. The amount ofblood was insufficient for complete analysis in0.08%. A total of 0.4% samples in the wards wereaccompanied by inappropriate requisition slips. Thehuman role in sample collection makes completeelimination of errors associated with laboratory test-ing unrealistic. However, good practise and compli-ance with the new strategies for error prevention canlead to a substantial reduction in pre-analyticalerrors. A practice of keeping a record of the errors atall stages of analysis and then devising correctivestrategies for their prevention can gradually free alaboratory from such errors.

P011UNAPRE\ENJE INTERNE

I EKSTERNE KOMUNIKACIJE U SLU@BI LABORATORIJSKE DIJAGNOSTIKE UVO\ENJEM

LABORATORIJSKOG INFORMACIONOG SISTEMA (LIS)

Jelena Trnavac, Danijela Ristovski-Kornic

Dom zdravlja Pan~evo

Komunikacija u okviru laboratorije je jedan odosnovnih vidova uspostavljanja kvalitetne saradnje iodnosa izme|u kolega, kao i odnosa sa drugim slu` -ba ma i deljenja informacija koje su bitne za samulaboratorijsku dijagnostiku pacijenata. Procedurom ointernoj i eksternoj komunikaciji usagla{ena su pravi-la u Domu zdravlja Pan~evo. Interna komunikacija seodvija izme|u zaposlenih u vezi sa informacijama opacijentima, uzorcima kao i dobijenim rezultatima inalazima za izdavanje. Vode se posebne sveske, kojeolak{avaju komunikaciju izme|u zaposlenih, kao {toje sveska hitnih pacijenata, sveska neusagla{enosti,sveska opisa seruma, sveska primopredaje smene.Standardizacijom i uvo|enjem procedure pobolj{avase kvalitet rada laboratorijske dijagnostike, putemevidentiranja podataka o pacijentima dobija se celo -kupna slika i me|usobna usagla{enost. Eksterna ko -mu nikacija se de{ava izme|u zaposlenih laboratorijesa pacijentima i lekarima. U Domu zdravlja Pan~evoona je redovna i regulisana procedurom, a odvija seu situacijama kada je potrebno izve{tava nje/tele fo -nom o alarmantnim laboratorijskim nalazima, kao ikad je uo~ena velika razlika izme|u dva uzastopnanalaza (delta ~ek). Svaki poziv se evidentira u svesciza obave{tavanje o alarmantnim vredno stima. Ovajvid komunikacije je usavr{en i pobolj{an uvo|enjemlaboratorijskog informacionog sistema (LIS-a), putemkojeg je mogu}a komunikacija lekara sa biohemi -~arem, uno{enjem napomena u elektronskom uputu.Na ovaj na~in i medicinski biohemi~ar mo`e komuni-cirati sa lekarom preko rezultata koji direktno sti`e uelektronski karton pacijenta posle va lidacije. Time jeubrzano dobijanje i izdavanje rezultata hitnih pacije-nata, pobolj{anje zadovoljstva pacijenata i efikasnoiz da vanje terapije od ordiniraju}eg lekara. Komuni ka -cija sa pacijentima se obavlja pu tem sveske evidenci-je o neusagla{enosti kao i putem napomena ukolikoje potrebno ponoviti va|enje krvi, uz mogu}u na po -menu ili sugestiju o pripremi pacijenta za ponovnouzorkovanje. Svakodnevna komunikacija izme|u pa -cijenata i laboratorijskog osoblja je utvr|ena eti~kimprincipima i pravima pacijenata, a to se evidentira uKnjizi utisaka koja je dostupna svim pacijentima i na -lazi se na vidnom mestu. Posle uvo |enja procedure injene primene, ura|ena je anketa zadovoljstva i po -bolj {anja komunikacije i saradnje izme|u biohemi -~ara i klini~ara na nivou Doma zdravlja Pan~evo, gde

P011IMPROVING INTERNAL

AND EXTERNAL COMMUNICATIONIN LABORATORY DIAGNOSTICS

BY INTRODUCING THELABORATORY INFORMATION

SYSTEM (LIS)

Jelena Trnavac, Danijela Ristovski-Kornic

Health Center Pan~evo

Communication within the laboratory is one ofthe basic aspects of establishing good cooperationand relationships between colleagues, as well as rela-tionships with other services and the sharing of infor-mation that are essential for the laboratory diagnos-tics of patients. The procedure for internal andexternal communication harmonized the rules in theHealth Center Pan~evo. Internal communicationtakes place between employees about patients, sam-ples, as well as the results obtained and the findingsfor the issue. Special notebooks are guided, whichfacilitate communication between employees, suchas emergency patients, non-compliant notebook,serum description notebook, shift handbook. By stan-dardizing and introducing the procedure, the qualityof laboratory diagnostics is improved, by recordingthe data on patients, the overall picture and mutualcompliance are obtained. External communicationoccurs between the laboratories with patients anddoctors. In the Pancevo Health Center it is commonand regulated by the procedure, and it occurs in sit-uations when reporting by phone about alarming lab-oratory findings, as well as when there is a big differ-ence between two successive findings (delta check).Each call is recorded in the Notification Alarm Alerts.This type of communication has been checked andimproved by the introduction of a laboratory informa-tion system (LIS), through which physician communi-cation with a biochemist is possible, by introducingnotes in an electronic instruction. In this way, a med-ical biochemist can communicate with a physicianthrough the results that directly arrive at the patient'selectronic card after validation. This is an acceleratedacquisition and release of the results of emergencypatients, improvement of patient satisfaction and effi-cient delivery of therapy by an ordinating physician.Communication with patient is done through a non-compliance record and as a reminder if blood extrac-tion is required, with a possible remark or suggestionon the patient's preparation for re-sampling. Dailycommunication between patients and laboratory staffis established by ethical principles and patients'rights, which is recorded in the Book of impressionsthat is available to all patients and is holded in a vis-ible place. Following the introduction of the proce-dure and its application, a survey of satisfaction andimprovement of communication and cooperation

212

J Med Biochem 2018; 37 (2) 213

su zabele`eni veoma pozitivni rezultati. Zaklju~ak jeda je standardizacija komunikacije u la bo ratorijamajedan od vrlo zna~ajnih faktora koji do prinosi obez be -|ivanju kvaliteta laboratorijskog ispitivanja i formira -nju baze podataka kao vida informacija koje su u naj -boljem interesu za kona~no zbrinjavanje pacijenata.

between biochemists and clinicians at the health cen-ter Pancevo was done and where very positive resultswere recorded. The conclusion is that standardiza-tion of communication in laboratories is one of themost important factors that contributes to ensuringthe quality of laboratory testing and the establish-ment of a database as the kind of information that isin the best interest for the ultimate care of patients.

P012PORE\ENJE VREDNOSTI

NATRIJUMA I KALIJUMA DOBIJENIHSA ANALIZATORA BECKMAN COULTER AU680® I ILYTE®

INSTRUMENTATION LABORATORY

Marija Andjelkovi}1,2, Ivana Nikoli}1, Marina Mitrovi}1, Ivanka Zelen1, Milan Zari}1,

Petar ^anovi}1, Sne`ana Radivojevi}2, Marijana Stanojevi} Pirkovi}1,2

1Univerzitet u Kragujevcu, Fakultet medicinskih nauka, Katedra za biohemiju

2Slu`ba za laboratorijsku dijagnostiku, Klini~ki centar Kragujevac, Srbija

Odre|ivanje elektrolitnog balansa je neophod-no za procenu ozbiljnih poreme}aja homeostaze.Na{ cilj je bio da se uporede vrednosti koncentracijanatrijuma i kalijuma dobijene sa analizatora AU680®

proizvodja~a Beckman Coulter i sistema Ilyte® proiz-vo|a~a Instrumentation Laboratory. Analizirano jeukupno 216 uzoraka pacijenata primljenih u Klini~kicentar Kragujevac. Za dobijanje seruma za svakogpacijenta kori{}ene su epruvete sa klot aktivatorom.Uzorci su prvo analizirani na Ilyte® sistemu, i odmahnakon toga na analizatoru AU680®. Oba analizatorakoriste isti metod – jon selektivne elektrode (ISE) kojiomogu}ava direktno merenje svakog jona od intere-sa u uzorku. Procena korelacije izme|u rezultata me -re nja je izvedena pomo}u Pearson-ovog testa ko re -lacije. Za pore|enje vrednosti ispitivanih parametaradobijenih sa razli~itih analizatora kori{}en je t-test zauparene uzorke. Vrednosti koncentracije natrijuma saoba analizatora koreliraju jedne sa drugim uz koefici-jent korelacije 0,639 (p<0,01). Koeficijent korelacijeza koncentracije kalijuma bio je vi{i 0,986 (p<0,01).Pore|enje vrednosti ispitivanih parametara dobijenihsa razli~itih analizatora ukazalo da nije bilo zna~ajnerazlike u koncentracijama natrijuma dobijenih saAU680® (M=136,44, SD=12,70) i Ilyte® (M=137,57,SD=8,01); t(215)=-1,704 (p=0,09). Rezultati kon-centracija kalijuma nisu pokazali zna ~aj nu razliku –AU680® (M=3,82, SD=1,18), Ilyte® (M=3,83,SD=1,23); t(215)=-0,258 (p<0,796). Na{i rezultatisu pokazali da izme|u rezultata koncentracija natriju-ma i kalijuma dobijenih sa analizatora AU680® iIlyte® sistema postoji slaganje.

P012COMPARISON OF SODIUM ANDPOTASSIUM LEVELS OBTAINED BY BECKMAN COULTER AU680®

ANALYZER AND ILYTE® SYSTEM BYINSTRUMENTATION LABORATORY

Marija Andjelkovic1,2, Ivana Nikolic1, Marina Mitrovic1, Ivanka Zelen1, Milan Zaric1,

Petar Canovic1, Snezana Radivojevic2, Marijana Stanojevic Pirkovic1,2

1University of Kragujevac, Faculty of MedicalSciences, Department of Biochemistry

2Department of Laboratory Diagnostics, Clinical Center Kragujevac, Serbia

Electrolytes balance is an essential measure-ment in the assessment of several homeostasis dis-ease. Our purpose was to compare the levels of sodi-um and potassium obtained by AU680® analyzer byBeckman Coulter, Inc. with the levels obtained usingILyte® system by Instrumentation Laboratory. A totalof 216 samples from the patients admitted in theClinical Center Kragujevac were analyzed. Serumsamples, using clot activator tubes, had been collect-ed from each participant. Samples were first pro -cessed by ILyte® system, and immediately thereafterby AU680® analyzer. Both analyzers used samemethod – Ion Selective Electrode (ISE) which offersdirect measurements for each ion of interest in thesample. The correlation study was performed usingPearson correlation test. A paired-samples t-test wasconducted to compare examined parametersobtained by different analyzers. Sodium levels quan-tified using both analyzers were significantly relatedwith correlation coefficient of 0.639 (p<0.01). Cor -relation coefficient for potassium was higher 0.986(p<0.01). The comparison of the values of examinedparameters obtained by two analyzers suggested thatthere was not a significant difference in the scores forNa+AU680® (M=136.44, SD=12.70) and Na+

ILyte® (M=137.57, SD=8.01); t(215)=-1.704(p<0.09). K+ results were not show significant dif -ference – K+ by AU680® (M=3.82, SD=1.18) andK+ by ILyte® (M=3.83, SD=1.23); t(215)=-0.258(p<0.796). Our results indicate acceptable concor-dance between the results of sodium and potassiumobtained from AU680® analyzer and ILyte® system.

214

P013NIVOI SERUMSKE KREATIN

KINAZE U HIPOTIREOZI

Mirjana Stanojkovi}, Ana Marinkovi}, Sne`ana Madi}, Tatjana Djordjevi}

Centar za medicinsku biohemiju, Klini~ki centar Ni{, Srbija

Cilj ovog istra`ivanja je da se utvrdi nivo se -rumske kreatin kinaze (CK) u izra`enoj i subklini~kojhipotireozi. Zatim da se istra`i promena nivoa CKnakon primenjene hormonske terapije i da se proceniodnos izme|u slobodnog trijodtironina (FT3), slobo-danog tiroksina (FT4) i tireotropnog hormona (TSH),kao i stepen o{te}enja skeletnih mi{i}a. Drugi uzrocipromene nivoa CK su bili isklju~eni. U istra`ivanje jebilo uklju~eno 26 pacijenata (24 `ene i 2 mu{karca,starosti 41,63 +/- 11,55 godina) sa izra`enom hipo -tireozom, 36 pacijenata (35 `ena, 1 mu{karac, sta -rosti 40,53 +/- 11,45 godina) sa subklini~kom hipo -tireozom, kao i 30 kontrola (27 `ena, 3 mu{karca,uzrasta od 39,71 +/- 11,10 godina). U serumu suodredjivani slede}i biohemijski parametri: TSH, FT4,FT3, i CK. Pove}an nivo serumske kreatin kinaze jeprona|en kod 17 pacijenata (58%) sa izra`enom hi -po tireozom i kod 4 pacijenta (10%) sa subklini~komhipotireozom. Iako je prona|eno statisti~ki zna~ajnopove}anje nivoa CK kod pacijenata sa izra`enomhipotireozom u pore|enju sa pacijentima sa subkli-ni~kom hipotireozom i kontrolama (p = 0,0001, p =0,01, respektivno), razlika je prona|ena izme|u sub-klini~ke hipotireoze i kontrolne grupe (p = 0,14).Kod hipotiroidnih pacijenata, utvr|ena je pozitivnakorelacija izme|u CK i TSH (R = 0,423; p = 0,04),i negativna korelacija izme|u CK i FT3 (R = 0,525;p = 0,002) i izme|u CK i FT4 (r = 0,435; p =0,04). Nivo serumske kreatin kinaze je smanjen nanormalan nivo posle primene adekvatne hormonsketerapije i funkcija {titne `lezde je normalizovana tret-manom. O{te}enje skeletnih mi{i}a je bilo izra`enijekod pacijenata sa izra`enom hipotireozom.

P013SERUM LEVELS OF CREATINEKINASE IN HYPOTHYROIDISM

Mirjana Stanojkovi}, Ana Marinkovi}, Sne`ana Madi}, Tatjana Djordjevi}

Centre for Medical Biochemistry, Clinical Centre of Nis, Serbia

The aim of this study was to determine serumlevels of creative kinase (CK) in overt and subclinicalhypothyroidism. To investigate the change in CK lev-els with treatment and to evaluate the relationshipbetween free triiodothyronine (FT3), free thyroxine(FT4), and thyrotropin (TSH) levels and the degree ofskeletal muscle involvement, as determined by serumCK levels. Patients with other causes of CK elevationwere excluded. We included 26 patients (24 womenand 2 men, ages 41.63 +/- 11.55 years) with overthypothyroidism, 36 patients (35 women, 1 man, ages40.53 +/- 11.45 years) with subclinical hypothy-roidism, and 30 age- and gender-matched controls(27 women, 3 men, ages 40.81 +/- 11.20 years) inthe study. Serum levels of TSH, FT4, FT3, and CKwere measured in all subjects. Creatine kinase eleva-tion was found in 17 patients (58%) with overt hypo -thyroidism and in 4 patients (10%) with subclinicalhypothyroidism. Although a statistically significantele vation of CK levels was found in patients with overthypothyroidism when compared with patients withsubclinical hypothyroidism and controls (p=0.0001,p = 0.01, respectively), no difference was foundbet ween the subclinical hypothyroidism and controlgroups (p = 0.14). In hypothyroid (overt and sub -clinical) patients, a positive corre lation was foundbetween CK and TSH (r = 0.423; p = 0.04), and anegative correlation between CK and FT3 (r = 0.525;p = 0.002) and between CK and FT4 (r =0.435; p= 0.04). Creatine kinase levels decreased to normallevels after thyroid function normalized with treat-ment. In conclusion, skeletal muscle is affected byhypothyroidism more profoundly in cases of overthypothyroidism, less so when subclinical hypothy-roidism is present.

