POSTERSKE SEKCIJE POSTER SESSIONS razlika ima manju zna~ajnost u odnosu na kontrolnu grupu (p

POSTERSKESEKCIJE

POSTERSESSIONS

A

KARDIOVASKULARNA OBOLJENJAI SRÂNI MARKERI

CARDIOVASKULAR DISEASESAND CARDIAC MARKERS

A1ZNAÂJ ADENOZIN DEZAMINAZE

KOD BOLESNIKA SA AKUTNIM INFARKTOM MIOKARDA

S. Kundali}, D. Stankovi}-Ferle`, V. ]osi}, L. Zvezdanovi}, J. Lali}, V. B. \or|evi}

Centar za medicinsku biohemiju, Ni{Medicinski fakultet, Ni{

Kod infarkta miokarda dolazi do varijabilnog obi-ma nekroze tkiva miokarda, kao posledica opstrukcijeateroskleroti~nih koronarnih arterija. Jedan od najve}ihfaktora rizika koji dovodi do ateroskleroze i koronarnebolesti srca je dijabetes melitus. Adenozin dezaminaza(EC.3.5.4.4) (ADA) enzim je koji katalizuje dezaminaci-ju adenozina do inozina i amonijaka. Adenozin, degra-dacioni produkt ATP-a, ima za{titu ulogu kod o{te}enjasrca u ishemiji-reperfuziji. Kod 55 bolesnika sa akutniminfarktom miokarda (AIM), koji su imali pozitivan tro-ponin I, ispitivali smo aktivnost ADA. Trideset devetzdravih osoba istih godina starosti ~inili su kontrolnugrupu. Zna~ajno pove}ana aktivnost ADA (p<0,001)na|ena je u serumu bolesnika sa AIM u odnosu nazdrave dobrovoljce. Tako|e bolesnici iz AIM grupe kojisu imali dijabetes (50%) pokazali su visoke nivoe ADA(p<0,01) u pore|enju sa bolesnicima koji nisu imalidijabetes. Rezultati pokazuju da porast aktivnosti ADAnakon akutnog infarkta miokarda dovodi do pada ade-nozina koji je endogeni celularni aktivator antioksi-danasa. Ovo istraìvanje pokazuje da je odre|ivanjeaktivnosti ADA va`no kod bolesnika sa AIM i moèimati vitalnu ulogu kod o{te}enja u ishemiji-reperfuziji.

A1THE SIGNIFICANCE OF ADENOSINE

DEAMINASE IN PATIENTS WITH ACUTEMYOCARDIAL INFARCTION

S. Kundali}, D. Stankovi}-Ferle`, V. ]osi}, L. Zvezdanovi}, J. Lali}, V. B. \or|evi}

Centre of Medical Biochemistry,University School of Medicine, Ni{

In myocardial infarction ischemic necrosis of avariable amount of myocardial tissue occurs as a resultof atherosclerotic coronary artery obstruction. One ofthe major risk factors leading to atherosclerosis andcoronary heart disease is diabetes mellitus. Adenosinedeaminase (EC.3.5.4.4) (ADA) is the enzyme whichcatalyzes the deamination of adenosine to inosine andammonia. Adenosine, a degradation product of ATP,has been attributed to exert a protective effect on injurycaused by ischemia-reperfusion in the heart. In 55patients with acute myocardial infarction (AMI) whowere troponin I positive, we investigated serum ADAactivity. Thirthy nine age-matched healthy personsserved as a control group. A highly significant increase(p<0.001) in the activity of ADA was found in the seraof AMI patients as compared to healthy donors. Also,AMI patients, with diabetes (50%) showed high levelsof ADA (p<0.01) when compared to non-diabeticpatients. The results indicate that increased levels ofADA activity in patients after AMI lead to the depletionof adenosine that is an endogenous activator of cellularantioxidants. These findings show that the estimation ofADA activity is important in patients with AMI and mayplay a vital role in ischemia-reperfusion injury.

JMB 2008; 27 (2) 223

UDK 577.1 : 61 ISSN 1452-8258

JMB 27: 223–235, 2008 Posterske sekcije

Poster sessions

A2THE ROLE OF CATALASE IN PATIENTS

WITH STABLE AND UNSTABLE ANGINA PECTORIS

D. Stankovi}-Ferle`, B. Kundali}, S. Kundali}, Z. Peri{i}, V. B. \or|evi}

Centre of Medical Biochemistry,Clinic of Cardiovascular Diseases, Clinical Centre Ni{

University School of Medicine, Ni{

Ischemic heart disease (IHD) – stable and unsta-ble angina pectoris and myocardial infarction – is themost frequent cause of morbidity and mortality today.Generation of reactive oxygen species-free radicals inIHD contributes to irreversible myocardial tissue injurywith concomitant changes of certain key endogenousantioxidant compounds, e.g. catalase, superoxide dis-mutase, reduced glutathione and glutathione peroxi-dase. The purpose of this study is to show in which waywe can use catalase activity in the evaluation ofischemic heart damage, in patients with stable (SAP)and unstable (USAP) angina pectoris. In the study weobserved the activity of catalase in the plasma and ery-throcytes (Er) of 40 patients with assessed SAP and 45patients with USAP. The control group consisted of 30healthy volunteers. The activity of catalase in Er wasdetermined according to the Beutler method, and inplasma it was measured by the method of Goth. Wenoted a significant increase in catalase activity in theplasma of patients with USAP (p<0.01), while in pa-tients with SAP we noted less significant increase com-pared with the control group (p<0.05). The catalaseactivity in Er in both groups did not show any significantdifference compared with healthy volunteers. Theseresults show an increase in catalase activity, which is aconsequence of chronic oxidative stress, so we believethat the follow-up of this enzyme could be useful in thediagnosis and treatment of patients with USAP and SAP.

A3CORRELATION BETWEEN INTIMA-MEDIA

THICKNESS AND THE LEVELS OF CHOLESTEROL AND TRIGLYCERIDES

IN PATIENTS WHO RECOVERED FROM ACUTE MYOCARDIAL INFARCTION

D. Miljkovi}, N. Kosti}

Health Centre Varvarin, Medical Centre Kru{evac

The growth of the intima-media complex is awidely accepted parameter in studies related to theprocess of atherosclerosis. A large number of clinicalstudies showed that the intima-media thickness (IMT)of the extracranial segments of arteriae carotis com-munis presents a measurable index of the presence ofatherosclerosis. The aim of this study was to investigate

A2ULOGA KATALAZE KOD PACIJENATA

SA STABILNOM I NESTABILNOM ANGINOM PEKTORIS

D. Stankovi}-Ferle`, B. Kundali}, S. Kundali}, Z. Peri{i}, V. B. \or|evi}

Centar za medicinsku biohemiju,Klinika za kardiovaskularne bolesti, Klini~ki centar Ni{

Medicinski fakultet, Ni{

Ishemijska bolest srca – stabilna i nestabilna angi-na pectoris i infarkt miokarda – jedan je od naj~e{}ih inajva`nijih uzroka morbiditeta i mortaliteta danas. Slo-bodni radikali nastali u ishemijskoj bolesti srca dovodedo ireverzibilnog o{te}enja funkcije srca uzrokuju}i pro-menu nivoa glavnih endogenih antioksidativnih kom-ponenata kao {to su katalaza, superoksid dizmutaza,redukovani glutation i glutation peroksidaza. Cilj ovograda bio je da se pokaè na koji na~in merenje aktivnos-ti katalaze moè da se iskoristi za procenu ishemijskogo{te}enja miokarda u pacijenata sa stabilnom (SAP) inestabilnom (NSAP) anginom pektoris. Ispitivana jeaktivnost katalaze u plazmi i eritrocitima (Er) u 40 paci-jenata sa potvr|enom SAP i 45 pacijenata sa NSAP.Kontrolnu grupu ~inilo je 30 zdravih dobrovoljaca.Aktivnost katalaze u Er merena je metodom poBeutleru, a u plazmi po Gothu. Utvr|eno je da postojizna~ajno pove}anje aktivnosti katalaze u plazmi u paci-jenata sa NSAP (p<0,01), dok kod pacijenata sa SAPta razlika ima manju zna~ajnost u odnosu na kontrolnugrupu (p<0,05). Aktivnost katalaze u Er u obe grupepacijenata nije pokazala zna~ajne razlike u odnosu nazdrave ispitanike. Ovi rezultati pokazuju zna~ajan porastaktivnosti katalaze {to je posledica hroni~nog oksi-dativnog stresa, stoga pra}enje nivoa ovog enzima mo-è biti korisno u dijagnostici i terapiji stabilne i nesta-bilne angine pektoris.

A3KORELACIJA IZME\U INTIMOMEDIJALNOG

KOMPLEKSA I NIVOA HOLESTEROLA I TRIGLICERIDA U BOLESNIKA SA

ZNAÂJNOM STENOZOM KORONARNIHARTERIJA KOJI SU PREBOLELI INFARKT

MIOKARDA

D. Miljkovi}, N. Kosti}

Dom zdravlja Varvarin, Zdravstveni centar Kru{evac

Debljina kompleksa intimomedije zida ekstrakar-nijalnih karotidnih arterija predstavlja merljiv pokazateljateroskleroze. Ukazano je na zna~ajnu udruènost ka-rotidne bolesti sa hiperlipidemijom. Ispitivanja su po-tvrdila povezanost ishemi~ne bolesti srca i karotidnebolesti. Ispitivana je povezanost debljine intimomedi-jalnog kompleksa (IMC) zida zajedni~ke karotidne arte-

224

rije (ACC) i nivoa holesterola i triglicerida u serumubolesnika koronarografski potvr|enom okluzivnombole{}u koronarnih arterija koji su preboleli infarktmiokarda. Ispitivanjem je obuhva}eno 28 bolesnika sainfarktom miokarda, prose~ne starosti x=60,28+ 7,39godina, 22 (78,58%) mu{karca i 6 (21,42%) èna.Svim ispitanicima je ura|en »color duplex« ehosono-grafski pregled karotidnih arterija. Ispitivanje je obavlje-no na ultrazvu~nom aparatu firme »SONOACE 8000SE« sa linearnom sondom od 7,5 MHz i dubine 5,0cm. Intimomedijalna debljina je merena na posteri-ornom zidu zajedni~ke karotidne arterije (ACC), 2 cmod vrha bifurkacije, i uzimana prose~na vrednost tri uza-stopna merenja tokom dijastole. Svim bolesnicima jeizmeren nivo ukupnog holesterola, LDL holesterola itriglicerida. Kod svih bolesnika je prethodno ura|enaselektivna koronarografija. Trosudovnu koronarnubolest imalo je 10 (35,7%), dvosudovnu 16 (57,1%) ijednosudovnu 2 (7,2%) bolesnika. Prose~na vrednostdebljine IMC zajedni~ke karotidne arterije iznosilaje x=1,67+0,13 mm. Prose~na vrednost ukupnog ho-lesterola iznosila je x=7,40+1,81 mmol/L, triglice-rida x=3,10+2,41 mL/L i LDL holesterola x=5,89+1,50 mmol/l. Postoji pozitivna, niska i nesignifikantnakorelacija izme|u debljine IMC-a i nivoa ukupnogholesterola (r=0,35, p>0,05). Tako|e postoji pozi-tivna, niska i nesignifikantna korelacija izme|u debljineIMC-a i nivoa LDL holesterola (r=0,31, p>0,05). Po-stoji pozitivna, niska i signifikantna korelacija izme|udebljine IMC-a i nivoa triglicerida (r=0,45, p<0,05).Dobijeni rezultati ukazuju na povezanost lipida i ate-roskleroze, kao i na to da su lipidi zna~ajan, ali ne i jedi-ni faktor rizika ateroskleroze merene debljinom intimo-medijalnog kompleksa.

A4NIVO IZOENZIMA KREATIN KINAZE

MB MASE U BOLESNIKA SA AKUTNIMINFARKTOM MIOKARDA

N. Kosti}, D. Miljkovi}

Zdravstveni centar Kru{evac, Dom zdravlja Varvarin

Odre|ivanje koncentracije CK MB mase isti~e sekao rani i veoma senzitivan marker akutnog infarktamiokarda (AIM), posebno kod bolesnika sa odsutnimEKG promenama ili malim infarktnim podru~jima. Po-rast koncentracije CK MB mase nakon AIM nastajeznatno ranije u odnosu na aktivnost CK MB izoenzimai aktivnost ukupne CK. Cilj rada je bio da se odredi nivoCK MB mase u serumu bolesnika sa AIM i sagleda nje-gov zna~aj u dijagnostici AIM. Ispitivanjem je obuhva-}eno 47 bolesnika sa AIM, prose~ne starosti x=61.7+9.0 godina, 12 (25.5%) èna i 35 (74.5%) mu{kara-ca, i 20 zdravih osoba, 10 èna i 10 mu{karaca, kaokontrolna grupa. Kod svih bolesnika krv za analizu uzi-mana je odmah po prijemu i odre|ivane su koncen-tracije CK MB mase, CK, CK MB, AST, LDH, hsCRP,holesterol, HDL, LDL, trigliceridi i glikemija. CK MBmase je odre|ivan na principu elektrohemiluminiscen-cije na aparatu »ELECSYS 1010«, »Roche« dijagnosti-ka, sa normalnim vrednostima 0,1–0,5 ng/mL. Pro-

the relation between the IMT of the arteriae carotiscommunis and the levels of cholesterol and triglyc-erides in patients who recovered from acute myocardialinfarction. The study involved 28 patients who recov-ered from acute myocardial infarction, 22 (78.6%)men and 6 (21.4%) women, average age x=60.3+7.4 years. IMT was measured in the end-diastole onthe back wall of a. carotis communis; for detection weused a high resolutive color-Doppler ultrasonic systemfrequency of 7.6 MHz. Total cholesterol, HDL choles-terol and triglycerides were determined. The averagevalue of the IMT of a. carotis communis in patientswho recovered from acute myocardial infarctionis x=1.67+0.13 mm. The average value total choles-terol is x=7.4+1.8 mmol/L, LDL x=5.89+1.50mmol/l and triglycerides x=3.10+2.40 mmol/L. Wefound a positive but insignificant corelation betweentotal cholesterol and IMT (r=0.35 p>0.05). We founda significant positive correlation between IMT andtriglycerides (r=0.45, p<0.05).

A4THE LEVEL OF CK-MB MASS ISOENZYMESIN PATIENTS WITH ACUTE MYOCARDIAL

INFARCTION

N. Kosti}, D. Miljkovi}

Medical Centre Kru{evac, Health Centre Varvarin

The determination of the level of CK-MB massisoenzymes is important in the early diagnosis of acutemyocardial infarction. The aim of this study was to deter-mine the levels of plasma CK-MB mass in patients withacute myocardial infarction. The study involved 47 pa-tients with acute myocardial infarction, 12 (25.5%) wo-men and 35 (74.5%) men, average age x=61.7+ 9.0years, and 20 healthy persons, as a control group. CK-MB mass, CK, CK-MB, LDH, AST, ALT, hsCRP, choles-terol, HDL, LDL and triglycerides were determined. CK-MB mass was determined using an electrochemilumi-nescent method on an Elecsys 1010, »Roche Diagno-stics«. Reference ranges were from 0.1 to 0.5 ng/mL.The average values of CK-MB mass in patients with acu-te myocardial infarction and persons control groupare: x=95.6 + 103.9 ng/mL and x=2.3 = 0.7 ng/mL(t=6.1, p<0.01); CK x=1930+1401.04 U/L and x=91.1+40.0 U/L (t=8.9, p<0.01); CK MB x=969.5

JMB 2008; 27 (2) 225

+140.2 U/L and x=10.6+3.5 U/L (t=8.0, p<0.01);AST X=188.0=176.1 U/L and x=20.7+5.4 U/L(t=6.5, p<0.01); LDH x=989.2+521.9 and x=303.8+80.7 U/L (t= 8.7, p<0.01). All patients withacute myocardial infarction have increased values ofCK-MB mass. On the basis of the presented data it canbe concluded that CK-MB mass is a sensitive marker forthe early diagnosis of acute myocardial infarction.

A5ANTHROPOMETRIC INDICES,

CRP AND CAD IN ADULT SERBIAN WOMEN

L. Memon1, N. Bogavac-Stanojevi}2, V. Spasojevi}-Kalimanovska1, D. Kalimanovska-O{tri}3,

S. Spasi}2, Z. Jeli}-Ivanovi}2, A. Zeljkovi}2

1Clinical center »Beànijska kosa«, Belgrade, Serbia

2Institute of Medical Biochemistry, Faculty of Pharmacy, Belgrade, Serbia3Institute for Cardiovascular Diseases,

Clinical Center of Serbia, Belgrade, Serbia

C-reactive protein (CRP) is an independent riskfactor for coronary artery disease (CAD) that is strong-ly associated with measures of obesity. Our purposewas to explore the relationship between CRP, bodymass index (BMI) and waist-to-hip ratio (WHR) inhealthy women and women with angiographicallyassessed CAD. A total of 63 patients and 111 healthysubjects, aged 31 to 75, were included in the study.Height, weight and waist and hip circumferences weremeasured. BMI and WHR were computed. CRP wasassayed using an ultra-sensitive immunoturbidimetricmethod. According to age, women were classified intofour groups: healthy 50 years of age (group 1),healthy 51 years (group 2), patients 50 years(group 3), patients 51 years (group 4). The resultsobtained by using Student's t-test have shown thatthere was no significant difference in BMI, but thatthere was a statistically significant difference in WHRand CRP between groups 1 and 2. Elderly healthywomen had significantly higher WHR (0.82 vs 0.77)and CRP (1.47 vs. 0.9 mg/L) compared to the youngergroup. On the other hand, there was no difference forany of the mentioned parameters between younger

se~ne koncentracije CK MB mase i drugih markerao{te}enja miokarda kod bolesnika sa AIM i kontrolnegrupe su: CK MB masa x = 95,6 + 103,9 ng/mL i x= 2,3 + 0,7 (t=6,1, p<0,01); CK x = 1930,5 +1401,0 U/L i x = 91,1 + 40,0 U/L (t=8,9, p<0,01);CK MB x = 969,5 + 140,2 U/L i 10,6 = 3,5 U/L(t = 8,0 p<0,01); AST x = 188 + 176,1 U/L i x =20,7 + 5,4 U/L (t = 6,5, p<0,01); LDH x = 989,2 +521,9 U/L i x=303,8 + 80,7 U/L (t=8,7, p<0,01).Relativni indeks koji se dobija iz odnosa CK MBmass/CK MB aktivnost iznosio je prose~no x=54,5%.Vrednosti vi{e od 2,5 % ukazuju pre na sr~ano poreklo,nego na poreklo iz skeletnih mi{i}a i na|ene su kod45/47 (95,7%) bolesnika sa AIM. Postoji vrlo visokapozitivna korelacija izme|u nivoa CK MB i CK MB mase(r=0,85, p<0,01). Postoji pozitivna i niska, ali statisti~kizna~ajna korelacija izme|u nivoa CK i CK MB mase(r=0,45, p<0,01). Postoji pozitivna i visoka korelacijaizme|u CK i CK MB (r=0,67, p<0,01). Odre|ivanjekoncentracije CK MB mase u serumu je zna~ajno udijagnostici pacijenata sa akutnim bolom u grudima iranoj dijagnostici akutnog infarkta miokarda.

A5ANTROPOMETRIJSKI INDEKSI, CRP IKORONARNA ARTERIJSKA BOLEST

KOD ODRASLIH @ENA U SRBIJI

L. Memon1, N. Bogavac-Stanojevi}2, V. Spasojevi}-Kalimanovska1, D. Kalimanovska-O{tri}3,

S. Spasi}2, Z. Jeli}-Ivanovi}2, A. Zeljkovi}2

1Klini~ko-bolni~ki centar »Beànijska kosa«, Beograd, Srbija

2Institut za medicinsku biohemiju, Farmaceutski fakultet, Beograd, Srbija

3Institut za kardiovaskularne bolesti, Klini~ki centar Srbije, Beograd, Srbija

C-reaktivni protein (CRP) nezavisni je faktor rizikaza koronarnu arterijsku bolest (KAB) i u sna`noj jekorelaciji sa gojazno{}u. Cilj ovog rada je bio da se ispi-ta veza izme|u CRP-a i antropometrijskih indeksa:odnosa struk/kuk (WHR) i indeksa telesne mase (BMI)kod zdravih èna i èna sa angiografski dokazanomKAB. Grupu ispitanika ~inilo je 111 zdravih èna i 63pacijentkinje starosti od 31 do 75 godina. Visina, teìnai obim struka i kuka su izmereni dok su BMI i WHRizra~unati. CRP je odre|en upotrebom ultrasenzitivneimunoturbidimetrijske metode. U zavisnosti od godina,sve ène su klasifikovane u ~etiri grupe: zdrave 50godina (grupa 1), zdrave 51 godina (grupa 2), paci-jentkinje 50 godina (grupa 3), pacijentkinje 51godina (grupa 4). Rezultati dobijeni upotrebom studen-tovog t-testa su pokazali da ne postoji statisti~kizna~ajna razlika u BMI izme|u mla|ih i starijih zdravihèna. Starije zdrave ène imale su zna~ajno ve}i WHR(0,82 prema 0,77) i CRP (1,47 prema 0,9 mg/L) uodnosu na mla|e zdrave ène. S druge strane, nije bilozna~ajne razlike ni za jedan ispitivani parametar izme|ustarijih i mla|ih pacijentkinja (p>0,05). Statisti~ki zna-~ajna pozitivna korelacija je dokazana izme|u koncen-

226

tracije CRP-a i mera gojaznosti kod zdravih èna (BMIp<0,001 i WHR p<0,01, linearna regresiona analiza),dok je statisti~ki zna~ajna korelacija izme|u pomenutihparamatara izostala kod pacijentkinja. Rezultati ovestudije ukazuju na to da se gojaznost nalazi u sna`nojvezi s koncentracijom CRP-a kod zdravih odraslih ènau populaciji Srbije. Na osnovu toga moè se zaklju~itida je gojaznost vaàn faktor koji uti~e na hroni~nu infla-maciju niskog stepena. Starije ène imaju vi{e vrednos-ti odnosa struk/kuk i CRP-a u pore|enju sa mla|imènama najverovatnije zbog odsustva protektivnogefekta estrogena.

A6VEZA IZME\U VISOKO-NORMALNOGKRVNOG PRITISKA I FAKTORA RIZIKA

ZA KARDIOVASKULARNU BOLEST

L. Memon1, N. Bogavac-Stanojevi}2, V. Spasojevi}-Kalimanovska2, S. Spasi}2,

Z. Jeli}-Ivanovi}2, A. Zeljkovi}2

1Klini~ko-bolni~ki centar »Beànijska kosa«, Beograd, Srbija

2Institut za medicinsku biohemiju, Farmaceutski fakultet, Beograd, Srbija

Prema klasifikaciji JNC VI i WHO-ISH, osobe bezhipertenzije koje imaju sistolni pritisak od 130 do 139mmHg, dijastolni pritisak od 85 do 89 mmHg, ili obakategorisane su u grupu visoko-normalnog krvnog pri-tiska. Nema dovoljno informacija o tome koliki je rizikza obolevanje od kardiovaskularne bolesti (KAB) kod tihosoba. Cilj rada je bio ispitivanje veze izme|u kategori-ja normalnog pritiska i faktora rizika za KAB. Grupu ispi-tanika je ~inilo 99 zdravih osoba, 53 mu{karca i 46èna. Ispitanici su klasifikovani u tri nehipertenzivnegrupe: optimalan (sistolni pritisak manji od 120 mmHgili dijastolni pritisak manji od 80 mmHg, grupa 1), nor-malan (sistolni pritisak od 120 do 129 mmHg ili dijas-tolni pritisak od 80 do 84 mmHg, grupa 2) i visoko-nor-malan (sistolni pritisak od 130 do 139 mmHg ili dijas-tolni pritisak od 85 do 89 mmHg, grupa 3). Informacijeo starosti, konzumiranju alkohola, statusu pu{enja ifizi~koj aktivnosti ispitanici su dali samoizja{njavanjem.Indeks telesne mase (BMI) i odnos struk/kuk (WHR) suizra~unati. U uzorcima su odre|eni hsCRP, mokra}nakiselina i lipidni parametri (ukupan holesterol, HDL-H,LDL-H, trigliceridi, Lp(a), apoA-I, apoB) standardnimlaboratorijskim metodama. Fibrinogen je izmerenKlausovom metodom. Statisti~ka analiza je ura|enaupotrebom ANOVE i dokazano je da postoji zna~ajnarazlika izme|u ispitivanih grupa za slede}e faktorerizika: WHR (grupa 2 prema grupi 3, Tuckey testp=0,008), HDL-H (grupa 1 prema grupi 2, p=0,024i grupa 1 prema grupi 3, p=0,034), mokra}na kiselina(grupa 1 prema grupi 3, p=0,039). Nije bilo zna~ajnerazlike ni za jedan drugi ispitivani parametar (p>0,05).Dobijeni rezultati ukazuju na to da sniènje HDL-holes-

and older patients (p>0.05). A strong positive rela-tionship was found between CRP and measures of obe-sity in healthy subjects (BMI p<0.001 and WHRp<0.01 by linear regression analysis), but the statisti-cally significant relationship disappeared in the groupof patients. The results of this study indicate that adi-posity is strongly associated with CRP in healthy adultwomen. Also, these data suggest that adipose tissue isan important determinant of a low level, chronicinflammatory state as reflected by CRP levels. Elderlywomen have higher WHR and CRP levels than youngerwomen probably because of a lack of the protectiveinfluences of estrogen.

A6THE ASSOCIATION BETWEEN HIGH-NORMAL

BLOOD PRESSURE AND RISK FACTORS FOR CARDIOVASCULAR DISEASE

L. Memon1, N. Bogavac-Stanojevi}2, V. Spasojevi}-Kalimanovska2, S. Spasi}2,

Z. Jeli}-Ivanovi}2, A. Zeljkovi}2

1Clinical Center »Beànijska kosa«, Belgrade, Serbia

2Institute of Medical Biochemistry, Faculty of Pharmacy, Belgrade, Serbia

According to the classification approaches devel-oped by the joint JNC VI and WHO-ISH, nonhyperten-sive persons with a systolic pressure of 130 to 139mmHg, diastolic pressure of 85 to 89 mmHg, or bothare categorized as having high-normal blood pressure.Information is limited regarding the risk of cardiovas-cular disease (CAD) in these subjects. We investigatedthe association between blood pressure category andthe risk factors for CAD in these persons. A total of 99healthy persons, 53 men and 46 women, were includ-ed in the study. The subjects were classified into one ofthe three nonhypertensive blood pressure categories:optimal (systolic pressure of less than 120 mmHg anddiastolic pressure of less than 80 mmHg, group 1),normal (systolic pressure of 120 to 129 mmHg or dias-tolic pressure of 80 to 84 mmHg, group 2), or high-normal (systolic pressure of 130 to 139 mmHg or dias-tolic pressure of 85 to 89 mmHg, group 3). We col-lected information such as age, smoking status, alco-hol consumption and exercise from self-reports. Bodymass index (BMI) and waist-to-hip ratio (WHR) werecalculated. In the blood samples, hsCRP, uric acid andlipid parameters (total cholesterol, HDL-H, LDL-H,triglycerides, Lp(a), apoA-I, apoB) were determined bystandard laboratory procedures. Fibrinogen was meas-ured by Clauss method. The results obtained by usingan ANOVA have shown that there was a significant dif-ference in WHR (group 2 vs. group 3, Tuckey testp=0.008), HDL-H (group 1 vs. group 2, p=0.024 andgroup 1 vs. group 3, p=0.034) and uric acid (group 1vs. group 3, p=0.039). There was no difference forany of the other examined risk factors (p>0.05). Thesedata suggest that the lowering of HDL-cholesterol and

JMB 2008; 27 (2) 227

a rise of uric acid level may lead to a subsequent devel-opment of hypertension. Also, we found a progressiveincrease in blood pressure with the increasing of adi-pose tissue. In primary care WHR is better tool thanBMI for screening and following obesity in subjects withoptimal to high-normal blood pressure.