J Med Biochem 2018; 37 (2) 215

P014LA@NO POVI[ENA

VREDNOST SLOBODNOG TIROKSINA IZMERENOG

ELEKTROHEMILUMINISCENTNOMMETODOM: PRIKAZ SLU^AJA

Tanja Stojakovic-Jelisavac1, Bosa Mirjanic-Azaric1,2, Ljiljana Grahovac3,

Svetlana Ignjatovic4,5

1Zavod za klini~ku laboratorijsku dijagnostiku,Univerzitetski klini~ki centar Republike Srpske,

Banja Luka, BiH 2Medicinski fakultet, Univerzitet u Banjoj Luci,

Banja Luka, BiH3Klini~ki zavod za nuklearnu medicinu i bolesti

{titne `ljezde, Univerzitetski klini~ki centar Republike Srpske, Banja Luka, BiH

4Katedra za medicinsku biohemiju, Farmaceutskifakultet, Univerzitet u Beogradu, Beograd, Srbija

5Centar za medicinsku biohemiju, Klini~ki centar Srbije, Beograd, Srbija

Prikazan je slu~aj `ene, u dobi od 65 godina, sadijagnozom Hypothyreosis supclinica, Thyroiditis chr.susp. i Struma multinodosa gl. thyroidea (dijagnozapostavljena prije devet godina), na redovnoj kontrolihormona tireotropina (TSH) i slobodnog tiroksina(FT4) u na{oj laboratoriji. Pacijentica je svih proteklihdevet godina imala uskla|ene nalaze hormona, saurednom klini~kom slikom. U serumu je izmerenavisoka koncentracija FT4 (>100 pmol/L), a normal-na vrednost TSH (3,67 mIU/L). Za merenje je ko -ri{ten elektrohemiluminiscentni immunoesej (ECLIA,Cobas e 601, Roche Diagnostics GmbH, Germany).U razgovoru sa ordiniraju}im lekarom dobijena jeinformacija da se na{ nalaz FT4 ne uklapa u klini~kusliku pacijenta. Tako|e, pacijentica nije uzimala bio -tin koji bi mogao interferirati u ECLIA metodi. Nakontri dana iz novog uzorka napravljen je kontrolni nalazkoji je bio isti. Uzorak smo sa~uvali, te naknadno ana -lizirali sa jo{ dve metode razli~itih proizvo|a~a (pre-poruka kod sumnje na interferencije). Dobijeni rez ul -tati za FT4 su bili na ARCHITECT ci8200 Abbott-u13,22 pmol/L, a na ADVIA Centaur-u 14,05 pmol/L(Siemens dijagnostika), dok nije bilo ve}ih promenau rezultatima TSH. Time smo potvrdili da se radi ointerferenciji u metodi ECLIA, najverovatnije uzroko-vanoj prisutno{}u heterofilnih antitela ili autoantitela.Interferencije koje se javljaju u imunohemijskim me -to dama neophodno je pravodobno uo~iti kako bi seizbeglo postavljanje pogre{ne dijagnoze kao i po gre{ -ne terapije.

Klju~ne re~i: slobodni tiroksin; interferencija;prikaz slu~aja.

P014FALSELY ELEVATED

VALUE OF FREE THYROXINE MEASURED BY

ELECTROCHEMILUMINESCENCEMETHOD: CASE REPORT

Tanja Stojakovic-Jelisavac1, Bosa Mirjanic-Azaric1,2, Ljiljana Grahovac3,

Svetlana Ignjatovic4,5

1Institute of Laboratory Diagnostic, University Clinical Centre of the Republic of Srpska,

Banja Luka, B&H 2Faculty of Medicine, University of Banja Luka, B&H

3Institute of Nuclear Medicine and Thyroid Gland Disease, University Clinical Centre

of the Republic of Srpska, Banja Luka, B&H

4Institute of Medical Biochemistry, Faculty ofPharmacy, University of Belgrade, Serbia

51Center for Medical Biochemistry, Clinical Centre of Serbia, Belgrade, Serbia

Here, it is shown the case of a 65-year-old fe -male patient diagnosed with Hypothyreosis subclini-cal, Thyroiditis chr. susp. and Struma multinodosa gl.thyroidea (this diagnosis was set nine years ago) onregular control of thyrotropin (TSH) and free thyrox-ine hormone (FT4) in our laboratory. The patient hashad harmonized hormonal findings with an orderlyclinical picture for the past nine years. The high con-centration of FT4 (> 100 pmol/L) and the normalvalue of TSH (3.67 mIU/L) were measured in theserum. Hormones were measured by electrochemi -luminiscence immunoassay (ECLIA, Cobas e 601,Roche Diagnostics, Mannheim, Germany. We gotinformation that our FT4 finding was inconsistentwith the clinical picture of the patient in conversationwith a doctor. Also, the patient did not take a biotinthat could interfere with the ECLIA. A control findingmade from a new sample was the same after threedays. We were preserved the sample and thenanalysed it with two other methods of different man-ufacturers (the recommendation when interference issuspected). The gained values for FT4 using ARCHI-TECT ci8200 Abbott Diagnostics were 13.22 pmol/Land the gained values for FT4 using ADVIA CentaurSIEMENS Diagnostics were 14.05 pmol/L while therewere no major changes in TSH results. Herewith, weconfirmed that this is interference most likely causeddue to the presence of heterophile antibodies orautoantibodies using the ECLIA method. It is neces-sary to perceive the interferences in immunochemi-cal methods on time in order to avoid misdiagnosisas well as wrong therapy.

Keywords: free thyroxine; interference; casereport.

216

P015PORE\ENJE DVA TESTA

ZA ODRE\IVANJE ANTI-XAAKTIVNOSTI

Nikola Milanovi}, Svetlana Obradovi}, Sne`ana Stevanovi}, Tatjana Stojakovi},

Ana Vuksanovi}

Zavod za laboratorijsku dijagnostiku BioMedica,Beograd, Srbija

Niskomolekularni heparini (LMWH) su lekovikoji se koriste u prevenciji i terapiji tromboembolijskihbolesti. Efekat ovih lekova se prati odre|ivanjem anti-Xa aktivnosti. U Zavodu za laboratorijsku dijagnos-tiku BioMedica su kori{}ena dva testa: Berichrom®

Heparin proizvo|a~a Siemens i STA®- LIQUID ANTI-Xa proizvo|a~a Stago. Anti-Xa aktivnost testom Beri -chrom® Heparin je odre|ivana end-point meto domna spektrofotometru StatFax, a STA®-LIQUID ANTI-Xa testom kineti~kom metodom na analizatoru STACompact Max. Kod 46 pacijenata koji primajuLMWH, odre|ivana je anti-Xa aktivnost i statisti~ki suobra|eni rezultati. Iz uputstava proizvo|a~a, anali -ti~ke karakteristike za Berichrom® Heparin i STA®-LIQUID ANTI-Xa su redom: analiti~ka osetljivost (zaoba testa je 0,10 IU/mL), merni opseg (zavisi od lotakalibratora, do 2 IU/mL) i preciznost (CV 4–7 %, CV2,9–3,7%). Neparametarskom korelacionom anali-zom je dobijen statisti~ki zna~ajan Spirmanov koefici-jent korelacije (r=0,776, p<0,01), {to ukazuje napozitivnu povezanost izme|u ova dva testa. Regre sio -nom analizom je dobijena jedna~ina prave: y= -0,01+ 0,79x (x: aktivnost dobijena Berichrom® Heparintestom, a y: aktivnost dobijena STA®-LIQUID ANTI-Xa testom). Statisti~ka analiza ukazuje da je anti-Xaaktivnost odre|ivana STA®-LIQUID ANTI-Xa testomproizvo|a~a Stago oko 20 % ni`a, {to je u dobroj ko -relaciji sa klini~kim efektom LMWH.

P015COMPARISON OF TWO ASSAYS

FOR DETERMINATION OF ANTI-XAACTIVITY

Nikola Milanovi}, Svetlana Obradovi}, Sne`ana Stevanovi}, Tatjana Stojakovi},

Ana Vuksanovi}

Institute for Laboratory Diagnostics BioMedica,Belgrade, Serbia

Low molecular weight heparins (LMWH) aremedicinal products used in prevention and treatmentof thromboembolic diseases. Pharmacological effectof these products is monitored by determining anti-Xa activity. At the Institute for Laboratory DiagnosticsBioMedica, two assays were used: Siemens Beri -chrom® Heparin and Stago STA®-LIQUID ANTI-Xa.Anti-Xa activity with Berichrom® Heparin assay wasdetermined using the endpoint method on theStatFax spectrophotometer, and with STA®-LIQUIDANTI-Xa assay using the kinetic method on the STACompact Max analyzer. The anti-Xa activity wasmeasured in 46 patients receiving LMWH and theresults were statistically processed. The analyticalperformance characteristics of Berichrom® Heparinand STA®-LIQUID ANTI-Xa are listed respectively:analytical sensitivity (for both assays is 0.10 IU/mL),measuring range (depending on the calibrator lot, upto 2 IU/mL) and precision (CV 4–7%, CV 2.9–3.7%).Non-parametric correlation analysis showed a statis-tically significant Spearman's correlation coefficient(r=0.776, p <0.01), indicating a positive correla-tion between these two assays. Comparison of assaysusing regression analysis obtained the followingequation: y= -0.01 + 0.79x (x: activity obtained byBerichrom® Heparin assay, and y: activity obtainedby STA®-LIQUID ANTI-Xa assay). Statistical analysisindicates that the anti-Xa activity obtained by theStago STA®-LIQUID ANTI-Xa assay is approximately20% lower, which correlates well with the clinicaleffect of LMWHs.

J Med Biochem 2018; 37 (2) 217

P016EFEKAT OLANZAPINA NA

MITOHONDRIJALNE KOMPLEKSE 1, 2 I 3 I ATP-SINTAZU

U TKIVU MOZGA

Ilker Taner Gumrukcuoglu, Birsen Bilgici, Ozgur Korhan Tuncel, Arzu Karaustaoglu

Katedra za klini~ku biohemiju, Medicinski fakultet,Univeritet Ondokuz Mayis, Samsun, Turska

Olanzapin pripada grupi lekova atipi~nih anti -psihotika. Antimani~ni i antipsihoti~ni efekti olanza -pina verovatno su posredovani blokadom dopamin-skih D2 i serotoninskih 5HT2A receptora, naro~ito umezolimbi~kom putu u mozgu. Pored toga, prime -}eni su i poreme}aji energetskog metabolizma ili dis-funkcije mitohondrija kod pacijenata sa bipolarnimporeme}ajem i {izofrenijom. Nije ta~no poznato da liolanzapin mo`e imati terapeutsko dejstvo na mito-hondrijalnu respiraciju u mo`danim }elijama. Iz ovograzloga, ova studija ima za cilj ispitivanje efekataolanzapina na mitohondrijalni kompleks 1, 2, 3 i 5ATP-sintaze u tkivima prednjeg desnog korteksamozga pacova. U na{oj studiji kori{}eno je 30 odra -slih vistar-albino pacova. Eksperimentalno, pacovi supodeljeni u dve grupe: kontrolnu i olanzapin grupu.Olanzapin i izotoni~ni fiziolo{ki rastvor davani suintra peritonealno tokom 30 dana. Na kraju eksperi-menta, pacovi su bili `rtvovani. Desni prefron talnikorteksi su potopljeni u SET pufer (250 mmol/L sa -ha roza, 2 mmol/L EDTA, 10 mmol/L TRIS pH 7,4).Homo genizacija je zatim izvr{ena u laboratoriji zabiohemiju. Frakcija supernatanta iz homogenata kojasadr`i delove mitohondrija uzeta je za biohemijskeanalize. Mitohondrijalni kompleksi 1, 2, 3 i ATP-sin-taza su odre|eni komercijalnim testovima, a proteiniLowry metodom. Prema rezultatima statisti~ke ana -lize, nije postojala statisti~ki zna~ajna promena zaparametre kompleksa 1, 2, 3 i aktivnosti ATP-sintazeizme|u kontrolne i olanzapinske grupe. U zaklju~ku,sada{nji rezultati pokazuju da tretman olanzapina nedovodi do promena u kompleksima 1, 2, 3 i ATP-sin-tazne aktivnosti transportnog lanca elektrona u tkivi-ma mozga pacova.

P016THE EFFECT OF OLANZAPİNE

ON MITOCHONDRIA COMPLEXES 1, 2, 3 AND ATP-SYNTASE

IN BRAIN TISSUE

Ilker Taner Gumrukcuoglu, Birsen Bilgici, Ozgur Korhan Tuncel, Arzu Karaustaoglu

Ondokuz Mayis University, School of Medicine,Department of Clinical Biochemistry, Samsun, Turkey

Olanzapine belongs to the group of medicinaldrugs called atypical antipsychotics. The primer anti-manic and antipsychotic effect of olanzapine are like-ly regulated by the blockade of dopamine D2 andserotonin 5HT2A receptors particularly in themesolimbic pathway in the brain. In addition, a dis-tubance of energy metabolism or disfunction of mito-chondria in patients with bipolar disorder and schizo-phrenia are observed frequently in these disorders. Itis exactly unknown whether olanzapine may havetheropeutic potential on mitochondrial respiration inthe brain cells. For this reason, the present study aimsat examining the effects of olanzapine on mitochon-drial complex 1, 2, 3 and 5 ATP-synthase in the brainright frontal cortex tissues of rats. In our study, 30adult wistar-albino rats were used. Experimentally,rats were divided into two groups: control and olan-zapine groups. Olanzapine and isotonic saline solu-tion were administered intraperitoneally for 30 days.At the end of the experiment, the rats were decapi-tated. Brain right prefrontal cortices were taken intoSET buffer (250 mmol/L sucrose, 2 mmol/L EDTA,10 mmol/L tris-base pH 7.4). Homogenization wasthen carried out in biochemistry laboratory. Super -natant fraction from homogenate containing mito-chondria parts were taken for biochemical analyzes.Mitochondria complex 1, 2, 3 and ATP-synthasewere measured by using kits and protein amount wasmeasured by using Lowry method. According to sta-tistical analyse results, no statistical alteration wasobserved for studied parameters complexes 1, 2, 3and ATP-synthase activities between the control andolanzapine groups. In conclusion the present resultsshow that olanzapine treatment does not lead to anychanges in complexes 1, 2, 3 and ATP-synthaseactivity of the electron transport chain in brain tissuesof rats.

218

P017PORE\ENJE NOVOG

POINT-OF-CARE INCLIXTM PCTTESTA I KOMPARATIVNE

IMUNOHEMIJSKE METODE ZAODRE\IVANJE PROKALCITONINA

Gordana Dmitra{inovi}1, Marija Sari}1,Branislava Coki}1, Sanja Obradovi}2,

Svetlana Ignjatovi}1

1Centar za laboratorijsku medicinu, Univerzitet u Beogradu – Farmaceutski fakultet, Srbija

2Slu`ba za laboratorijsku dijagnostiku, Klini~ko-bolni~ki centar »Zvezdara«, Beograd, Srbija

Prokalcitonin (PCT) se smatra osetljivim biomar -kerom za rano utvr|ivanje postojanja bakterijske infek-cije i sepse, jer u prisustvu sistemske infekcije do lazi dozna~ajnog i naglog porasta njegove koncentracije.Poslednjih godina posebna pa`nja se po klanja razvojubrzih testova za odre|ivanje PCT koji zna~ajno skra -}uju vreme dobijanja rezultata, {to omo gu}ava br`epostavljanje dijagnoze i zapo~injanje tretmana pacijen-ta. INCLIXTM PCT test se koristi na point-of-careINCLIXTM analizatoru (Sugentech, Inc., Korea) gdese kvantifiacija nivoa PCT vr{i analiziranjem slika dobi-jenih primenom imunohromatografske metode. Ciljovog rada je pore|enje rezultata INCLIXTM PCT testasa ve} postoje}om imunohemijskom metodom koja seprimenjuje u laboratoriji. Uzorci pacijenata (n = 47) susakupljeni u laboratoriji Klini~ko-bolni~kog centra»Zvezdara«, gde je nivo PCT izmeren upotrebomElecsys BRAHMS PCT testa ECLIA metodom (RocheDiagnostics). Odre|ivanje nivoa PCT uz pomo}INCLIXTM PCT testa izvr{eno je u Centru za laborato -rijsku medicinu Farmaceutskog fakulteta u Beogradu.Prema uputstvu proizvo|a~a, redom su navedene zapoint-of-care i imunohemijsku metodu analiti~ka oset -ljivost ( 0,13 ng/mL i 0,02 ng/mL) i dinami~ki opseg(0,25–40 ng/mL i 0,02–100 ng/mL). Pore|enje me -to da je izvr{eno uporednom analizom uzoraka ~ije sukoncentracije utvr|ene upotrebom obe navedene me -tode, isklju~uju}i uzorke PCT van dinami~kog opsega.Neparametarskom korelacionom analizom dobijen jestatisti~ki zna~ajan koeficijent korelacije (r=0,96,p<0,01), {to ukazuje na linearnu povezanost ispiti-vanih metoda. Primenom Passing i Bablok regresioneanalize dobijena je slede}a jedna~ina prave: y =0,02+1,19x (95%CI za odse~ak je -0,29–0,26,95%CI za nagib je 0,84–1,53). Dobijeni interval pouz-danosti od 95% za nagib prave i odse~ak ukazuju danema konstantnog i proporcionalnog odstupanja umerenju dve metode. Cusum-ovim testom linearnostinije utvr|eno zna~ajno odstupanje od linearnosti(p=1.00). Na osnovu prikazanih podataka mo`e sezaklju~iti da nema sistematskog odstupanja izme|urezultata dobijenih primenom INCLIXTM PCT testa iElecsys BRAHMS PCT zbog ~ega je mogu}a uporednaupotreba ispitivanih metoda.

P017COMPARISON OF A NEW

POINT-OF-CARE INCLIXTM PCT TESTAND COMPARATIVE IMMUNOASSAY

FOR PROCALCITONIN QUANTIFICATION

Gordana Dmitra{inovi}1, Marija Sari}1,Branislava Coki}1, Sanja Obradovi}2,

Svetlana Ignjatovi}1

1Center for Laboratory Medicine, University of Belgrade – Faculty of Pharmacy, Serbia

2Department of Laboratory Diagnostics, UniversityMedical Center »Zvezdara«, Belgrade, Serbia

Prokalcitonin (PCT) is considered as a sensitive bio-marker for early diagnose of bacterial infection andsepsis, due to its significant and quick increase in sys-tematic infection. During last several years there is agrowing interest for development of rapid tests forPCT determination, which would considerably short-en turnaround time, allowing faster diagnose andtreatment of patients. INCLIXTM PCT test is usedwith the point-of-care INCLIXTM Analyzer (Sugen -tech, Inc., Korea) which gives quantitative assess-ment of PCT by analyzing images obtained withimmunochromatographic method. The aim of thiswork was to compare the results of INCLIXTM PCTtest with the immunoassay currently used in labora-tory. Patient samples (n = 47) were collected in thelaboratory of University Medical Center »Zvezdara«,where PCT concentrations were measured usingElecsys BRAHMS PCT test with ECLIA method(Roche Diagnostics). Measurements of PCT usingINCLIXTM PCT test were conducted in The Centerfor laboratory medicine at The Faculty of pharmacyin Belgrade. According to manufacturers’ instruc-tions for point-of-care and immunochemistry assay,analytical sensitivity ( 0.13 ng/mL and 0.02 ng/mL)and dynamic range (0.25–40 ng/mL and 0.02–100ng/mL) are given respectively. Comparison of meth-ods was performed based on PCT results availablefrom both methods, excluding samples with PCT out-side the reportable range. Nonparametric correlationanalyses showed statistically significant coefficient ofcorrelation (r=0.96, p<0.01), indicating linear cor-relation of examined methods. The Passing andBablok regression analysis gave the following regres-sion equation: y=0,02+1,19x (95%CI for interceptwas -0.29–0.26, 95%CI for slope was 0.84–1.53).Based on 95%CI for intercept and slope, it can beconcluded that there is neither constant nor propor-tional difference between two methods. Cusum testfor linearity indicates no significant deviation from lin-earity (p=1.00). According to presented data, it canbe concluded that there is no systematic deviationbetween results determined by INCLIXTM PCT testand Elecsys BRAHMS PCT, so both methods can beused interchangeably.

XXI Srpski kongres medicinske i laboratorijske medicine

sa me|unarodnim u~e{}em

XXI Serbian Congress of Medical Biochemistry and Laboratory Medicine

with international participation

Posteri studenata/Students Posters

220

P001 ODRE\IVANJE

ANTIOKSIDATIVNE AKTIVNOSTIDODATAKA ISHRANI NA

BAZI GLJIVA

Dimitrije Mitrovi}, Jelena Pavlovi}, Bojana Vidovi}, Nevena Dabeti}

Katedra za bromatologiju, Univerzitet u Beogradu – Farmaceutski fakultet, Srbija

Gljive imaju dugu istoriju medicinske upotrebe.Cenjene su zbog svog izvrsnog ukusa i medicinskihsvojstava. Divlje ili kultivisane, pored svojih nutritivnihsvojstava, povezane su sa zna~ajnim antioksidativnimosobinama zbog sadr`aja bioaktivnih jedinjenja, kao{to su polisaharidi, polifenoli, vitamini i minerali. Ciljovog rada bio je da se ispita antioksidativna aktivnostdodataka ishrani na bazi gljiva na na{em tr`i{tu, kojise koriste primarno kao izvor beta glukana saimunomodulatornim svojstvima. Istra`ivanje je obuh-vatilo 8 komercijalnih dodataka ishrani i 5 razli~itihsirovina na bazi gljiva koje slu`e za proizvodnju do da -taka ishrani. Pripremane su tri vrste ekstrakata: vo -deni, metanolni (80%) i etanolni (50%). Odre|ivan jesadr`aj ukupnih polifenolnih jedinjenja, kao i anti -oksi dativna aktivnost FRAP, ABTS i CUPRAC spektro-fotometrijskim metodama. Antioksidativni kompozitniindeks (ACI) dobijen je ra~unskim putem. Sadr`ajukupnih polifenolnih jedinjenja je bio najve}i uuzorcima dodataka ishrani na bazi ^aga i Reishi glji-va. Najvi{e vrednosti ukupnih polifenolnih jedinjenjautvr|ene su u etanolnim (50%) ekstraktima i kretalesu se u rasponu 382,8–1508,0 mgGAE/g. Sva tritesta za ispitivanje antioksidativne aktivnosti supokazala konzistentne rezultate, pri ~emu su uzorcisa ve}im sadr`ajem polifenolnih jedinjenja pokazali inajve}u antioksidativnu aktivnost. Utvr|ene su sta-tisti~ki zna~ajno vi{e vrednosti ACI za 80% metanolneekstrakte u odnosu na vodene i 50% etanolneekstrak te (p<0,05). U sve tri vrste analiziranih ekstra -kata, utvr|ena je pozitivna Spearman-ova korelacijaizme|u ukupnog sadr`aja polifenolnih jedinjenja ivrednosti ACI (r=0,883 za vodu, r=0,889 za 80%metanol i r=0,950 za 50% etanol, p<0,0001 za svatri). Rezultati istra`ivanja ukazuju na postojanje anti -oksidativnog potencijala dodataka ishrani na bazi glji-va kao i na njihove mogu}e zdravstveno protektivneefekte.

P001 DETERMINATION OF

ANTIOXIDATIVE ACTIVITY OFDIETARY SUPPLEMENTS BASED

ON MUSHROOMS

Dimitrije Mitrovi}, Jelena Pavlovi}, Bojana Vidovi}, Nevena Dabeti}

Department of Bromatology, University of Belgrade – Faculty of Pharmacy, Serbia

Mushrooms have a long history of medical use.They are appreciated for their excellent taste andmedical properties. Wild or cultivated, in addition totheir nutritional properties, they are associated withsignificant antioxidant properties due to the contentof bioactive compounds, such as polysaccharides,polyphenols, vitamins and minerals. The aim of thisstudy was to investigate the antioxidant activity ofnutritional supplements based on mushrooms on ourmarket, used primarily as a source of beta glucanwith immunomodulatory properties. The studyincluded 8 commercial nutritional supplements and5 different mushroom-based raw materials for theproduction of nutritional supplements. Three types ofextracts were prepared: aqueous, methanolic (80%)and ethanolic (50%). The content of total polypheno-lic compounds as well as the antioxidative activity ofFRAP, ABTS and CUPRAC was determined with spec-trophotometric methods. The antioxidative compos-ite index (ACI) was obtained by calculation. The con-tent of total polyphenolic compounds was the highestin the samples of dietary supplements based onChaga and Reishi mushrooms. The highest values oftotal polyphenolic compounds were found ineth nolic (50%) extracts in the range of 382.8-1508.0 mgGAE/g. All tests for antioxidant activitytesting showed consistent results, where sampleswith higher content of polyphenolic compoundsshowed the highest antioxidant activity. Statisticallysignificantly higher ACI values for 80% methanolextracts relative to aqueous and 50% ethanolicextracts (p<0.05) were determined. In all three typesof extracts, a positive Spearman correlation betweenthe total content of the polyphenolic compounds andthe ACI value was observed (r=0.883 for water,r=0.889 for 80% methanol and r=0.950 for 50%ethanol, p<0.0001 for all three). The results of thestudy indicate the existence of an antioxidant poten-tial of nutritional supplements based on mushrooms,as well as their possible health-protective effects.

UDK 577.1 : 61 ISSN 1452-8258

J Med Biochem 37: 220–237, 2018 Poster sessionsPosterske sekcije

J Med Biochem 2018; 37 (2) 221

P002 PROTEKTIVNI EFEKAT

POVR[INSKI MODIFIKOVANIHNANO^ESTICA TIO2 KAFEINSKOM

KISELINOM NA O[TE]ENJA DNK IN VITRO

Autori: Milica Nemoda, Tamara Trajkovi} e-mail: milican92ªgmail.com Mentori: Prof. dr Lada @ivkovi},

dr Vesna Lazi}

Katedra za fiziologiju, Farmaceutski fakultetUniverzitet u Beogradu

Uvod: Kafeinska kiselina (KK) je prirodni antiok-sidant i zahvaljuju}i sposobnosti uklanjanja slobodnihradikala ostvaruje svoj antigenotoksi~ni efekat.Upotreba antigenotoksi~nih jedinjenja posredstvomnano~estica je namenjena pove}anju njihove bio-raspolo`ivosti uz odr`anje njihovih karakteristika.

Cilj rada: Cilj ove studije je bio da se, po prviput, uporede antigenotoksi~ni efekti KK i ST kom-pleksa (TiO2/KK), pod identi~nim eksperimentalnimuslovima.

Materijal i metode: Nano~estice 45-Å TiO2 save}im udelom kristali~nosti anataza oblika su cinteti-zovane hidrolizom titanijum (IV) hlorida. Sinteti zo - vane nano~estice TiO2 su okarakterisane pomo}upre nosne elektronske mikroskopije i rentgenske di -frak cione analize. Formiranje kompleksa sa preno-som naelektrisanja (charge transfer (CT) kompleks)izme|u povr{inskih Ti atoma i KK identifikovano jeneposrednom pojavom crvene boje. Spektro foto me -trijska D@ob metoda – metod kontinuiranih varijacija- primenjena je za odre|ivanje sastava ST kompleksa.Prilikom evaluacije antigenotoksi~nosti, pra}ena jesposobnost rastvora KK, nano~estica TiO2 i CT kom-plesa da smanje broj }elija sa H2O2 indukovanimDNK o{te}enjima na leukocitima pune krvi, in vitro.Stepen o{te}enja DNK procenjen je primenomkomet testa.

Rezultati: Kao {to je o~ekivano, rezultati supokazali da je prisustvo KK izazvalo zna~ajno sman-jenje o{te}enja DNK u opsegu upotrebljenih koncen-tracija. Sli~ni rezultati su dobijeni i za ST kompleks.

Zaklju~ak: Mo`emo zaklju~iti da povr{inskimodifikovane nano~estice TiO2 sa KK ili sli~nim jedi -njenjima mogu biti iskori{}ene za pobolj{anje njihovebioraspolo`ivosti uz odr`anje korisnih karakteristika,jer zna~ajne razlike izme|u antigenotoksi~nih svojsta-va KK i ST kompleksa nisu prime}ene.

Klju~ne re~i: kafeinska kiselina; nano~esticeTiO2; antigenotoksi~nost; antioksidativna svojstva

P002 PROTECTIVE EFFECT OF SURFACE-MODIFIED TIO2

NANOPARTICLES WITH CAFFEICACID AGAINST DNA DAMAGE

IN VITRO

Authors: Milica Nemoda, Tamara Trajkovi}e-mail: milican92ªgmail.com

Mentors: Assoc. Prof. dr Lada @ivkovi}, dr Vesna Lazi}

Department of Physiology, Faculty of PharmacyUniversity of Belgrade

Inroduction: The caffeic acid (CA) is a naturalantioxidant and its radical scavenging properties alsoappear to be responsible for its antigenotoxic proper-ties. Nanoparticle-mediated delivery of antigenotoxiccompounds is intended to increase their bioavailabil-ity while maintaining their effectiveness.

The Aim: The aim of this study was to compare,for the first time, antigenotoxic effects of free andbound CA to the surface of TiO2 NPs under identicalexperimental conditions.

Material and Methods: Colloids consisting ofthe 45-Å TiO2 nanoparticles (NPs) with anatase crys-tal structure were prepared by acidic hydrolysis ofTiCl4. The synthesized TiO2 NPs were characterizedusing transmission electron microscopy and X-ray dif-fraction analysis. The interstitial charge transfer (ICT)complex formation between surface Ti atoms and CAis indicated by immediate appearance of red color.The spectrophotometric Job’s method the continualvariations method was applied for determination ofthe composition of ICT complex. During the evalua-tion of antigenotoxicity, the ability of free CA, TiO2NPs and ICT complex to reduce the number of cellswith H2O2-induced DNA damage in leukocytes ofwhole blood cells was observed, in vitro. The level ofDNA damage was evaluated by comet assay method.

Results: As expected, the data indicated thatthe presence of free CA induced significant reductionof DNA damage in entire investigated concentrationrange. Similar results were also obtained for surface-modified TiO2 NPs with CA.

Conclusion To summarize, we suggest that sur-face-modified TiO2 NPs with CA and/or similar com-pounds can be used to improve their bioavailabilitywhile maintaining its beneficial activities, since nosignificant differences between the antigenotoxicproperties of free and bound CA to the TiO2 NPswere noticed.