A7INCIDENCE OF HYPERHOMOCYSTEINEMIA

AND MTHFR C677T POLYMORPHISM AMONG YOUNG PATIENTS WITH ACUTE

MYOCARDIAL INFARCTION

A. Beleti}1, D. Mirkovi}1, N. Antonijevi}2,V. \or|evi}3, V. [ango4, B. Jakovljevi}5, J. Peruni~i}1,

M. Ili}1, Z. Vasiljevi}1, N. Majki}-Singh1

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

2Institute of Cardiovascular Diseases,Clinical Center of Serbia, Belgrade, Serbia

3Institute of Molecular Genetics and GeneticEngineering, Belgrade, Serbia

4Institute of Biology and Human Genetics, School of Medicine, Belgrade, Serbia

5Institute of Hygiene and Medical Ecology, School of Medicine, Belgrade, Serbia

Hyperhomocysteinemia (HHcy) is considered anindependent risk factor for premature cardiovasculardiseases. The reduction of the methylentetrahydropho-late reductase (MTHFR) activity, due to the presence ofC677T mutation, is one of the causes of HHcy. Weassessed the HHcy incidence, homocysteine (Hcy) leveland distribution of all the three MTHFR 677 genotypes(C/C, C/T and T/T) in a group of young patients withacute myocardial infarction (AMI) and compared themwith those obtained in healthy persons, matched forage. Thes study involved 86 patients younger than 45years of age (77 men and 9 women) and 35 healthycontrols. Homocysteine was measured in serum, usingan HPLC method with fluorescent detection. PCRamplification and digestion with restrictive endonucle-ase Hinf I were employed for the determination ofMTHFR 677 genotype. Statistical analyses included chi-square and Mann-Whitney U tests. HHcy, defined asHcy higher than 12 mmol/L, was present in 32.6%patients and in 14.3% controls, which was a significantdifference (p=0.038). Medians of Hcy in young adultswith AMI (10.4 mmol/L) and in healthy young adults(9.6 mmol/L) were significantly different (p=0.035).Among the patients, 50.0% had C/C, 41.9% had C/Tand the remaining 8.1% had T/T genotype. No signifi-cant difference in the distribution of MTHFR genotypeswas observed between the patients and controls.MTHFR genotype had no influence on HHcy incidenceand Hcy level, both in patients and controls. We con-clude that young patients with AMI have higher inci-dence of HHcy and a higher Hcy level than healthyyoung adults, while there is no significant difference inthe distribution of MTHFR C677T genotypes.

terola, pove}anje mokra}ne kiseline i gojaznost moguda dovedu do razvoja hipertenzije. U primarnoj preven-ciji, odnos struk/kuk je bolji parametar od BMI zadetektovanje i pra}enje stepena gojaznosti kod osobabez povi{enog krvnog pritiska.

A7INCIDENCA HIPERHOMOCISTEINEMIJE I MTHFR C677T POLIMORFIZAM KOD

MLADIH BOLESNIKA SA AKUTNIM INFARKTOM MIOKARDA

A. Beleti}1, D. Mirkovi}1, N. Antonijevi}2,V. \or|evi}3, V. [ango4, B. Jakovljevi}5, J. Peruni~i}1,

M. Ili}1, Z. Vasiljevi}1, N. Majki}-Singh1

1Institut za medicinsku biohemiju, Klini~ki centar Srbije, Beograd

2Institut za kardiovaskularne bolesti, Klini~ki centar Srbije, Beograd

3Institut za molekularnu genetiku i geneti~ko inènjerstvo, Beograd

4Institut za biologiju i humanu genetiku, Medicinski fakultet, Beograd

5Institut za higijenu i medicinsku ekologiju, Medicinski fakultet, Beograd

Hiperhomocisteinemija (HHcy) smatra se neza-visnim faktorom rizika za preuranjeni razvoj kardio-vaskularnih bolesti. Snièna enzimska aktivnost metilen-tetrahidrofolat reduktaze (MTHFR), usled prisustvaC677T mutacije, jedan je od uzroka HHcy. Odre|eni suincidenca HHcy, koncentracija homocisteina (Hcy) iraspodela MTHFR 677 genotipova (C/C, C/T i T/T)kod grupe mladih bolesnika sa akutnim infarktommiokarda (AIM) i upore|eni sa vrednostima dobijenimkod zdravih osoba odgovaraju}e starosti. Studija jeobuhvatila 86 bolesnika sa AIM, mla|ih od 45 godina(77 mu{karaca i 9 èna), i kontrolnu grupu od 35 oso-ba. Homocistein je odre|ivan u serumu, HPLC me-todom uz fluorescentnu detekciju. PCR amplifikacija idigestija sa restriktivnom endonukleazom Hinf I kori{-}ene su za genotipizaciju MTHFR. Podaci su statisti~kiobra|eni Mann-Whitney U i Chi-square testom. HHcy,definisana kao nivo Hcy vi{i od 12 mmol/L, bila je pri-sutna kod 32,6% bolesnika i kod 14,3% kontrola, {to jepredstavljalo zna~ajnu razliku (p=0,038). Medijane Hcymladih ljudi sa AIM (10,4 mmol/L) i zdravih mladih ljudi(9,6 mmol/L) bile su zna~ajno razli~ite (p=0,035).Genotip C/C je bio prisutan kod 50,0%, C/T kod 41,9%i T/T kod 8,1% bolesnika. Nije uo~ena zna~ajna razlikau raspodeli genotipova MTHFR izme|u bolesnika i kon-trola, kao ni u incidenci HHcy i nivou Hcy izme|ugenotipova MTHFR. Moè se zaklju~iti da mladi bolesni-ci imaju vi{u incidencu HHcy i vi{i nivo Hcy nego zdraveosobe iste starosti, pri ~emu nema zna~ajne razlike uraspodeli genotipova MTHFR C677T.

228

A8PARAMETRI FIBRINOLIZE KOD PACIJENATA

SA INFARKTOM MIOKARDA

D. Vukosavljevi}1, N. Antonijevi}2, T. Vodnik1, I. Vujo{evi}1, A. Beleti}1

1Institut za medicinsku biohemiju, Klini~ki centar Srbije, Beograd, Srbija2Institut za kardiovaskularne bolesti, Klini~ki centar Srbije, Beograd, Srbija

U nastanku infarkta miokarda (IM) izvesnu ulogumogu imati i poreme}aji fibrinoliti~kog sistema, pa seprocena njegove funkcije smatra veoma zna~ajnom zaprognozu. U studiji su upore|ene aktivnosti inhibitoraaktivatora plazminogena (PAI) i plazminogena, kao inivoi fibrinogena izme|u grupe od 50 bolesnika sa IM,mla|ih od 50 godina (30 mu{karaca i 20 èna), i kon-trolne grupe od 50 zdravih osoba, odgovaraju}e sta-rosti (30 mu{karaca i 20 èna). Navedeni parametri suodre|eni u uzorcima plazme, standardnim metodama,a rezultati upore|eni Mann-Whitney U testom. Aktivno-sti PAI i plazminogena nisu pokazale statisti~ki zna~ajnurazliku, dok su nivoi fibrinogena bili zna~ajno vi{i(p=0,000) kod bolesnika nego u kontrolnoj grupi.Mu{karci sa IM su imali zna~ajno vi{e vrednosti fibrino-gena (p=0,000) u odnosu na zdrave mu{karce, a ta-kva razlika nije uo~ena kod èna. Moè se zaklju~iti daizme|u bolesnika sa IM i zdravih osoba nema razlike uaktivnostima PAI i plazminogena, a bolesnici imaju vi{enivoe fibrinogena nego zdrave osobe. Mladi mu{karcisa IM imaju ve}u koncentraciju fibrinogena u odnosuna zdrave mu{karce odgovaraju}e starosti, dok se takvarazlika ne uo~ava kod èna.

A9NIVO C-REAKTIVNOG PROTEINA

NEPOSREDNO PO PRIJEMU UINTERNISTI^KU AMBULANTU KOD

PACIJENATA KOD KOJIH SE SUMNJA NA INFARKT MIOKARDA

B. Mirjani}-Azari}1, M. \eri}2, S. Avram1, N. Ra{eta3

1Klini~ki centar »Banja Luka«,Banja Luka, Republika Srpska

2Klini~ki centar Vojvodine, Centar za laboratorijsku medicinu, Novi Sad, Srbija

3Medicinski fakultet, Banja Luka, Rep. Srpska

C-reaktivni protein (CRP) jedan je od najosetljivijihmarkera inflamacije a istovremeno i biomarker atero-skleroze, koja se danas smatra hroni~nom inflama-tornom bole{}u. Cilj rada je da se odredi nivo CRP-a kodpacijenata kod kojih se sumnja na akutni infarkt miokar-da i izvr{i ispitivanje korelacije izme|u inflamatornog

A8THE PARAMETERS OF FIBRINOLYSIS IN

PATIENTS WITH MYOCARDIAL INFARCTION

D. Vukosavljevi}1, N. Antonijevi}2, T. Vodnik1,I. Vujo{evi}1, A. Beleti}1

1Institute of Medical Biochemistry, Clinical Centre of Serbia, Belgrade, Serbia

2Institute of Cardiovascular Diseases, Clinical Centre of Serbia, Belgrade, Serbia

The disturbances in various components of the fi-brinolytic system may account for the development ofmyocardial infarction (MI) and are considered as valu-able prognostic factors. Our study compared plasmi-nogen activator inhibitor (PAI), plasminogen and fibri-nogen levels between the group of 50 patients with MI,younger than 50 (30 men and 20 women), and a con-trol group of 50 healthy persons (30 men and 20women), matched for age. Plasma samples were ana-lyzed by standard methods. The results were comparedusing Mann-Whitney U test. PAI and plasminogen lev-els showed no statistically significant difference, whilefibrinogen levels were significantly higher (p=0.000) inpatients than in controls. Men with myocardial infarc-tion had significantly higher fibrinogen levels(p=0.000) than healthy men, while such differencesdid not exist between healthy women and women withmyocardial infarction. We conclude that there was nodifference in PAI and plasminogen levels, while fibrino-gen levels were higher in all patients with myocardialinfarction compared with the control group. Youngmen with myocardial infarction have higher fibrinogenlevels than healthy young men, while there is no suchdifference for women.

A9LEVELS OF C-REACTIVE

PROTEIN AT ADMISSION TO AN INTERNIST OUT-PATIENT CLINIC

IN PATIENTS SUSPECTED OF HAVINGACUTE MYOCARDIAL INFARCTION

B. Mirjani}-Azari}1, M. \eri}2, S. Avram1, N. Ra{eta3

1Clinical Center »Banja Luka«, Banja Luka, Republic of Srpska

2Clinical Center of Vojvodina, Dept. of Laboratory Medicine, Novi Sad, Serbia

3Medical Faculty, Banja Luka, Republic of Srpska

C-reactive protein (CRP) is one of the most sensi-tive markers of inflammation and at the same time abiomarker of atherosclerosis, that is considered a chro-nic illness. The aim of this project is to determine theCRP level in patients suspected of having acute myo-cardial infarction, and also to establish the presence of

JMB 2008; 27 (2) 229

correlation between an inflammatory response express-ed in C-reactive protein and acute myocardial infarc-tion and to establish the presence of correlation bet-ween C-reactive protein and the lipid profile. The proj-ect involved 94 patients who were admitted into theInternist Out-Patient Clinic of the Clinical Centre inBanjaluka, with suspected myocardial infarction. Thevalues of total cholesterol, HDL and LDL cholesterol,aminotransferase (AST, ALT), creatine kinase (CPK),and glucose were determined by standard laboratorymethods. C-reactive protein was measured by a high-sensitivity immunoturbidimetric method (Latex) on a»Hitachi 902« analyzer using »Roche« diagnostic re-agents. The average age of the patients was 59 years.There were 71 men and 23 women in the group. Aftera diagnose for each patient was established, two groupswere formed: a group of 22 patients with the diagno-sis of myocardial infarction and a group of 72 withoutmyocardial infarction. In the group of patients withmyocardial infarction the mean value of C-reactive pro-tein was 12.89 mg/L, total cholesterol 5.13 mmol/L,HDL cholesterol 1.02 mmol/L, LDL cholesterol 3.17mmol/L, triglycerides 2.00 mmol/L. In the group ofpatients without myocardial infarction the mean valueof C-reactive protein was 8.50 mg/L, total cholesterolwas 5.34 mmol/L; HDL cholesterol 1.26 mmol/L, LDLcholesterol 3.16 mmol/L and triglycerides 2.03mmol/L. Statistical analyses showed that there is a sta-tistically significant relation between CRP values andHDL cholesterol, and also between CRP and total cho-lesterol. No connection was found between the con-centrations of CRP within the two examined groups.There is a statistically important difference in the HDLvalues of the examinees with or without myocardialinfarction (p<0.01).

A10COMPARISON OF THREE DIFFERENT

RISK SCORES FOR A 10-YEAR CORONARYHEART DISEASE RISK EVALUATION

N. Eremi}, M. \eri}, S. Koji}-Damjanov

Clinical Centre of Vojvodina, Dept. of Laboratory Medicine, Novi Sad, Serbia

Considering that atherosclerosis is a multifactorialdisease and that risk factors have multiplicative effects,in the everyday routine identification and treatment ofasymptomatic individuals at high risk for developingcoronary heart disease (CHD) different risk scoringschemes are used. The aim of this study was to com-pare the estimated 10-year CHD risk according toFramingham (FHS), PROCAM and SCORE risk scoresin 110 asymptomatic individuals of both sexes, 30–85years old, at first examination and after a one-year lipiddisorder treatment. The risk factors involved in the cal-

odgovora izraènog serumskom koncentracijom C-reak-tivnog proteina i akutnog infarkta miokarda, te korelacijeizme|u C-reaktivnog proteina i lipidskih parametara.Studija je obuhvatila 94 pacijenta, neposredno po prije-mu u internisti~ku ambulantu Klini~kog centra »Ba-njaluka« kod kojih se sumnjalo na infarkt miokarda.Standardnim biohemijskim metodama odre|ivane suvrednosti ukupnog, HDL i LDL holesterola, aminotrans-feraza (AST, ALT), kreatin kinaze (CK), i glikemija. C-reaktivni protein odre|ivan je visokosenzitivnom imuno-turbidimetrijskom metodom na aparatu »Hitachi 902«,reagensima firme »Roche«. Prose~na starost pacijenatabila je 59 godina. U grupi je bio 71 mu{karac i 23 ène.Na osnovu kasnije postavljene dijagnoze za svakog paci-jenta, oformljene su grupe od 22 pacijenta sa infarktommiokarda i 72 pacijenta bez infarkta miokarda. U grupipacijenata koji su imali infarkt miokarda, srednja vred-nost C-reaktivnog proteina bila je 12,89 mg/L, ukupnogholesterola 5,13 mmol/L, HDL holesterola 1,02mmol/L, LDL holesterola 3,17 mmol/L, triglicerida 2,00mmol/L, a u grupi bez infarkta srednja vrednost C-reak-tivnog proteina bila je 8,50 mg/L, ukupnog holesterola5,34 mmol/L, HDL holesterola 1,26 mmol/L, LDLholesterola 3,16 mmol/L, triglicerida 2,03mmol/L. Sta-tisti~kom obradom podataka utvr|eno je da postoji vrlozna~ajna povezanost (p<0,01) izme|u vrednosti C-reak-tivnog proteina i HDL holesterola, kao i CRP-a i ukupnogholesterola. Nije na|ena korelacija izme|u koncentraci-ja CRP-a unutar dve grupe ispitanika. Postoji statisti~kiveoma zna~ajna razlika izme|u vrednosti HDL kod ispi-tanika sa i bez infarkta miokarda (p<0,01).

A10PORE\ENJE TRI RAZLIÎTA BODOVNA SISTEMA ZA PROCENU 10-GODI[NJEG

RIZIKA ZA NASTANAK KORONARNE BOLESTI SRCA


Klini~ki centar Vojvodine, Centar za laboratorijsku medicinu, Novi Sad, Srbija

Budu}i da je ateroskleroza multifaktorijalna bolesti da faktori rizika imaju multiplikativno dejstvo, u ciljuidentifikacije i le~enja asimptomatskih osoba sa visokimrizikom za razvoj koronarne bolesti srca (KBS) u praksise koriste razli~iti bodovni sistemi za procenu rizika. Ciljistraìvanja bio je pore|enje procenjenog 10-godi{njegrizika za KBS prema Framinghamskom (FHS), PRO-CAM i SCORE bodovnom sistemu kod 110 asimpto-matskih osoba oba pola, starosti 30–80 godina, pri-likom prvog pregleda i godinu dana posle le~enja lipid-skog poreme}aja. Faktori rizika koji su uklju~eni u izra-

230

~unavanje su pol, starost, ukupni, LDL i HDL holes-terol, sistolni krvni pritisak, pozitivnost porodi~ne anam-neze, kao i podatak o uzimanju antihipertenzivne te-rapije i pu{a~kom statusu. Od 110 ispitanika, nizak rizikimalo je prema FHS 60 osoba (54,55%), PROCAM-u86 (78,18%) i SCORE-u 48 (43,64%), srednji rizikprema FHS 35 (31,82%), PROCAM-u 15 (13,64%) iSCORE-u 37 (33,64%), i visok rizik prema FHS 15(18,18%), PROCAM-u 9 (8,18%) i SCORE-u 25(22,73%) osoba. Posle godinu dana le~enja lipidskogporeme}aja niskorizi~nih je bilo prema FHS 86(78,18%), PROCAM-u 97 (88,18%) i SCORE-u 61(55,45%), srednjerizi~nih prema FHS 17 (15,45%),PROCAM-u 11 (10,00%) i SCORE-u 36 (32,73%), avisokorizi~nih prema FHS 7 (6,36%), PROCAM-u 2(1,82%) i SCORE-u 13 (11,82%). Postoji statisti~kizna~ajna razlika (p<0,05) u pripadnosti odre|enoj ka-tegoriji rizika i to prilikom prvog pregleda izme|u FHSi PROCAM-a, kao i izme|u PROCAM-a i SCORE-a, zasve kategorije rizika, a posle godinu dana izme|u FHSi PROCAM-a, FHS i SCORE-a za kategorije s niskim isrednjim rizikom, a izme|u PROCAM-a i SCORE-a zasve kategorije rizika. Razlike u pripadnosti odre|enojkategoriji rizika koje se dobijaju kori{}enjem tri razli~itabodovna sistema mogu se objasniti razli~itim faktorimarizika koji su uklju~eni u svaki algoritam kao i statisti-~kim hazard modelima koji su u osnovi svakog od njih.

A11PARAMETRI AKTIVACIJE TROMBOCITA

KOD PACIJENATA PODVRGNUTIHPERKUTANOJ TRANSLUMINALNOJ

KORONARNOJ ANGIOPLASTICI

V. Subota, Z. Miju{kovi}, E. Stankovi}, J. Pejovi}, S. Obradovi}

Institut za medicinsku biohemiju, Klinika za urgentnu medicinu, Klinika za kardiologiju,

Vojnomedicinska akademija, Beograd, Srbija

Trombociti igraju va`nu ulogu u razvoju i formi-ranju tromba kod pacijenata koji su podvrgnuti perku-tanoj transluminalnoj koronarnoj angioplasici (PTCA),zbog efekta PTCA na }elije endotela na mestu implan-tacije stenta. Shodno tome, antikoagulantna i anti-trombocitna terapija se ~esto koriste kod tih pacijenata.Cilj ove studije je bio da se ispita novi parametar trom-bocitne aktivacije (MPC, srednja gustina trombocita,g/L), za utvr|ivanje aktivacije tormbocita i njihovogodgovora na antiagregacionu terapiju (klopidogrel){est meseci posle PTCA intervencije. Parametri trom-bocita MPC, srednji volumen trombocita (MPV, fL),distribucija {irine trombocitne gustine (PCDW, g/L),srednja masa trombocita (MPM, pg) i broj trombocita(PC × 109/L) analizirani su na automatskom hemato-lo{kom sistemu »ADVIA 120.« Za merenje ADP (%),kao parametra agregabilnosti trombocita, kori{}en jekomercijalni test firme »Dade Behring«. Krv je saku-

culation are gender, age, total cholesterol, HDL andLDL cholesterol, systolic blood pressure, treatment ofhypertension and positive family history. Among 110individuals, there were 60 persons at low risk accordingto FHS (54.55%), PROCAM 86 (78.18%) and SCORE48 (43.64%); intermediate according to FHS 35(31.82%), PROCAM 15 (13.64%) and SCORE 37(33.64%) and high risk according to FHS 15 (18.18%),PROCAM 9 (8.18%) and SCORE 25 (22.73%). After aone-year lipid disorder treatment 86, persons were atlow risk according to FHS (78.18%), PROCAM 97(88.18%) and SCORE 61 (55.45%); intermediateaccording to FHS 17 (15.45%), PROCAM 11(10.00%) and SCORE 36 (32.73%) and high riskaccording to FHS 7 (6.36%), PROCAM 2 (1.82%) andSCORE 13 (11.82%). There were significant differen-ces (p<0.05) in the risk category assignment at firstexamination between FHS and PROCAM as well asbetween PROCAM and SCORE, and after a one-yearlipid disorder treatment between FHS and PROCAMand FHS and SCORE in low and intermediate risk ca-tegories and between PROCAM and SCORE for all riskcategories. The differences in risk category assignmentusing the three different risk scoring schemes can beexplained by the use of different risk factors involved inalgorithm calculation as well as different statistic hazardmodels used for their construction.

A11PLATELET ACTIVATION PARAMETERS

IN PATIENTS UNDERGOING PERCUTANEOUS TRANSLUMINAL

CORONARY ANGIOPLASTY

V. Subota, Z. Miju{kovi}, E. Stankovi}, J. Pejovi}, S. Obradovi}

Institute of Medical Biochemistry, Emergency Medicine Clinic, Clinic of Cardiology,

Military Medical Academy, Belgrade, Serbia

Platelets play a major role in the development ofthrombus formation in patients undergoing percuta-neous transluminal coronary angioplasty (PTCA),because PTCA has an effect of damaging the endothe-lial cells on the spot when the stent is present. Con-sequently, anticoagulant and antiplatelet therapies arewidely used for those patients. The aim of the studywas to investigate the usefulness of new activatingplatelet parameters (MPC, mean platelet componentconcentration, g/L) for evaluating the platelet activa-tion and the platelet response to antiplatelet therapy(clopidogrel) in patients six months after PTCA. Theplatelet research parameters MPC and other relatedparameters i.e., the mean platelet volume (MPV, fL),platelet component distribution width (PCDW, g/L),mean platelet mass (MPM, pg) and platelet count (PC× 109/L) were analyzed on an automated ADVIA 120Hematology System. A commercial Dade Behring test

JMB 2008; 27 (2) 231

was used for measuring the platelet aggregometryparameter, ADP (%). Blood samples were collectedfrom 26 patients before (baseline) and after an ergo-metric test. A decrease in platelet density, measured byreduction in MPC (means SD, 242.8 13.56, 225.8

15.37, p<0.001) is indicative of the platelet activa-tion. The values of MPV increased significantly(9.7 1.11, 10.25 1.08, p<0.001), reflecting a sha-pe change. ADP values was mildly decreased(52.8 10.48, 48.9 12.22) reflecting the response inpatients on antiaggregation therapy which modifiedthe platelet activation. Our data show that the use ofplatelet activation parameters in patients undergoingthe PTCA may be helpful in detecting the risk ofplatelet activation, and hence as an indicator of pa-tients at risk of thrombosis.

A12CASPASE-3 ACTIVITY IN THE

LYMPHOCYTES OF PATIENTS WITHISCHEMIC HEART DISEASE

T. Risti}1, V. ]osi}1, P. Vlahovi}1, M. Delanin-Ili}3, D. Stankovi}- Ferle`1, L. Zvezdanovi}1, B. V. \or|evi}2

1Centre for Medical Biochemistry, Clinical Centre, Ni{, Serbia

2Institute of Biochemistry, Medical Faculty, Ni{, Serbia3Institute for Cardiovascular and Rheumatic Diseases,

Ni{ka Banja, Serbia

Apoptotic cell death may play a critical role in avariety of cardiovascular diseases and may occur inresponse to ischemia, toxins and physical stimuli. A keyphenomenon of apoptotic cell death is the activation ofone of the effector caspases – caspase-3, which maybe activated by both extrinsic and intrinsic apoptoticpathways. The caspase-3 activation leads to DNA frag-mentation and cell death. In this study, the activity ofcaspase-3 was determined in the lymphocytes of pe-ripheral blood isolated using a lymphocyte separationmedium. Caspase-3 activity was measured by a colori-metric commercially available ELISA kit based on thedegradation of synthetic tetrapeptide DEVD-pNa.Enzyme activity was determined in the lymphocytes ofpatients with stable angina (SAP, 30), with unstableangina pectoris (USAP, 26), and with acute myocardialinfarction (AMI, 38). The obtained results were compa-red to the caspase-3 values in the control group (he-althy individuals). In the lymphocytes of patients withSAP the enzyme activity was 0.093±0.035 mmol/mgprotein, but in patients with AMI 0.106±0.061 mmol/mgprotein, and both values were insignificantly higher incomparison with controls (0.092±0.022 mmol/mgprotein). In the lymphocytes of NSAP patients the cas-pase-3 activity (0.118±0.061 mmol/mg protein) wassignificantly higher (p<0.05) compared to the controlas well as to both other patient groups. In conclusion,the caspase-3 activity may be a valid parameter forassessing atherosclerotic plaque activity, and a new tar-get for therapeutic intervention.

pljena od 26 pacijenata pre (bazalno) i posle ergo-metrije. Sniènje u gustini trombocita, mereno reduk-cijom u MPC (srednja vrednost SD, 242,8 13,56,225,8 15,37, p<0,001) je znak trombocitne akti-vacije. Vrednost MPV je zna~ajno pove}ana (9,71,11, 10,25 1,08, p<0,001), reflektuju}i promenuoblika trombocita. ADP je blago snièn (52,8 10,48,48,9 12,22) kao odgovor pacijenata na anti-agrega-cionu terapiju koja modifikuje aktivaciju trombocita.Na{i podaci pokazuju da upotreba parametara akti-vacije trombocita kod pacijenata podvrgnutih PTCAmoè biti korisna alatka u otkrivanju rizika za aktivacijutrombocita i sledstveno tome rani indikator rizika zarazvoj tromboze.

A12AKTIVNOST KASPAZE-3 U LIMFOCITIMA

BOLESNIKA SA ISHEMIJSKOM BOLE[]U SRCA

T. Risti}1, V. ]osi}1, P. Vlahovi}1, M. Delanin-Ili}3, D. Stankovi}- Ferle`1, L. Zvezdanovi}1, B. V. \or|evi}2

1Centar za medicinsku biohemiju, Klini~ki centar, Ni{, Srbija

2Institut za biohemiju, Medicinski fakultet, Ni{, Srbija3Institut za kardiovaskularne i reumatske bolesti,

Ni{ka Banja, Srbija

Smrt }elija apoptozom moè imati kriti~nu uloguu razli~itim kardiovaskularnim oboljenjima. Apoptozase moè indukovati kao odgovor na ishemiju, toksine ifizi~ke stimuluse. Klju~ni fenomen u apoptozi je akti-vacija jedne od efektorskih kaspaza – kaspaze-3, kojase moè aktivisati indukcijom i spolja{njeg i unutra{-njeg puta apoptoze. Njena aktivacija vodi fragmentaci-ji DNK i smrti }elije. U ovom radu odre|ivana je aktiv-nost kaspaze-3 u limfocitima periferne krvi, koji su izo-lovani pomo}u separacionog medijuma. Aktivnost kas-paze-3 merena je komercijalnim, kolorimetrijskimELISA testom koji se bazira na razgradnji sintetskogtetrapeptida DEVD-pNa. Aktivnost enzima je odre|i-vana u limfocitima bolesnika sa stabilnom anginom(SAP, 30), sa nestabilnom anginom pektoris (NSAP, 26)i sa akutnim infarktom miokarda (AIM, 38). Dobijenirezultati su pore|eni sa vrednostima kaspaze-3 u kon-trolnoj grupi (zdravi ispitanici). U limfocitima bolesnikasa SAP aktivnost enzima je bila 0,093±0,035 mmol/mg proteina, a kod bolesnika sa AIM 0,106±0,061mmol/mg proteina, (to su nezna~ajno vi{e vrednosti uodnosu na one u kontolnoj grupi (0,092±0,022mmol/mg proteina). U limfocitima bolesnika sa NSAPaktivnost kaspaze-3 (0,118±0,061 mmol/mg pro-teina) bila je zna~ajno povi{ena (p<0,05) u odnosu nakontrolnu i ostale dve grupe bolesnika. Moè se za-klju~iti da aktivnost kaspaze-3 moè biti validan para-metar u proceni aktivnosti ateroskleroti~nog plaka, ali inova mogu}nost za terapijsku intervenciju.

232

JMB 2008; 27 (2) 233

A13THE UTILITY OF BIOMARKERS AND

CLINICAL ANALYTES AS PREDICTORS IN CARDIAC SURGERY

V. Maravi}-Stojkovi}1, D. Brunner-Pavlovi}2, T. Spasi}1, A. Rankovi}1, M. Jovi}1, Z. Bobar2

1»Dedinje« Cardiovascular Institute, Belgrade, Serbia2»Inter-Light« Laboratory, Belgrade, Serbia

The cardiac hormone BNP is a valuable diagnos-tic marker in patients with congestive heart failure(CHF), but BNPis also an important prognostic indica-tor for patients undergoing open heart surgery. Openheart surgery is asssociated with significant risks forpostoperative cardiac events and an inflammatory pha-se with different degrees of severity is always present.The ability to effectively predict postoperative compli-cations, the duration of hospital stay and mortality is yetuncertain, and no gold standard currently exists. Thepurpose of the present study was to propose a varietyof multifactor indexes for preoperative risk assessmentin patients undergoing cardiac surgery. This part of thestudy focuses on the following tests: BNP and the»Inflammatory index« based on procalcitonin (PCT)and C-reactive protein (CRP) analysis, and the relationto renal function of the patients. This choice offers animportant addition to the preoperative quantitative eva-luation. One hundred and two patients with CHF(13.7% female, 86.3% male; ages: 62.9 7.8 vs 599.7 years) were divided into three groups, on the basisof the surgical intervention: CABG – group 1, valvereconstruction – group 2, and the combination of both,CABG+valve procedures – group 3. We introduce anddefine the »inflammatory index« established on thebasis of the PCT-ILMA method and CRP immunotur-bidimetric analysis, BNP (P) by a direct immunochemi-luminiscent automatic assay, measured on anACS180Plus (Bayer) analyzer. GFR was calculated withMDRD. The analyses were performed preoperatively, 6hours, 24 hours and 48 hours after the intervention.Results for BNP, the Inflammatory index, creatinine andGFR are:

The inflammatory process predicts both short-term and long-term cardiac risks. The mean Infla-mmatory index was lowest in group 2 and highest inthe group 3. Postoperatively, all the inflammatory indexvalues were increased. The inflammatory markers PCTand CRP have significant predictive values. This studyshows that the inflammatory index is a stronger pre-

A13KORISNOST BIOMARKERA

I KLINI^KIH ANALITAKAO PREDIKTORA U KARDIOHIRURGIJI

V. Maravi}-Stojkovi}1, D. Brunner-Pavlovi}2, T. Spasi}1, A. Rankovi}1, M. Jovi}1, Z. Bobar2

1Kardiovaskularni institut »Dedinje«, Beograd, Srbija2Laboratorija »Inter-Light«, Beograd, Srbija

Kardiohormon BNP je dragocen dijagnosti~kimarker kod pacijenata sa kongestivnom sr~anom sla-bo{}u (HF), ali BNP je i vaàn prognosti~ki indikator zapacijente koji podleù kardiohirurgiji na otvorenomsrcu. Hirurgiju na otvorenom srcu prate zna~ajni post-operativni kardio-doga|aji, a prisutna je uvek i inflama-torna faza sa razli~itim stepenom teìne. Mogu}nostpredvi|anja efektivnih postoperativnih komplikacija,duìne hospitalizacije i morbiditeta jo{ uvek je nesigur-na, jer zasad ne postoji »zlatni standard«. Cilj ove studi-je bio je da predloì varijetet »multifaktornih indeksa« zapreoperativni rizik pacijenata kod kojih treba da se izvr{ikardiohirur{ki zahvat. Za ovaj deo studije izabrani suslede}i testovi: BNP i »inflamatorni indeks« zasnovan naanalizama prokalcitonina (PCT) i C-reaktivnog proteina(CRP) u odnosu na renalnu funkciju pacijenta. Takavizbor pruà zna~ajan doprinos za preoperativnu kvanti-tativnu evaluaciju. Stodvoje pacijenata sa CHF (13,7%èna, 86,3% mu{karaca starosne dobi 62,9 7,8 vs.59 9,7 godina) podeljeni su u tri grupe na bazi hirur-{ke intervencije: CABG – grupa 1 (coronary artery by-pass grafting), valve rekonstrukcija – grupa 2, i kombi-nacija obe, CABG+valve – grupa 3. Za sve testove sukori{}ene imunohemijske metode: PCT (ILMA), CRP(imunoturbidimetrijska), BNP direktna imunohemilu-miniscenca (»Bayer ACS180Plus«). Analize su izvo|e-ne preoperativno, 6 ~asova, 24 ~asa i 48 ~asova posleoperacije. Dobijeni rezultati su slede}i:

Inflamatorni proces predvi|a i kartkoro~ne i du-goro~ne kardio-rizike. Srednja vrednost inflamatornogindeksa bila je najnià u grupi 2, a najvi{a u grupi 3.Postoperativno su sve vrednosti Inflamatornog indeksabile povi{ene. Inflamatorni markeri PCT i CRP imajuzna~ajnu prediktivnu vrednost. Ova studija pokazuje daje Inflamatorni indeks ja~i prediktor za rizike komp-likacija. ̂ etrdesetosam ~asova posle operacije oni osta-ju jo{ veoma visoki (cutoff < 10) u sve tri grupe. To po-kazuje da inflamatorni proces jo{ uvek nije zavr{en, ikorisno ih je primeniti za izbor adekvatne terapije i

Klini~ki statusCABG (group 1) VALVE (group 2) CABG+VALVE (group 3)

Infl. BNP GFR creat(S) Infl. BNP GFR creat(S) Infl. BNP GFR creat(S)

index (odnos) Index ng/L mmol/L Index ng/L mmol/L Index ng/L mmol/L

srednje vrednosti srednje vred. srednje vred.

pre-op 54,4 585 61,9 118,7 36,7 515 72,2 99 90 346 63,7 109,8

0 day postop 120,2 181,2 211,3

24h postop 354,1 379,3 399,7

48h postop 389,6 381,1 458,8

range: (Cl: 95%) range: (Cl: 95%) range: (Cl: 95%)

GFR mL/min/1.73 m2 (MDRD) Infl.index =

Clinical statusCABG (group 1) VALVE (group 2) CABG+VALVE (group 3)

Infl. BNP GFR creat(S) Infl. BNP GFR creat(S) Infl. BNP GFR creat(S)

ratio Index ng/L mmol/L Index ng/L mmol/L Index ng/L mmol/L

mean values mean values mean values

pre-op 54.4 585 61.9 118.7 36.7 515 72.2 99 90 346 63.7 109.8

0 day postop 120.2 181.2 211.3

24h postop 354.1 379.3 399.7

48h postop 389.6 381.1 458.8

range: (Cl: 95%) range: (Cl: 95%) range: (Cl: 95%)

GFR mL/min/1.73 m2 (MDRD) Infl.index =

234

dalju kontrolu iste. BNP srednje (mean) vrednosti, anaro~ito srednje vrednosti koreliraju sa inflamatornimindeksom. GFR i kreatinin (i mean i median) vrednos-ti (uklju~eni zbog kontrole renalne funkcije pacijenata)bile su < 90 mL/min /1,73 m2 u sve tri grupe. BNPsrednje vrednosti bile su najvi{e u grupi 2, a najniè ugrupi 1. Odnos izme|u niskih i visokih BNP cutoff vred-nosti stratificira preporu~ene GFR (na{e slede}e sa-op{tenje, drugi deo). Gornje i donje grani~ne vrednos-ti inflamatornog indeksa su se striktno i stalno zna~ajnopomerale u toku celog 48-~asovnog postoperativnogperioda. Shifting grani~nih limita ukazuje na pobolj{a-nje maksimalnog scattering-a analiza (kompaktnostpostoperativnih zbivanja). Predloèni set prediktora jeveoma pouzdan, ekonomski efikasan, {tedi vreme i pri-menjiv je u urgentnoj slu`bi.

A14PRIMENA REYNOLDS RIZIK SKORA U PROCENI KARDIOVASKULARNOG

RIZIKA KOD @ENA

S. Jovi~i}1, S. Ignjatovi}2, N. Majki}-Singh2

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

2Institut za medicinsku biohemiju, Klini~ki centar Srbijei Farmaceutski fakultet, Beograd, Srbija

Starost, hipertenzija, pu{enje, dijabetes i hiperlipi-demija su uklju~eni u globalne rizik skorove za procenukardiovaskularnog rizika. Me|utim, kod èna, do 20%svih koronarnih bolesti se javlja u odsustvu svih ovihglavnih faktora rizika, a kod mnogih èna sa prisutnimtradicionalnim faktorima rizika ne dolazi do koronarnihkomplikacija. Tako|e, skoro 70% svih kardiovaskular-nih bolesti se javlja kod osoba sa intermedijarnim rizi-kom (rizik od 5–20% odre|en Framingham rizik sko-rom (FRS) ili rizik od 1–5% odre|en na osnovu SCOREsistema). Reynolds rizik skor (RRS), novi model proce-ne globalnog kardiovaskularnog rizika, razvijen samoza ène, uklju~uje visoko osetljivi C-reaktivni protein(hsCRP) i pojavu infarkta miokarda pre 60. godine kodjednog od roditelja, pored glavnih faktora rizika – sis-tolnog krvnog pritiska, pu{enja, ukupnog i HDL hole-sterola. Cilj ovog rada je bio pore|enje kardiovasku-larnog rizika odre|enog na osnovu FRS, SCORE i RRSkod 138 zdravih, asimptomatskih èna, starih 30–85godina. Ukupan i HDL holesterol i hsCRP su odre|enina analizatoru »Olympus AU2700«, u serumima izd-vojenim iz uzoraka krvi uzetih posle 12h gladovanja.FRS je izra~unavan pomo}u programa ATP III Risk Esti-mator, rizik na osnovu SCORE-a je dobijen pomo}uHeartScore programa, a RRS je odre|en kori{}enjem

dictor for risks of complications. Forty-eight hours afteroperation, they remain very high (cutoff < 10) in allthree groups. This proves that the inflammatory pro-cess is not finished after 48 hours which is very usefulfor the choice of adequate therapy as well controlmeasures for this therapy. The mean BNP levels, andparticularly the median levels, correlate with the inflam-matory indexes. GFR and creatinine (included for thecontrol of renal dysfunction) mean and median levelswere < 90 mL/min/m2 in all three groups. The medi-an BNP level was the highest in group 2, the lowest ingroup 1. Nevertheless, the relation between the lowerand higher BNP cutoffs stratifies the recommendedGFR (our next communication, part 2 of this study).The upper and lower range limits of the inflammatoryindex shifted strikingly and continuously during thecomplete 48-hour postoperative study phase. Theshifting of the range limits points at an improvement ofthe maximal analysis scattering. The proposed set ofpredictors is very reliable, cost-effective, time-efficientand applicable in emergency services.

A14APPLICATION OF THE REYNOLDS RISK

SCORE IN CARDIOVASCULAR RISK ASSESSMENT IN WOMEN

S. Jovi~i}1, S. Ignjatovi}2, N. Majki}-Singh2

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

2Institute of Medical Biochemistry, Clinical Center of Serbia and Faculty of Pharmacy, Belgrade, Serbia

Age, hypertension, smoking, diabetes and hyper-lipidemia are incorporated into global risk scores forassessment of cardiovascular risk. However, in women,up to 20% of all coronary events occur in the absenceof these major risk factors and many women with tra-ditional risk factors do not experience coronary events.Also, almost 70% of all cardiovascular disease (CVD)events occur among individuals at intermediate risk (5-20% of risk determined using Framingham Risk Score(FRS) or 1–5% of risk determined using SCORE). Rey-nolds Risk Score (RRS), the new risk prediction modelfor global cardiovascular risk, developed exclusively forwomen, includes high sensitivity C-reactive protein(hsCRP) and parental history of myocardial infarctionbefore age 60 next to major risk factors: systolic bloodpressure, smoking, total and HDL cholesterol. The aimof this study was to compare CVD risk assessed usingFRS, SCORE and RRS in 138 healthy, asymptomaticwomen, 30–85 years old. Total and HDL cholesteroland hsCRP were determined in sera collected after a12h fasting period using an Olympus AU2700 analy-zer. FRS was calculated using the ATP III Risk Estimatorand risk status according to SCORE was obtained usingthe HeartScore Programme for populations at highrisk. RRS was determined using Reynolds Risk Score

JMB 2008; 27 (2) 235

Calculator. With RRS, 6 (5%) participants were movedto a higher and 7 (5%) were moved to a lower risk cat-egory compared with their FRS. In comparison withtheir SCORE results, 25 (18%) participants were reclas-sified into a lower risk category and only 3 (2%) werereclassified into a higher risk category according totheir RRS. The difference between risk distribution inthe selected population was significant when SCOREand RRS were compared (c2= 21.53; p<0.001;df=2). Risk distributions according to FRS and RRSshowed no statistical difference (c2=3.08; p=0.214;df=2), which is in disagreement with published data.The reason for this might be the large number of par-ticipants with low initial risk (<5% 10-year risk basedon FRS or <1% according to SCORE) in the selectedpopulation.

A15PATIENTS WITH END STAGE RENAL

DISEASE AND FACTORS WHICH INCREASE THE RISK OF

CARDIOVASCULAR MORBIDITY

M. Perovi}1, M. Novkovi}2

1General Hospital, Clinical Biochemical Laboratory, Vrbas, Serbia

2General Hospital, Department of Internal Medicine, Vrbas, Serbia

Renal failure is accompanied with significantlyincreased risk of cardiovascular morbidity and mor-tality. Previous studies have revealed that progressiverenal failure is accompanied by abnormalities of plas-ma tryglyceride, and a decrease in HDL cholesterolconcentrations with smaller change in the levels ofcholesterol rich lipoproteins. Also, markers of inflam-mation (CRP, fibrinogen), malnutrition (albumin) andhipercoagulability (fibrinogen) have been linked to anexcessive morbidity and mortality. The concentrationsof total cholesterol (TC), low density lipoprotein cho-lesterol (LDL), tryglycerides (TG), apolipoprotein AI(ApoAI), apolipoprotein B (Apo B), lipoprotein a(Lpa), CPR, fibrinogen, albumin were measured inthe samples of 100 patients (65 hemodialysed (HD),35 with chronic renal failure (CRF)) and 100 age andsex-matched healthy patients as controls. Parameterswere examined by standard automated assays usingcommercial reagents. LDL was calculated accordingto the Friedewald formula. Atherogenic risk factorswere calculated as TC/HDL-c and LDL/HDL-c ratio.TC, TG, CRP, fibrinogen, LDL cholesterol, Apo B lev-els were significantly higher (p>0.05) and HDL-cand ApoAI were lower than in healthy persons(p<0.05). Cardiovascular events were noted in 21patients. Prevalence of hypertension and the CRPconcentration were also higher in CRF and HD pa-tients with CVD.

Reynolds Risk Score kalkulatora. Upotrebom RRS 6(5%) u~esnica je pomereno u vi{u, a 7 (5%) u~esnicaje pomereno u niù kategoriju rizika u odnosu na rizikodre|en po FRS-u. U pore|enju sa svojim rizikom poSCORE-a, 25 (18%) u~esnica je klasifikovano u niùkategoriju rizika, a samo 3 (2%) u~esnica je klasifi-kovano u vi{u kategoriju rizika na osnovu RRS-a. Raz-lika izme|u raspodela rizika u odabranoj populaciji jezna~ajna kada se uporede SCORE i RRS (c2=21,53;p<0,001; df=2). Raspodele rizika na osnovu FRS iRRS nisu statisti~ki zna~ajno razli~ite (c2=3,08;p=0,214; df=2), {to se ne slaè sa objavljenim poda-cima. Mogu}i razlog je veliki broj u~esnica sa niskiminicijalnim rizikom (desetogodi{nji rizik <5% na osnovuFRS-a, odnosno <1% na osnovu SCORE-a) u odabra-noj populaciji.

A15PACIJENTI SA BUBRE@NOM

INSUFICIJENCIJOM I FAKTORI KOJI POVE]AVAJU RIZIK OD

KARDIOVASKULARNIH BOLESTI

M. Perovi}1, M. Novkovi}2

1Op{ta bolnica, Biohemijska laboratorija, Vrbas, Srbija

2Op{ta bolnica, Interno odeljenje, Vrbas, Srbija

Oboljenja bubrega su udruèna sa zna~ajnopove}anim rizikom za nastanak kardiovaskularnihobolenja. Mnoga istraìvanja su pokazala da je sma-njena bubre`na funkcija udruèna sa poreme}ajemkoncentracije triglicerida, a zapaà se i sniàvanjeHDL holesterola. Promene u koncentraciji markerainflamacije (CRP, fibrinogen), malnutricije (albumin) ikoagulacije (fibrinogen) tako|e su udruène sa pove-}anim mobiditetom i mortalitetom usled kardiovasku-larnih bolesti kod bubre`nih pacijenata. Koncentra-cije ukupnog holesterola (TC), LDL holesterola, trigli-cerida (TG), apolipoproteina AI (Apo AI), apolipopro-teina B (Apo B), lipoprotein a (Lpa), CRP, fibrinogen,albumin odre|ivani su u 100 uzoraka pacijenata sao{te}enom funkcijim bubrega (65 hemodijaliziranihpacijenata i 35 pacijenata sa hroni~nom bubre`nominsuficijencijom) i 100 zdravih pacijenata. Parametrisu odre|ivani komercijalnim testovima automatizo-vanim metodama. LDL holesterol je izra~unat Friede-waldovom formulom. Aterogeni rizik je izra~unat krozodnos TC/ HDL-c i LDL/HDL-c. TG, CRP, fibrinogen,TC, LDL, Apo B su zna~ajno vi{i kod bubre`nih paci-jenata, dok su HDL-c, Apo AI niì nego u kontrolnojgrupi. Prevalenca hipertenzije kao faktora rizika zanastanak koronarnih bolesti tako|e je prisutna kodpacijenata sa bubre`nom insuficijencijom.

B

METODE U KLINI^KOJ HEMIJI

METHODS IN CLINICAL CHEMISTRY

JMB 2008; 27 (2) 239

B16THE DEVELOPMENT OF A METHOD FOR

THE SIMULTANEOUS MEASUREMENT OF GLUTATHIONE AND GLUTATHIONE

DISULFIDE IN BLOOD WITHOUT DERIVATIZATION EMPLOYING CAPILLARY

ELECTROPHORESIS AND INCLUDING A FACTORIAL DESIGN

G. @uni}1, S. Spasi}2

1Institute for Medical Research, Military Medical Academy, Belgrade2Institute of Medical Biochemistry,

Faculty of Pharmacy, University of Belgrade, Belgrade

Glutathione (GSH) has important roles in theoverall metabolism, including cell homeostasis, radio-protection and antioxidant defense. In the presence offree oxygen radicals it is oxidized to the disulfide form(GSSG). Thus, their measurement is useful for deter-mining the oxidative/antioxidative status in variousphysiological and patophysiological conditions. Sinceblood glutathione concentrations may reflect glu-tathione status in other less accessible tissues, meas-urement of both GSH and GSSG in blood has beenconsidered essential as an index of the whole subjectoxidative status and as a useful indicator of disease riskin humans. Usually, GSH is measured, while GSSG iscalculated. The capillary electrophoresis permits theirseparation, but needs their derivatization. Using a fullfactorial design for screenning important variables (i.e.carrier electrolyte pH, concentration and separationtemperature), we developed a simple and rapidmethod for the simultaneous analysis of GSH andGSSG without any derivatization procedures for theexamined compounds. The obtained results indicatedthat their separation was mainly influenced by the pHof the running electrolyte, while its concentration aswell as temperature had little effect upon their resolu-tions. The best separations of the examined com-pounds were obtained with a 10 mmol/L phosphatebuffer, pH from 2.90 to 2.95, 180 °C temperature and15kV voltage. It was achieved within less than 30 mi-nutes. A linear relationship between peak area andconcentrations (r=0.995 and 0.999) with good preci-sions (9.7 and 12.1% for GSH and GSSG, respective-ly) were obtained. The method was used to analyzehuman capillary blood needing only its hemolyses. Theobtained results have shown that in the blood ofhealthy subjects (n=10) GSH (1340 ±292 mmol/L)was almost fifteen times greater than the GSSG (80 ±

B16RAZVIJANJE METODE ZA ISTOVREMENO

ODRE\IVANJE REDUKOVANOG I OKSIDOVANOG GLUTATIONA U KRVI BEZ DERIVATIZACIJE KORI[]ENJEM

KAPILARNE ELEKTROFOREZE I FAKTORSKOG DIZAJNA

G. @uni}1, S. Spasi}2

1Institut za medicinska istraìvanja, Vojnomedicinska akademija, Beograd

2Institut za medicinsku biohemiju, Farmaceutski fakultet,

Univerzitet u Beogradu, Beograd

Glutation (GSH) ima va`ne uloge u celokupnommetabolizmu, uklju~uju}i hemostazu }elije, radiopro-tekciju i antioksidativnu odbranu. U prisustvu slobod-nih radikala kiseonika on se oksiduje u disulfidnuformu (GSSG). Stoga je njegovo merenje korisno zaodre|ivanje oksidativno/anitoksidativnog statusa uraznim fiziolo{kim i patofiziolo{kim stanjima. Kakokoncentracija glutationa u krvi moè da odraàva glu-tationski status u drugim tkivima manje dostupnim zaanalizu, odre|ivanje GSH i GSSG u krvi se smatra kaodobar pokazatelj oksidativnog statusa osobe, koji pred-stavlja koristan indikator rizika za razvoj bolesti. Dosada kori{}ene metode uglavnom mere samo GSH,dok se nivo GSSG izra~unava. Kapilarna elektroforezapruà mogu}nost za njihovo istovremeno odre|ivanje,a dosad razvijene metode obi~no zahtevaju njihovuderivatizaciju. Kori{}enjem faktorijalnog dizajna za vari-ranje odgovaraju}ih parametara (pH i koncentracijaelektrolita, temperatura separacije), mi smo razvili jed-nostavnu i brzu metodu za istovremeno analiziranjeGSH i GSSG bez njihove prethodne derivatizacije.Dobijeni rezultati su pokazali da njihovo razdvajanjezavisi od pH elektrolita u kom se vr{i separacija, a umanjoj meri od koncentracije rastvora ili temperature.Najbolje razdvajanje ispitivanih komponenti je posti-gnuto u 10 mmol/L fosfatnom puferu, pH 2,90–2,95,na temperaturi od 180 °C i pri naponu od 15 kV. Podoptimalnim uslovima analiza traje 30 minuta. Dobijenje linearan odnos izme|u povr{ine pika i koncentracije(r=0,995 i 0,999) uz dobru preciznost odre|ivanja(9,7 i 12,1% za GSH i GSSG). Metoda je testiranaanaliziranjem hemolizata humane krvi. Dobijeni rezul-tati pokazuju da je u krvi zdravih odraslih osoba(n=10) nivo GSH (1340 ± 292 mmol/L) skoro pet-naest puta ve}i od nivoa GSSG (80 ± 36 mmol/L).Rezultati na{ih ispitivanja su tako|e pokazali da je fak-

UDK 577.1 : 61 ISSN 1452-8258


Poster sessions

240

torijalni dizajn korisan postupak za definisanje pro-menljivih koje omogu}avaju najbolje razdvajanje kom-ponenti iz sme{e pomo}u kapilarne elektroforeze.

B17PORE\ENJE DVE METODE ZA

KVANTITATIVNO ODRE\IVANJE FERITINA U HUMANOM SERUMU

D. ]uji}, O. Nedi}, S. Golubovi}, Lj. Vi}ovac

Institut za primenu nuklearne energije – INEP, Zemun, Srbija

Feritin je veliki makromolekul (450kDa) koji sesinteti{e u jetri i drugim tkivima i ima glavnu ulogu udeponovanju gvo`|a. Feritin je osetljiv pokazatelj ne-dostatka gvo`|a u organizmu, jer koncentracija feritinaopada pre smanjenja nivoa serumskog gvo`|a, hemo-globina ili veli~ine eritrocita. Stoga je osnovni klini~kizna~aj odre|ivanja feritina dijagnostikovanje anemije.Koncentracija feritina moè biti pove}ana usled pre-optere}enja organizma gvo`|em (kod hemohromatozaili hemosideroza), infekcija ili inflamacija, malignihoboljenja i o{te}enja tkiva jetre. U ovom radu izvr{enoje pore|enje dva komercijalna testa za odre|ivanjekoncentracije feritina, zasnovana na razli~itim principi-ma: 1) lateks test »FERITINA LATEX L«, »Globe Diag-nostics«, Italija i 2) ELISA test »Ferritin«, »DIMA Diag-nostika«, Getingen, Nema~ka. Koncentracija feritinaodre|ena je prema uputstvu proizvo|a~a u 30 uzorakaseruma. Dve metode su upore|ene primenom linearneregresione i korelacione analize, a razlika izme|u dvemetode je testirana t-testom za parove odre|ivanja.Dobijeni rezultati pokazali su dobro slaganje izme|udve metode (r2=0,9950). Razlika izme|u dve gruperezultata nije bila statisti~ki zna~ajna. Koncentracije fe-ritina odre|ene lateks testom bile su generalno niè odvrednosti dobijenih ELISA testom. U opsegu koncen-tracija feritina niìh od referentnih vrednosti, ELISA testje osetljiviji od lateks testa, koji daje vi{e vrednosti uopsegu niskih koncentracija. Rezultati pokazuju da ispi-tani ELISA test ima bolju analiti~ku osetljivost od ispi-tanog lateks testa i da je kao takav osetljiviji u otkrivan-ju pacijenata sa nedostatkom gvo`|a, kod kojih seobi~no o~ekuju vrednosti feritina u serumu niè od re-ferentnih.

36 mmol/L) level. This study also confirms factorial de-sign as a suitable procedure in screening importantvariables leading to the best separation of the com-pounds present in a mixture employing capillary elec-trophoresis.