Keywords: ascorbic acid; TiO2 nanoparticles;antigenotoxic properties; antioxidative properties

222

P003 ANTIOKSIDATIVNA AKTIVNOST

KOMERCIJALNIH ^AJEVA IPROIZVODA NA BAZI ^AJA

Autori: Andrea Aleksi}, Marija Mladenovi},Du{an Jovanovi}

e-mail: mimadeaªhotmail.com marija.a.mladenovicªgmail.com

duschan951ªgmail.com Mentori: Prof. dr Bri`ita \or|evi},

Asistent Vanja Todorovi}

Katedra za Bromatoligiju, Farmaceutski fakultetUniverziteta u Beogradu

Uvod: Oksidativni stres, kao poreme}aj ravno -te`e oksidanasa i antioksidanasa, igra va`nu ulogu upatogenezi razli~itih hroni~nih i degenerativnih bo -lesti. Odre|eni dijetetski suplementi privla~e sve ve}upa`nju kao potencijalni agensi za prevenciju mnogihbolesti uzrokovanih oksidativnim stresom. Osim toga,postoji zna~ajno interesovanje za utvr|ivanje ukup -nog sadr`aja fenola i antioksidativnog kapacitetarazli~itih vrsta ~ajeva kao namirnica koje zna~ajno do -prinose ukupnom dijetarnom unosu antioksidanasa.

Cilj rada: Cilj rada je odre|ivanje antioksidativneaktivnosti ~ajeva i biljnih dijetetskih suplemenata nabazi ekstrakta lista zelenog ~aja. Tako|e, odre|ivan jesadr`aj ukupnih polifenola i flavonoida koji zna~ajnou~estvuju u antioksidativnosti ovih proizvoda.

Materijali i metode: U eksperiment je bilo uklju -~eno sedam vrsta biljnih i pet vrsta vo}nih ~ajeva, kaoi tri vrste biljna suplementa. Za procenu sadr`ajaukupnih polifenola i flavonoida kori{}eno je spektofo-tometrijsko odre|ivanje na polistirenskim mikrotitra-cionim plo~ama. Za odre|ivanje antioksidativnog ka -paciteta kori{}eni su DPPH, ABTS, FRAP i CUPRACtest, te da bi se upotpunio profil antioksidativnogkapaciteta ~ajeva i suplemenata na bazi zelenog ~aja,primenjena su dva matemati~ka modela – ukupniantioksidativni skor i relativni indeks antioksidativnogkapaciteta.

Rezultati: Rezultati eksperimenta su pokazali davo}ni ~ajevi imaju ve}i sadr`aj polifenola i flavonoida,kao i antioksidativni kapacitet po dozi u odnosu nabiljne ~ajeve i analizirane suplemente.Me|utim, anal-izom rezultata antioksidativne aktivnosti, kori{}enjemdva pomenuta modela, pokazano je da je ukupniantioksidativni skor suplemenata ve}i od ukupnogantioksidativnog skora biljnih ~ajeva.

Zaklju~ak: Shodno dobijenim rezultatima mo`ese izvesti zaklju~ak da doza vo}nih ~ajeva ima protek-tivnije dejstvo u smislu antioksidativnog kapaciteta isadr`aja polifenola i flavonoida, u odnosu na analizi -rane biljne ~ajeve i suplemente. Osim toga, analizomrezultata dobijenih upotrebom novih mate mati~kih

P003 ANTIOXIDATIVE ACTIVITY OF

COMMERCIAL TEAS AND DIETARYSUPPLEMENTS BASED ON TEA

Authors: Andrea Aleksi}, Marija Mladenovi},Du{an Jovanovi}

e-mail: mimadeaªhotmail.com marija.a.mladenovicªgmail.com

duschan951ªgmail.com Mentors: Full Prof. Bri`ita \or|evi},

TA Vanja Todorovi}

Department of Bromatology, Faculty of PharmacyUniversity of Belgrade

Introduction: Oxidative stress plays an impor-tant role in pathogenesis of chronic and degenerativediseases. Certain dietary supplements are drawingattention based on their antioxidant potential, whichsignifies that they can be used in prevention of dis-eases caused by oxidative stress. Besides that, thereis growing interest in determination of total polyphe-nol and flavonoid content and antioxidant activity innumerous types of tea, as foods that significantlycontribute to total dietary antioxidants intake.

The Aim: Determine antioxidant activity of vari-ous tea types and dietary supplements based ongreen tea leaves, along with total polyphenol andflavonoid content, which partially participate in theirantioxidant potential.

Material and Methods: This study was conduct-ed on seven types of herbal, five types of fruit tea andthree dietary supplements based on green tea leaves.Total polyphenol and flavonoid content were deter-mined using spectrophotometry in polystyrenemicrotitration plates. DPPH, ABTS, FRAP and CUP -RAC assays were used to investigate antioxidantcapacity and two more mathematical models wereapplied to provide a more complete antioxidant pro-file – relative antioxidant capacity index (RACI) andglobal antioxidant score (GAS).

Results: In comparison to herbal teas andanalysed supplements, results from this study showedthat fruit teas have had higher polyphenol andflavonoid content, as well as antioxidant capacity perone dose. However, the results from applying men-tioned mathematical models show higher GAS valuesin supplements than the herbal teas.

Conclusion: From the results of this study it canbe concluded that a dose of fruit tea (in comparisonto the herbal teas and analysed supplements) has amore protecting effect in regard to antioxidantcapacity, along with antioxidant protection frompolyphenols and flavonoids. Also, higher GAS valuesin analysed supplements bring us to conclusion that,

J Med Biochem 2018; 37 (2) 223

modela koji otvaraju nove mogu}nosti istra`ivanjaantioksidativnog kapaciteta hrane i pi}a, dolazi se dozaklju~ka da kod analiziranih suplemenata postojiverovatno}a prisustva drugih antioksidativnih jedin-jenja u uzorcima suplemenata.

Klju~ne re~i: ~ajevi; suplementi; antioksidativnaaktivnost; polifenoli; flavonoidi

probably, those supplements a presence of anotherantioxidans.

Keywords: tea; supplements; antioxidant activi-ty; polyphenols; flavonoids

P004 ODRE\IVANJE NIVOA OLOVA

KOD STANOVNIKA BEOGRADA:PILOT ISTRA@IVANJE HUMANOG

BIOMONITORINGA

Autori: Lidija Kostadinov, Sne`ana Kuzmanovi}e-mail: lidijavkostadinovªyahoo.com,snezana.kuzmanovic10ªgmail.com

Mentor: doc. dr Danijela \uki}-]osi}, mag. farm.–med. biohem. Katarina Barali}

Katedra za toksikologiju »Akademik Danilo Soldatovi}«,

Farmaceutski fakultet Univerziteta u Beogradu

Uvod: Humani biomonitoring je metod kojim seprocenjuje izlo`enost ljudi hemikalijama odre|iva -njem njihovog sadr`aja u biolo{kim uzorcima. Inten -zivno se sprovodi na podru~ju Evropske unije kako bise utvrdili referentni nivoi za razli~ite zemalje. Me|uhemikalijama od zna~aja za humani biomonitoring sutoksi~ni metali, posebno olovo (Pb), koje zbog svojih{tetnih efekata predstavlja op{ti zdravstveni problem.Prema Svetskoj zdravstvenoj organizacije ne postojebezbdeni nivoi Pb u krvi jer su brojne studije pokazalepovezanost Pb i prevalence za nastanak o{te}enja naskoro svih organskim sistemima, posebno nervnomsistemu i kardiovaskularnom sistemu.

Cilj rada: Odrediti sadr`aj Pb kod stanovnikaBeograda i uporediti dobijene vrednosti sa nivoimatoksi~nog metala u evropskim zemljama.

Materijal i metode: Istra`ivanje je uklju~ilo 52osobe, oba pola, starosti izme|u 30 i 74 godine koje`ive na teritoriji grada Beograda. Uzorkovana je ven-ska krv i izdvojeni eritrociti (99% Pb se nalazi u eritro -citima). Nakon mineralizacije uzoraka u mikrotalas-noj pe}nici (START D, Milestone, SAD) u prisustvuHNO3 i H2O2 (7:1; v/v), koncentracija Pb je odre -|ena metodom ICP-MS (masena spektrometrija s in -duktivno spregnutom plazmom).

Rezultati: Izmerene vrednosti Pb su se kretale uopsegu od 0,4 do 34 mg/dL, {to je zna~ajno vi{e uodnosu na nivoe toksi~nog metala u populacijama

P004 DETERMINATION OF LEAD LEVELS

IN BELGRADE POPULATION: A PILOT STUDY OF HUMAN

BIOMONITOING

Authors: Lidija Kostadinov, Sne`ana Kuzmanovi}e-mail: lidijavkostadinovªyahoo.com,snezana.kuzmanovic10ªgmail.com

Mentors: Assist. Prof. Danijela \uki}-]osi}, M. Pharm.-Med. Biochem. Katarina Barali}

Department of Toxicology »Akademik Danilo Soldatovi}«,

Faculty of Pharmacy University of Belgrade

Introduction: Humane biomonitoring is amethod for assessing human exposure to chemicalsby determining their content in biological samples. Itis intensively implemented in European Union todetermine reference levels for different countries.Among important chemicals for human biomonitor-ing are toxic metals, especially lead (Pb). Its harmfuleffects are a general health problem. There are nosafe levels of Pb in the blood according to the WorldHealth Organization. Numerous studies have shownthe association of Pb and the prevalence of damageto almost all organ systems, especially the nervoussystem and cardiovascular system.

The aim: To determine Pb content in Belgradepopulation and to compare the obtained values withthe levels of Pb in European countries.

Methods: The study included 52 individuals, ofboth sexes, aged between 30 and 74 years living inBelgrade. Venous blood samples were taken anderythrocytes were separated (99% Pb is found inerythro cytes). After mineralization of samples in amicro wave oven (START D, Milestone, SAD) in thepresence of HNO3 and H2O2 (7:1; v/v), the concen-tration of Pb was determined by ICP-MS (inductivelycoupled plasma mass spectrometry).

Results: Pb values ranged from 0.4 to 34 mg/dL,which is significantly higher than the levels of toxicmetals in the populations of European countries

224

evropskih zemalja (2,47–11,7 mg/dL u Belgiji, 2,48-22,14 mg/dL u ^e{koj, 4,11–12,39 mg/dL u Poljskojitd). Nivoi Pb kod stanovnika Beograda su bliski dobi-jenim vrednostima u Hrvatskoj, {to je u saglasnosti sa~injenicom da su Hrvatska i Srbija me|u poslednjimzemljama u Evropi zabranile upotrebu olovnog benz-ina. Sadr`aj Pb kod mu{karaca je zna~ajno vi{i negokod `ena (p<0,05). Osim toga, uo~ene su zna~ajnerazlike u sadr`aju Pb kod `ena razli~ite starosne dobi(p<0,05).

Zaklju~ak: Sadr`aj Pb kod osoba koje `ive nateritoriji Beograda varira u odnosu na pol i starost ivi{i je u odnosu na nivoe toksi~nog metala u zemlja-ma Evropske unije.

Klju~ne re~i: humani biomonitoring; olovo;Beograd

(2.47–11.7 mg/dL in Belgium, 2.48–22.14 mg/dL inthe Czech Republic, 4.11–12.39 mg/dL in Poland,etc.). Pb levels in the Belgrade population are closeto the values obtained in Croatia, probably becauseCroatia and Serbia among the last countries bannedthe use of lead gas. Pb content in male group wassignificantly higher compared to female group(p<0.05). In addition, there were significant differ-ences in Pb level in women of different age groups(p<0.05).

Conclusion: Pb content in Belgrade populationvaries in relation to gender and age and is higher inrelation to the levels of toxic metals in the countriesof European Union.

Keywords: human biomonitoring; lead; Bel -grade

P005UTICAJ IZLO@ENOSTI

FTALATIMA NA RAZVOJ GOJAZNOSTI – ANALIZA

TOKSIKOGENOMI^KIH PODATAKA

Autor: Katarina @ivan~evi} e-mail: kajaziv93ªgmail.com

Mentori: doc. dr Danijela \uki}-]osi}, mag. farm-med. biohem. Katarina Barali}

Katedra za toksikologiju »Akademik Danilo Soldatovi}«,

Farmaceutski fakultet Univerziteta u Beogradu

Uvod: Gojaznost je jedan od zna~ajnih proble-ma javnog zdravlja i predstavlja faktor rizika za razvojmnogih bolesti. Zna~ajan mehanizam u nastankugojaznosti predstavlja promena ekspresije i aktivnostigena prouzrokovana supstancama koje ometajufunkcije endokrinog sistema. U ove supstance ubra-jaju se i ftalati, kojima su ljudi izlo`eni svakodnevnimkori{}enjem razli~itih proizvoda.

Cilj rada: Ispitati povezanost izme|u izlo`enostiftalatima i razvoja gojaznosti analizom toksikogeno-mi~kih podataka.

Metode: Ispitivani ftalati podeljeni su u dvegrupe. Prvu grupu ftalata ~ine-di(2-etilheksil) ftalat(DEHP), dibutil ftalat (DBP), butilbenzil ftalat (BBP),kojima su ve}a ograni~enja za upotrebu, a drugu- di-izo-nonil ftalat (DINP), di-izo-decil ftalat (DIDP) i di-n-oktil ftalat (DNOP). Kori{}ena je CTD (Compa -rative Toxicogenomic Database) baza podataka zadobijanje informacija o interakcijama ftalata i genapovezanih sa gojazno{}u, a podaci o funkciji genapreuzeti su iz GeneCards i STITCH baza podataka.

P005 THE INFLUENCE OF PHTHALATES

EXPOSURE ON OBESITY DEVELOPMENT – ANALYSIS OF

TOXICOGENOMIC DATA

Author: Katarina @ivan~evi}e-mail: kajaziv93ªgmail.com

Mentor: Assist. Prof. Danijela \uki}-]osi}, TA Katarina Barali}

Department of Toxicology »Academic Danilo Soldatovi}«,

Faculty of Pharmacy University of Belgrade

Introduction: Obesity is one of the significantpublic health issues and is considered a risk factor forthe development of many diseases. An importantpathogen mechanism in the onset of obesity ischange in the expression and activity of genes causedby chemicals that interfere with the functions of theendocrine system. These chemicals also includephthalates.

The Aim: To examine a correlation betweenexposure to phthalates and the development of obe-sity by analyzing toxicogenomic data.

Methods: The investigated phthalates weredivided into two groups. The first group of phthalateconsists of di(2-ethylhexyl) phthalate (DEHP), dibutylphthalate (DBP) and butylbenzyl phthalate (BBP),which are more potent and have greater restrictionsin use, and di-iso-nonyl phthalate (DINP), diisodecylphthalate (DIDP) and di-n-octyl phthalate (DNOP).Comparative Toxicogenomic Database (CTD) wasused to obtain information on phthalate and geneinteractions (gene products)/proteins and their asso-ciation with obesity. Additional used tools for studying

J Med Biochem 2018; 37 (2) 225

Rezultati: Prva grupa ftalata (DEHP, DBP, BBP)u~estvuje u 87% ispitivanih interakcija, pri ~emuDEHP intereaguje sa 59, DBP sa 82, a BBP sa 20gena (dobijeno pomo}u CTD Batch Query alata).Pomo}u MyVenn alata izdvojeno je 13 gena na kojesva tri ftalata deluju. Ovi ftalati intereaguju sa genimaodgovornim za sintezu PPAR peceptora. PPARA imauticaj na metabolizam lipida i ugljenih hidrata, dokPPARG ima klju~nu ulogu u adipogenezi i vi{e je zas-tupljen od PPARA. Ova tri ftalata dejstvom na INS3gen remete ekspresiju insulinskog receptora, {totako|e mo`e pogodovati razvoju gojaznosti. Ftalati izdruge grupe me|usobno deluju samo na jedan gen– PPARA.