B17A COMPARISON OF TWO METHODS FOR

THE QUANTITATIVE DETERMINATION OF FERRITIN IN HUMAN SERUM

D. ]uji}, O. Nedi}, S. Golubovi}, Lj. Vi}ovac

Institute for the Application of Nuclear Energy – INEP, Zemun, Serbia

Ferritin is a large macromolecule (450kDa) whichis synthesized in the liver and other tissues and plays amajor role in iron storage. Ferritin is a sensitive indica-tor of iron deficit in the body, since ferritin concentra-tion falls prior to a decrease in serum iron level, haemo-globin or size of erythrocytes. Thus, the main clinicalapplication of ferritin measurement is in the diagnosisof anaemia. The ferritin concentration may increase inthe case of iron overload (haemochromatosis or hae-mosiderosis), infection or inflammation, malignanciesand destruction of liver tissue. In this work, two com-mercial kits for the determination of ferritin concentra-tion, based on different immunochemical principles,were compared: 1) a latex test »FERITINA LATEX L«,»Globe Diagnostics«, Italy and 2) ELISA test »Ferritin«,»DIMA Diagnostika«, Goettingen, Germany. The ferri-tin concentration was determined in 30 serum samples,according to the manufacturer's instructions. Linearregression and correlation analysis were used to com-pare the two methods and the difference between themethods was tested by a t-test: paired two sample formeans analysis. The obtained results exhibited a goodcorrelation between the two methods (r2=0.9950).There was no significant difference between two gro-ups of results. A latex test gives generally lower valuesthen an ELISA test. However, in the ranges bellow thereference limit, an ELISA test is more sensitive than alatex test, which gives higher values in the low concen-tration range. The results presented here show that theanalysed ELISA test has better analytical sensitivity thanthe analysed latex test and as such is more sensitive indetecting patients with iron deficiency, whose ferritinlevels in serum are often expected to be lower than thereference range.

JMB 2008; 27 (2) 241

B18QUALITY CONTROL OF LABORATORY

TESTING IN INEP

Lj. Hajdukovi}, S. Golubovi}, D. Cveji}, S. Savin, J. Miloradovi}

Institute for the Application of Nuclear Energy – INEP, Zemun, Belgrade

The Institute for the Application of Nuclear Ener-gy (INEP) is a multidisciplinary research organizationcontributing to biological, medical and agriculturalresearch through projects both in pure and applied sci-ence. Scientific results in INEP are used for the deve-lopment and production of a range of immunodiag-nostic sets, together with various laboratory examina-tions, such as endocrinological, immunological, bio-chemical and hematological analyses, the monitoringof contamination by radionuclides, and agroecology.Such diverse activity at the Institute demands a perma-nent concern about the quality of both diagnostic kitsand laboratory testing. In order to attain such a goal, aquality management system according to standard ISO9001:1994 was established at INEP during 2001 andrecertification according to standard ISO 9001:2000was done in 2004. The certification was performed bythe international organization »IQNet OQS« (Austria).The laboratory was accredited according to standardISO/IEC 17025 during 2003. The analytical quality oflaboratory testing is provided by applying continuousinternal (IQA) and external (EQA) quality assesment.IQA includes all activities of laboratory staff in the qual-ity control of analyses: control sera are tested with eachgroup of patients sera, and the results are statisticalyprocessed. A program for an automatic graphical ana-lyses of control sera concentrations (Levey-Jenningscontrol cards) is implemented in the gamma countercomputer, and the obtained data are regularly ana-lysed. A validation of the methods developed at INEP isnecessary in order to confirm that the new methodsfulfill all demands for the intended application. Forevery new analysis, the uncertainty of measurementwas determined. INEP participates in the EQA organ-ized by DMBS and KCS (SNEQAS – medical biochem-istry), and also took part in the project IMEP 17 »Traceelements and the minor constituents of human se-rum«, by measuring the concentration of the only hor-mone included in this project, T4. The result of T4determination at INEP was by less than 5% differentfrom the exact value determined by the refferentmethod, mass spectrometry. Due to the lack of suchorganized EQA for measuring concentrations of hor-mones, tumor markers and other analytes by immu-noassays in Serbia, INEP has been organizing interla-boratory testing of the same analytes with laboratoriesof a similar profile. The institute is orientated towardsconstant improvement of the quality system.

B18KONTROLA KVALITETA LABORATORIJSKOG

ISPITIVANJA U INEP-U

Lj. Hajdukovi}, S. Golubovi}, D. Cveji}, S. Savin, J. Miloradovi}

Institut za primenu nuklearne energije – INEP, Zemun, Beograd

Institut za primenu nuklearne energije – INEP jemultidisciplinarna nau~noistraìva~ka organizacija, kojase, pored istraìvanja kroz projekte u osnovnim i prime-njenim naukama, ve} godinama bavi razvojem i pro-izvodnjom imunodijagnosti~kih kompleta, kao i broj-nim laboratorijskim ispitivanjima, koja uklju~uju endo-krinolo{ke, imunolo{ke, biohemijske i hematolo{keanalize, kao i ispitivanje kontaminacije radionuklidima iagroekologiju. Takva opredeljenost Instituta zahtevastalnu brigu o kvalitetu proizvoda, kao i pouzdanostirezultata ispitivanja. Zbog toga je u INEP-u tokom2001. godine uspostavljen sistem upravljanja kvalite-tom prema zahtevu standarda ISO 9001:1994, a u2004. godini i prema zahtevu standarda ISO9001:2000. Sertifikaciju je obavila me|unarodnaorganizacija »IQNet OQS« (Austrija). Laboratorija zaispitivanje je akreditovana prema zahtevu standardaISO/IEC 17025 u toku 2003. godine. Analiti~ki kvalitetrada laboratorije obezbe|uje se kontinuiranim spro-vo|enjem unutra{nje i spolja{nje kontrole kvaliteta.Unutra{nja kontrola kvaliteta (IQA) obuhvata sve aktiv-nosti koje redovno sprovodi osoblje u laboratoriji zaproveru rada, kako bi se omogu}ila pouzdanost rezul-tata koji se izdaju korisniku usluga. Sa svakom serijomuzoraka paralelno se testiraju kontrolni serumi i izvodistatisti~ka obrada rezultata. Program za automatsku igrafi~ku analizu vrednosti koncentracija kontrolnih se-ruma (Levey-Jennings kontrolne karte) implementiranje u kompjuteru vezanom za gama broja~, a dobijenipodaci se redovno analiziraju. U INEP-u postoji velikibroj interno razvijenih metoda, koje pre primene pro-laze kroz postupak validacije, kako bi se potvrdilo dametoda zadovoljava sve pojedina~ne zahteve za pred-vi|enu primenu. Za svaku novu metodu ura|ena je pro-cena merne nesigurnosti. INEP redovno u~estvuje uspolja{njoj kontroli kvaliteta (EQA) koju organizujeDMBS i KCS (SNEQAS – medicinska biohemija), au~estvovao je i u projektu IMEP 17 »Elementi u tragu iminorni konstituenti humanih seruma« ispitivanjem je-dinog predvi|enog hormona, T4. Dobijeni rezultat me-renja T4 u INEP-u je za manje od 5% odstupao odta~ne vrednosti odre|ene referentnom metodom, ma-senom spektrometrijom. Kako u Srbiji nisu uspostav-ljene EQA za odre|ivanje hormona, tumorskih markerai drugih analita koji se odre|uju imunotestovima, INEPredovno organizuje me|ulaboratorijska uporedna ispiti-vanja sa laboratorijama sli~nog profila. Institut se opre-delio za stalno pobolj{anje postoje}eg sistema kvaliteta.

242

B19UPOREDNO ODRE\IVANJE HEMATOLO[KIHPARAMETARA NA ANALIZATORIMA ADVIA

120® I HmX-AL®

I. Vujo{evi}, D. Vukosavljevi}, A. Beleti}, T. Vodnik, M. Ili}, N. Majki}-Singh

Institut za medicinsku biohemiju, Klini~ki centar Srbije, Beograd

U radu hematolo{kih laboratorija ve}eg kapacite-ta moè se pojaviti potreba za kori{}enjem analizatorarazli~itih proizvo|a~a. Cilj rada je bio da se uporedevrednosti osnovnih hematolo{kih parametara dobijenena analizatorima ADVIA 120-BAYER® i HmX-AL-BECKMAN COULTER®. U 180 uzoraka venske krvi,prikupljenih sa K

2EDTA kao antikoagulansom, paralel-

no su odre|eni: leukociti (WBC), eritrociti (RBC), he-moglobin (HGB), hematokrit (HCT), srednji volumeneritrocita (MCV) i trombociti (PLT). Statisti~ka obradapodataka je obuhvatila Studentov t-test i regresionu ikorelacionu analizu. Na osnovu dobijenih vrednostimoè se zaklju~iti da pri odre|ivanju leukocita, eritroci-ta, hemoglobina, hematokrita i trombocita nema sta-tisti~ki zna~ajnih razlika. Pore|enjem vrednosti MCVuo~eno je neslaganje izme|u analizatora, koje je uka-zalo na ~injenicu da u uzorcima gde je MCV<83 fLADVIA 120-BAYER® daje vi{e rezultate od HmX-AL®,dok je iznad te vrednosti odnos obrnut.

B20MERENJE KONCENTRACIJE

HEMOGLOBINA IZ KAPILARNE I VENSKE KRVI

M. Stanojkovi}, A. Marinkovi}

Centar za medicinsku biohemijuKlini~ki centar Ni{, Srbija

Koncentracija hemoglobina odre|uje se kako izkapilarne tako i iz venske krvi. Upore|ivana je koncen-tracija hemoglobina iz uzoraka kapilarne krvi hemoglo-binometrom sa parametrima uzoraka iz venske krvi ra-|enih na automatizovanom hematolo{kom analizato-ru. Ispitivanje je ra|eno na uzorku od 90 zdravih dobro-voljnih davalaca krvi. Uzorci su uzimani neposrednopre davanja krvi. Kapilarna krv ispitivana je na aparatuHaemoCue a uzimana je iz prsta, a iz venske krvi ra|e-ne su slede}e analize: koncentracija hemoglobina, brojeritrocita i hematokrit iz uzorka u epruveti sa K

2EDTA

na aparatu Nihon Kohden AVL i odre|ivanje nivoagvo`|a, TIBC, UIBC i feritina iz seruma na aparatuOlympus AU400. Vrednosti hemoglobina kretale su seod 128 g/L do 168 g/L. Iz uzoraka venske krvi, broj

B19PARALLEL DETERMINATION OF

HAEMATOLOGICAL PARAMETERS ON ADVIA 120® AND HmX-AL® ANALYZERS

I. Vujo{evi}, D. Vukosavljevi}, A. Beleti}, T. Vodnik, M. Ili}, N. Majki}-Singh

Institute of Medical Biochemistry, Clinical Center of Serbia, Belgrade

Haematology laboratories with a high throughputof samples may need analyzers from different manufac-turers. The aim of this study was to compare values ofbasic haematological parameters determined by ADVIA120-BAYER® and HmX-AL-BECKMAN COULTER®analyzers. In 180 samples of venous blood, collectedwith K

2EDTA as an anticoagulant, the following param-

eters were simultaneously measured: leukocytes (WBC),erythrocytes (RBC), haemoglobin (HGB), haematocrit(HCT), mean cell volume (MCV) and platelets (PLT).The results were compared using a Student's t-test andregression correlation analysis. On the basis of theobtained values we can conclude that there is no statis-tically significant difference in the determination ofleukocytes, erythrocytes, haemoglobin, haematocritand platelets on the analyzers mentioned above. Acomparison of MCV values revealed a significant dis-crepancy between the obtained results, which drove usto the conclusion that in the samples with microcytosis(MCV<83 fL), the values obtained on the ADVIA 120® are higher than those measured on the HmX-AL.

B20A COMPARISON BETWEEN THE

CONCENTRATION OF HEMOGLOBIN FROMCAPILLARY AND VENOUS BLOOD

M. Stanojkovi}, A. Marinkovi}

Center for Medical Biochemistry Clinical Center Ni{, Serbia

The measurement of the hemoglobin concentra-tion has been done from capillary blood taken from thepulp of the finger or from samples of venous blood. Acomparison of the concentration of hemoglobin fromcapillary blood by hemoglobin meter with parametersfrom venous blood using an automated hematologicalanalyzer was made. The analysis was done on a sampleof 90 healthy voluntary blood donors. The samples hadbeen taken immediately before blood giving. Capillaryblood was examined on a HemoCue device taken fromthe pulp of the finger, and from venous blood we did thefollowing analysis: concentration of hemoglobin, num-ber of erythrocytes, and hematocrite from samples intest tubes with K

2EDTA on a Nihon Kohden AVL device

and determination of the level of iron, TIBC, UIBC and

JMB 2008; 27 (2) 243

ferritine from sera on an Olympus AU 400 analyzer. Thevalues of hemoglobin were from 128 g/L to 168 g/L.From samples of venous blood, the number of erythro-cytes was close to the limits of from 3.8 x 1012/L to 5.5x 1012/L, and in three cases the values were below thenormal limits. The values of hemoglobin from venousblood were from 120 to 165 g/L, and in two cases theywere under normal limits. In these two persons, iron wasbelow the normal limits, and ferritin was within the nor-mal limits. There was no significant difference betweenthe results of hemoglobin concentration from capillaryand venous blood. With adequate training and monitor-ing, the value of hemoglobin from capillary blood sup-plies valid results of hemoglobin in urgent states, sur-gery states and in patients with malignant diseases inwhom we need to preserve the veins for later therapy,and in all patients who have bad veins or veins that aredifficult to reach.

B21THE SIGNIFICANCE OF LDL CHOLESTEROL

MEASUREMENT BY USE OF THE DIRECTMETHOD IN PRIMARY HEALTH CARE

M. [ekerovi}

Dom zdravlja Ba~ka Palanka

The determination of the ratio between LDL andHDL cholesterol according to Friedewald, i.e. by thecalculation of LDL cholesterol concentration on thebasis of triacylglycerol, total and HDL cholesterol con-centration, was for 40 years the basic method for theevaluation of the risk for atherosclerosis in primaryhealth care. From the late nineties, when a directmethod for the measurement of LDL concentrationwas developed, many authors reported significant dif-ferences between the directly measured concentra-tions and those calculated according to Friedewald.Also, several better methods were introduced for thecalculation of LDL cholesterol on the basis of triacyl-glycerol concentration. The aim of this study was toestablish how big are these differences in the condi-tions of primary health care, and also whether theydemand abandoning of Friedewald's method and themeasurement of LDL cholesterol by the direct method,or its replacement by some better method of calcula-tion. To establish that, measurements of serum con-centrations of total cholesterol, HDL cholesterol andtriacylglycerol were carried out in 140 outpatients, andon the basis of the obtained results the concentrationsof LDL cholesterol were calculated by the use ofFriedewald’s, and a newer method of Anandaraya:LDLCh=(0.9×Total Ch) – (0.9×TG/2.2) – 0.73. Thevalues obtained by both methods were then comparedwith the values of LDL cholesterol measured in thesame samples by use of the direct enzymatic method,and the differences and correlations between themwere calculated. The results are shown in the table:

eritrocita je bio na granici od 3,8 x 1012/L do 5 x1012/L, a u tri slu~aja vrednosti su bile ispod normalnihvrednosti. Vrednosti hemoglobina iz venske krvi bile suod 120 do 165 g/L a u dva slu~aja su bile ispod nor-malne granice. Kod ove dve osobe gvo`|e je bilo ispodgranica normale, a feritin je bio normalan. Ne postojizna~ajana razlika izme|u rezultata koncentracije he-moglobina, bilo da je krv uzeta kapilarno ili venski.Adekvatnom analizom i pra}enjem vrednosti hemoglo-bina iz kapilarne krvi obezbe|eni su validni rezultatihemoglobina za urgentna stanja, tokom operacija, kodbolesnika sa malignim bolestima kod kojih se vene~uvaju za primenu terapije i svih ostalih pacijenata kodkojih je te{ko do}i do povr{inskih vena ili su oneosetljive.

B21ZNAÂJ ODRE\IVANJA

LDL HOLESTEROLA DIREKTNOM METODOMU PRIMARNOJ ZDRAVSTVENOJ ZA[TITI

M. [ekerovi}

Dom zdravlja Ba~ka Palanka

Odre|ivanje odnosa izme|u LDL i HDL holes-terola po Friedewaldu, tj. izra~unavanjem koncen-tracije LDL holesterola na osnovu koncentracije triacil-glicerola, ukupnog i HDL holesterola, predstavlja ve}skoro 40 godina osnovnu metodu za procenu rizika zaaterosklerozu u primarnoj zdravstvenoj za{titi. Kada sekrajem devedesetih godina pojavila mogu}nost zadirektno merenje koncentracije LDL-a, mnogi suautori utvrdili zna~ajne razlike izme|u direktno izme-rene koncentracije i one izra~unate po Friedewaldu.Tako|e su se pojavile i druge, ta~nije metode izra~u-navanja koncentracije LDL holesterola na osnovu kon-centracije triacilglicerola. Cilj ovoga rada je bio da seutvrdi kolike su navedene razlike u uslovima primarnezdravstvene za{tite, i da li one iziskuju napu{tanjeFriedewaldove metode i direktnog odre|ivanja LDLholesterola ili njenu zamenu nekom boljom metodomizra~unavanja. U tu svrhu izvr{eno je kod 140 ambu-lantnih bolesnika odre|ivanje serumskih koncentraci-ja ukupnog holesterola, HDL holesterola i triacil-glicerola, na osnovu kojih je izra~unata koncentracijaLDL holesterola upotrebom Friedewaldove metode inovije metode po Anandaraji, LDLCh=(0,9× UkupniCh) – (0,9×TG/2,2) – 0,73. Vrednosti dobijene po-mo}u obe metode upore|ene su zatim sa vrednosti-ma koncentracija LDL holesterola odre|enih u istimtim uzorcima seruma direktnom enzimatskom meto-dom, i izra~unate razlike i korelacija izme|u njih.Rezultati su prikazani u tabeli:

244

Iz dobijenih rezultata se vidi da se ra~unskimmetodama, i po Friedewaldu i po Anandaraji, dobijajuniè vrednosti za LDL holesterol, ali da one u obemetode nisu statisti~ki zna~ajne. Vrednosti izra~unatepomo}u obe metode imaju dobru korelaciju sa vred-nostima LDL holesterola izmerenim direktnom me-todom, me|utim, pri vrednostima triacilglicerola iznad6,0, odnosno iznad 12,0 mmol/L pojavljuju se neprih-vatljivo velike razlike, preko 100%. Iz ovih rezultataproizilazi da se u primarnoj zdravstvenoj za{titi mogu idalje upotrebljavati ra~unske procene koncentracijeLDL holesterola po Friedewaldu ili po Anandaraji (dru-ga metoda je ne{to ta~nija i ne iziskuje odre|ivanjeHDL holesterola), ali da se pri znatnije povi{enim vred-nostima triacilglicerola kao i u sloènijim tipovima hi-perlipoproteinemija mora vr{iti direktno odre|ivanjekoncentracije LDL holesterola pri proceni rizika zaaterosklerozu.

B22LABORATORIJSKI INFORMACIONI

SISTEM – POMO] ILI SMETNJA

M. Krsnik

Univerzitetski Medicinski Centar Ljubljana, Institut za klini~ku hemiju i biohemiju,

Ljubljana, Slovenija

Glavni princip laboratorijskog informacionog sis-tema (LIS) je da se stavi informacija u red. LIS moraobezbediti podr{ku i ne sme biti smetnja u dnevnomradu laboratorijskih radnika. Instituti primaju oko 2000uzoraka ili 15000 analiza dnevno. Starija verzija LIS jeaùrirana u 2000. godini. Mi smo po~eli sa razvojemLIS, koji je produkt bliske saradnje izme|u na{e labo-ratorije i kompjuterskih eksperata. Glavni cilj i o~eki-vanja od aùriranja LIS su slede}a: kra}e TAT, manjegre{aka u administraciji, pove}anje u broju elektron-skih zahteva od strane odeljenja i klinika, bolja sledivostuzoraka i rezultata, kao i cilj grafi~kog prezentovanjarezultata, povezivanje ve}ine analizatora (bi- ili mono-usmereni), posedovanje {to je vi{e mogu}e informaci-ja o pacijentu i njegovim/njenim zahtevima u jednomelektronskom zapisu. Neki ciljevi su postignuti na sa-mom po~etku, neki od njih su kasnije razvijeni sa deli-mi~nom reorganizacijom na{eg instituta. Te{ko da jeiznena|uju}e da nismo na samom po~etku bili u mo-

From the obtained results it is visible that lowervalues for LDL cholesterol were obtained by calculationwith both Friedewald and Anandaraya methods, butthese differences were not statistically significant inboth methods. The values calculated by the use of bothmethods showed good correlation with the correspon-ding values of LDL cholesterol measured by the directmethod, meanwhile at values of triacylglycerol concen-tration above 6.0 and respectively 12.0 mmol/L, unac-ceptably high differences, above 100%, were found.From these results it can be concluded that calculationsof LDL cholesterol according to Friedewald of Anan-daraya (which is more accurate and does not need thedetermination or HDL cholesterol) may be further usedin primary health care, but at remarkably elevated tria-cylglycerol concentrations, as well as in complex typesof hyperlipoproteinemias, direct measurement of LDLconcentrations should be used for the evaluation of therisk for atherosclerosis.

B22LABORATORY INFORMATION SYSTEM

– SUPPORT OR NUISANCE

M. Krsnik

University Medical Centre Ljubljana, Institute of Clinical Chemistry and Biochemistry,

Ljubljana, Slovenia

The main principle of the laboratory informationsystem (LIS) is to put information in order. LIS mustprovide support and not be a nuisance in the daily workof laboratory workers. The institute receives around2000 samples or 15000 analyses daily. The older ver-sion of LIS was updated in the year 2000. We startedto develop LIS, which is a product of close collabora-tion between our laboratory and computer experts. Themain goals and expectations from the updated LISwere as follows: a shorter TAT, fewer errors at theadmission, an increase in the number of electronicorders from wards and clinics, better traceability of thesamples and results, as well as the aim to presentresults graphically, to connect most of the analysers (bi-or mono-directional), to have as much information aspossible from the patient and his/her orders in oneelectronic record. Some goals were attained at the verybeginning, some of them were developed later with apartial reorganisation of our institute. It is hardly sur-

Parameter Friedewald Anandaraya

Difference of the mean comparedto the direct method

–0.40 mmol/L –0.20 mmol/L

t-test 0.21 (NS) 0.11 (NS)

Coefficient of correlation compared to the direct method

0.824019 0.902479

Influence of TG concentration Iznad 6.0 mmol/L Iznad 12 mmol/L

Pokazatelj Po Friedewaldu Po Anandaraji

Razlika srednje vrednosti u odnosu na izmereni LDLCh

–0,40 mmol/L –0,20 mmol/L

t-test 0,21 (NS) 0,11 (NS)

Koeficijent korelacije u odnosuna izmereni LDLCh

0,824019 0,902479

Veliki uticaj koncentracije TG1. grupa2. grupa

Iznad 6,0 mmol/L Iznad 12 mmol/L

JMB 2008; 27 (2) 245

prising that at the very outset we were unable to pre-dict all the problems which were encountered duringthe implementation of the new LIS. During those peri-ods excellent communication between the laboratoryand computer experts was essential, as was the devel-opment of the 'missing' parts of LIS in the shortest pos-sible time. But the most important thing of all was thepersistent effort to keep laboratory workers motivated.After 9 years of experience and the use of LIS in ourdaily work we have changed the focus of developmentand have started to work on autoverification, autovali-dation, and automation on all the levels (based onEvidence-based Laboratory Medicine). There is a pos-sibility to receive orders or send results in electronicform to other health centers in Slovenia and abroad.Clinical chemistry and biochemistry is constantly devel-oping scientific field where progress cannot bestopped. At the same time, if we want to pass on ourknowledge to the patients, also LIS has to progress lestit should turn into a nuisance to laboratory people,which is a very common occurance in many medicallaboratories.

B23LABORATORY SERVICE INFORMATION SYSTEM AS PART OF THE HOSPITAL

INFORMATION SYSTEM

A. \or|evi}, M. Ljuji}-Gli{i}

Institute of Neonatology, Belgrade, Serbia

The information system of the laboratory servicehas been functioning as a part of the information sys-tem of the Institute of Neonatology since 1996. All thepieces of information are stored in the server whichconnects all six clinical departments of the Institute, theultrasound and X-ray units, units for enteral and par-enteral nutrition, out-patient department for consulta-tion and laboratory. The system has applications forelectronic patient records, case history, condition atadmission, laboratory results, CRIB score I and II andthe ponderal index, the calculation of meal preparationfor newborns, of the volume of meals and the calcula-tion of energetic and protein intake per child. At admis-sion, each patient receives a unique number to be usedby all units. When a sample of biologic material is deli-vered to the laboratory, the patient's number is enteredand simultaneously the basic demographic dataappear, time of sampling is recorded, the sample getsa laboratory number and on the available list of analy-ses the ones that are requested are marked. Uponmanual entering of the results they can be directly seenby a physician. There is a possibility of printing of theresults of urgent analyses only and for the particulardepartments, of the orders for individual work posi-tions, or a possibility to receive a report with all the

gu}nosti da predvidimo sve probleme na koje se na-ilazilo za vreme implementacije novog LIS. U toku ovihperioda odli~na komunikacija izme|u laboratorije ikompjuterskih eksperata je bila neophodna, kao irazvoj »nedostaju}ih« delova LIS u najkra}em mogu-}em vremenu. Ali najva`nija stvar od svega je bio sta-lan napor da se odrì motivacija laboratorijskih radnika.Posle 9 godina iskustva i primene LIS u na{em dnev-nom radu, promenili smo fokus razvoja i po~eli smo daradimo na autoverifikaciji, autovalidaciji i automatizacijisvih nivoa (zasnovanim na Laboratorijskoj medicini zas-novanoj na dokazima). Postoji mogu}nost za prijemzahteva ili slanje rezultata u elektronskoj formi u drugezdravstvene centre u Sloveniji i u inostranstvo. Klini~kahemija i biohemija konstantno razvijaju nau~no poljegde progres ne moè biti zaustavljen. U isto vreme, akoèlimo da predamo na{e znanje pacijentima, LIS moratako|e da napreduje kako se ne bi pretvorio u smetnjuza laboratorijske ljude, {to je veoma ~est doga|aj umnogi medicinskim laboratorijama.

B23INFORMACIONI SISTEM LABORATORIJSKE

SLU@BE KAO DEO BOLNI^KOG INFORMACIONOG SISTEMA

A. \or|evi}, M. Ljuji}-Gli{i}

Institut za neonatologiju, Beograd, Srbija

Informacioni sistem laboratorijske slu`be funkcio-ni{e kao deo informacionog sistema Instituta za neona-tologiju od 1996. godine. U serveru su deponovanesve informacije i preko njega se ostvaruje veza izme|usvih {est klini~kih odeljenja u Institutu, kabineta zaultrazvuk, rentgen, enteralnu i parenteralnu ishranu,konsultativne ambulante i laboratorije. Sistem ima apli-kacije za elektronsku istoriju bolesti pacijenta, anam-nezu, status na prijemu, laboratorijske rezultate, CRIBskor I i II i ponderalni indeks, prera~unavanje pripremeobroka za novoro|en~ad, volumena obroka i izra~una-vanje energetskog i proteinskog unosa po detetu. Svakipacijent na prijemu dobija jedinstveni broj koji koristesve slu`be. Kada se uzorak biolo{kog materijala predau laboratoriju, unosi se broj pacijenta te se istovremenopojavljuju osnovni demografski podaci, memori{e sevreme uzimanja uzorka, uzorak dobija laboratorijskibroj i na ponu|enoj listi analiza markiraju se one kojesu traène. Posle manuelnog uno{enja rezultata, oni semogu neposredno videti od strane lekara. Postoji mo-gu}nost {tampanja samo rezultata urgentnih analiza ipo odeljenjima, naloga za pojedina radna mesta, ilidobijanja izve{taja sa svim rezultatima jednog pacijen-ta u definisanom vremenu za èljenu grupu analiza.Mogu}e je dobiti razli~ite statisti~ke izve{taje u vezi sa

246

svim vrstama usluga, dijagnoza, morbiditeta i mortali-teta, ura|enih laboratorijskih analiza odre|enog tipa,razvrstane po odeljenjima, po smenama, o broju paci-jenata i zdravstvenim filijalama iz kojih pacijenti dolaze.Izve{taji se koriste na zahtev zdravstvenog fonda.