Zaklju~ak: Analiza toksikogenomi~kih podatakaukazuje da se izlo`enost ispitivanim ftalatima mo`edovesti u vezu sa razvojem gojaznosti, posebno zaprvu grupu ftalata. Dobijene in silico rezultateneophodno je potvrditi ispitivanjima kod ljudi, odnos-no odre|ivanjem nivoa ftalata kod gojaznih osoba iispitivanjem ekspresije i aktivnosti gena od zna~aja.

Klju~ne re~i: ftalati; gojaznost; toksikogenomika

interactions are Batch Query, MyVenn, VennViewer,MyGeneVenn. Data on the function of genes wereobtained from GeneCards: The Human GeneDatabase and STITCH databases.

Results: he first group of phthalates (DEHP,DBP, BBP) was involved in 87% of the studied inter-actions (295 interactions with 161 genomes). hesephthalates interacted with the peroxisome prolifera-tor activated receptors (PPAR) (a particularly impor-tant interaction with peroxisome proliferator receptoralpha (PPAR )-the greatest effect on the carbohy-drate and lipid metabolism and peroxisome prolifera-tor receptor gamma (PPAR )-a key role in adipogen-esis) and disturb the expression of insulin receptor 3by interacting with INS3 gene. This gene participatesin the regulation of insulin sensitivity and glucosehomeostasis, which can lead to metabolic dysfunc-tion and favor obesity development.

Conclusion: The toxicogenomic analysis indi-cates that exposure to the examined phthalates canbe linked to the development of obesity. Theobtained in silico results need to be confirmed inhuman studies.

Keywords: phthalates; obesity; toxicogenomics

P006 UTICAJ OKSIDATIVNOG I

INFLAMATORNOG SKORA (OI SKOR)NA STANJE PACIJENATA NAKONKARDIOHIRUR[KE OPERACIJE

An|ela [aponji}1, Jelena Kotur-Stevuljevi}2,Petar Vuki}evi}3, Nata{a Bogavac-Stanojevi}2,

Aleksandar Miki}4,5

1Farmaceutski fakultet Univerziteta u Beogradu2Katedra za medicinsku biohemiju,

Farmaceutski fakultet Univerziteta u Beogradu3Katedra za kardiohirurgiju,

Vojnomedicinska akademija u Beogradu4Klinika za kardiohirurgiju, Klini~ki centar Srbije

5Katedra za hirurgiju sa anesteziologijom, Medicinski fakultet Univerziteta u Beogradu

Pra}enje oksidativnog i inflamatornog statusakod pacijenata je pokazano kao va`an parametar upra}enju progresije razli~itih bolesti i/ili efikasnostiterapije. Cilj rada je bio da se stekne uvid u nivo OIskorova kod pacijenata i kontrolne grupe, kao i testi-ranje uticaja ranije prele`anog infarkta miokarda, pu -{enja i vrste kardiohirur{ke operacije na skor. Studijaje obuhvatila 63 pacijenta koji su podvrgnuti operacijiugradnje koronarnog bajpasa na Klinici za kardio-hirurgiju Vojnomedicinske akademije u Beogradu.

P006 INFLUENCE OF OXIDATIVE AND

INFLAMMATION SCORE (OI SCORE)ON PATIENTS’ CONDITION AFTER

CARDIAC SURGERY

An|ela [aponji}1, Jelena Kotur-Stevuljevi}2,Petar Vuki}evi}3, Nata{a Bogavac-Stanojevi}2,

Aleksandar Miki}4,5

1Faculty of Pharmacy University of Belgrade,2Department of Medical Biochemistry,

Faculty of Pharmacy University of Belgrade3Department for Cardiac Surgery,

Military Medical Academy in Belgrade4Clinic for Cardiac Surgery, Clinical Centre of Serbia,

5Department of Surgery with Anesthesiology, Faculty of Medicine University of Belgrade

Monitoring of oxidative and inflammation statusin patients has shown to be important parameter inmonitoring of progression of various diseases and/oreffectiveness of therapy. The aim was to gain insightof patients’ and control group’s levels of OI scoresand to test influence of previously survived myocar-dial infarction, smoking and type of cardiac surgeryon that score. Study included 63 patients who under-go aortocoronary bypass surgery at the Departmentfor Cardiac Surgery at Medical Military Academy in

226

Parametri su analizirani kori{}enjem SPSS programa(Statistical package for Social sciences 11.5) iMajkro soft Eksela (Microsoft Excel 2007) Od 63pacijenta bilo je 13 `ena (21%) i 50 mu{karaca(79%), sa rasponom godina 41–78 godina (u pros-eku 61,8 godina), a od 289 zdravih ispitanika bilo je168 `ena (43%) i 221 mu{karac (57%), sa rasponomgodina 41–80 godina (u proseku 59,4 godine). OIskor za pacijene je izra~unat u 6 razli~itih perioda i topre, po samom zavr{etku operacije i 6, 24, 48 i 96sati kasnije. Prose~na vrednost skorova se kretala urasponu 15,43 (11,6-19,37) do 33,41 (25,98–39,31) i u svakom trenutku su te vrednosti bilezna~ajno razli~ite od kontrolne grupe ~ija je srednjavrednost OI skorova 1,23 (-0,22 do 5,89) pa je iprona|ena je zna~ajna razlika u OI skorovima(r<0,001) izme|u pacijenata i kontrolne grupe. Pro -na|ena je i zna~ajna razlika izme|u pacijenata sa ibez prele`anog infarkta miokarda u preoperativnomOI skoru (p<0,05) gde su ve}e skorove imali paci-jenti koji su prele`ali infarkt miokarda, dok razlika iz -me |u pu{a~a i nepu{a~a i izme|u vrsta izvr{enih ope -racija nije uo~ena. Konstantno pove}anje OI skoraposle operacije ukazuje na masovno generisanje slo-bodnih radikala i ~inilaca inflamacije nakon kardio-hirur{kih operacija zbog ~ega je neophodno du`epra}enje ovih faktora kako bi se uo~ilo kada dolazi donjihovog sni`avanja od ~ega i zavisi brzina oporavkapacijenata.

Belgrade. Parameters were analyzed by SPSS(Statistical Package for Social Sciences 11.5) andMicrosoft excel 2007. Out of 63 patients, 13 werefemales (21%) and 50 were males (79%), in the 41–78 age range (average 61.8 years), and out of 289healthy participants 168 were females (43%) and221 were males (57%), in the 41–80 age range(average 59.4 years). Patients’ scores were calculat-ed in 6 different periods – before, immediately afterand 6, 24, 48 and 96 hours after surgery. Mean val-ues of scores were ranging from 15.43 (11.6–19.37)to 33.41 (25.98–39.31) and in every moment val-ues were significantly different from control groupwhose median of OI scores was 1.23 (from -0.22 to5.89) and patients’ OI scores were significantly differ-ent (p<0.001) from control group. Statistically signif-icant difference was found between patients who hadsurvived myocardial infarction and those who hadn’thad it in preoperative OI score (p<0.05) wherepatients, who had survived myocardial infartion, hadhigher OI scores, difference was not observedbetween smokers and non-smokers and betweentypes of surgeries. Constant increase of OI scoresafter surgery indicates massive generation of freeradicals and inflammation factors after cardiac sur-geries which is why is needed to longer observe thisfactors to determine when it comes to their reduc-tions which depends on the speed of recovery ofpatients.

P007 ANALIZA TOKSIKOGENOMI^KIHPODATAKA O UTICAJU OLOVA

NA GOJAZNOST

Autori: Ana Ivanovi}, Monika Radovanovi}e-mail: ivanovica62ªgmail.com,

monikasvetozarªgmail.comMentori: doc. dr Danijela \uki}-]osi},

mag. farm. Dragica Jovanovi}

Katedra za toksikologiju »Akademik Danilo Soldatovi}«,

Farmaceutski fakultet Univerziteta u Beogradu

Uvod: Etiologija mnogih hroni~nih bolesti uklju -~uje interakcije izme|u faktora spolja{nje sredine igena koji moduli{u fiziolo{ke procese. Razu mevanjeinterakcija izme|u hemikalija koje se nalaze u `ivot-noj sredini i gena, odnosno proteina, mo`e pru`itiuvid u mehanizme hemijskih reakcija, osetljivost pre -ma bolestima, toksi~nost i inetrakcije sa lekovima.Brojnim istra`ivanjima je pokazano da olovo (Pb), kaojedan od najzna~ajnijih zaga|iva~a `ivotne sredine,uti~e na razvoj gojaznosti, ali su mehanizami nastan-ka ovih promena nedovoljno poznati.

P007 TOXICOGENOMICS ANALYSIS

OF LEAD INFLUENCE ON OBESITY

Authors: Ana Ivanovi}, Monika Radovanovi}e-mail: ivanovica62ªgmail.com,

monikasvetozarªgmail.comMentors: Assist. Prof. Danijela \uki}-]osi},

M. Pharm. Dragica Jovanovi}

Department of Toxicology »Akademik Danilo Soldatovi}«,

Faculty of Pharmacy University of Belgrade

Introduction: The etiology of many chronic dis-eases involves interactions between the environmen-tal factors and genes that modulate physiologicalprocesses. Understanding of interactions betweenenvironmental chemicals and genes/proteins mayprovide insights into the mechanisms of chemicalactions, disease susceptibility, toxicity, and therapeu-tic drug interactions. Numerous studies have shownthat lead (Pb), as one of the most significant environ-mental pollutants, affects the development of obesity,

J Med Biochem 2018; 37 (2) 227

Cilj rada: Cilj ovog rada bio je da se pomo}uanalize toksikogenomi~kih podataka (pronala`enjerelacija hemikalija-gen-bolest) ispita povezanostizme|u izlo`enosti olovu i nastanka gojaznosti.

Metode: Komparativna toksikogenomi~ka bazapodataka (Comparative Toxicogenomics Database-CTD) je kori{}ena da bi se ispitali metaboli~ki putevipovezani sa ekspozicijom Pb i razvojem gojaznosti.Interakcije izme|u Pb sa razli~itim genima preuzetesu kori{}enjem izvoznih i »batch query« alatki u CTDbazi. Dobijena lista gena analizirana je na protein/bo l est povezanost i uklju~enost u razli~ite meta -boli~ke puteve.

Rezultati: Dobijeno je da Pb stupa u interakcijusa ~ak 3 049 gena. Od tog broja, daljom analizom jeutvr|eno da Pb ispoljava dejstvo na 43 gena koji supovezani sa razvojem gojaznosti. Identifikovana 43gena su uklju~ena u 41 molekularni put, uklju~uju}imetabolizam lipida i lipoproteina, metabolizam mas-nih kiselina, triglicerida i ketonskih tela, metabolizamugljenih hidrata, itd. Od najve}eg zna~aja su geni kojiu~estvuju u signalnom putu adipocitokina i metabo-lizmu lipida i lipoproteina, kao {to su: acetil koenzimA karboksilaza beta (ACACB), leptin (LEP), faktornekroze tumora (TNF) i receptor aktiviran proliferaci-jom peroksizoma – gama (PPARG).

Zaklju~ak: Dobijeni rezultati ukazuju da Pbuti~e na gene od zna~aja za razvoj gojaznosti. Osimtoga, ovi rezultati ukazuju na dalja ispitivanja da sekod gojaznih osoba odrede nivoi toksi~nog metala iispita aktivnost gena od zna~aja u cilju potvrde ovihrezultata i poja{njenja mehanizama nastanka gojaz -nosti pri izlo`enosti olovu.

Klju~ne re~i: gojaznost; olovo; toksikogenomika

although mechanism of these changes is not suffi-ciently explained.

The aim: The aim of this study was to examinea correlation between Pb exposure and obesity onthe bases of toxicogenomic analysis (chemical–gene–disease testing).

Methods: The Comparative ToxicogenomicsDatabase-CTD was used to obtain metabolic path-ways associated with lead exposure and obesitydevelopment. Interactions between lead with differ-ent genes were downloaded using export and »batchquery« tools on the CTD base. The gene list was ana-lyzed for the »KEGG« annotation of pathways and theprotein/disease association.

Results: It was found that lead interacts with 3049 genes. A further analysis of this gene set wasaimed at identifying those associated with the devel-opment of obesity. Lead was found to interact with43 such genes involved in 41 molecular pathway,including metabolism of lipids and lipoproteins, fattyacid, triglycerids and keton bodies metabolism, car-bohydrate metabolism, etc. Of particular importanceare the genes involved in the signal pathway ofadipocytokines and metabolism of lipids and lipopro-teins, such as: acetyl coenzyme A carboxylase beta-ACACB, leptin-LEP, tumor necrosis factor-TNF andperoxisome-gamma proliferation receptor activated-PPARG.

Conclusion: The obtained results indicate thatPb affects the obesogenic genes. In addition, theseresults indicate further studies in obese individuals(determination of Pb level and activity of identificatedgenes) in order to confirm these results and clarifythe mechanisms of obesity occurrence in lead expo-sure.

Keywords: obesity; lead; toxicogenomics

228

P008 ODRE\IVANJE KONCENTRACIJE

VISOKO MOLEKULARNOGADIPONEKTINA KOD PACIJENATA

SA KOLOREKTALNIM KARCINOMOM

Jelena Novakovi}1, Milica Stevanovi}1, Milica Miljkovi}1, Aleksandra Stefanovi}1,

@eljka Stjepanovi}2, Dejan Zeljkovi}3, Bratislav Trifunovi}3,4, Vesna Spasojevi}Kalimanovska1, Aleksandra Zeljkovi}1,

Nata{a Bogavac-Stanojevi}1

1Katedra za medicinsku biohemiju, Farmaceutski fakultet Univerziteta u Beogradu2Op{ta bolnica «Medigroup«, Beograd, Srbija

3Klinika za op{tu hirurgiju, Vojno Medicinska akademija, Beograd, Srbija

4Medicinski fakultet Vojno Medicinske akademije,Ministarstvo odbrane, Beograd, Srbija

Uvod: Adiponektin je protektivni adipokin, ~ijase koncentracija u kolorektalnom karcinomu zna -~ajno smanjuje. Na{a studija se bavila pra}enjemvisokomolekularne frakcije.

Cilj rada: Odre|ivanje koncentracije visoko-molekularnog adiponektina kod pacijenata sakolorektalnim karcinomom.

Materijal i metode: Za odre|ivanje koncen-tracije visokomolekularnog adiponektina kori{}en je«dvostruki sendvi~ imunotest« (The Quantikine ELISAHuman HMW Adiponectin/Acrp30 Immunoassay),dok su koncentracije glukoze, triglicerida i HDL hole -sterola odre|ene standarnim enzimskim metodama.

Rezultati: Detektovane su zna~ajno ni`e vred-nosti visokomolekularnog adiponektina kod ispitani-ka sa kancerom kolona (p=0,039) i kancerom rektu-ma (p=0,033) u odnosu na zdrave osobe. Dokazanaje statisti~ki zna~ajna negativna korelacija visoko-molekularnog adiponektina sa indeksom telesnemase (ITM), R=-0,457, koncentracijom glukoze(R=-0,257) i triglicerida (R=-0,308) kao i pozitivnakorelacija sa HDL holesterolom (R=0,584) i staro{}upacijenata (R=0,392) u kontrolnoj grupi. U grupi pa -cijenata sa kancerom kolona utvr|ena je negativnakorelacija visokomolekularnog adiponektina saglukozom (R=-0,382). Kod pacijenata sa kanceromrektuma nije utvr|ena sli~na zavisnost. Tako|e, rezul-tati pokazuju da vrednosti visokomolekularnog adipo -nektina (OR=0,955 za kolon i OR=0,935 za rek-tum), godine pacijenata (OR=1,147 za kancerkolona i OR = 1,116 za kancer rektuma) i vrednostHDL holesterola (OR=0,191 za kolon i OR=0,172za rektum) predvi|aju rizik za nastanak kolorektalnogkarcinoma. Nezavisni prediktori kako kancera kolonatako i kancera rektuma su starost pacijenata i kon-centracija visokomolekularnog adiponektina.

P008HIGH MOLECULAR WEIGHT

ADIPONECTIN IN PATIENTS WITH

COLORECTAL CANCER

Jelena Novakovi}1, Milica Stevanovi}1, Milica Miljkovi}1, Aleksandra Stefanovi}1,

@eljka Stjepanovi}2, Dejan Zeljkovi}3, Bratislav Trifunovi}3,4, Vesna Spasojevi}Kalimanovska1, Aleksandra Zeljkovi}1,

Nata{a Bogavac-Stanojevi}1

1Department of Medical Biochemistry, Faculty ofPharmacy, University of Belgrade, Belgrade, Serbia

2Medigroup General Hospital, Belgrade, Serbia3Clinic of General Surgery, Military Medical

Academy, Belgrade, Serbia4Faculty of Medicine of the Military Medical

Academy, University of defence, Belgrade, Serbia

Introduction: Adiponectin has a protective rolein the human metabolism, however its concentra-tions seem to be significantly lower in patients withcolorectal cancer. Our study examined the highmolecular form of adiponectin in colorectal patients.

The Aim: Determining concentrations of highmolecular weight (HMW) adiponectin in colorectalcancer patients.

Material and methods: HMW adiponectin wasmeasured by ELISA Human HMW AdiponectinImmunoassay. Glucose, HDL cholesterol and triglyc-eride levels were measured using standard enzymemethods.

Results: Our study detected significantly lowerHMW adiponectin concentrations in patients withcolon cancer (CC) (p=0.039) and rectal cancer (RC),p=0.033 compared to the control group. The ob -tained results indicated a statistically significant neg-ative correlation between HMW adiponectin andbody mass index R=-0.457, glucose (R=0.257) andtriglycerides (R=0.308) levels, and positive correla-tion with HDL cholesterol concentrations (R=0.584)and patients’ age (R=0.392) in control group. In theCC group there was a negative correlation of HMWadiponectin with glucose concentrations (R=0.382).Also, the results indicated that HMW adiponectin-concentrations (OR=0.995 in CC group, andOR=0.935 in RC group), patients’ age (OR=1.147in CC group, and OR=1.116 in RC group), and HDLcholesterol concentrations (OR=0.191 in CC groupand OR=0.172 in RC group) predicted the risk forcolorectal cancer. The independent predictors for CCand RC were patients’ age and HMW adiponectinconcentrations.

J Med Biochem 2018; 37 (2) 229

Zaklju~ak: Kod pacijenata sa kolorektalnim kar-cinomom pokazano je zna~ajno smanjenje koncen-tracije visokomolekularnog adiponektina, kao i nje-gova sposobnost da zajedno sa HDL holesterolom igodinama pacijenata predvi|a rizik za nastanak istog.

Klju~ne re~i: kolorektalni karcinom; adiponektin

Conclusion: Patients with colorectal cancer hadsignificantly lower HMW adiponectin concentrationsthan subjects in control group. The results alsoshowed that HMW adiponectin, HDL cholesterolconcentrations and the patients’ age were independ-ent predictors for colorectal cancer.

Keywords: adiponectin; colorectal cancer

P009 VEZA IZME\U DISLIPIDEMIJE,

OKSIDATIVNOG STRESA I INFLAMACIJE KOD PACIJENATA

SA KANCEROM PLU]A

Tijana Ili}1, Nikola Ranisavljevi}1, Marina Roksandi}-Milenkovi}2,3,

Vesna ]eriman2, Natalija Samard`i}2, Milica Beli}1, Vesna Spasojevi}-Kalimanovska1, Jelena Kotur-Stevuljevi}1, Dragana Jovanovi}2,3

1Katedra za Medicinsku biohemiju, Farmaceutskifakultet, Univerzitet u Beogradu, Srbija

2Institut za plu}ne bolesti, Klini~ki centar Srbije,Beograd, Srbija

3Medicinski fakultet, Univerzitet u Beogradu, Srbiija

Uvod: Stvaranje reaktivnih kiseonikovih jedinje -nja remeti redoks ravnote`u u respiratornom sistemu{to pove}ava produkciju medijatora inflamacije plu}ai promovi{e kancerogenezu. Plu}a su izlo`ena oksi-dansima koji se endogeno ili egzogeno stvaraju(zaga|enje, pu{enje cigareta).

DOI (dislipidemija, oksidativni stres, inflamacija)skor je kombinacija tri razli~ita faktora rizika uklju -~ena u razvoj mnogih bolesti kao u kanceru plu}a iizra~unava se primenom Z-skor statistike.

Materijal i metode:Na{a studija ja uklju~ila 29zdravih ispitanika kao kontrolnu grupu (KG) i 91 pa -cijenta sa kancerom plu}a (KP): 14 sa mikrocelular -nim karcinomom i 77 sa nemikrocelularnim karci-nomom (46 sa adenokarcinomom i 31 pacijent saskvamocelularnim karcinomom, prema patohisto -lo{koj klasifikaciji)

Rezultati: Izra~unati skorovi: DOI skor, kao i nje-govi elementi Oksi skor (OS) i Inflamatorni skor (IS)su bili zna~ajno vi{i kod pacijenata u pore|enju saKG (P<0,001). ROC analizom, sva tri skora su poka -zala odli~nu (IS, AUC=0,884, P<0,001) i izvanred-nu dijagnosti~ku ta~nost (OS i DOI, AUC=0,994,P<0,001 i AUC=0,940, P<0,001, redom) za pre -dikciju pripadnosti zdravim osobala ili bolesnicima.Pore|enjem skorova izme|u razli~itih patohistolo{kiklasifikovanih kancera uo~ili smo zna~ajno pove}an

P009 RELATIONSHIP BETWEEN

DYSLIPIDEMIA, OXIDATIVE STRESSAND INFLAMMATION IN LUNG

CANCER PATIENTS

Tijana Ili}1, Nikola Ranisavljevi}1, Marina Roksandi}-Milenkovi}2,3,

Vesna ]eriman2, Natalija Samard`i}2, Milica Beli}1, Vesna Spasojevi}-Kalimanovska1, Jelena Kotur-Stevuljevi}1, Dragana Jovanovi}2,3

1Department for Medical Biochemistry, Faculty of Pharmacy, University of Belgrade, Serbia

2Institute for Pulmonary Disease, Clinical Center of Serbia, Belgrade, Serbia

3Faculty of Medicine, University of Belgrade, Serbia

Introduction: Reactive oxygen species genera-tion disturbs redox balance in the respiratory systemand thus increases the production of mediators ofpulmonary inflammation and promote carcinogene-sis. The lungs are exposed to oxidants generatedeither endogenously or exogenously (pollution, ciga-rette smoking).

DOI (dyslipidemia, oxidative stress and inflam-mation) score is a combination of three different riskfactors, involved in many disease development suchas lung cancer, calculated with Z score statistics.

Material and Methods: Our study included 29healthy control group (CG) subjects and 91 lung can-cer (LC) patients: 14 small cells lung cancer – SCLCand 77 non-small cells lung cancer – NSCLC patients(46 adenocarcinoma and 31 squamocellular accord-ing to pathohystological classification).

Results: Calculated scores: DOI score, so as itselements Oxy score (OS) and Inflammatory score (IS)were significantly higher in patients compared to CG(P<0.001). According to ROC analysis all threescores showed excellent (IS, AUC=0.884, P<0.001)and outstanding diagnostic accuracy (OS and DOI,AUCs= 0.944, P<0.001 and 0.940, P<0.001,respectively) in control-patients status prediction.Comparing scores between different pathohistologi-cal tumor classes we noticed significantly increased

230

IS kod podgrupe pacijenata sa skvamocelularnim uodnosu na adenokarcinom (P<0,05) a tako|e kodKP sa vi{im stadijumom bolesti (IV u odnosu na IIstadijum, P<0,05) i kod preminulih u odnosu na `ivepacijente, tokom jednogodi{nje studije. [to se ti~egeneralnog odgovora na hemioterapiju zabele`ilismo najni`i OS, IS i ukupan DOI skor kod pacijenatasa potpunim odgovorom u odnosu na pacijente saprogresijom bolesti, stabilnom bole{}u i pacijente sadelimi~nim odgovorom na hemioterapiju.

Korelaciona analiza je pokazala zna~ajnu pozi-tivnu korelaciju izme|u OS i IS skora (r=0,660,P<0,001) i negativnu korelaciju izme|u IS i ukupnogvremena pre`ivljavanja (r= –0,306, P<0,05).

Zaklju~ak: Na{i rezultati su dokazali zajedni~kou~e{}e oksidativnog stresa i inflamacije u patogenezikancera plu}a, u predikciji statusa pacijenata i nji-hovom pre`ivljavanju, a tako|e i u odgovoru na pri-menjenu terapiju.

Klju~ne re~i: dislipidemija; oksidativni stres;inflamacija; DOI skor; kancer plu}a (KP)

IS in squamocellular vs. adenocarcinoma NSCLCpatients subgroups (P<0.05), as well as in LCpatients in higher disease stadium (IV vs. II stadium,P<0.05), so as in deceased compared to livepatients during one year of the study duration.Regarding patients’ overall response rate to hemo-therapy (ORR-HT) we reported the lowest OS, IS andsummary DOI score in patients with completeresponse than in patients with progressive disease,stable disease and partial response.

Correlation analysis showed significant positivecorrelation between OS and IS (r=0.660, P<0.001)and negative correlation between IS and overall sur-vival time (r= –0.306, P<0.05).

Conclusion: Our results explicitly evidencedmutual involvement of oxidative stress and inflamma-tion in lung cancer pathogenesis, even in patients’status/survival prediction and also in response toimplemented therapy.

Keywords: dyslipidemia; oxidative stress; infla-mation; DOI score; lung cancer (LC)

P010 GENSKA EKSPRESIJA

ADIPONEKTINSKIH RECEPTORA U MONONUKLEARNIM ]ELIJAMA

KRVI KOD PACIJENATA SAKOLOREKTALNIM KARCINOMOM

Jelena Pavlovi}1, Dimitrije Mitrovi}1, Ana Nini}1, Marija Mihajlovi}1, Miron Sopi}1,Vesna Spasojevi}-Kalimanovska1, Aleksandra

Stefanovi}1, Milica Miljkovi}1, Tamara Gojkovi}1,Jelena Jana}1, Dejan Zeljkovi}2, BratislavTrifunovi}2, Nata{a Bogavac-Stanojevi}1,

@eljka Stjepanovi}3, Aleksandra Zeljkovi}1

1Katedra za medicinsku biohemiju, Univerzitet u Beogradu-Farmaceutski fakultet, Srbija

2Klinika za Op{tu hirurgiju, Vojno-medicinska akademija, Beograd, Srbija3Medigroup Op{ta bolnica, Beograd, Srbija

Kolorektalni karcinom (eng. Colorectal carcino-ma, CRC) je u vrhu liste u~estalosti malignih tumoradigestivnog trakta. Niske vrednosti adiponektina supovezane sa insulinskom rezistencijom i rizikom zarazvoj CRC-a. Zbog svojih antionkogenih i antiinfla -matornih osobina, smatra se da adiponektin mo`eimati ulogu u smanjenju proliferacije tumora. Cilj ovogistra`ivanja je bio da se ispita genska ekspresija adi po -nektinskih receptora (AR) kod pacijenata sa CRC-omu odnosu na zdravu populaciju i da se utvrde razlikeu genskoj ekspresiji AR izme|u razli~itih stadijuma i

P010ADIPONECTIN RECEPTORS GENE EXPRESSION LEVELS

IN PERIPHERAL BLOOD MONO-NUCLEAR CELLS IN PATIENTS

WITH COLORECTAL CARCINOMA

Jelena Pavlovi}1, Dimitrije Mitrovi}1, Ana Nini}1, Marija Mihajlovi}1, Miron Sopi}1,Vesna Spasojevi}-Kalimanovska1, Aleksandra

Stefanovi}1, Milica Miljkovi}1, Tamara Gojkovi}1,Jelena Jana}1, Dejan Zeljkovi}2, BratislavTrifunovi}2, Nata{a Bogavac-Stanojevi}1,

@eljka Stjepanovi}3, Aleksandra Zeljkovi}1

1Department for Medical Biochemistry, University of Belgrade – Faculty of Pharmacy, Serbia

2Clinic for General Surgery, Military Medical Academy, Belgrade, Serbia

3Medigroup General Hospital, Belgrade, Serbia

Colorectal carcinoma (CRC) is in the top of inci-dence list of the digestive tract’s malignant tumors.Low adiponectin levels are associated with insulinresistance and the risk for CRC development. Due toits anti-oncogenic and anti-inflammatory characteris-tics, adiponectin is considered to play a role in reduc-ing the proliferation of tumors. The aim of this studywas to determine adiponectin receptors (AR) geneexpression levels in patients with CRC and healthypopulation and to determine differences in AR geneexpression levels between different stages and grades

J Med Biochem 2018; 37 (2) 231

gradusa bolesti. U istra`ivanju je bilo uklju~eno 85pacijenata sa CRC-om (64,87±10,76 godina) i 109zdravih osoba (54,73±7,65 godina). Pacijenti supodeljeni prema Dukes-ovoj klasifikaciji na stadijumeB, C, D i prema stepenu diferentovanja tumorskih }e -lija na graduse 1, 2, 3. Nivoi informacione ribonuk-leinske kiseline RNK (iRNK) AR1 i AR2 su mereni umononuklearnim }elijama periferne krvi metodomlan ~ane reakcije polimeraze u realnom vremenu(Real time PCR). Nivoi iRNK AR1 su bili statisti~kizna~ajno ni`i kod osoba sa CRC-om u odnosu nazdravu populaciju (p<0,001), dok su nivoi iRNK AR2bili na granici statisti~ke zna~ajnosti (p=0,0510).Utvr|ena je statisti~ki zna~ajna razlika u nivoimaiRNK AR1 izme|u svih stadijuma bolesti i zdravepopulacije (p<0,001), a najni`i nivoi iRNK AR1uo~eni su kod pacijenata u stadijumu C. Tako|e, pos-tojala je statisti~ki zna~ajna razlika u nivoima iRNKAR1 izme|u zdravih ispitanika i pacijenata u razli -~itim gradusima bolesti (p<0,001), a najni`i nivoiiRNK AR1 su pokazani kod pacijenata u gradusu 3.Pozitivna korelacija uo~ena je izme|u nivoa iRNKAR1 i AR2 (r=0,282, p<0,001). Rezultati ovog istra -`ivanja su pokazali da je kod pacijenata sa CRC-omsmanjena genska ekspresija AR1 {to verovatno mo`ebiti razlog zbog ~ega adiponektin vezivanjem za njihne mo`e u potpunosti da ispolji svoja antiinflama -torna i antiproliferativna dejstva kao kod zdravihosoba.

of disease. The study included 85 patients with CRC(64.87±10.76 years) and 109 healthy controls(54.73±7.65 years). The patients were dividedaccording to the Dukes classification to stages B, C,D and to the degree of tumor cells differentiation togrades 1, 2, 3. AR1 and AR2 messenger ribonucleicacid’s (mRNA) levels were measured in peripheralblood mononuclear cells using polymerase chainreaction in real time (Real time PCR). AR1 mRNAlevels w r significantly lower in patients with CRCthan in healthy population (p<0.001), while AR2mRNA levels didn’t differ significantly between testedpopulations (p=0.0510). There were significant dif-ferences in the AR1 mRNA levels between all stagesof the disease and the healthy population(p<0.001), the lowest AR1 mRNA levels wereobserved in patients in C stage. Also, there were sig-nificant differences in AR1 mRNA levels betweenhealthy population and patients in different grades ofdisease (p<0.001), where grade 3 AR1 mRNA levelswere the lowest. A positive correlation was observedbetween AR1 and AR2 mRNA levels (r=0.282,p<0.001). Results of this study showed that patientswith CRC had decreased AR1 expression which couldbe the reason why adiponectin binding to them can-not completely express its anti-inflammatory andantiproliferative effects as expected.