B24ODRE\IVANJE BAKRA METODOM AAS

I SPEKTROFOTOMETRIJSKOM METODOM

M. Baturan, J. Su|i, S. Doj~inovi}

Zavod za zdravstvenu za{titu radnika, Novi Sad, Srbija

Za odre|ivanje bakra u serumu do sada se koris-tila metoda atomske apsorpcione spektrofotometrije(AAS), {to je preporu~ena metoda za odre|ivanje me-tala u biolo{kom materijalu. Po{to takav ure|aj ve}inalaboratorija ne poseduje smatrali smo da je potrebnouporediti rezultate dobijene AAS-om i kolorimetrijskimtestom odre|ivanja bakra u serumu bez deproteinizaci-je. Ta~nost i preciznost smo proverili i potvrdili kori{}e-njem kontrolnih seruma. Upore|ivanjem eksperimen-talnih rezultata pomo}u obe metode utvrdili smo da jekolorimetrijski test uporediv sa metodom AAS tako dase moè koristiti u svim laboratorijama koje posedujuspektrofotometar koji pokriva talasnu duìnu od 580nm. Odre|ivanje smo vr{ili u serumu, dok se za odre-|ivanje bakra u urinu preporu~uje metoda AAS zbogniskih referentnih vrednosti (0,40–1,10 mmol/L).

B25PORE\ENJE ÊTIRI METODE ZA ODRE\IVANJE FERITINA

M. Dajak1, R. Kangrga1, I. Draga{evi}1, I. Obradovi}1, S. Ignjatovi}2

1Institut za medicinsku biohemiju, Klini~ki centar Srbije,

2Institut za medicinsku biohemiju, Klini~ki centar Srbije i Farmaceutski fakultet,

Univerzitet u Beogradu, Beograd, Srbija

Koncentracija feritina u serumu odraàva telesnerezerve gvo`|a i zbog toga se imunotestovi za odre|i-vanje koncentracije feritina ~esto koriste u dijagnostiko-vanju poreme}aja vezanih za gvo`|e i u pra}enju sta-tusa gvo`|a u populaciji. Cilj ovog rada je bio da seuporede metode za odre|ivanje serumskog feritinakoje se zasnivaju na ~etiri razli~ita principa: CMIA (che-miluminescent microparticle immunoassay), ECLIA(electrochemiluminescence immunoassay), PEINA

results of a patient in a defined period for the desiredgroup of analyses. It is possiblSe to receive differentstatistic reports regarding all types of services, diag-noses, morbidity and mortality, conducted laboratoryanalyses of certain type, classified per departments, pershifts, on the number of patients and health affiliatesfrom which the patients come. The reports are used atthe request of the Health Insurance Fund.

B24COPPER DETERMINATION BY THE

AAS METHOD AND SPECTROPHOTOMETRY

M. Baturan, J. Su|i, S. Doj~inovi}

Institute of Occupational Medicine Novi Sad

The method of atomic absorption spectropho-tometry (AAS) was used so far for the determinationof copper in serum, and was the method of choice forthe determination of metals in biological material.Since such an apparatus is not available to most labo-ratories we considered it necessary to compare theresults obtained by AAS and by a colorimetric test forthe determination of copper in serum without depro-teinisation. Accuracy and precision were checked andconfirmed by use of control serums. Comparing theexperimental results of both methods we have con-cluded that the colorimetric test is comparable to theAAS method and can be used in all laboratories thathave a spectrophotometer, covering a wavelength ran-ge of 580 nm. These analyses were conducted in se-rum while for the determination of copper in urine theAAS method is recommended due to low referencevalues (0.40–1.10 mmol/L).

B25COMPARISON OF FOUR METHODS

FOR FERRITIN MEASUREMENT

M. Dajak1, R. Kangrga1, I. Draga{evi}1, I. Obradovi}1, S. Ignjatovi}2

1Institute of Medical Biochemistry, Clinical Center of Serbia,

2Institute of Medical Biochemistry, Clinical Center of Serbia and School of Pharmacy,

University of Belgrade, Belgrade, Serbia

The concentration of serum ferritin reflects bodyiron stores and therefore ferritin immunoassays havebeen widely used in diagnosing iron related disordersand in population surveys of iron status. The aim of thisstudy was to compare the methods for the determina-tion of serum ferritin which were based on four differ-ent principles: CMIA (chemiluminescent microparticleimmunoassay), ECLIA (electrochemiluminescenceimmunoassay), PEINA (particle-enhanced immunone-

JMB 2008; 27 (2) 247

phelometric assay) and ITA (immunoturbidimetricassay). These methods are used on the following sys-tems: CMIA on Architect ci8200 (Abbott Diagnostics,Wiesbaden, Germany), ECLIA on Elecsys 2010 (RocheDiagnostics, Mannheim, Germany), PEINA on BNIInephelometer (Dade Behring, Marburg, Germany) andITA on Olympus AU2700 (Olympus Diagnostica,Hamburg, Germany). In imprecision studies at increas-ing serum ferritin concentration, intra-assay and inter-assay CVs ranged: 3.4–5.0%, 1.8–6.0%, 1.9–3.9%and 2.7–4.5%, for Abbott, Olympus, Roche and DadeBehring methods, respectively. The correlation of fer-ritin values, determined with these methods, was eval-uated using least-square regression analysis andabsolute difference plot according to Bland andAltman. Obtained correlation coefficients (slope, inter-cept) were: Dade Behring vs. Abbott method, 0.9954(1.051, 7.415); Dade Behring vs. Olympus method,0.9948 (0.935, 1.195); Abbott vs. Olympus method,0.9933 (1.004, 1.996); Olympus vs. Roche method,0.9958 (1.334, 2.736); Dade Behring vs. Rochemethod, 0.9979 (1.341, 12.928) and Abbott vs.Roche method, 0.9947 (1.250, 9.319). Mean (SD)absolute differences from the Bland-Altman plot were:21.0 (43.8) mg/L, –7.5 (13.1) mg/L, 2.5 (13.3) mg/L,47.8 (40.5) mg/L 91.1 (108.6) mg/L and 70.8 (92.7)mg/L, for the comparison of: Dade Behring and Abbottmethods, Dade Behring and Olympus methods,Abbott and Olympus methods, Olympus and Rochemethods, Dade Behring and Roche methods andAbbott and Roche methods, respectively. Ferritin con-centrations measured with Roche method were25–33% higher than the ferritin concentrations meas-ured with the other three methods, which correspond-ed with higher reference intervals given by Roche man-ufacturer. The results of this study showed that therewas good agreement among ferritin values determinedwith these four methods.

B26COMPARISON OF TWO METHODS FOR

PROCALCITONIN DETERMINATION

D. Pejak1, S. Jovi~i}1, S. Ignjatovi}2


2Institute of Medical Biochemistry,Clinical Center of Serbia and Faculty of Pharmacy,

Belgrade, Serbia

Procalcitonin (PCT) is the prohormone of calci-tonin and PCT is secreted by the C-cells of the thyroidgland. PCT can be produced by numerous cell types,especially liver cells, after proinflammatory stimulation.Elevated PCT levels indicate bacterial infection accom-panied by a systemic inflammatory reaction. One ma-jor advantage of PCT compared to other parameters isits early and specific increase in response to severe sys-

(particle-enhanced immunonephelometric assay) andITA (immunoturbidimetric assay). Ove metode sekoriste na slede}im sistemima: CMIA na »Architectci8200« (Abbott Diagnostics, Vizbaden, Nema~ka),ECLIA na Elecsys 2010 (Roche Diagnostics, Mann-heim, Nema~ka), PEINA na BNII nefelometru (DadeBehring, Marburg, Germany) i ITA na »OlympusAU2700« (Olympus Diagnostica, Hamburg, Nema-~ka). U ispitivanju nepreciznosti pri rastu}oj koncen-traciji feritina u serumu, koeficijenti varijacije unutar iizme|u serija su bili u opsegu: 3,4–5,0%, 1,8–6,0%,1,9–3,9% i 2,7–4,5%, za Abbott, Olympus, Roche iDade Behring metode ponaosob. Korelacija vrednostiferitina odre|enih tim metodama je procenjena pri-menom regresione analize najmanjih kvadrata i dija-gramom apsolutnih razlika prema Bland i Altmanu.Dobijeni koeficijenti korelacije (nagib, odse~ak) bili su:Dade Behring vs Abbott metodi, 0,9954 (1,051,7,415); Dade Behring vs Olympus metodi, 0,9948(0,935, 1,195); Abbott vs. Olympus metodi, 0,9933(1,004, 1,996); Olympus vs. Roche metodi, 0,9958(1,334, 2,736); Dade Behring vs. Roche metodi,0,9979 (1,341, 12,928) i Abbott vs. Roche metodi,0,9947 (1,250, 9,319). Srednje vrednosti (Sd) apso-lutnih razlika iz Bland-Altmanovog dijagrama su bile:21,0 (43,8) mg/L, –7,5 (13,1) mg/L, 2,5 (13,3) mg/L,47,8 (40,5) mg/L 91,1 (108,6) mg/L i 70,8 (92,7) mg/L,pri pore|enju: Dade Behring i Abbott metoda, DadeBehring i Olympus metoda, Abbott i Olympus metoda,Olympus i Roche metoda, Dade Behring i Roche meto-da i Abbott i Roche metoda. Koncentracije feritinaodre|ene Roche metodom su bile 25–33% vi{e odkoncentracija feritina odre|enih sa ostale tri metode,{to odgovara vi{em referentom intervalu datom odstrane Roche proizvo|a~a. Rezultati ovog ispitivanja supokazali da postoji dobro slaganje izme|u vrednosti fe-ritina odre|enih sa ove ~etiri metode.

B26PORE\ENJE DVE METODE

ODRE\IVANJA PROKALCITONINA

D. Pejak1, S. Jovi~i}1, S. Ignjatovi}2


2Institut za medicinsku biohemiju Klini~ki centar Srbijei Farmaceutski fakultet, Beograd, Srbija

Prokalcitonin (PCT) jeste prohormon kalcitoninakoji lu~e C-}elije tireoidee. Nakon proinflamatorne sti-mulacije izazvane bakterijskom infekcijom, PCT produ-kuju mnoge }elije razli~itih tkiva, posebno }elije jetre.Povi{ene vrednosti PCT ukazuju na bakterijsku infekci-ju. Velika prednost PCT u odnosu na druge parametresepse je rano i specifi~no pove}anje vrednosti kod sis-temskih bakterijskih infekcija i stanja sepse. Koncentra-

248

cije PCT < 0,5 ng/mL ukazuju na nizak rizik od nastan-ka sistemske infekcije, vrednosti 0,2–2,0 ng/mL pred-stavljaju umereni rizik za razvoj sepse, dok koncen-tracije PCT >2,0 ng/mL predstavljaju visok rizik, odno-sno ukazuju na veliku verovatno}u za prisustvo sepse.Vrednosti PCT su normalne kod virusnih infekcija iautoimunih poreme}aja. Rana detekcija i adekvatnaklini~ka terapija su od velikog vitalnog zna~aja za paci-jente obolele od sepse. Cilj ovog rada je bio pore|enjevrednosti PCT dobijenih pomo}u dve razli~ite metode -ELFA (Enzyme Linked Fluorescent Assay), primenjenena analizatoru »mini VIDAS®« (bioMérieux Industry,Hajzelvud, Misuri, SAD), i TRACE tehnologije (TimeResolved Amplified Cryptate Emission) na analizatoru»KRYPTOR®« (BRAHMS Aktiengesellschaft, Henigs-dorf, Nema~ka). Na oba anlizatora su kori{}eni origi-nalni testovi: BRAHMS PCT sensitive na »KRYP-TOR®«-u i VIDAS BRAHMS PCT na mini »VIDAS®«-u. Nepreciznost u seriji (N=15) odre|ena je pomo}ukomercijalnih kontrolnih uzoraka: BRAHMS PCT sensi-tive Kryptor - Control 1 (K1, koncentracija PCT 1,32-2,10 ng/mL) i VIDAS BRAHMS PCT – Control 2 (K2,koncentracija PCT 0,22–0,34 ng/mL), i dobijeni koefi-cijenti varijacije su bili za K1 2,12% i 0,79%, a za K27,86% i 7,10% na »mini VIDAS®«-u i »KRYPTOR®«-u redom. Za ispitivanje slaganja dve metode, koncen-tracije PCT su odre|ene u sveìm serumima 50 pacije-nata na oba anlizatora simultano. Dobijene su slede}eregresione jedna~ine: y = 1,200x – 0,096 linearnomregresionom analizom, i y = 1,045x – 0,025 Passing-Bablok regresionom analizom. U oba slu~aja koefici-jent korelacije je bio r = 0,99. Srednja vrednost razlikeizme|u metoda dobijena analizom po Altmanu i Blan-du je bila 0,636 (Sd = 0,322). Na osnovu dobijenihrezultata moè se zaklju~iti da su nepreciznosti obemetode i njihova me|usobna korelacija zadovoljava-ju}e i da se oba aparata mogu koristiti za odre|ivanjePCT-a u ranom otkrivanju bakterijskih infekcija i sepse.

B27LEKOVI KAO VA@AN FAKTOR

U PREANALITI^KOJ FAZI

S. \. Jovanovi}, K. Birinji

Hemolab, Sremska Kamenica (Novi Sad), SrbijaDom zdravlja, Irig, Srbija

Lekovi se smatraju va`nim faktorima pri stvaranjupogre{nih laboratorijskih laboratorijskih rezultata. Iakoje ta mogu}nost stalno prisutna, lekovi su dosta ~estovan kontrole. Oni mogu da uti~u na biohemijske idruge analize per se ili preko svojih derivata ili meta-bolita, deluju}i sami ili u kombinaciji sa jednim, dva ilivi{e lekova. Ovaj rad na primeru kreatinifokinaze (dejst-vo diklofenaka), kreatinina (dejstvo askorbinske kise-line) i natrijuma (dejstvo triglicerida) i samih pacijenata

temic bacterial infections and sepsis. PCT levels are lowin viral infections or autoimmune processes. PCT con-centrations < 0.5 ng/mL indicate low risk for progres-sion to severe systemic infection, concentrations0.2–2.0 ng/mL represent moderate risk for progres-sion to severe sepsis, while PCT concentrations >2.0ng/mL represent high risk and indicate high likelihoodof sepsis. Early detection and specific clinical interven-tion has been shown to be crucial for patients with sep-sis. The aim of this study was the comparison of PCTvalues obtained using two different methods – ELFA(Enzyme Linked Fluorescent Assay), used by miniVIDAS® analyzer (bioMérieux Industry, Hazelwood,Missouri, USA), and TRACE technology (Time Resol-ved Amplified Cryptate Emission) used by KRYPTOR®analyzer (BRAHMS Aktiengesellschaft, Hennigsdorf,Germany). Both analyzers used original test kits:BRAHMS PCT sensitive for KRYPTOR® and VIDASBRAHMS PCT for mini VIDAS®. Within-run impreci-sion (N=15) was determined using commercial con-trol samples: BRAHMS PCT sensitive Kryptor – Control1 (K1, PCT concentration 1.32–2.10 ng/mL) andVIDAS BRAHMS PCT – Control 2 (K2, PCT concen-tration 0.22-0.34 ng/mL), and obtained coefficients ofvariation were: for K1 2.12% and 0.79%, and for K27.86% and 7.10%, on mini VIDAS® and KRYPTOR®,respectively. For method comparison study, PCT con-centrations were determined in 50 fresh serum sam-ples on both analyzers, simultaneously. Obtained re-gressions were: y = 1.200x – 0.096 with linear regres-sion analysis, and y = 1.045x – 0.025 with Passing-Bablok regression analysis. The coefficients of correla-tion in both cases were r = 0.99. Mean bias and SDfor Bland-Altman plot were 0.636 and 0.322, respec-tively. Based on the obtained results we can concludethat both methods showed satisfactory imprecision andgave comparable results. Therefore, both analyzers canbe used for PCT determination in the early detection ofbacterial infection and sepsis.

B27DRUGS AS IMPORTANT FACTORS

IN THE PREANALYTICAL PHASE

S. \. Jovanovi}, K. Birinji

Hemolab, Sremska Kamenica (Novi Sad), SrbijaHealth Center Irig, Irig, Srbija

Although they have an important role in the cre-ation of potentially misleading laboratory results,whichis a constant possibility, drugs are often beyond control.They could influence biochemical and other analysesper se or through their derivates and metabolites actingalone or in combination with one, two or more drugs.The paper presents a few parameters (creatine phos-phokinase – influenced by diclofenac, creatinine theinfluenced by ascorbic acid, sodium – influenced by

JMB 2008; 27 (2) 249

triglycerides) and some patients with impressive inter-fering problems with the idea to take more attention tothe commenting of pathological or even »normal« la-boratory results. No general approach to the solutionof this problem does not give the right to neglect it.

B28SENSITIVITY AND SPECIFICITY OF A LATEX

AGGLUTINATION TEST FOR THEDETERMINATION OF D-DIMER

CONCENTRATION IN THE DIAGNOSIS OF PULMONARY EMBOLISM

R. \or|evi}1, M. \or|evi}2

1Special Hospital for Lung Diseases »Ozren«, Sokobanja, Serbia

2Special Hospital for Lung Diseases »Sokobanja«, Serbia

The diagnosis of pulmonary embolism remains acomplicated issue for physicians because the signs andsymptoms of this condition are not specific. Tests thatallow confident exclusion of pulmonary embolism arevery useful. The concentration of D-dimer in citrateblood was determined with a latex agglutination test,n.v. 0.5 g/L. We determined the D-dimer in 81 patients(64% men) with pulmonary embolism (30–64 years).Median concentration of D-dimer was 4.2 g/L. Thesensitivity of this method is 93.0 % and specificity is88.6%. The latex agglutination method is very specificand sensitive and the D-dimer assay should thereforebe incorporating as an algorithm for the diagnosis ofpulmonary embolism.

B29COMPARISON OF TWO INTACT

PARATHYROID HORMONEIMMUNOASSAYS

R. Kangrga1, M. Dajak1, S. Jovi~i}1, I. Draga{evi}1, S. Ignjatovi}2

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

2Institute of Medical Biochemistry, Clinical Center of Serbia and School of Pharmacy,

University of Belgrade, Belgrade, Serbia

The determination of parathyroid hormone(PTH) is of great clinical relevance in the assessmentof calcium metabolic disorders, as it provides directinformation about parathyroid activity and indirectinformation about bone turnover. Although PTH wasone of the first hormones measured by immunoassay,there are still many difficulties in its determination,

sa problemima interferencije èli da skrene pa`nju natuma~enje patolo{kih ili ~ak »normalnih« laboratorij-skih rezultata. Ne smemo uzeti pravo da osporavamozna~aj ovog problema u vreme kada ne postoji usvoje-na strategija za njegovo re{avanje.

B28OSETLJIVOST I SPECIFI^NOST LATEKS

AGLUTINACIONOG TESTA ZA ODRE\IVANJE KONCENTRACIJE D-DIMERA

U DIJAGNOSTICI PLU]NE EMBOLIJE

R. \or|evi}1, M. \or|evi}2

1Specijalna bolnica za plu}ne bolesti »Ozren«– Sokobanja, Srbija

2Specijalna bolnica za nespecifi~ne plu}ne bolesti»Sokobanja« – Sokobanja, Srbija

Dijagnostikovanje plu}ne embolije je veomakomplikovan proces, jer znaci i simptomi nisu strogospecifi~ni. Testovi koji isklju~uju plu}nu emboliju su vrlokorisni u njenoj dijagnostici. Koncentracija D-dimeraodre|ivana je u citratnoj krvi pomo}u lateks aglutina-cionog testa, norve{ke firme NYCOMED. Referentnavrednost D-dimera je do 0,5 g/L. Kod 81 pacijenta(64% mu{karci), sa plu}nom embolijom (30–64 godi-ne), odre|ivali smo D-dimer. Srednja vrednost koncen-tracije D-dimera je bila 4,2 g/L. Osetljivost ove metodeje 93,9% a specifi~nost 88,6%. Ovaj lateks aglutina-cioni test za odre|ivanje koncentracije D-dimera u cit-ratnoj krvi je vrlo specifi~an i osetljiv i treba ga usvojitikao algoritam u dijagnostici plu}ne embolije.

B29PORE\ENJE DVE IMUNOHEMIJSKE

METODE ZA ODRE\IVANJE INTAKTNOGPARATIREOIDNOG HORMONA

R. Kangrga1, M. Dajak1, S. Jovi~i}1, I. Draga{evi}1, S. Ignjatovi}2


2Institut za medicinsku biohemiju, Klini~ki centar Srbije i Farmaceutski fakultet,

Univerziteta u Beogradu, Beograd, Srbija

Odre|ivanje paratireoidnog hormona (PTH) imaveliki klini~ki zna~aj u proceni poreme}aja metaboli-zma kalcijuma, daje direktnu informaciju o aktivnostiparatireoidnih `lezda i indirektnu informaciju o ko{ta-nom metabolizmu. Iako je PTH jedan od prvih hor-mona koji je odre|ivan imunohemijskim metodama,postoje mnogi problemi, pre svega velike razlike izme-

250

|u komercijalnih testova za odre|ivanje intaktnog PTH(iPTH) i sledstvena neadekvatnost primene algoritamaiz vodi~a klini~ke prakse kod pacijenata sa sekundarn-im paratireoidizmom. Cilj ovog rada je bio da se odre-di varijabilnost u rezultatima PTH, dobijenim pomo}udve metode druge generacije: CMIA (chemilumines-cent microparticle immunoassay) i ECLIA (electro-chemiluminescence immunoassay). Ove metode sekoriste na slede}im sistemima: CMIA na »Architectci8200« (Abbott Diagnostics, Vizbaden, Nema~ka),ECLIA na Elecsys 2010 (Roche Diagnostics, Manhajm,Nema~ka). U ispitivanju nepreciznosti iz dana u dan,koriste}i tri nivoa kontrole (PreciControl Bone, Roche iIntactPTH Controls, Abbott) koeficijenti varijacije subili u opsegu: 3,9–5,5% i 2,5–6,7% za Roche i Abbottmetode, redom. Pore|enje metoda je odre|enoPassing-Bablok regresionom analizom, kao i vizuelno,dijagramom apsolutnih razlika po Blandu i Altmanu.Rezultati dva imunotesta su u dobroj korelaciji, (korela-cioni koeficijent je 0,995, N=83). Nagib od 1,225 iodse~ak od 3,0 nisu bili neo~ekivani usled razlika ustandarizaciji imunotestova, zbog prisustva razli~itihPTH fragmenata i promenljivog afiniteta antitela za tePTH fragmente. Srednja vrednost apsolutnih razlika izBland-Altmanovog dijagrama je bila 62,3 pg/mL (95%interval pouzdanosti: 47,9 do 76,7) sa jasnim doka-zom pozitivnog odstupanja CMIA metode koje zavisiod koncentracije. Neprihvatljiva posledica velike varija-bilnosti izme|u metoda moè biti razli~ita klini~ka i te-rapijska odluka kod istoga pacijenta zavisno od ko-ri{}enog iPTH testa. Prema tome, rezultati iPTH suzavisni od metode i potrebno je nastaviti napore da seimunotestovi harmonizuju. Dostupnost internacio-nalnog referentnog materijala moè doprineti tim nas-tojanjima.

B30PORE\ENJE TRI AUTOMATIZOVANE

METODE ZA ODRE\IVANJE TIREOSTIMULIRAJU]EG HORMONA

N. Gligorovi}, P. Radovi}, D. Boljevi}

Centar za klini~ko-laboratorijsku dijagnostiku, Klini~ki centar Crne Gore, Vojni zdravstveni centar,

Privatna zdravstvena ustanova »Sine Morbo«,Podgorica, Crna Gora

Odre|ivanje koncentracije tireostimuliraju}eghormona (TSH) ima superiornu dijagnosti~ku ose-tljivost za detekciju primarnog hiper- i hipotireoidizmakod ambulantne populacije pacijenata, terapeutski je»end-point« za titriranje supstitucione doze L-tiroksi-na (L-T4) kod primarnog hipotireoidizma i monitoringsupresione terapije kod diferenciranog karcinoma{titne `lezde. Komparabilnost razli~itih metoda kojese koriste za odre|ivanje koncentracije ovog parame-tra od velikog je zna~aja kako za lekare tako i za paci-jente. Mi smo poredili rezultate za TSH dobijene

especially due to large differences between commer-cial iPTH assays and, consequently, the inappropriateuse of algorithms from Clinical Practice Guidelines onpatients with secondary hyperparathyroidism. The aimof this study was to determine the variability in resultsobtained with two second generation assays for PTH:CMIA (chemiluminescent microparticle immunoassay)and ECLIA (electrochemiluminescence immunoassay).These methods are used on the following systems:CMIA on the Architect ci8200 (Abbott Diagnostics,Wiesbaden, Germany), ECLIA on Elecsys 2010 (RocheDiagnostics, Mannheim, Germany). The imprecisionstudy yielded between-days CVs of 3.9–5.5% and2.5–6.7% for Roche and Abbott methods, respective-ly, using three control levels (PreciControl Bone, Rocheand IntactPTH Controls, Abbott). Method comparisonwas performed using the Passing-Bablok regressionmethod and visually, using the Bland-Altman plot. Theresults of two immunoassays correlated well accordingto the correlation coefficient, which was 0.995. Theslope of 1.225 and intercepts of 3.0 were not unex-pected due to differences in immunoassay standardi-zation, and due to the presence of different PTH frag-ments and a variable affinity of the used antibodies tothese PTH fragments. Mean of the absolute differ-ences from the Bland-Altman plot was 62.3 pg/mL(95% CI 47.9 to 76.7) with clear evidence of concen-tration related positive bias for CMIA method. Theunacceptable consequence of this is that oppositetherapeutic attitudes may be reached in a singlepatient depending on the PTH assay used. Thus, iPTHresults are method-dependent, and continuing effortsto harmonize assays are needed. The availability of theinternational reference material could facilitate thiseffort.

B30COMPARISON OF THREE AUTOMATED

METHODS FOR THYROID SIMULATING HORMONE IMMUNOASSAYS

N. Gligorovi}, P. Radovi}, D. Boljevi}

Center for Clinical-Laboratory Diagnostics, Clinical Center of Montenegro,

Military Medical Center, Private Health Institution»Sine Morbo«, Podgorica, Crna Gora

Thyroid Stimulating Hormone (TSH) measure-ment is the most diagnostically sensitive test fordetecting primary hypo- or hyperthyroidism in ambu-latory patients and it is a therapeutic endpoint fortitrating the L-thyroxin (L-T4) replacement dose forprimary hypothyroidism and monitoring LT4 suppres-sion therapy for differentiated thyroid carcinoma.Between-method comparability for this parameter isof great importance for physicians and patients. Wecompared the TSH results of three different meth-ods: acridinium derived chemiluminescence (»ARCHI-

JMB 2008; 27 (2) 251

TECT i2000sr«), electrochemiluminescence basedon the use of ruthenium(II)-tris(bypiridil) complexand trpropylamin (»ELECSYS 2010«) and microparti-cle enzyme immunoassay using 4-methyl-umbellife-ryl phosphate as substrate (AxSYM). Precision wasassayed during 30 days, using three levels of com-mercial control serums for ARCHITECT i2000sr(Ar1, Ar2, Ar3), »ELECSYS 2010« (PUTSH, PU1,PU2) and »AxSYM« (Ax1, Ax2 and Ax3). TSH wasmeasured in 157 serum samples. CV for Ar1 (0.10mU/L) was – 6.6%, Ar2 (6.00 mU/L) – 6.4% andAr3 (30.00 mU/L) – 4.3%. CV for PUTSH (0.20mU/l) was – 5.3%, PU1 (1.28 mU/L) – 6.9% and PU2(8.05 mU/L) – 6.3%. CV for Ax1 (0.10 mU/L) was –7.0%, Ax2 (3.00 mU/L) – 5.6% and Ax3 (29.00 mU/L)– 7.2%. Linear regression analysis gave the followingresults ARCHITECT=0.89, ELECYS-0.20 (r=0.996),ARCHITECT=0.78 AxSYM-0.23 (r=0.994) and ELE-CYS=0.98, AxSYM-0.70 (r=0.996). This evaluationstudy confirms the overall good correlation of theresults obtained with assays for TSH.

trima razli~itim metodama: imunohemiluminiscenci-jom koja poti~e od akridinijuma (»ARCHITECTi2000sr«), imunoelektrohemiluminiscencijom koja sezasniva na upotrebi rutenium(II)-tris(bipiridil)-a itripropilamina (»ELECYS 2010«) i mikro~esti~nomenzimimunotehnikom koja koristi 4-metil-umbeliferilfosfat kao supstrat (»AxSYM«). Koncentracija TSHodre|ivana je u 157 uzoraka. Preciznost metodatestirana je u periodu od 30 dana, upotrebom po trinivoa komercijalno dostupnih kontrolnih seruma: za»ARCHITECT i2000sr« (Ar1, Ar2, Ar3), »ELECSYS2010« (PUTSH, PU1, PU2) i za »AxSYM« (Ax1, Ax2and Ax3). CV za Ar1 (0,10 mU/L) bio je – 6,6%, Ar2(6,00 mU/L) – 6,4% i Ar3 (30,00 mU/L) – 4,3%. CVza PUTSH (0,20 mU/L) bio je – 5,3%, PU1 (1,28mU/L) – 6,9% i PU2 (8,05 mU/L) – 6,3%. CV za Ax1(0,10 mU/L) bio je – 7,0%, Ax2 (3,00 mU/L) – 5,6%i Ax3 (29,00 mU/L) – 7,2%. Linearnom regresionomanalizom dobili smo slede}e rezultate: ARCHI-TECT=0,89ELECYS-0,20 (r=0,996), ARCHITECT=0,78 AxSYM-0,23 (r=0,994) i ELECYS=0,98AxSYM-0,70 (r=0,996). Ova evaluaciona studija po-kazala je dobru me|usobnu korelaciju metoda kojese {iroko koriste za odre|ivanje koncentracija TSH.