P011 PROFIL LIPOPROTEINSKIH

SUBFRAKCIJA U SINDROMUOPSTRUKTIVNE APNEJE U SNU

Aleksandra Miri}1, Tijana Vu~kovi}1, Pavol Joppa2, Radovan Tisko2, Pavol Pobeha2,

Zuzana Strbova2, Zuzana Kuklisova2, Jelena Veki}1, Jasmina Ivani{evi}1,

Aleksandra Zeljkovi}1, Zorana Jeli}-Ivanovi}1,Vesna Spasojevi}-Kalimanovska1,

Ruzena Tkacova2

1Katedra za medicinsku biohemiju, Univerzitet u Beogradu – Farmaceutski fakultet

2Odeljenje za respiratornu medicinu i tuberkulozu,Univerzitetska bolnica L. Pasteur,

Ko{ice, Slova~ka

Opstruktivna apneja u snu (obstructive sleepapnea, OSA) je poreme}aj spavanja koji se karak-teri{e ponavljanim kolapsom gornjih disajnih putevatokom sna, koji mo`e biti potpun (pra}en apnejom)ili parcijalan (pra}en hipopnejom). Poreme}aj u kon-centraciji lipidnih parametara doprinosi pove}anomriziku za nastanak kardiovaskularnih bolesti (KVB)kod pacijenata sa OSA. U OSA se mogu javiti i kval-itativne promene lipoproteinskih ~estica, stoga je cilj

P011 LIPOPROTEIN SUBFRACTIONS

PROFILE IN OBSTRUCTIVE SLEEP APNEA

Aleksandra Miri}1, Tijana Vu~kovi}1, Pavol Joppa2, Radovan Tisko2, Pavol Pobeha2,

Zuzana Strbova2, Zuzana Kuklisova2, Jelena Veki}1, Jasmina Ivani{evi}1,

Aleksandra Zeljkovi}1, Zorana Jeli}-Ivanovi}1,Vesna Spasojevi}-Kalimanovska1,

Ruzena Tkacova2

1Department of Medical Biochemistry, University of Belgrade – Faculty of Pharmacy

2Department of Respiratory Medicine andTuberculosis, L. Pasteur University Hospital,

Kosice, Slovakia

Obstructive sleep apnea (OSA) is a sleeping dis-order, characterised by a repeated upper airway col-lapse during sleep that can be total (apnea) or partial(hypopnea). The disorder in lipid parameters concen-trations contributes to the increased risk of cardiovas-cular diseases development (CVD). Qualitativechanges of lipoprotein particles may occur in OSApatients as well, therefore the aim of our study was todetermine size and distribution of low-density (LDL)

232

na{eg istra`ivanja bio ispitati veli~inu i raspodelu sub-frakcija lipoproteina niske (LDL) i visoke (HDL) gus-tine kod ovih pacijenata, i to u zavisnosti od prisustvaKVB.

Ispitivanje je obuhvatilo 105 pacijenata sa OSA,od ~ega 40 pacijenata sa KVB i 65 pacijenata bezKVB. U~esnici u studiji su podvrgnuti testu poli-somnografije u Univerziteteskoj bolnici »LouisPasteur« (Ko{ice, Slova~ka). Razdvajanje LDL i HDLsubfrakcija je vr{eno metodom vertikalne elektro-foreze na poliakrilamidnom gradijent gelu.Koncentracije parametara lipidnog statusa (triglieridi,ukupan i holesterol u lipoproteinima niske i visokegustine - LDL-h i HDL-h) i glukoze su odre|ene stan-dardnim laboratorijskim metodama.

Statisti~kom analizom rezultata uo~eno je da sevrednosti lipidnih parametara nisu razlikovale izme|ugrupa, izuzev koncentracije LDL-h, koja je bila ni`a ugrupi sa KVB (P<0,01). Relativni udeo LDL I je bio ni`i(P<0,05), a LDL III subfrakcija zna~ajno vi{i (P<0,001)kod pacijenata kod kojih je OSA udru ̀ena sa KVB. Kodpacijenata sa KVB dominantni LDL dijametar je biozna~ajno manji (P<0,001). Do datno, dijametar LDL~estica je negativno korelirao sa koncentracijamatriglierida (P<0,05) i glukoze (P<0,001).

Razmatraju}i sve rezultate na{eg ispitivanja,mo ̀emo zaklju~iti da je razvoj KVB u OSA pra}enpro menama u profilu LDL subfrakcija, koje seogledaju u pove}anju udela malih gustih LDL ~estica,pri ~emu je te`i stepen bolesti pra}en ve}im smanje -njem dijametra LDL ~estica, jo{ izra`enijim u grupusa KVB. Dalje ispitivanje distribucija LDL i HDL sub-frakcija kod pacijenata sa OSA mo`e imati zna~aj uproceni rizika za KVB.

and high-density (HDL) lipoprotein subfractions inpatients with OSA, depending on the presence ofCVD.

Study included 105 patients with OSA, of which40 with and 65 without CVD. Participants in thestudy were subjected to polysomnography test at theUniversity Hospital ‘Louis Pasteur’ (Kosice, Slovakia).Vertical electrophoresis on polyacrylamide gradientgel was used for separation of lipoprotein subfrac-tions. Lipid parameters (triglycerides, total and cho-lesterol in low-density (LDL-C) and high-density(HDL-C) lipoproteins) and glucose concentrationswere determined with standard laboratory methods.

Using statistical analysis it was observed thatlipid parameters did not differ between the groups,except LDL-C concentration, which was lower in thegroup with CVD (P<0.01). Relative proportion ofLDL I subfractions was significantly lower (P<0.05),but the proportion of LDLIII particles was higher(P<0.001) in OSA patients with CVD. Also, the dom-inant LDL diameter was lower (P<0.001) in patientswith CVD. LDL diameters showed negative correla-tions with triglycerides (P<0.05) and glucose con-centrations (P<0.001).

Considering results of our study, we can con-clude that LDL subfractions profile was significantlyaltered in OSA, which is more pronounced as the dis-ease progresses and in the group with CVD. Furtheranalysis of LDL and HDL subfractions in OSApatients may be significant in estimating the risk ofCVD.

J Med Biochem 2018; 37 (2) 233

P012 HB-EGF KAO INFLAMATORNI

PARAMETAR KOD KOLOREKTALNOGKARCINOMA ODRE\EN DUOSET

ELISA METODOM

Sandra Civi}1, Milica Stevanovi}1, Milica Miljkovi}1, Aleksandra Stefanovi}1,

@eljka Stjepanovi}2, Dejan Zeljkovi}3, Bratislav Trifunovi}3,4, Vesna Spasojevi}Kalimanovska1, Aleksandra Zeljkovi}1,

Nata{a Bogavac-Stanojevi}1

e-mail: sandra_civicªyahoo.com

1Katedra za medicinsku biohemiju, Farmaceutski fakultet Univerziteta u Beogradu,

Beograd, Srbija 2Op{ta bolnica »Medigroup«, Beograd, Srbija

3Klinika za op{tu hirurgiju, Vojno Medicinska akademija, Beograd, Srbija

4Medicinski fakultet Vojno Medicinske akademije,Ministarstvo odbrane, Beograd, Srbija

Uvod: Heparin-vezuju}i epidermalni faktorsli~an faktoru rasta (HB-EGF), igra klju~nu ulogu umnogim fiziolo{kim i patolo{kim }elijskim procesima,uklju~uju}i proliferaciju, apoptozu, migraciju }elija,inva ziju, diferencijaciju, morfogenezu i razvoj. U ovojstudiji, merena je koncentracija HB-EGF u uzorcimadobijenih od zdravih dobrovoljaca i pacijenata sakolo rektalnim karcinom razli~itog stadijuma.

Cilj rada: Odre|ivanje koncentracije HB-EGFkod pacijenata sa kolorektalnim karcinomom.

Materijali i metode: Istra`ivanje je obuhvatilo 44pacijenata sa dijagnozom kolorektalnog karcinoma i50 osoba bez istorije malignih bolesti. Na osnovupatohistolo{kog nalaza odre|en je stepen diferent-nosti kancera i izvr{ena je podela pacijenata na trigrupe (slaba, srednja i dobra diferentovanost tumo-ra). Za odre|ivanje nivoa HB-EGF kori{}ena je Duo -Set sendvi~ ELISA ( DY008, DuoSet ELISA AncillaryReagent Kit 2) metoda.

Rezultati: U odnosu na diferentnost tumoranajvi{e su bili zastupljeni pacijenti sa srednje diferen-tovanim adenokarcinomom (61%), zatim pacijenti sadobro (27%) pa oni sa slabo diferentovanim karci-nomom (12%), p<0,001. Postoji razlika u vrednosti-ma HB-EGF izme|u pacijenata i kontrolne grupe(p<0,001), vrednosti su vi{e kod pacijenata 10,93mg/mL (7,76–16,42) u odnosu na kontrolnu grupu1,91 mg/mL (0,73–4,82). Dokazana je razlika u od -nosu na diferentnost tumora (p=0,030). VrednostiHB-EGF-a kod pacijenata sa dobro diferentovanimkarcinomom bile su zna~ajno ni`e 7,42 mg/mL(5,40–11,58) u pore|enju sa vrednostima kod paci-jenata sa slabo diferentovanim karcinomom 13,92

P012HEPARIN-BINDING

EGF-LIKE GROWTH FACTOR IN PATIENTS WITH

COLORECTAL CANCER

Sandra Civi}1, Milica Stevanovi}1, Milica Miljkovi}1, Aleksandra Stefanovi}1,

@eljka Stjepanovi}2, Dejan Zeljkovi}3, Bratislav Trifunovi}3,4, Vesna Spasojevi}Kalimanovska1, Aleksandra Zeljkovi}1,

Nata{a Bogavac-Stanojevi}1

e-mail: sandra_civicªyahoo.com

1Department of Medical Biochemistry, Faculty of Pharmacy, University of Belgrade,

Belgrade, Serbia 2Medigroup General Hospital, Belgrade, Serbia

32Clinic of General Surgery, Military Medical Academy, Belgrade, Serbia4Faculty of Medicine of the Military Medical

Academy, University of defence, Belgrade, Serbia

Introduction: Heparin-binding EGF-like GrowthFactor (HB-EGF), plays the key role in many physio-logical and pathological cellular processes includingproliferation, apoptosis, cell migration/invasion, dif-ferentiation, morphogenesis and development. Inthis study, we measured HB-EGF concentrations inserum samples obtained from healthy volunteers andpatients with colorectal carcinoma with differentstages.

The Aim: Determining concentrations of HB-EGFin colorectal cancer patients.

Mate rials and methods: The study included 44patients who were diagnosed colorectal cancer and50 people without a history of malignant disease.Based on the pathohistological findings, the degreeof differentiation of cancer is determined and thepatients were divided into three groups (poorly, medi-um, and a high differentiated tumor). The levels ofHB-EGF were quantified with DuoSet sandwichELISA (DY008, Duos ELISA Ancillary Reagent Kit 2)method.

Results: In relation to differentiation of tumorsthe most patients were with medium differentiatedadenocarcinoma (61%), followed by patients withhigh differentiated (27%) then those with poorly dif-ferentiated carcinoma (12%), p <0.001. There wasa difference in the values of HB-EGF among thepatients and the control group (p <0.001), the val-ues were higher in patients 10.93 mg/mL (7.76 to16.42) compared to the control group 1.91 mg/mL(0.73–4.82). The difference in relation to differenti-ation of the tumors was proven (p = 0.030). The val-ues of HB-EGF in patients with high-differentiated

234

mg/mL (12,76–32,23), p=0,037 i zna~ajno vi{e uodnosu na vrednosti u kontrolnoj grupi (p=0,001).

Zaklju~ak: HB-EGF mo`e biti koristan dijagnos-ti~ki serolo{ki biomarker za kolorektalni karcinom.Vrednosti HB-EGF zna~ajno su vi{e kod slabo, sred-nje i dobro diferentovanog karcinoma u odnosu nakontrolnu grupu {to je potvrdilo va`nost njegovogdaljeg ispitivanja kao prognosti~kog markera.

Klju~ne re~i: HB-EGF; kancer; serolo{ki bio-markeri; DuoSet ELISA

carcinoma were significantly lower 7.42 mg/mL(5.40 to 11.58) compared to values in patients withpoorly differentiated carcinoma 13.92 mg/mL(12.76 to 32.23) , p = 0.037, and significantly high-er compared to the values in the control group (p =0.001).

Conclusion:HB-EGF may be a useful diagnosticserological biomarker for colorectal cancer. The val-ues of HB-EGF were significantly higher in poorly,medium and high-differentiated carcinoma as com-pared to the control group which confirmed theimportance of its examination as prognostic marker.

Keywords: HB-EGF; Cancer; Serological bio-marker; DuoSet ELISA

P013E-KADERIN U

KOLOREKTALNOM KARCINOMU

Autor: Natalija Stojanovi}1, Sandra Vladimirov1,Jasmina Ivani{evi}1, Milica Miljkovi}1,

Marija Mihajlovi}1, Aleksandra Zeljkovi}1, Dejan Zeljkovi}2, Aleksandra Stefanovi}1,

Vesna Spasojevi}-Kalimanovska1

e-mail: natalija.atpªgmail.com

1Katedra za medicinsku biohemiju, Farmaceutski fakultet Univerziteta u Beogradu

2Klinika za op{tu hirurgiju, Vojnomedicinska akademija, Beograd

Uvod: E-kaderin je kalcijum-zavisni transmem-branski glikoprotein koji se sastoji iz ekstracelularnog,transmembranskog i intracelularnog domena i nalazise na bazolateralnoj membrani epitelijalnih }elija.U~estvuje u me|u}elijskom povezivanju tako {to seekstracelularni domen E-kaderina jedne }elije vezujeza ekstracelularni domen E-kaderina druge }elije gra -de}i homodimer. Funkcija E-kaderina je usko pove -zana sa citoplazmatskim proteinima kateninima. Poddejstvom razli~itih faktora dolazi do destabilizacijekompleksa E-kaderin-katenin i posledi~no do akti-vacije gena za deobu i rast }elije i tumorigeneze.

Cilj rada: Cilj ovog rada je odre|ivanje koncen-tracije E-kaderina i ispitivanje potencijalne veze iz -me|u E-kaderina i oksidativno-stresnog statusa kodpacijenata sa kolorektalnim karcinomom.

Materijal i metode: U studiji je u~estvovalo 105pacijenata sa kolorektalnim karcinomom i 109 zdra -vih ispitanika. Osnovni biohemijski parametri su odre -|eni komercijalnim biohemijskim testovima. Para -metri oksidativno-stresnog statusa (totalni oksidativnistatus, totalni antioksidativni status i prooksidativno-antioksidativni balans) su odre|eni spektrofotometrij -skim metodama. E-kaderin je odre|en sendvi~ ELISAmetodom.

P013E-CADHERIN IN

COLORECTAL CARCINOMA

Autor: Natalija Stojanovi}1, Sandra Vladimirov1,Jasmina Ivani{evi}1, Milica Miljkovi}1,

Marija Mihajlovi}1, Aleksandra Zeljkovi}1, Dejan Zeljkovi}2, Aleksandra Stefanovi}1,

Vesna Spasojevi}-Kalimanovska1

e-mail: natalija.atpªgmail.com

1Department of Medical Biochemistry, Faculty of Pharmacy University of Belgrade

2Clinic for General Surgery, Military Medical Academy, Belgrade

Introduction: E-cadherin is calcium-dependenttransmembrane glycoprotein composed of extracel-lular, transmembrane and intracellular domain and islocalised at the basolateral surface of epithelial cells.It is involved in cell-cell adhesion so the extracelluardomain of one E-cadherin binds the extracellulardomain of the E-cadherin from neighboring cellforming homodimer. E-cadherin function is tightlyconnected with cytoplasmatic proteins known as thecatenins. Due to variety of factors, destabilisation ofE-cadherin-catenin complex may occur leading to theactivation of gens for cell growth and tumorigenesis.

The Aim: The aim of this study was to determineconcentrations of E-cadherin and to examine the con-nection between E-cadherin and oxidative stress statusparemeters in patients with colorectal carcinoma.

Material and methods: 105 patients with col-orectal carcinoma and 109 healthy volunteers partic-ipated in this study. Basic biochemical parameters weredetermined using commercial biochemical tests.Oxidative stress status parameters (prooxidant-antioxi-dant balance, total oxidant status and total antioxidantstatus) were measured by spectrophotometric assays.E-cadherin was measured by ELISA method.

J Med Biochem 2018; 37 (2) 235

Rezultati: Mann-Whitney U-testom je dokazanoda je koncentracija E-kaderina kod pacijenata zna -~ajno vi{a u odnosu na kontrolnu grupu (p<0,0001).Koncentracija prooksidativno-antioksidativnog ba lansa(PAB) je zna~ajno ve}a kod pacijenata (p<0,0001),a pokazana je i zna~ajna korelacija izme|u vrednostiE-kaderina i PAB-a (p<0,05). Kon cenracija E-kaderi-na se pokazala zna~ajnim parametrom koji razdvajakontrolne ispitanike i pacijente, nezavisno od uticajastarosti i pola.

Zaklju~ak: Ova studija je potvrdila vezu izme|ukoncentracije E-kaderina i kolorektalnog karcinoma,kao i zna~ajnu povezanost ovog parametra sa para-metrima oksidativno-stresnog statusa.

Klju~ne re~i: E-kaderin; kolorektalni karcinom;oksidativno-stresni status.

Results: Mann-Withney U test showed that con-centrations of E-cadherin were significantly higher inCRC patients compared with the control group(p<0.0001). Concentration of prooxidant-antioxi-dant balance (PAB) was significantly higher inpatients group (p<0.0001), and there was a signifi-cant correlation between E-cadherin and PAB(p<0.05). E-cadherin concentrations showed signifi-cant ability in discrimination control group andpatients, independent of gender and age.

Conclusion: This study confirmed the relationbetween concentrations of E-cadherin and colorectalcarcinoma, as well as significant link between thisparameter and parameters of oxidative stress andantioxidant defens.

Keywords: E-cadherin; colorectal carcinoma;oxidative stress status.

P014REZISTIN: KONCENTRACIJA,

GENSKA EKSPRESIJA I POLIMORFIZAM RETN RS1862513

KOD PACIJENATA SA KOLOREKTALNIM KANCEROM

Autor: Ana \or|evi}1, Marija Mihajlovi}1, Ana Nini}1, Miron Sopi}1, Aleksandra

Stefanovi}1, Milica Miljkovi}1, Dejan Zeljkovi}2,Bratislav Trifunovi}2,3, Jelena Veki}1,

Tamara Gojkovi}1, Aleksandra Zeljkovi}1, Vesna Spasojevi}-Kalimanovska1

e-mail: anadjordjevicmbªgmail.com

1Katedra za medicinsku biohemiju, Univerzitet u Beogradu-Farmaceutski fakultet

2Klinika za op{tu hirurgiju, Vojnomedicinska akademija, Beograd

3Medicinski fakultet Vojnomedicinske akademije,Univerzitet odbrane, Beograd

Uvod: Kolorektalni kancer (KRK) je tre}i naj -~e{}i tip kancera kod mu{karaca i drugi naj~e{}i tipkancera kod `ena u svetu, ~ime je odgovoran za 10%ukupnih slu~ajeva kancera kod oba pola. Smatra seda je rezistin, adipokin kojeg u humanom organizmulu~e monociti i tkivne makrofage, povezan sa razvo-jem ove bolesti.

Cilj: Ispitati promene serumske koncentracije igenske ekspresije rezistina kod pacijenata sa kolorek-talnim kancerom u odnosu na zdrave osobe, izvr{itigenotipizaciju kod u~esnika u studiji i ispitati uticajprisustva G alela u RETN C-420G polimorfizmu nagensku ekspresiju i koncentraciju rezistina.

P014RESISTIN: CONCENTRATION,

GENE EXPRESSION AND POLYMORPHISM RETN RS186251

IN PATIENS WITH COLORECTAL CANCER

Author: Ana \or|evi}1, Marija Mihajlovi}1, Ana Nini}1, Miron Sopi}1, Aleksandra

Stefanovi}1, Milica Miljkovi}1, Dejan Zeljkovi}2,Bratislav Trifunovi}2,3, Jelena Veki}1,

Tamara Gojkovi}1, Aleksandra Zeljkovi}1, Vesna Spasojevi}-Kalimanovska1

e-mail: anadjordjevicmbªgmail.com

1Department of Medical biochemistry, Faculty of Pharmacy, University of Belgrade

2Clinic for General Surgery of the Military Medical Academy, Belgrade

3Faculty of Medicine of the Military MedicalAcademy, University of Defence, Belgrade

Introduction: Colorectal cancer (CRC) is theworld third most common type of cancer in men andthe second most common in women, which makes itresponsible for 10% incidence of cancer in bothsexes. Resistin, an adipokine secreted by monocytesand tissue macrophages, is considered to be associ-ated with this cancer.

The Aim: To examine changes in concentrationand gene expression of resistin in patients with CRCversus healthy individuals, as well as to examine theeffect of the presence of G alleles in the RETN C-420G single nucleotide polymorphism on geneexpression and protein concentration.

236

Pacijenti i metode: U istra`ivanju je u~estvovalo94 pacijenta Klinike za op{tu hirurgiju Vojno medi -cinske akademije sa dijagnostifikovanim KRK, a kon-trolnu grupu je ~inilo 106 zdravih osoba. Koncen tra -cija rezistina u serumu odre|ena je primenom ELISAtesta. Genska ekspresija je izvr{ena kori{}enjem7500 Real-Time PCR analizatora i TaqMan hemije.Detekcija SNP-a (-420 C/G) u promotorskomregionu RETN gena je izvr{ena reakcijom zasnovanojna 5’- egzonukleaznoj aktivnosti DNK polimeraze pri-menom specifi~nih VIC/FAM obele`enih MGB proba.

Rezultati: Serumska koncentracija rezistina jepovi{ena u grupi pacijenata sa KRK u odnosu nakontrolnu grupu (P<0,001). Genska ekspresija zarezistin u M PK u grupi pacijenata je sni`ena u odno-su na kontrolnu grupu (P<0,001). Kroz sva tri gra -dusa bolesti koncentracija rezistina ostaje pribli`nokonstantna, nasuprot genske ekspresije rezistina uperifernoj krvi koja ima trend opadanja. Geno tipi -zacija za SNP (-420 C/G) pokazuje da CC genotipdominira u obe grupe.

Zaklju~ak: Na{e istra`ivanje je potvrdilo prisust-vo povi{enih koncentracija rezistina kod pacijenata saKRK. Istovremeno, genska ekspresija u M PK bila jesni`ena, {to ukazuje na mogu}u nishodnu regulacijuovog gena u perifernoj krvi u prisustvu povi{enih kon-centracija proteina sintetisanog i sekretovanog umalignom tkivu. Potrebne su opse`nije budu}e studi-je sa ve}im brojem ispitanika razli~itih etni~kih grupada bi se preciznije definisala veza izme|u rezistina,njegove ekspresije i kolorektalnog kancera.

Klju~ne re~i: koncentracija rezistina; ekspresijaRETN gena; polimorfizam RETN rs1862513;kolorektalni kancer

Patients and Methods: We recruited 94 patientswith CRC at Clinic of General Surgery, MilitaryMedical Academy in Belgrade. The control groupconsisted of 106 healthy individuals. Serum resistinconcentration was determined by ELISA test. ThePCR experiment was done using 7500 Real-TimePCR Analyzer and TaqMan chemistry. SNP detection(-420C/G) in the promoter region of RETN gene wasperformed by the reaction based on the 5'-exonucle-ase activity of DNA polymerase using specificVIC/FAM labeled MGB probes.

Results: Resistin concentration was increased inCRC patients compared to the control group (P<0.001). Resistin gene expression in the patientswas lower in relation to the control group (P<0.001). Through cancer grades, concentration ofresistin remains approximately constant, opposed tothe gene expression that has a declining trend.Genotyping indicates that the CC genotype domi-nates both groups.

Conclusion: Our results confirmed increasedresistin serum concentration in patients with CRC. Incontrast, expression of resistin in PBMC wasdecreased, suggesting downregulation of peripheralblood resistin synthesis in the presence of increasedresistin secretion in malignant tissue. More extensivestudies, with a larger number of respondents of dif-ferent ethnic groups are needed to define a deeperlink between resistin concentration, gene expressionand colorectal cancer.

Keywords: resistin serum levels; gene expres-sion; SNP RETN rs1862513; colorectal cancer

P015 PORE\ENJE ANTIOKSIDATIVNOG

EFEKTA PREPARATA N-ACETILCISTEINA SA

KOMBINOVANIM PREPARATOM N-ACETILCISTEINA

I PROPOLISA

Autori: Biljana Radi{i}, Teodora Obradovi}Mentor: prof. dr Jelena Kotur-Stevuljevi}

e-mail: radisicbiljana307ªgmail.com ooteodoraªgmail.com

Katedra za medicinsku biohemiju, Univerziteta u Beogradu

Uvod: Oksidativni stres predstavlja stanje na -ru{ene ravnote`e izme|u prooksidanasa i endogenihsistema antioksidativne za{tite. N-acetilcistein (NAC) je

P015COMPARISON OF THE ANTIOXIDANT

EFFECT OF N-ACETYLCISTEINEPREPARATION WITH THE COMBINED SUPPLEMENT

PRODUCT OF N-ACETYLCISTEINEAND PROPOLIS

Authors: Biljana Radi{i}, Teodora Obradovi}Mentor: Assoc. Prof. Jelena Kotur-Stevuljevi}

e-mail: radisicbiljana307ªgmail.comooteodoraªgmail.com

Department of Medical Biochemistry, University of Belgrade

Introduction: Oxidative stress reflects a state ofdisturbed balance between pro-oxidants andendogenous systems of antioxidative protection. N-

J Med Biochem 2018; 37 (2) 237

dobro poznat antioksidans zbog svojih redukcionihosobina, koje poti~u od sulfhidrilnih grupa cisteina.Mukoliti~ko dejstvo ostvaruje raskidaju}i disuflidnemostove mukopolisaharida. Olak{ava iska{ljavanje,ubla`ava disanje i smanjuje viskoznost bronhijalnogsekreta. Propolis je sme{a smola koje sakupljaju p~e -le sa biljaka, koristi se za razli~ite svrhe zbog svog anti -bakterijskog i antiinflamatornog efekta. Kom plek snasme{a razli~itih jedinjenja iz pro po lisa sinergisti~ki do -pri nosi njegovom ukupnom efektu, a bioflavonoidi kojiulaze u sastav propolisa, ispoljavaju antioksidativniefekat.

Cilj: Ispitivanje antioksidativnog efekta propolisapore|enjem oksidativno – stresnog statusa ispitanikapre i posle suplementacije NAC-om ili kombinovanimpreparatom NAC-a i propolisa.

Materijal i metode: Studija obuhvata 20 zdravihispitanika (18 pu{a~a i 2 nepu{a~a), podeljenih nadve grupe. Prva grupa ~ini 10 ispitanika opredeljenihna kombinovani preparat NAC-a sa propolisom, adruga grupa obuhvata 10 ispitanika opredeljenih najednokomponentni preparat NAC-a. Krv je uzorko va -na pre i nakon desetodnevne suplementacije pre pa -ratima. Odre|eni su slede}i parametri prooksida-tivnog efekta u serumu: produkti uznapredovaleoksi dacije proteina (AOPP), malondialdehid (MDA),totalni oksidativni status (TOS), prooksidativno –antioksidativni balans (PAB) i parametri antioksida-tivne za{tite: totalni antioksidativni status (TAS),ukupne sulfhidrilne grupe (SHG), aktivnost enzimasuperoksid – dizmutaze (SOD) i paraoksonaze – 1(PON1).

Rezultati: Nakon suplementacije, kombinovanipreparat je statisti~ki zna~ajno pove}ao parametreantioksidativne za{tite SOD, PON i TAS kod svih ispi-tanika, dok je jednokomponenti zna~ajno uticao napove}anje SOD, SHG i smanjenje AOPP, tako|e kodsvih ispitanika u grupi.

Zaklju~ak: Oba preparata pobolj{avaju antioksi -dativnu za{titu i dokazan je zna~ajan doprinos efektapropolisa u kombinaciji sa NAC-om.

Klju~ne re~i: oksidativni stres; antioksidativnaza{tita; N-acetilcistein; propolis.

acetylcysteine (NAC) is well-known antioxidant for itsreduction properties which originate from sulfhydrylgroup of cysteine. It is used as a mucolytic because itreduces viscosity of the bronchial secretion. Also, italleviates expectoration and hard breathing by dis-rupting disulfide bonds of mucopolysaccharide.Propolis is used for various purposes due to its anti-bacterial and anti-inflammatory effects. Complexmixture of various compounds synergistically con-tributes its overall effect, and bioflavonoids, whichare part of propolis, manifest antioxidant effects.

Aim: The goal was to examine antioxidativeeffect of propolis by comparing the oxidative stressstatus of the respondents before and after supple-mentation of NAC or combined supplement productof NAC and propolis.

Material and Methods: The study includes 20healthy respondents (18 smokers and 2 nonsmoker),divided into two groups. The first group consists of10 respondents defined on the combined supple-ment product of NAC and propolis and the secondgroup consists of 10 respondents defined on one-component preparation of NAC. The blood sampleswere taken before and after a ten-day supplementa-tion of preparations. The following parameters of theprooxidative effect in the serum were determined:products of advanced protein oxidation (AOPP), mal-ondialdehyde (MDA), total oxidative status (TOS),protoxidative-antioxidant balance (PAB). Also, theparameters of antioxidant protection were deter-mined: Total antioxidant status (TAS), total sulfhydrylgroups (SHG), activity of superoxide-dismutaseenzyme (SOD) and paraoxonase 1 (PON1).

Results: After supplementation, the combinedpreparation has significantly increased the parame-ters of antioxidant protection: SOD, PON and TASfor all subjects in the group. Moreover, one-compo-nent preparation of NAC has significantly influencedon the increase of SOD, SHG and on decrease ofAOPP.

Conclusion: Both preparations improve antioxi-dant protection and it is showed significant contribu-tion of effect of propolis in combination with NAC.

Keywords: oxidative stress; antioxidant protec-tion; N-acetylcysteine; propolis.