C

PROTEINI I ENZIMI

PROTEINS AND ENZYMES

C31ZNAÂJ ODRE\IVANJA HUMANOG

PLACENTALNOG LAKTOGENA U TRE]EM TRIMESTRU TRUDNO]E

J. Durkovi}, B. Mandi}

Odsek za genetiku – Bolnica SuboticaMegaLab biohemijska laboratorija

Subotica

Humani placentalni laktogen (hPL) jeste hor-mon koji produkuje placenta i regulator je feto-pla-centalnog rasta. Odre|ivanje koncentracije hPL-a userumu trudne ène koristi se kao pokazatelj placen-talne funkcije. U radu su prikazani rezultati odre|i-vanja hPL-a u tre}em trimestru trudno}e, sa ciljem dase ispita senzitivnost tog biohemijskog markera zaotkrivanje poreme}aja funkcije placente, fetalnogvitaliteta i rizika za lo{ ishod. hPL se odre|uje ELISAmetodom testovima Bioserv Diagnostics, a rezultatise o~itavaju na rideru STAT-FAX 303+. Ispitivanje jeobavljeno na uzorku od 370 rizi~nih trudno}a izme|u20 i 36 nedelje trudno}e. Indikacije za odre|ivanjehPL su bila stanja rizi~na za preeklampsiju, hiperten-zija, dijabetes melitus, toksemiju, edeme, bubre`nuinsuficijenciju kao i ultrazvu~no uo~en intrauterini za-stoj rasta ploda, placentalni trofoblasni tumor, krvare-nje i prete}i poba~aj. Rezultate hPL markera poredilismo sa standardnim referentnim vrednostima ponedeljama trudno}e. U normalnim okolnostima kon-centracija hPL u serumu trudnice raste sa napredo-vanjem trudno}e. Uo~eno je zna~ajno opadanje hPLkod prete}ih poba~aja, krvarenja, kalcifikacije pla-cente, intrauterinog zastoja rasta, intrauterine smrtiploda i trudno}ama iz kojih je ro|eno dete sa fetal-nim distresom i asfiksijom. Nagli porast hPL utvr|enje kod trudnica sa dijabetes melitusom trudnice ifetalne makrosomije. Rezultati istraìvanja su pokaza-li da hPL u tre}em trimestru trudno}e moè da se ko-risti kao pokazatelj placentalne insuficijencije i pore-me}aja fetalnog vitaliteta. Budu}i da je disfunkcijaplacente povezana sa poreme}ajem oksigenacije plo-da, odre|ivanje hPL u tre}em trimestru trudno}e mo-glo bi da ima izvesna prediktivna svojstva.

C31THE IMPORTANCE OF DETERMINING

HUMAN PLACENTAL LACTOGEN IN THETHIRD TRIMESTER OF PREGNANCY

J. Durkovi}, B. Mandi}

Department of Genetics – Town Hospital, SuboticaMegaLab, Biochemical Laboratory

Subotica

Human placental lactogen (hPL) is a hormoneproduced by the placenta and a regulator of fetopla-cental growth. The determination of hPL concentra-tion in the serum of pregnant women is used as anindicator of placental function. In this paper, theresults of the determination of hPL in the third trime-ster of pregnancy are presented with the aim of test-ing the sensitivity of this biochemical marker for de-tecting placental dysfunction decline in, fetal vitalityand risk of bad outcome. hPL is determined by anELISA method using Bioserv Diagnostics tests andthe results are read by a STAT-FAX 303+ reader. Thetests have been done on 370 women with high riskpregnancy, between the 20th and 36th week of preg-nancy. The indications for determining hPL werestates associated with a risk of preeclampsia, hyper-tension, diabetes mellitus, toxemia, edema, kidneyinsufficiency, as well as an ultrasonic detection ofintrauterine fetal growth halt, placental trophoblasttumor, bleeding and a threatened abortion. The re-sults of hPL marker were compared to the standardreference values according to the weeks of pregnan-cy. Normally, the concentration of hPL in the serumof a pregnant woman increases as the pregnancyprogresses. A significant decrease of hPL has beenidentified in threatened abortions, bleeding, placen-tal calcification, intrauterine fetal growth halt,intrauterine letal death and in pregnancies whichresulted in children born with fetal distress andasphyxia. A rapid increase in hPL was determined inpregnant women suffering from diabetes mellitusand in cases of fetal macrosomia. The results ofresearch showed that hPL in the third trimester ofpregnancy can be used as an indicator of placentalinsufficiency and disturbed fetal vitality. Since placen-tal dysfunction is related to a disturbed oxygenationof the fetus, the determination of hPL in the thirdtrimester of pregnancy could have some predictivecharacteristics.

JMB 2008; 27 (2) 255

UDK 577.1 : 61 ISSN 1452-8258


Poster sessions

C32PROCALCITONIN AS A MARKER OF SEPSIS

L. Doì}, J. Oros, Z. Sto{i}, A. Nikoli}

Klini~ki centar Vojvodine, Novi Sad, Centar za laboratorijsku medicinu

Urgent and adequate diagnostic procedures arenecessary to achieve early diagnosis and correct ther-apeutic treatment of sepsis. Among the latest bio-markers of sepsis, procalcitonin (PCT) has been pro-ved to be one of the most useful. After an infectionwith systemic manifestations the level of PCT signifi-cantly increases in the first 6–12h. The physiological,namely pathological role of this protein in the process-es of systemic infection, as well as in posttraumaticconditions remains still unknown. The aim of our studywas to compare the value of the levels of PCT inpatients with clinically proved sepsis to that of patientswith local infection. The goal was to prove the impor-tance and reliability of PCT as a biomarker. For obtain-ing the PCT value, a standard procedure of takingblood from 20 patients, divided into two groups wasused. The first group consisted of 10 patients withlocal infection chosen by the following criteria: leuko-cytosis, raised CRP, clinical signs of local infection andbacteriologically tested. The second group were thepatients with laboratory and microbiological confirma-tion of a septic state of different origin. PCT was meas-ured on a B.R.A.H.M.S. KRYPTOR, by a TRACE met-hod. The mean value of PCT in the first group was0.0574 ng/mL 0,0196, and in the second 0.7542ng/mL 0.6578. The difference between the meanvalues of the two groups was 0.6968 ng/mL, which isstatically significant (p<0.01). On the basis of theobtained results we can deduce that there is a notice-able difference between the two groups of patients.The mean value was between the expected levels,though on a lower level than was expected. We believethat the reason for the lower level is antibiotic therapy.PCT was recognized as a good marker of sepsis, aswell as an indicator of therapeutic monitoring.

C33RECEPTORS AND BINDING PROTEINS FOR

INSULIN AND INSULIN-LIKE GROWTHFACTORS IN THE PLACENTA OF HEALTHY

AND DIABETIC MOTHERS

M. Weber1, I. Bari~evi}-Jones1, R. Masnikosa1, D. Filimonovi}2, @. Mikovi}2, O. Nedi}1

1Institute for the Application of Nuclear Energy – INEP,Zemun, Belgrade, Serbia

2GAK »Narodni front«, Belgrade, Serbia

The IGF system in the human placenta consists ofinsulin-like growth factors (IGF) -I i -II, their receptors(IGF1R i IGF2R), and binding proteins (IGFBP-1 to -6).

C32PROKALCITONIN KAO MARKER SEPSE

L. Doì}, J. Oros, Z. Sto{i}, A. Nikoli}

Klini~ki centar Vojvodine, Novi Sad, Centar za laboratorijsku medicinu

Brze i adekvatne dijagnosti~ke procedure ne-ophodne su za postizanje rane dijagnoze i odgovara-ju}e terapije sepse. Me|u najnovijim biomarkerimasepse prokalcitonin (PCT) pokazao se kao jedan odnajkorisnijih. Nakon infekcije sa sistemskim posledica-ma, unutar 6–12h, nivo PCT-a zna~ajno raste. Fiziolo-{ka, odnosno patofiziolo{ka uloga tog proteina u pro-cesima sistemskih infekcija, kao i u posttraumatskimstanjima, ostaje jo{ uvek nepoznanica. Cilj na{e studi-je bio je pore|enje vrednosti nivoa PCT-a kod pacije-nata sa klini~ki dokazanom sepsom i pacijenata salokalnom infekcijom, radi provere zna~aja i stepenapouzdanosti PCT-a kao biomarkera. Za odre|ivanjevrednosti PCT-a kori{}en je standardno izdvojen serumiz krvi 20 pacijenata podeljenih u dve grupe. Prvugrupu je ~inilo 10 pacijenata sa lokalnom infekcijom,odabranih prema slede}em kriterijumu: povi{eni leu-kociti, povi{en CRP, klini~ki znaci lokalne infekcije ibakteriolo{ka potvrda. Drugu grupu su ~inili pacijentisa klini~ko-laboratorijskom i mikrobiolo{kom potvrdomsepti~kog stanja razli~itog uzroka. PCT je odre|ivan naBrahms Kriptoru-u, tehnologijom TRACE. Prose~navrednost PCT-a u prvoj grupi pacijenata bila je 0,0574ng/mL 0,0196, a u drugoj grupi 0,7542 ng/mL 0,6578. Razlika u srednjim vrednostima izme|u dvegrupe je 0,6968 ng/mL, {to je statisti~ki zna~ajno(p<0,01). Na osnovu dobijenih rezultata moè sezaklju~iti da postoji zna~ajna razlika izme|u dve grupepacijenata, mada je ona bila na donjoj granici o~eki-vanih vrednosti. Na{a je pretpostavka da je antibiotskaterapija razlog sniènih nivoa PCT-a. Pored njegoveuloge kao markera sepse, PCT se u na{im istraìvanji-ma pokazao kao dobar potencijalni pokazatelj teìnebolesti, njenog toka i uspe{nosti terapije.

C33RECEPTORI I VEZUJU]I PROTEINI ZAINSULIN I INSULINU SLI^NE FAKTORE

RASTA U PLACENTI ZDRAVIH I DIJABETI^NIH MAJKI

M. Weber1, I. Bari~evi}-Jones1, R. Masnikosa1, D. Filimonovi}2, @. Mikovi}2, O. Nedi}1

1Institut za primenu nuklearne energije – INEP,Zemun, Beograd, Srbija

2GAK »Narodni front«, Beograd

IGF sistem humane placente sa~injavaju insulinusli~ni faktori rasta (IGF) - I i -II, receptori za koje se onivezuju (IGF1R i IGF2R), kao i njihovi vezuju}i proteini

256

JMB 2008; 27 (2) 257

Due to many structural and metabolic similarities withinsulin, the IGF system cannot be examined separatelyfrom insulin and its receptor. In this study gel filtrationwas used to detect solubilised membrane proteins ofplacenta obtained from 3 healthy and 3 diabetic moth-ers. Solubilised placental membranes were preincubat-ed with three 125I- ligands: 125I-IGF-I, 125I-IGF-II and125I-insulin, and further chromatographed on a»Sephadex G-100« column. Radioactivity was measu-red in each collected fraction. Gel filtration of the solu-bilised membranes incubated with 125I-IGF or 125I-IGF-II resolved in comparable elution profiles, whereby thefirst eluted peak represented complexes of radioligandswith receptors, the second peak representeds radioli-gand bound to IGFBP, while the third maximum repre-sented free radioligands. To the contrary, gel filtrationof the solubilised membranes incubated with 125I-insulin produced a profile lacking a peak that includedIGFBP. By measuring the area under peaks it was foundthat there was no significant difference in the contentof IGF and insulin receptors in the solubilised placentalmembranes of healthy and diabetic mothers, while theamount of IGFBP in the placenta of healthy motherswas significantly lower. Based on these results, differentregulatory mechanisms of binding proteins in the pla-centa of healthy and diabetic mothers can be sugge-sted.

C34S-100b PROTEIN IN PATIENTS

WITH SEVERE SEPSIS

M. Vu~elji}1, M. [urbatovi}2, S. Vujani}3

1Biochemical Laboratory »Galathea«, 2Clinic of Anesthesiology and Intensive Therapy,

3Institute of Medical Biochemistry, Military Medical Academy, Belgrade, Serbia

The effects of sepsis on the brain have not beenfully elucidated. The interplay between direct effectsresulting from toxic mediators and indirect effects,such as hypotension, hyperthermia and increasedintracranial pressure, contribute to the unclear imageof the brain during sepsis. This study investigated theserum levels of S100b protein in patients with severesepsis, as a biomarker of brain damage. The aim of thestudy was to determine whether the levels of S100b areincreased early, at the onset of sepsis, and if this pro-tein is a good early predictor of outcome. We studied30 patients with severe sepsis, treated in the IntensiveCare Unit. The patients were divided into survivors(n=8) and norsurvivors (n=22). Venous blood wassampled within the first 24h after the onset of symp-toms. The concentrations of S100b protein weremeasured using an electrochemiluminescence immu-noassay on an Elecsys 2010 analyzer (Roche Diagno-

(IGFBP-1 do -6). Tako|e, usled strukturnih i metaboli-~kih sli~nosti sa insulinom, IGF sistem se ne moèizu~avati izdvojeno od insulina i insulinskog receptora.U ovom radu gel filtracija je kori{}ena kako bi se detek-tovali membranski proteini iz solubilizata membranadobijenih iz terminskih placenti tri zdrave i tri dijabe-ti~ne majke. Pojedina~ni uzorci solubilizata preinkubi-rani su sa tri 125I- liganda: 125I-IGF-I, 125I-IGF-II i 125I-insulinom. Nakon toga uzorci su razdvajani na koloni»Sephadexa G-100«, a hromatografija je pra}enamerenjem radioaktivnosti 125I-liganada u frakcijamaeluiranim sa kolone. Gel filtracijom solubilizata inku-biranog sa 125I-IGF ili 125I-IGF-II dobijaju se me|usob-no sli~ni elucioni profili, pri ~emu prvi pik sa hroma-tograma poti~e od kompleksa radioliganda sa odgo-varaju}im receptorom, drugi pik od kompleksa radioli-ganda sa IGFBP, dok tre}i pik poti~e od slobodnih radi-oliganada. Gel filtracijom solubilizata inkubiranog sa125I-insulinom dobija se hromatogram koji, za razlikuod prethodnih, ne sadrì pik koji bi odgovarao kom-pleksu sa IFGBP. Integracijom povr{ina ispod pika utvr-|eno je da se koli~ine IGF i insulinskih receptora u so-lubilizatima iz membrana placente zdravih i dijabeti~nihmajki neznatno razlikuju, dok je koli~ina IGFBP u solu-bilizatu zdrave, u odnosu na dijabeti~nu majku, zna-~ajno manja. Na osnovu ovih rezultata moè se pret-postaviti da se mehanizmi regulatorne funkcije vezu-ju}ih proteina u placenti zdrave i dijabeti~ne majkeme|usobno razlikuju.

C34PROTEIN S-100b KOD PACIJENATA

SA TE[KOM SEPSOM

M. Vu~elji}1, M. [urbatovi}2, S. Vujani}3

1Biohemijska laboratorija »Galathea«, 2Klinika za anesteziju i intenzivnu terapiju,

3Institut za medicinsku biohemiju, Vojnomedicinska akademija, Beograd, Srbija

Uticaj sepse na mozak nije u potpunosti razja-{njen. Direktni efekti posredstvom delovanja toksi~nihmedijatora i indirektni efekti kao {to su hipotenzija,povi{ena telesna temperatura, povi{en intrakranijalnipritisak, doprinose konfuznoj slici o mozgu u tokusepse. U ovom radu, ispitivali smo vrednosti proteinaS100b, kao biomarkera o{te}enja mozga, kod pacije-nata obolelih od te{ke sepse. Cilj rada je bio da se utvr-di da li su vrednosti proteina S100b povi{ene na sa-mom po~etku bolesti i da li se na osnovu njih moèpredvideti ishod. Ispitano je 30 pacijenata sa te{komsepsom, koji su le~eni u jedinici intenzivne terapije.Pacijenti su bili podeljeni u dve grupe: preìveli (n=8) iumrli (n=22). Za analizu je uzimana venska krv u prvih24h od pojave simptoma. Koncentracija proteinaS100b merena je pomo}u imunolo{kog testa meto-dom elektrohemiluminiscencije na aparatu Elecsys2010 (Roche Diagnostics). Od 30 ispitanih pacijenata,

258

74,4% (n=22) imalo je povi{ene vrednosti proteinaS100b (6 preìvelih i 16 umrlih), dok je 8 pacijenata (2preìvela i 6 umrlih) imalo vrednosti u okviru refe-rentnog opsega. Izme|u preìvelih i umrlih nije bilo sta-tisti~ki zna~ajne razlike u srednjoj vrednosti proteinaS100b (0,390±0,515 vs. 0,415±0,508 mg/L). Povi-{ene vrednosti tog proteina ukazuju na verovatna difuz-na okultna o{te}enja mozga, koja mogu biti i reverzi-bilna. Osim toga, protein S100b nije dovoljno pouzdanmarker za ranu predikciju ishoda kod pacijenata obo-lelih od te{ke sepse.

C35IMUNSKI KOMPLEKSI I KOMPLEMENT U SERUMU I SINOVIJALNOJ TE^NOSTI KOD BOLESNIKA SA REUMATOIDNIM

ARTRITISOM

Z. Miju{kovi}1, V. Subota1, J. Pejovi}1, Lj. Rackov2, Z. Kova~evi}3

1Institut za medicinsku biohemiju2Klinika za reumatologiju i klini~ku imunologiju

3Klinika za nefrologiju, Vojnomedicinska akademija, Beograd, Srbija

Reumatoidni artritis (RA) jeste predominantnointraartikularna zapaljenska i autoimunska bolest kojauklju~uje razli~ita autoantitela i efektorne mehanizme.Cilj ispitivanja je bio da se ustanovi zna~aj cirkuli{u}ihimunskih kompleksa (CIK) i komponenti komplementa(C3, C4), kao pokazatelja stepena aktivnosti RA za lab-oratorijsku dijagnostiku. U studiji preseka stanja je ispi-tano 59 bolesnika koji su prema klini~kim kriterijumimaza aktivnost RA podeljeni u dve grupe: grupa saumerenom (UA, n=24) i grupa sa visokom aktivno{}u(VA, n=35) RA. Koncentracije CIK, C3c i C4 u serumui sinovijalnoj te~nosti (ST) odre|ivane su imunone-felometrijskom metodom (»DADE – Behring«) u obegrupe ispitanika i upore|ene sa vrednostima u kontrol-noj grupi od 15 pacijenata sa povredama meniskusa.Rezultati su pokazali da nije bilo statisti~ki zna~ajnihrazlika u koncentracijama za C3c i C4 u oba biolo{kauzorka izme|u ispitivanih grupa. Statisti~ki zna~ajnerazlike na|ene su u koncentracijama CIK testiranjemvrednosti (x SD, IU/mL), a dobijene vrednosti u se-rumu grupe sa VA i grupe sa UA RA bile su:7,43 13,40; 3,01 2,92 (p<0,05) i za vrednosti uST: 13,47 21,1, 5,33 7,53 (p<0,001). Te razlike subile jo{ vi{e izraène izme|u grupe sa VA RA i KG.Rezultati koncentracija CIK su bili zna~ajno vi{i u ST uodnosu na serum u obe grupe bolesnika: u grupi saUA za 77% a u grupi sa VA RA za 82%. Ovi podaci iduu prilog potvrdi hipoteze o lokalnoj, intraartikularnojprodukciji autoantitela, odnosno CIK. Moè se zaklju-~iti da je laboratorijsko odre|ivanje koncentracije CIKkorisan pokazatelj stepena aktivnosti RA, {to se neodnosi na ispitivane komponente komplementa. To ne

stics). Out of 30 examined patients, 74.4% (n=22)had increased levels of S100b protein (6 survivors and16 nonsurvivors), while 8 patients (2 survivors and 6nonsurvivors) had values within the reference range.The mean value of S100b protein did not differ signif-icantly between the survivors and nonsurvivors(0.390±0.515 vs. 0.415±0.508 mg/L). The increa-sed levels of this protein indicate possible occult diffusebrain injury, that can be reversible. Moreover, the studyshowed that S100b protein is not a good early predic-tor of outcome in severe sepsis.

C35IMMUNE COMPLEXES AND COMPLEMENT IN THE SERUM AND SYNOVIAL FLUID OF

RHEUMATOID ARTHRITIS PATIENTS

Z. Miju{kovi}1, V. Subota1, J. Pejovi}1, Lj. Rackov2, Z. Kova~evi}3

1Institute of Medical Biochemistry2Clinic of Rheumatology and Clinical Immunology

3Clinic of Nephrology, Military Medical Academy, Belgrade, Serbia

Rheumatoid arthritis (RA) is predominantly anintraarticular inflammatory and autoimmune disease inwhich different auto antibodies and effector mecha-nisms are involved. The aim of the study was to deter-mine the utility of circulating immune complexes (CIC)and complement components (C3c, C4) as possiblemarkers for disease activity in laboratory diagnostics. Ina cross-section study 59 patients were categorized intotwo groups according to the clinical criteria: a groupwith moderate (MA, n=24), and a group with severe(SA, n=35) RA activity. The concentrations of CIC, C3cand C4 in the sera (S) and synovial fluids (SF) wereexamined by the immunonephelometric (»DADEBehring«) method in both groups and compared withthe values in the control group (n=15) of patients withlesions of the menisci. The obtained results showedthat there was no statistical significance of the values ofC3c and C4, in both biological fluids, in all testedgroups. Significant differences were found for the le-vels of CIC in both fluids, testing the parameters(x SD, IU/mL) in the sera of the groups with SA andMA of RA: 7.43 13.40; 3.01 2.92 (p<0.05) and SF:13.47 21.1, 5.33 7.53 (p<0.001), respectively.These differences were more pronounced in the groupwith SA and CG. The results for the concentrations ofCIC were significantly higher in the SF compared to thesera: in the RA group with SA by 77% and in the groupwith MA by about 82%. These data could provide aconfirmation of the hypothesis about the local, intraar-ticular autoantibodies and the subsequent CIC produc-tion. It could be concluded that the examination of theCIC concentration in serum, and where it is possible in

JMB 2008; 27 (2) 259

SF, is a useful marker of disease activity in RA patients,in contrast to the tested components of the comple-ment. This statement does not exclude their activity asimmune effector mechanisms, but suggests the possi-bility that lower molecular fragments (C3a, C4a) couldbe better disease activity markers in RA patients.

C36THE DIAGNOSTIC SIGNIFICANCE OF

DEFINING PROSTATE SPECIFIC ANTIGEN(PSA) AND PROSTATIC ACID PHOSPHATASE(PAP) WHEN AN ILLNESS OF THE PROSTATE

GLAND IS DIAGNOSED

V. Markovi}, N. Kosti}, S. \or|evi}-Cvetkovi}, S. Ra{ovi}, S. Kova~evi}

Health Centre Kru{evac, Kru{evac, Serbia

Prostate gland cancer is the most common malig-nant illness in males aged fifty and more. Thereforeearly detection is crucial for the prevention and treat-ment of this illness. The main role in this belongs totumor markers for prostate gland illness: PSA and PAP.The aim of this study is to establish the relation of thelevel of PSA and activity of PAP between a group ofpatients suffering from adenoma and prostate glandcancer and a group of healthy males, as well as toexamine the correlation of PSA and PAP in thesegroups. Test subjects are males aged from 56 to 78divided into groups diagnosed with prostate gland can-cer (32) and adenoma (47) and a group of healthymales (30). PSA was determined on an immunologicalanalyzer »IMMULITE-1000«, while PAP was determi-ned by standard biochemical methods (»Biosystems«).The results were statistically processed and it was de-termined that there is a statistically significant diffe-rence in the concentrations of PSA in the group ofpatients suffering from prostate gland cancer, and inthe group of patients suffering from adenoma that con-centration is considerably higher. The values of PAPactivity are considerably higher in the group of patientssuffering from cancer, while in the group of patientswith adenoma that difference is not statistically signifi-cant. Correlation analysis of these parameters was alsodone in all three groups. In the group with cancer thiscorrelation is the highest, while in the control group itis low, that is, there is no connection between PAP andPSA. Based on the obtained data, it can be concludedthat there is a significant connection between PSA andPAP in cases of prostate gland cancer, that it is slightlylower in cases of adenoma, and therefore that it is use-ful to determine both parameters in the diagnostic pro-cedure.

isklju~uje njihovu aktivnost u okviru efektornog imun-skog mehanizma, ali ukazuje na to da bi anafilakti~ki,manji molekulski fragmenti (C3a, C4a) mogli biti boljipokazatelji stepena aktivnosti RA.

C36DIJAGNOSTI^KI ZNAÂJ ODRE\IVANJA

PROSTATI^NOG SPECIFI^NOG ANTIGENA(PSA) I PROSTATI^NE KISELE FOSFATAZE

(PAP) KOD OBOLJENJA PROSTATE

V. Markovi}, N. Kosti}, S. \or|evi}-Cvetkovi}, S. Ra{ovi}, S. Kova~evi}

Zdravstveni centar Kru{evac, Kru{evac, Srbija

Karcinom prostate je naj~e{}a maligna bolestmu{karaca starijih od 50 godina pa je rano otkrivanjeod vitalnog zna~aja za prevenciju i terapiju bolesti. Glav-nu ulogu u tome imaju tumorski markeri za oboljenjaprostate: PSA i PAP. Cilj ovog rada je da se utvrdi odnosnivoa PSA i aktivnosti PAP izme|u grupa pacijenata saadenomom i karcinomom prostate u odnosu na zdravemu{karce, kao i da se ispita korelacija PSA i PAP udatim grupama. Ispitanici su mu{karci starosti od 56do 78 godina, svrstani u grupe sa verifikovanim karci-nomom prostate (32), adenomom prostate (47), i gru-pa zdravih mu{karaca (30). PSA je odre|ivan na imu-nolo{kom analizatoru »IMMULITE-1000«, a PAP stan-dardnom biohemijskom metodom (»Biosystems«).Statisti~kom obradom dobijenih rezultata utvr|eno jeda postoji statisti~ki zna~ajna razlika u koncentracijamaPSA u grupi pacijenata sa karcinomom prostate, a i ugrupi sa adenomom ta koncentracija je zna~ajno vi{a.Vrednosti aktivnosti PAP zna~ajno su vi{e u grupi paci-jenata sa karcinomom, dok kod pacijenata sa ade-nomom ta razlika nije statisti~ki zna~ajna. Ura|ena je ikorelaciona analiza tih parametara kod sve tri grupeispitanika. U grupi sa karcinomom takva korelacija jenajve}a (r=0,632), u grupi sa adenomom ne{to nià(r=0,421), dok je u kontrolnoj grupi ona niska(r=–0,222), to jest, nema povezanosti izme|u PSA iPAP. Na osnovu dobijenih podataka moè se zaklju~itida postoji zna~ajna povezanost PSA i PAP kod karcino-ma prostate, da je ne{to manja kod adenoma, i da jekorisno odre|ivati oba parametra u dijagnosti~koj pro-ceduri.

260

C37KLINI^KI ZNAÂJ ODRE\IVANJA

FERITINA U HEMATOLO[KIM MALIGNIM OBOLJENJIMA

M. Ple}a{, D. Begovi}, S. Kapetanovi}, M. Radojkovi}, S. Risti}

Slu`ba za laboratorijsku dijagnostiku, KBC »Dr Dragi{a Mi{ovi}-Dedinje«, Beograd, Srbija

Serumski feritin je sekretorna komponenta intra-celularnog feritina sintetisanog na endoplazmati~nomretikulumu. Kod hematolo{kih malignih oboljenja, pojednoj hipotezi, povi{ena koncentracija serumskog fer-itina je posledica sinteze neoplasti~nih }elija, ali tako|emoè biti uzrokovana o{te}enjem tkiva bogatih feriti-nom (jetra, slezina i ko{tana sr`). Cilj rada je da se odre-di povezanost nivoa feritina i enzima zna~ajnih za pro-cenu funkcije jetre kod limfoma, plazmocitoma i hro-ni~ne limfocitne leukemije. Kod ovih pacijenata supored feritina ispitivani jetreni enzimi (ALT, AST, GGT),kao i Fe. Feritin je odre|ivan hemiluminiscentnom me-todom (testom firme »DPC« na analizatoru »Immulite1000«), pri ~emu su referentne vrednosti 6–195 ng/mL.ALT, AST i GGT odre|ivani su enzimskim metodamana biohemijskom analizatoru »Dimension Rxl«, dok jeFe odre|ivano metodom sa ferenom. Visoke vrednostiferitina na|ene su kod plazmocitoma i limfoma, dok jekod HLL koncentracija feritina blago povi{ena. Rezul-tati pokazuju da postoji statisti~ki zna~ajna korelacijaizme|u feritina i GGT (p<0,01), dok nije uo~ena sta-tisti~ki zna~ajna korelacija feritina sa nivoima AST i ALT(p>0,05). Budu}i da GGT moè posluìti kao markermaligne infiltracije jetre, pove}anje koncentracije feriti-na moè nastati usled destrukcije hepatocita. Tako|e jeuo~eno da ne postoji statisti~ki zna~ajna korelacijaizme|u gvo`|a i feritina (p>0,05), {to ide u prilog hi-potezi da je feritin u tim oboljenjima sintetisan od stra-ne malignih }elija.

C38ODRE\IVANJE AKTIVNOSTI ALKALNE

FOSFATAZE KOD PACIJENATA SA ANEURIZMOM ABDOMINALNE AORTE

S. Kova~evi}, V. Markovi}, N. Kosti},S. Ra{ovi}, S. \or|evi}-Cvetkovi}

Laboratorijska slu`ba, ZC Kru{evac, Srbija

Alkalna fosfataza (EC 3.1.3.1) {iroko je raspros-tranjen enzim i nalazi se u svim }elijama a posebno naili u membranama. Cilj istraìvanja je da se utvrdi da lise na osnovu odre|ivanja nekih biohemijskih para-metara moè uspe{nije proceniti rizik od rupture aneur-izme abdominalne aorte (AAA). Parametri koji se za

C37CLINICAL IMPORTANCE OF FERRITINDETERMINATION IN HEMATOLOGIC

MALIGNANCIES

M. Ple}a{, D. Begovi}, S. Kapetanovi}, M. Radojkovi}, S. Risti}

Center for Laboratory Diagnostics, Centar »Dr Dragi{a Mi{ovi}-Dedinje«, Belgrade, Serbia

Serum ferritin is a secretory component of intra-cellular ferritin synthesized on the endoplasmic reticu-lum. A recent hypothesis suggested that an increasedconcentration of serum ferritin in hematologic malig-nancies is the result of synthesis by neoplastic cells, butit can also be caused by damage to ferritin-rich tissues(liver, spleen and bone marrow). The aim of this studywas to determine the relationship between serum fer-ritin and enzymes important for the estimation of liverfunction in lymphomas, plasmacytomas and chroniclymphocytic leukemia. In all patients we measured theconcentrations of ferritin, iron and liver enzymes (ALT,AST, GGT). Ferritin was examined using a chemilumi-nescent assay (»DPC« reagents applied on a »Immulite1000« analyzer), with a reference range of 6–195ng/mL. ALT, AST, GGT were determined using enzymemethods applied on a »Dimension Rxl« biochemicalanalyzer, and iron was determined using the Ferenmethod. High levels of serum ferritin were found inplasmacytomas and lymphomas, while the concentra-tion of serum ferritin in CLL was slightly increased. Theresults showed a significant correlation between thelevels of ferritin and GGT (p<0.01). No significant cor-relation was found between ferritin levels and ALT orAST (p>0.05). Considering that GGT can be used asa marker of liver malignant infiltration, high levels ofserum ferritin might be caused by the destruction ofhepatocytes. Results also showed that there was no sig-nificant correlation between the levels of ferritin andiron (p>0.05), which supports the hypothesis that anincrease in ferritin is the result of its production bymalignant tissues.

C38DEFINING THE ACTIVITY OF ALKALINE

PHOSPHATASE IN PATIENTS WITH ABDOMINAL AORTIC ANEURYSM

S. Kova~evi}, V. Markovi}, N. Kosti}, S. Ra{ovi}, S. \or|evi}-Cvetkovi}

Laboratory Service, HC Kru{evac, Serbia

Alkaline phosphatase (EC 3.1.3.1) is a widelyspread enzyme. It can be found in all cells, especiallyin or on all cell membranes. The aim of this researchwas to establish whether some biohemical parameterscould be certain more successfully for the estimation ofthe risk of aneurisym rupture. Parameters used so far,

JMB 2008; 27 (2) 261

such as aneurysm size, have not been proved in theanticipation of a rupture of AAA. It has been assumedthat the activity of endothelial enzymes in plasma, thatis serum, could aid in the estimation of the risk of AArupture. The activity of AP as an endothelial enzymewas defined, considering that a common cause ate-riosclerosis is, and in base of ateriosclerosis, the integri-ty of the cells of the intimate matter, that is endobodzcells,is endangered. No activity of AP in the group ofpatients with a ruptured aneurysm was found. Controlgroup was a group of healthy persons with values of APwithin reference limits. The activity was expressed inU/mL and per mg of total serum proteins. First, theactivity of AP in serum was defined with the IFCCmethod and 4-phenylphosphate as substrate. ARandox Kit 132 for AP was used. Absorbent was readat 405 nm at 37 °C on a spectrophotometer Evolution3000. The concentration of total proteins in the samesamples was thendetermined with a biuretic method(modification by Benedict) No statistically significantdifference was found on neither level of significance(p>0.05) between the patients with ruptured AA andthe patients who had no such rupture. There are nosignificant differences between each of these groupsand the control group, as well. According to these datawe came to a conclusion that the activity of AP cannotbe used as a diagnostic parameter for the anticipationof the risk of AAA rupture.

C39THE IMPORTANCE OF ACID PHOSPHATASE

AS A MARKER OF RENAL OSTEODISTROPHYIN PATIENTS ON DIALYSIS TREATMENT

A. Kostre{evi}

Public Health Center »Novi Sad«, Novi Sad, Serbia

Acid phosphatase (AP) encloses a group ofenzymes which catalyse the hydrolitic process of manyphosphoric acid monoestars. It is present in every hu-man cell. Serum AP comes from blood cells, osteo-clasts and from prostate cells in males. There are atleast five isoenzymes in the human body. This studyaims: firstly to prove the clinical importance of AP as abiochemical marker of osteoclastic activity which israised in all patients with increased bone destruction(patients with chronic kidney failure, CKF) and second-ly to define the ratio of the correlation of AP and themarker of hyperparathyroidism HBI parathormone(PTH) level in patients with CKF. Ca+ metabolism isseriously changed in CKF and clinically manifests asrenal osteodistrophy. Renal osteodystrophy presentswith increased or decreased bone Ca

+exchange

which demands different therapeutic approaches. Thesample used was: a group of 40 patients on dialysistreatment in the Center for dialysis of the P.H.C. »Dr M.Stojanovi}« and a control group of 30 healthy persons.Total AP was measured by the photometric method

sada koriste u praksi, kao {to su veli~ina anurizme,godine ìvota i dr, nisu se pokazali sigurnim u pred-vi|anju prete}e rupture AAA. Po{lo se od pretpostavkeda aktivnost endotelnih enzima u plazmi, odnosno userumu, moè biti pomo}ni dijagnosti~ki kriterijum uproceni rizika od rupture AA. Odre|ivana je aktivnostAP u grupi pacijenata sa rupturiranom aneurizmom AAi u grupi operisanih pacijenata kod kojih nije do{lo dorupture aneurizme. Kontrolnu grupu predstavljajuzdave osobe sa vrednostima AP u referentnim granica-ma. Aktivnost AP izraèna je u U/mL i na mg ukupnihproteina seruma. Prvo je odre|ivana aktivnost AP userumima pomo}u IFCC metode i sa 4-fenilfosfatomkao supstratom, kori{}en je »Randoks Kit 132« za AP.Apsorbanca je ~itana na 405 nm na 37 °C na spektro-fotometru »Evolution 3000«. Potom je odre|ena kon-centracija ukupnih proteina istih uzoraka biuretskommetodom (modifikacija po Benediktu) i aktivnost APizraèna na mg ukupnih proteina. Pojedina~ni t-testovisu pokazali da nema statisti~ki zna~ajne razlike ni najednom nivou zna~ajnosti (p>0,05) izme|u grupe pa-cijenata sa rupturiranom AA i grupe pacijenata kodkojih nije do{lo do rupture AA, kao ni zna~ajne razlikeizme|u svake te grupe ponaosob i kontrolne grupe. Naosnovu dobijenih podataka izvodi se zaklju~ak da seaktivnost AP ne moè koristiti kao dijagnosti~ki para-metar za procenu rizika od rupture AAA.

C39ZNAÂJ KISELE FOSFATAZE KAO

MARKERA RENALNE OSTEODISTROFIJEKOD DIJALIZIRANIH PACIJENATA

A. Kostre{evi}

Dom zdravlja »Novi Sad«, Novi Sad, Srbija

Renalna osteodistrofija predstavlja patolo{ke pro-mene ko{tanog tkiva izazvane poreme}ajem metabo-lizma kalcijuma tj. sekundarnim hiperparatireoidizmomu hroni~noj bubre`noj insuficijenciji (HBI). Ispoljava sestanjima sa poja~anom i smanjenom ko{tanom izme-nom od kojih svako zahteva odgovaraju}i terapijskipristup uz nagla{en zna~aj dijagnostike. Ovoj studiji jebio cilj da utvrdi mesto kisele fosfataze (AP) u moni-toringu renalne osteodistrofije kod dijaliziranih pacije-nata. Utvr|ena je laboratorijska pouzdanost fotometrij-skog odre|ivanje aktivnosti AP na automatskom anali-zatoru, i to: preciznost u seriji uzoraka u opsegu nor-malnih i visokih vrednosti (izraèna sa CV=6,9% odn.CV=3,5%), ta~nost izraèna kao procenat odstupanja(BIAS) od ciljne vrednosti primarnog standarda(CV=3,1%) i recovery testom (odstupanje do 1,24%),te ukupna gre{ka merenja od 3,89%. Potom je izvr{enopore|enje srednjih vrednosti AP u serumu ispitivanegrupe (40 pacijenata na dijalizi) i kontrolne grupezdravih osoba primenom studentovog t-testa sa rezul-tatom t=8,768, p=0,000 (p<0,001) i koreliranje

262

vrednosti AP i vrednosti parathormona (rutinski test umonitoringu dijaliziranih pacijenata) sa koeficijentomkorelacije r=0,750 (rezultat na granici srednje i jakekorelacije, a testiranjem nulte hipoteze po Fisheru sat=6,9, ova korelacija je visoko signifikantna sap=0,01). Zlatni standard za procenu oba pore|enamarkera (histolo{ki nalaz) nije bio dostupan. Ve}inastudija ukazuje na nedostatke u dijagnostici ako sekoriste izolovani biohemijski markeri ko{tanog metabo-lizma u komplikovanim i promenljivim metaboli~kimuslovima HBI, a shodno potrebi za izbegavanjem agre-sivnog dijagnosti~kog pristupa (biopsija kosti sa histo-morfometrijom), preporu~uje se uporedno kori{}enjeve}eg broja markera me|u kojima ima mesto i kiselafosfataza.

C40NIVO CISTATINA C KOD PACIJENATA

SA ENDEMSKOM NEFROPATIJOM

R. Obrenovi}1, D. Petrovi}3, B. Stojimirovi}2, N. Majki}-Singh1

1Institut za medicinsku biohemiju, KCS, Beograd2Klinika za urologiju i nefrologiju,KBC »Kragujevac«, Kragujevac

3Institut za urologiju i nefrologiju, Nefrolo{ka klinika, KCS,

Beograd, Srbija

Balkanska endemska nefropatija (BEN) jeste hro-ni~na tubulointersticijalna bolest koja dovodi do pro-gresivnog o{te}enja bubrega. BEN se javlja u izolova-nim selima Balkana oko reke Dunav. Maksimalan brojpacijenata zapaèn je u periodu izme|u 1965. i 1970.godine. Bolest zahvata ne vi{e od dve generacije. Uzrokje jo{ uvek nepoznat. Epidemiolo{ki podaci koji su do-bijeni nakon biopsije govore o primitivnim glomerulimai opstrukciji tubula. Ja~ina glomerulske filtracije (JGF)najbolji je indeks bubre`ne funkcije. Koncentracija cis-tatina C u serumu uglavnom zavisi od ja~ine glome-rulske filtracije. Shodno tome, cistatin C se name}e kaodobar interni parametar za procenu ja~ine glomerulskefiltracije. Cilj rada je bio da istraì nivoe i povezanostizme|u koncentracije cistatina C, koncentracije krea-tinina i klirensa kreatinina kod pacijenata sa BEN.Ispitivanje je obuhvatilo 40 osoba oba pola (17 èna i23 mu{karca) sa BEN ~ija je srednja vrednost starosnedobi bila 47,76±9,93 godina. Cistatin C u serumu jeodre|ivan PENIA metodom na laserskom nefelometrutestovima firme »Dade-Behring«. Srednja vrednost cis-tatina C bila je 3,69±1,39 mg/L. Srednja vrednost kre-atinina je bila 309,68±145,28 mmol/L a srednja vred-nost klirensa kreatinina 38,8±24,3 mL/min. Statisti~kaanaliza je obuhvatila srednju vrednost, Studentov t-testi Spirmanov test korelacije. Kod pacijenata sa BEN pos-toji statisti~ki veoma zna~ajna pozitivna korelacija izme-

with 1-naftil phosphate as a substrate on an automaticanalyzer Mira Cobas Plus. The compared results of themedian values of AP in the sera of two groups showedadifference which was highly statistically important(p<0.001). The values of AP and PTH are between amedium and a strong correlation measured by correla-tion coefficient r=0.750, tested by the Fisher hypothe-sis and is highely significant, p=0.01. Cytology as thegold standard for the valuation of both marker was notavailable. The study proved that AP has its place as aselfsufficient biomarker of increased osteoclastic activi-ty but (acording to a number of studies) usage ofother markers of osseal metabolism is highely recom-mended in cases of complicated and easily changedmetabolism in CKF patients. This comes with the needto avoid aggressive a diagnostic aproach (osseal biop-sy).

C40CYSTATIN C LEVEL IN PATIENTS

WITH BALKAN ENDEMIC NEPHROPATHY

R. Obrenovi}1, D. Petrovi}3, B. Stojimirovi}2, N. Majki}-Singh1

1Institute of Medical Biochemistry2Institute of Urology and Nephrology,

Clinical Center of Serbia, Belgrade3Clinic of Urology and Nephrology,

Clinical Hospital Center »Kragujevac«,Kragujevac, Serbia

Balkan endemic nephropathy (BEN) is a chronictubulointerstitial disease which leads to progressiverenal failure. BEN occurs in isolated villages in the Bal-kan region around the Danube river. Maximum num-ber of patients were seen during 1965–1970. The dis-ease affects no more than two generations. The etiolo-gy of BEN is still unknown. The epidemiological datashow primitive glomeruli and obstructed tubules afterkidney biopsy. GFR is traditionally considered the bestoverall index of kidney function. Cystatin C concentra-tion in serum depends mainly upon the glomerular fil-tration rate. Accordingly, serum cystatin C is a favorableinternal (endogenous) parameter for the measurementof glomerular filtration rate (GFR). The aim of thisstudy was to examine the association of serum cystatinC concentration, creatinine concentration and creati-nine clearance in BNE patients. The study included 40individuals of both sexes (17 females and 23 males),mean age 47.76±9.93 years, who had BEN. CystatinC in serum was determined by a PENIA method, DadeBehring. The mean value of cystatin C was 3.69±1.39mg/L. The mean value of serum creatinine was309.68±145.28 mmol/L and the mean value of crea-tinine clearance was 38.8±24.3 mL/min. Statisticalanalysis of the results was performed by the average,student t-test and Spearman test for rang correlation.In BNE, patients there is a statistically highly significant

JMB 2008; 27 (2) 263

positive correlation between the creatinine concentra-tion and cystatin C concentration (p<0.01) and thereis a statistically highly significant negative correlationbetween the creatinine clearance and cystatin C con-centration (p<0.01). Cystatin C is a good parameter todetermination the glomerular filtration rate in BNEpatients.

C41BONE ALKALINE PHOSPHATASE

AND ESTRADIOL AS MARKERS OF BONE REMODELING IN POSTMENOPAUSAL

OSTEOPOROSIS

C. V. Gurban1, M. Cojocaru2, F. Sfrijan1, D. Drugarin3

1»Victor Babes« University of Medicine and Pharmacy,Biochemistry Department, Timisoara, Romania

2Colentina Clinical Hospital, Clinical ImmunologyDepartment, Bucharest, Romania

3»Victor Babes« University of Medicine and Pharmacy,Immunology Department, Timisoara, Romania

The bone remodeling imbalance in post-menopausal women with osteoporosis is produced bydecreased bone formation and increased bone resorp-tion. The aim of this study is to determine whether se-rum levels of bone specific alkaline phosphatase (Bone-ALP) and estradiol (E2) are elevated in postmenopausalwomen with osteoporosis. We determined the serumlevels of Bone-ALP and E2 in postmenopausal womenwith osteoporosis (two cohorts of patients, depending onthe estrogenic deprivation) as compared to controls(menopausal women without osteoporosis). The serumlevels of the markers were measured by the ELISA tech-nique and the evaluation of Bone Mineral Density(BMD) as analyzed using the DEXA methods assess-ment of T spine scores (sT spine). In cohort 1 (under 15years of estrogenic deprivation) serum levels of Bone-ALP were 13.76±0.6 mg/mL, serum levels of E2 were28.32±1.78 pg/mL, and sT spine values were –3.63±0.65. In cohort 2 (over 15 years of estrogenic depriva-tion) serum levels of Bone-ALP were 11.88±0.38mg/mL, serum levels of E2 were 19.66±1.23 pg/mL,and sT spine values were –3.78±0.36. In the controlgroup the serum levels of Bone-ALP were 8.68±0.44mg/mL, serum levels of E2 were 43.07±4.04 pg/mL,and sT spine values were –1.78±0.11. Increased serumlevels of Bone-ALP demonstrate osteoblast activation,which increases significantly bone turnover, and whenassociated with low BMD demonstrates increased boneresorption in postmenopausal women with osteoporosiscompared to menopausal women without osteoporosis.The serum levels of E2 are significantly decreased anddemonstrate decreased functions of the estrogen recep-tor ERa as expressed in the osteoblasts.

|u koncentarcije kreatinina i cistatina C, kao i statisti~kivisoko zna~ajna negativna korelacija izme|u cistatina Ci klirensa kreatinina (p<0,01). Cistatin C je dobar para-metar za procenu ja~ine glomerulske filtracije.

C41KO[TANA ALKALNA FOSFATAZA

I ESTRODIOL KAO MARKERI KO[TANOGREMODELOVANJA U POSTMENOPAUZALNOJ

OSTEOPOROZI

C. V. Gurban1, M. Cojocaru2, F. Sfrijan1, D. Drugarin3

1Univerzitet za medicinu i farmaciju »Victor Babes«Odeljenje za biohemiju, Timisoara, Rumunija

2Bolnica »Colentina«, Odeljenje za klini~ku imunologiju, Bukure{t, Rumunija

3Univerzitet za medicinu i farmaciju »Victor Babes«Odeljenje za imunologiju, Timisoara, Rumunija

Poreme}aj ko{tanog remodelovanja kod èna upostmenopauzi sa osteoporozom nastaje usled sma-njenja ko{tanog formiranja i pove}anja ko{tane resorp-cije. Cilj studije bio je da se utvrdi da li su serumski nivoialkalne fosfataze specifi~ne za kosti (Bone-ALP) i estra-diola (E2) povi{eni kod èna u postmenopauzi kojeimaju osteoporozu. Odredili smo serumske nivoeBone-ALP-a i E2 kod èna u postmenopauzi sa osteo-porozom (dve grupe pacijenata, u zavisnosti od gubit-ka estrogena) i uporedili ih sa kontrolnim uzorcima(èna u menopauzi bez osteoporoze). Serumski nivoimarkera mereni su ELISA tehnikom dok je gustina min-erala u kostima (BMD) analizirana DEXA metodamaprocenom T-spine skora (sT spine). U grupi 1 (manjeod 15 godina gubljenja estrogena) serumski nivoiBone-ALP bili su 13,76 0,6 mg/ml, serumski nivoi E2bili su 28,32±1,78 pg/mL, a vrednosti sT spine bile su–3,63 0,65. U grupi 2 (preko 15 godina gubljenjaestrogena) serumski nivoi Bone-ALP bili su 11,880,38 mg/mL, serumski nivoi E2 19,66±1,23 pg/mL, avrednosti sT spine –3,78 0,36. U kontrolnoj grupi,serumski nivoi Bone-ALP bili su 8,68 0,44 mg/mL,serumski nivoi E2 43,07 4,04 pg/mL a vrednosti sTspine 1,78 0,11. Povi{eni serumski nivoi Bone-ALPukazuju na aktivaciju osteoblasta, {to znatno poja~avako{tani promet, a kada su oni povezani sa niskim BMDukazuju na pove}anu ko{tanu resorpciju kod èna upostmenopauzi sa osteoporozom u pore|enju saènama u menopauzi bez osteoporoze. Serumski nivoiE2 su zna~ajno povi{eni i ukazuju na smanjenu funkci-ju receptora za estrogen Era kao {to je izraèno uosteoblastima.

264

C42KONCENTRACIJE UKUPNOG ANTIGENA

SPECIFI^NOG ZA PROSTATU I SLOBODNOGANTIGENA SPECIFI^NOG ZA PROSTATU,

AKTIVNOSTI KISELE FOSFATAZE,PROSTATI^NE KISELE FOSFATAZE I ALKALNE FOSFATAZE U SERUMU

PACIJENATA OBOLELIH OD KARCINOMA PROSTATE

S. Stojkovi}, I. Ivanovi}, B. Bosiljci}, R. Ponjavi}, S. Jagodi}, J. Rankovi}

Op{ta bolnica Uìce, Uìce, Srbija

Kod 70 pacijenata starosti 50–80 godina, obo-lelih od karcinoma prostate odre|ivane su koncentra-cije ukupnog antigena specifi~nog za prostatu (tPSA),slobodnog antigena specifi~nog za prostatu (free PSA),aktivnosti kisele fosfataze (ACP, EC 3.1.3.2), prosta-ti~ne kisele fosfataze (pACP) i alkalne fosfataze (AP, EC3.1.3.1) u serumu, sa ciljem utvr|ivanja zna~aja tihbiohemijskih parametara u proceni prisustva karcino-ma prostate i pra}enja daljeg toka maligne bolesti.Odre|ivanja su vr{ena kod 34 pacijenta obolelih odkarcinoma prostate, kod kojih jo{ nije primenjenanijedna vrsta terapije, kod 19 pacijenata posle ura|eneradikalne prostatektomije (pacijenti u ranijim stadijumi-ma bolesti, I i II) i 17 pacijenata posle primljene radio-terapije ili hormonalne terapije (pacijenti u kasnijimstadijumima bolesti). Tumori su dokazani histopatolo-{ki, koncentracije tPSA i freePSA odre|ene su elektro-hemiluminiscentnim imunoodre|ivanjem (»ECLIA«) naimunohemijskom analizatoru »Roche Elecsys 2010«,aktivnost ACP i neprostati~ne ACP odre|ena je modi-fikovanom kolorimetrijskom metodom koju je opisaoHillman, aktivnost pACP je izra~unata (pACP=ACP-neprostati~na ACP), aktivnost AP je odre|ena IFCCkineti~kom metodom. Kontrolnu grupu je sa~injavalo60 zdravih mu{karaca starosti 50–65 godina. ANOVAtestom su upore|eni nivoi ispitivanih parametara kon-trolne grupe i pacijenata obolelih od karcinoma, prepo~etka le~enja, posle ura|ene radikalne prostatek-tomije i posle primljene radioterapije ili hormonalneterapije. Koncentracije tPSA i freePSA zna~ajno su bilevi{e kod sve tri grupe obolelih od karcinoma prostate.Aktivnosti ACP, pACP i AP zna~ajno su bile vi{e kodpacijenata pre po~etka le~enja i posle primljene terapi-je ili hormonalne terapije. Koncentracije tPSA i freePSAbile su iznad gornjih granica o~ekivanih vrednosti kodnormalnih zdravih mu{karaca, a bile su prisutne i ve}eindividualne razlike. Aktivnosti ACP, pACP i AP su po-nekad bile iznad gornjih granica o~ekivanih vrednostikod normalnih zdravih mu{karaca. Dobijeni rezultatipokazuju da povi{ene koncentracije PSA i freePSAmogu biti zna~ajan pokazatelj karcinoma prostate.Aktivnosti ACP, pACP i AP mogu biti od koristi pri dijag-nostici i pra}enju daljeg toka karcinoma prostate.

C42CONCENTRATIONS OF TOTAL

PROSTATE-SPECIFIC ANTIGEN AND FREE PROSTATE-SPECIFIC ANTIGEN,ACTIVITES OF ACID PHOSPHATASE,

PROSTATIC ACID PHOSPHATASE AND ALKALINE PHOSPHATASE IN THE

SERUM OF PATIENTS SUFFERING FROMPROSTATE CARCINOMA

S. Stojkovi}, I. Ivanovi}, B. Bosiljci}, R. Ponjavi}, S. Jagodi}, J. Rankovi}

General Hospital Uìce, Uìce, Serbia

In 70 patients, aged 50-80 years, with prostatecarcinoma, the concentrations of total prostate- specif-ic antigen (tPSA) and free prostate-specific antigen(free PSA), activities of acid phosphatase (ACP, EC3.1.3.2), prostatic acid phosphatase (pACP) and alka-line phosphatase (AP, EC 3.1.3.1) in the serum weredetermined. The aim of the study was to emphasizethe important role of these biochemical parameters inthe evaluation of prostate carcinoma. These bioche-mical values were determined in 34 patients with pro-state carcinoma so far without any therapy, in 19patients after radical prostatectomy (patients in earlierstages of diseases, I and II) and in 17 patients receiv-ing radiotherapy or hormonal therapy (patients in laterstages of diseases). Prostate carcinoma were identifiedhistopathologically. The tPSA and free PSA concentra-tions were determined by an electrochemilumines-cence immunoassay (»ECLIA«), intended for use onthe »Roche Elecsys 2010«. The ACP activity and non-prostatic ACP activity were determined with a modifi-cation of the colorimetric method described by Hill-mann, the pACP activity was calculated (Activity pACP= Activity ACP-Activity non-prostatic ACP). The APactivity was determined by IFCC kinetic method. Thecontrol group consisted of 60 healthy males, aged50–65 years. ANOVA test was used to compare thevalues of the control group and the patients with pro-state carcinoma without any therapy, the patients afterradical prostatectomy and the patients receiving radio-therapy or hormonal therapy. The concentrations oftPSA and free PSA in patients in all three groups weresignificantly increased. The ACP, pACP and AP activityin patients with prostate carcinoma without therapyand in patients receiving radiotherapy or hormonal the-rapy were significantly increased. The values of tPSAand free PSA were above the upper limits of expectedvalues in normal healthy males and great individual dif-ferences were also noted. The activities of ACP, pACPand AP were occasionally above the upper limits ofexpected values in normal healthy males. The obtainedresults indicate that the tPSA values and free PSA va-lues may be a sign of the presence of prostate carci-noma. The activity of ACP, pACP and ALP may be usedin the evaluation of prostate carcinoma.

JMB 2008; 27 (2) 265

C43PROCALCITONIN IN THE DIAGNOSIS

OF ABDOMINAL SEPSIS

T. Vodnik1, N. Majki}-Singh1, N. Ivan~evi}2


2Department of Emergency Surgery, Clinical Center of Serbia, Belgrade, Serbia

Procalcitonin (PCT) is an established marker forsevere systemic bacterial infection and sepsis.Autoimmune, allergic and viral diseases, locally limitedbacterial infections and chronic inflammation do notinduce a rise in PCT. Bacterial endotoxins play a crucialrole in the induction process. Levels of procalcitoninare elevated in many conditions leading to a systemicinflammatory response syndrome (SIRS) such as bac-terial infection, pancreatitis, burns or polytrauma.Under normal metabolic conditions, the plasma con-centration of PCT is very low but is raised in severalinfections, where increased levels of PCT (>0.5 ng/ml)have been detected. The aim of our study was to iden-tify the diagnostic significance of procalcitonin in acuteabdominal conditions. The study included 50 patientswith acute abdominal conditions, divided into sepsisand SIRS group. Group were formed after the opera-tions depending on the positive or negative microbio-logical evidence of intraabdominal infection. Bloodsamples for determination were obtained before surgi-cal procedures. PCT was measured using an immuno-luminometric assay, BRAHMS Diagnostica, Berlin,Germany. Concentrations of PCT on admission weresignificantly higher in the sepsis group (2.45±7.32ng/mL) than in the SIRS group (0.45±2.52 ng/mL,p<0.001). The mean value of PCT in septic patientswas about five times higher than the PCT values innon-infected patients. PCT values in the septic patientswere increased, while values in non-septic patientswere still within the normal range. Our results suggestthat the PCT measurement may be useful for early,preoperative diagnosis of abdominal sepsis and PCThas been established as an early indicator and usefulmarker for the diagnosis and therapy monitoring ofsepsis, severe sepsis and septic shock.

C43PROKALCITONIN U DIJAGNOZI

ABDOMINALNE SEPSE

T. Vodnik1, N. Majki}-Singh1, N. Ivan~evi}2


2Centar za urgentnu hirurgiju, Klini~ki Centar Srbije, Beograd, Srbija

Prokalcitonin (PCT) jeste potvr|eni marker te{kihsistemskih bakterijskih infekcija i sepse. Autoimuna,alergijska i virusna oboljenja kao i lokalizovane bakteri-jske infekcije i hroni~na inflamacija ne dovode do pove-}anja prokalcitonina. Klju~nu ulogu u procesu indukci-je PCT imaju bakterijski endotoksini. Nivo prokalcito-nina pove}ava se u stanjima poznatim pod imenomsindrom sistemskog inflamatornog odgovora (engl.SIRS) kao {to su bakterijska infekcija, pankreatitis,groznica ili politrauma. U normalnim metaboli~kumstanjima, koncentracija prokalcitonina u plazmi je veo-ma mala (< 0,1 ng/mL) i raste u te{kim infekcijama,gde se mogu detektovati i nivoi prokalcitonina iznad0,5 ng/mL. Cilj ove studije bio je da se utvrdi dijag-nosti~ki zna~aj prokalcitonina u akutnim abdominalnimstanjima. U studiju je bilo uklju~eno 50 pacijenata, sadijagnozom akutnog abdominalnog stanja, podeljenihu dve grupe zavisno od toga da li su se kod njih razviliSIRS ili sepsa. Grupe pacijenata su oblikovane nakonoperativnog zahvata u zavisnosti od pozitivnog tj. neg-ativnog mikrobiolo{kog nalaza intra-abdominalneinfekcije. Uzorci krvi u kojima je odre|ivan nivo PCTuzimani su pre operativnog zahvata. Prokalcitonin jeodre|ivan imunoluminometrijskim testom firme»BRAHMS Diagnostica« (Berlin, Nema~ka). Dobijenevrednosti za prokalcitonin, po prijemu pacijenta, bile sustatisti~ki zna~ajno vi{e u grupi pacijenata kod kojih serazvila sepsa (2,45±7,32 ng/mL) u odnosu na paci-jente sa dijagnozom SIRS (0,45±2,52 ng/mL,p<0,001). Srednja vrednost za PCT kod septi~nihpacijenata bila je pet puta ve}a od vrednosti PCT kodneinfektivnih pacijenata. Nivoi prokalcitonina kod paci-jenata sa sepsom su bili pove}ani, dok su vrednosti zaneinfektivne pacijente i pored pove}anja ostale u opse-gu normalnih vrednosti. Na{i rezultati pokazali su veli-ku korist od odre|ivanja PCT za ranu, preoperativnudijagnozu abdominalne sepse, gde je prokalcitonin ipotvr|en kao rani indikator i vrlo koristan marker i zadijagnozu kao i za terapiju sepse i septi~nog {oka.

266

C44PRODUKTI UZNAPREDOVALE OKSIDACIJE

PROTEINA KOD PACIJENATA NA HEMODIJALIZI

T. Ivani}-]orlomanovi}1, S. Samardì}2, S. ]uk3, A. Stefanovi}4, A. Vujovi}4, V. Dopsaj4,

S. Simi}-Ogrizovi}5, S. Spasi}4, V. Kalimanovska-Spasojevi}4, J. Kotur-Stevuljevi}4

1Biohemijska laboratorija, Dom zdravlja Obrenovac2Biohemijska laboratorija,

Dom zdravlja Novi Beograd3Biohemijska laboratorija, Poliklinika »Vizim«

4Institut za medicinsku biohemiju, Farmaceutski fakultet, Beograd

5Nefrolo{ka klinika, Institut za urologiju i nefrologiju,Klini~ki centar Srbije, Beograd

Oksidativni stres se smatra zna~ajnim faktoromrizika za razvoj kardiovaskularnih bolesti i zajedni~kimmehanizmom delovanja drugih faktora rizika zarazvoj koronarne bolesti. Pacijenti na hemodijalizi suzbog visokog nivoa oksidativnog stresa pod visokimrizikom za razvoj kardiovaskularnih bolesti. U ovojstudiji je meren oksidativni stres odre|ivanjem kon-centracija produkata uznapredovale oksidacije pro-teina (engl. advanced oxidation protein products,AOPP) zajedno sa drugim klini~kim i biohemijskimparametrima. Sakupljani su serumi pripadnika kon-trolne grupe (n=102), kao i serumi pacijenata nahemodijalizi (n=63), pre same procedure dijaliziran-ja. Koncentracije AOPP, malondialdehida (MDA),lipidnih hidroperoksida (LOOH), superoksidnoganjona (O

2

.–), ukupnih SH grupa i aktivnosti enzima

superoksid-dizmutaze (SOD) odre|ivane su odgova-raju}im metodama. Zna~ajno pove}anje koncentra-cija AOPP uo~eno je kod pacijenata na hemodijaliziu odnosu na zdrave osobe (p<0,001). Tako|e jeuo~eno pove}anje MDA (p<0,01), O

2

.–(p<0,05) i

smanjenje sadràja ukupnih SH grupa (p<0,001) iaktivnosti enzima SOD (p<0,05) kod pacijenata.Zabeleèna je zna~ajna korelacija izme|u koncen-tracija AOPP i ukupnih proteina (r=0,465, p<0,001)kao i AOPP i koncentracija triglicerida (r=0,682,p<0,001) kod pacijenata. Ova studija je pokazala daje kod pacijenata na hemodijalizi evidentno pove}anoksidativni stres i smanjena sposobnost antioksida-tivne za{tite i takvo stanje izlaè pacijente pove}anomriziku od kardiovaskularnih bolesti.

C44ADVANCED OXIDATION PROTEIN

PRODUCTS (AOPP) IN HEMODIALYSIS PATIENTS

T. Ivani}-]orlomanovi}1, S. Samardì}2, S. ]uk3, A. Stefanovi}4, A. Vujovi}4, V. Dopsaj4,

S. Simi}-Ogrizovi}5, S. Spasi}4, V. Kalimanovska-Spasojevi}4, J. Kotur-Stevuljevi}4

1Biochemistry Laboratory, Health Center Obrenovac2Biochemistry Laboratory,

Health center Novi Beograd 3Biochemistry Laboratory, Polyclinic »Vizim«

4Institute of Medical Biochemistry, School of Pharmacy, Belgrade

5Clinic of Nephrology, Institute of Urology and Nephrology,


Oxidative stress is considered a significant car-diovascular risk factor and the unifying mechanismfor other cardiovascular risk factors. Hemodialysis pa-tients develop high levels of oxidative stress and the-refore are at high risk for cardiovascular disease(CVD). In the present study oxidative stress wasassessed using advanced oxidation protein products(AOPP) along with other clinical and biological pa-rameters. Sera of a control group (n=102) as well assera of patients (n=63) before hemodialysis sessionwere collected. Serum AOPP, malondialdehyde(MDA), lipid hydroperoxide (LOOH), superoxideanion (O

2

.–), total SH groups and superoxide dismu-

tase (SOD) activity were estimated by appropriateassays. A significant increase in AOPP value wasobserved in hemodialysis patients as compared tohealthy subjects (p<0.001). Also, a significant incre-ase was observed for MDA (p<0.01), O

2

.– (p<0.05)

and decrease in SH groups content (p<0.001) andSOD activity (p<0.05) in the patients. Significantcorrelation was noted for AOPP and total proteincontent (r=0.465, p<0.001) and AOPP and triglyc-eride concentration (r=0.682, p<0.001) in a pa-tient group. This study shows that the increasedoxidative stress in hemodialysis patients is evident aswell as diminished antioxidative capability and sucha situation exposes patients to a higher risk of car-diovascular disease.

D

LIPIDI I LIPOPROTEINI

LIPIDS AND LIPOPROTEINS

D45LIPID PARAMETER VALUES IN PATIENTS

WITH AGE-RELATED MACULAR DEGENERATION

E. ôlak1, S. Stankovi}1, N. Kosanovi}-Jakovi}2, A. Radosavljevi}2, D. Terzi}1, J. Stojanovi},

N. Majki}-Singh1

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

2Institute of Eye Diseases, CCS, Belgrade, Serbia

The peroxidation of lipids, especially polyunsatu-rated lipids, is a frequent occurence in stressed tissues,being particularly common as a response to increasedoxidative stress and free radical production. The aim ofthis study was to analyse the concentration of serumlipoproteins (total cholesterol, triglycerides, HDL- andLDL-cholesterol), as well as apolipoproteins A1, B andLp(a), in age-related macular degeneration, in order toexplore the role of lipid parameters as risk factors for thedevelopment of age-related macular degeneration(AMD). A total of 48 AMD patients aged 70.7±6.76and 17 age-matched healthy control subjects wereincluded in the study. The patients underwent a com-plete ophtalmological examination, including visualacuity, color fundus photography and fluoresceinangiography. The patients’ lipid status was estimated anOlympus AU400 biochemical analyzer. Statistical analy-ses were performed by an SPSS v10.0 statistical pack-age using a Mann-Whitney U test, Chi-Square andANOVA tests. A statistical processing of the datarevealed significantly higher values of total cholesterol,triglycerides and LDL-cholesterol (p<0.05), as well asapolipoproteins A1 and B (p<0.001) in AMD patients,compared to healthy control subjects. Seventy eightpercent of the tested AMD patients had lipid status dis-orders: 44% had type 2a hyperlipoproteinemia, 28%had type 2b and 6% type 4 hyperlipoproteinemia. Ba-sed on the obtained results, it may be concluded thatplasma lipid disorders could play an important role inthe development of age-related macular degenerationin elderly subjects.

D45KONCENTRACIJA LIPIDNIH PARAMETARA

KOD PACIJENATA SA SENILNOM DEGENERACIJOM MAKULE

E. ôlak1, S. Stankovi}1, N. Kosanovi}-Jakovi}2, A. Radosavljevi}2, D. Terzi}1, J. Stojanovi},

N. Majki}-Singh1

1Institut za medicinsku biohemiju2Institut za o~ne bolesti, KCS,

Beograd, Srbija

Lipidna peroksidacija, naro~ito polinezasi}enihmasti, ~esta je pojava u tkivima, koja predstavlja odgo-vor na pove}an oksidativni stres i produkciju slobodnihradikala. Cilj ovog rada je bio da se analiziraju koncen-tracije serumskih lipoproteina, i to ukupnog holestero-la, triglicerida, HDL- i LDL-holesterola, kao i koncen-tracije apolipoproteina A1, B i Lp(a) kod pacijenata sasenilnom degeneracijom makule (SDM), kako bi seispitala uloga lipidnih parametara kao faktora rizika zanastanak i razvoj bolesti. U ispitivanje je uklju~eno 48pacijenata sa senilnom degeneracijom makule, starosti70,7±6,76 godina, i 17 zdravih ispitanika iste sta-rosne dobi, koji su ~inili kontrolnu grupu. Pacijenti supro{li kompletan oftalmolo{ki pregled uklju~uju}i pre-gled fundusa i fluorescentnu angiografiju. Lipidni sta-tus je analiziran na biohemijskom analizatoru OlympusAU400. Obrada statisti~kih podataka vr{ena je po-mo}u statisti~kog paketa SPSS v10.0, koriste}i Mann-Whitney U test, c2 i ANOVA test. Statisti~kom obra-dom podataka utvr|eno je da su vrednosti ukupnogholesterola, triglicerida i LDL-holesterola (p<0,05),kao i vrednosti apolipoproteina A1 i B (p<0,001) bilezna~ajno vi{e kod pacijenata sa SDM u pore|enju sakontrolnom grupom zdravih ljudi. Od ukupnog brojapacijenata sa SDM, 78% je imalo poreme}aj lipidnogstatusa: 44% je imalo tip 2a, 28% tip 2b a 6% tip 4hiperlipoproteinemije. Na osnovu dobijenih rezultatamoè se zaklju~iti da poreme}aj serumskih lipida moèigrati zna~ajnu ulogu u nastanku i razvoju senilne de-generacije makule kod starijih ljudi.

JMB 2008; 27 (2) 269

UDK 577.1 : 61 ISSN 1452-8258


Poster sessions

270

D46SMALL DENSE LDL AND HDL PARTICLESARE ASSOCIATED WITH HIGHER SERUM

URIC ACID LEVELS AND LOW-GRADEINFLAMMATION

J. Veki}1, Z. Jeli}-Ivanovi}1, V. Spasojevi}-Kalimanovska1, L. Memon2,

A. Zeljkovi}1, N. Bogavac-Stanojevi}1, S. Spasi}1

1Institute of Medical Biochemistry, Faculty of Pharmacy, Belgrade2Clinical Chemistry Laboratory,

Clinical Centre »Beànijska kosa«, Belgrade

This study investigates the interrelationships bet-ween serum uric acid (UA), high sensitivity C-reactiveprotein (hsCRP) and fibrinogen concentrations withLDL and HDL sizes in healthy middle-aged subjects.Serum UA, hsCRP and plasma fibrinogen concentra-tions were measured by standard laboratory methodsin a sample of 194 healthy volunteers (112 men and82 women). The diameters of LDL and HDL particleswere determined by gradient gel electrophoresis. Theparticipants in the highest UA tertile had significantlysmaller LDL and HDL particle sizes (P<0.05 andP<0.01, respectively) and higher concentrations of fib-rinogen and hsCRP (P<0.05 and P<0.01, respective-ly). Elevated UA was a significant predictor of smaller,denser LDL and HDL particles (OR = 3.09; P<0.01and OR = 4.40; P<0.001, respectively). The obser-ved relationship of higher UA with smaller HDL sizepersisted after adjustment for conventional cardiovas-cular risk factors. Higher hsCRP concentration correla-ted with smaller LDL size (P<0.05), while fibrinogenconcentration was inversely related to HDL size(P<0.05). In conclusion, higher serum UA and low-grade inflammation are closely linked to alterations inlipoprotein metabolism, which may represent an earlysign of atherosclerosis in asymptomatic subjects.

D47THE INFLUENCE OF DIFFERENT

APOLIPOPROTEIN (a) ISOFORMS AND LOW DENSITY LIPOPROTEIN SUBCLASSES

ON LIPOPROTEIN (a) CONCENTRATION

A. Zeljkovi}1, N. Bogavac-Stanojevi}1, V. Spasojevi}-Kalimanovska1, Z. Jeli}-Ivanovi}1, J. Veki}1, S. Spasi}1, D. Kalimanovska-O{tri}22

1Institute of Medical Biochemistry, Faculty of Pharmacy, Belgrade

2Institute of Cardiovascular Diseases, Clinical Centre of Serbia, Belgrade

An elevated concentration of lipoprotein (a) Lp(a)is a risk factor for the development of coronary arterydisease (CAD). Lp(a) consists of a low density lipopro-

D46MALE GUSTE LDL I HDL ÊSTICE SU UDRU@ENE SA POVI[ENOM

KONCENTRACIJOM MOKRA]NE KISELINE I INFLAMACIJOM NISKOG STEPENA

J. Veki}1, Z. Jeli}-Ivanovi}1, V. Spasojevi}-Kalimanovska1, L. Memon2,

A. Zeljkovi}1, N. Bogavac-Stanojevi}1, S. Spasi}1

1Institut za medicinsku biohemiju, Farmaceutski fakultet, Beograd2Klini~ko-hemijska laboratorija,

Klini~ki centar »Beànijska kosa«, Beograd

U ovoj studiji ispitivana je me|usobna povezanostkoncentracija mokra}ne kiseline (MK), visoko osetljivogC-reaktivnog proteina (hsCRP) i fibrinogena saveli~inom LDL i HDL ~estica kod zdravih sredove~nihosoba. Koncentracije MK, hsCRP i fibrinogena sumerene standardnim laboratorijskim metodama uuzorcima seruma i plazme 194 zdrava dobrovoljca(112 mu{karaca i 82 ène). Veli~ina LDL i HDL ~esti-ca je odre|ena gradijent gel elektroforezom. Ispitanicisa vrednostima MK u najvi{em tercilu imali su manjeLDL i HDL ~estice (P<0,05 i P<0,01) i vi{e koncen-tracije fibrinogena i hsCRP (P<0,05 i P<0,01).Povi{ena MK je bila zna~ajan prediktor prisustva malih,gustih LDL i HDL ~estica (OR = 3,09; P<0,01 i OR =4,40; P<0,001). Utvr|ena veza izme|u povi{ene kon-centracije MK i manje veli~ine HDL-a ostala je sta-tisti~ki zna~ajna i u prisustvu tradicionalnih faktorarizika za KVB. Pove}ane koncentracije hsCRP su bile ukorelaciji sa manjom veli~inom LDL ~estica (P<0,05),dok su vi{e koncentracije fibrinogena bile u korelaciji samanjim dijametrima HDL ~estica (P<0,05). Rezultatistudije ukazuju da su povi{ena koncentracija MK i infla-macija niskog stepena povezane sa poreme}ajima umetabolizmu lipoproteina, {to moè biti rani pokazateljateroskleroze kod asimptomatskih osoba.

D47UTICAJ RAZLIÎTIH IZOFORMI

APOLIPOPROTEINA (a) I SUBKLASALIPOPROTEINA MALE GUSTINE NA

KONCENTRACIJU LIPOPROTEINA (a)

A. Zeljkovi}1, N. Bogavac-Stanojevi}1, V. Spasojevi}-Kalimanovska1, Z. Jeli}-Ivanovi}1, J. Veki}1, S. Spasi}1, D. Kalimanovska-O{tri}2

1Institut za medicinsku biohemiju,Farmaceutski fakultet, Beograd,

2Institut za kardiovaskularne bolesti, Klini~ki centar Srbije, Beograd

Povi{ena koncentracija lipoproteina (a) Lp(a) je-dan je od faktora rizika za razvoj koronarne arterijskebolesti (KAB). Taj lipoprotein se sastoji od apolipopro-

JMB 2008; 27 (2) 271

tein (LDL) particle to which a glycoprotein apolipopro-tein (a) apo(a) is attached. Smaller apo(a) isoformsand LDL subclasses are associated with an elevated riskfor CAD. It has been documented that the presence ofsmaller apo(a) isoforms is related to a high Lp(a) level,while it is unclear if changes in the size and distributionof circulating LDL particles have any impact on Lp(a)concentration. In this study, we examined the presenceof smaller apo(a) isoforms and LDL subclasses in cir-culation with respect to the concentration of Lp(a) in109 patients with CAD and 102 controls. The plasmaconcentration of Lp(a) was measured using immuno-turbidimetry. Apo(a) size isoforms were determined byhigh resolution SDS-agarose gel electrophoresis fol-lowed by immunoblotting. LDL subclasses were deter-mined by polyacrylamide gradient (3–31%) gel elec-trophoresis. Mean LDL and apo(a) sizes were lower inpatients than in controls (P<0.001 and P<0.01,respectively). The prevalence of small apo(a) isoformswas significantly higher among the patients (P<0.01).However, we found no differences in the prevalence ofsmall LDL particles. Next, we examined the predomi-nance of smaller apo(a) isoforms and LDL subclasseswith respect to the concentration of Lp(a) ( 300 mg/Lor<300 mg/L). The obtained results showed signifi-cantly higher presence of smaller apo(a) isoforms inindividuals with Lp(a) level 300 mg/L, in CAD(P<0.01), as well as in the control group (P<0.01).The highest proportion of small LDL particles was seenin CAD patients with Lp(a) levels 300 mg/L, but withno statistically significant difference (P>0.05) whencompared to the other groups. Our results indicatedthat CAD patients with elevated Lp(a) concentrationhad an abundance of smaller apo(a) isoforms andsmall LDL subclasses in the circulation. This topicshould be further elucidated by future studies.

D48THE DETERMINATION OF LIPID STATUS

IN THE ROMANI POPULATION OF IN\IJA MUNICIPALITY

D. Milo{evi}, V. Petrovi}, S. Mili}, M. Crn~evi}, B. Mili~evi}

Health Centre »Dr Milorad Mika Pavlovi}«, In|ija

The Health Centre of In|ija has been actively par-ticipating in the project called »The Decade of Roma«with its own project »Monitoring and Secondary Pre-vention of Cardiac Diseases in the Romani Populationof In|ija«. The research included 174 Romani subjectsfrom the territory of the town of In|ija. Thirty three sub-jects were chosen the method of free selection, 18women and 15 men average age 50±7. A controlgroup included healthy adults, average age 49±7. Inthe blood samples we determined triglycerides, total,HDL and LDL cholesterol using standard enzymatic

teina (a) apo(a) koji je vezan za ~esticu lipoproteinamale gustine (LDL). Poznato je da su manje izoformeapo(a) i male, guste LDL ~estice povezane sa pove-}anim rizikom za razvoj KAB. Tako|e je poznato da jepovi{ena koncentracija Lp(a) u cirkulaciji pra}ena pris-ustvom malih apo(a) izoformi. Me|utim, jo{ uvek nijejasno da li promene u veli~ini i raspodeli LDL ~estica ukrvi imaju uticaja na promene u koncentraciji Lp(a). Uovoj studiji smo ispitivali povezanost prisustva malihapo(a) izoformi i malih LDL subklasa sa promenama ukoncentraciji Lp(a) u krvi 109 pacijenata sa KAB i 102zdrava ispitanika. Koncentracija Lp(a) u plazmi je mer-ena imunoturbidimetrijski. Izoforme apo(a) su odre|e-ne elektroforetski na SDS-agaroznom gelu, nakon~ega je primenjen imunobloting. Subklase LDL ~esticasu odre|ene gradijent (3–31%) gel elektroforezom napoliakrilamidnom gelu. Veli~ina LDL i apo(a) ~esticabila je zna~ajno manja kod pacijenata (P<0,001 iP<0,01). Kod pacijenata je uo~eno zna~ajno ve}e pris-ustvo malih apo(a) izoformi (P<0,01), dok se zastu-pljenost malih LDL ~estica nije razlikovala u ispitivanimgrupama. Dalje smo ispitivali zastupljenost malih izo-formi apo(a) i malih subklasa LDL u odnosu na kon-centraciju Lp(a) ( 300 mg/L ili <300 mg/L). Dobijenirezultati pokazali su zna~ajno ve}e prisustvo malihapo(a) izoformi u krvi ispitanika sa koncentracijomLp(a) 300 mg/L, kako u KAB (P<0,01), tako i u kon-trolnoj grupi (P<0,01). S druge strane, najve}a zastu-pljenost malih LDL ~estica bila je u grupi KAB pacije-nata sa Lp(a) koncentracijom 300 mg/L, ali bez sta-tisti~ki zna~ajne razlike u odnosu na zastupljenost tihsubklasa u ostalim ispitivanim grupama. Na{i rezultatipokazuju da je kod pacijenata sa KAB povi{ena kon-centracija Lp(a) pra}ena pove}anim prisustvom malihizoformi apo(a) i malih, gustih LDL ~estica u krvi.Odnos izme|u komponenti Lp(a) treba dalje analiziratiu budu}im studijama.

D48LIPIDNI STATUS U POPULACIJI

ROMA OP[TINE IN\IJA

D. Milo{evi}, V. Petrovi}, S. Mili}, M. Crn~evi}, B. Mili~evi}

Dom zdravlja »Dr Milorad Mika Pavlovi}«, In|ija

U okviru projekta »Dekada Roma«, Dom zdravljaIn|ija radio je na samostalnom projektu »Monitoring isekundarna prevencija koronarne bolesti kod popu-lacije Roma u op{tini In|ija«. Tim istraìvanjem obuh-va}eno je 174 pripadnika romske populacije SO In|ija,od kojih je metodom slu~ajnog izbora izdvojeno njihukupno 33–18 èna i 15 mu{karaca prose~ne starosti50±7 godina. Kontrolnu grupu je obuhvatila 31 zdra-va osoba prose~ne starosti 49±7. Iz uzoraka krviodre|ivana je koncentracija triglicerida, ukupnog ho-lesterola, HDL holesterola i LDL holesterola primenom

272

methods, calculating the atherosclerosis index (AI). Li-pid concentration was examined using »Roche Diagno-stics GmbH« reagents on a discrete biochemical auto-mat »Roche/Hitachi 912« analyzer. The results showeda statistically significant difference (P<0.01) in the con-centrations of triglycerides, total and LDL cholesterol(LDL-C) and atherosclerosis index between the exam-ined population and control group. No statistically sig-nificant difference (P>0.05) was found in the values ofHDL holesterol (HDL-C) in both groups (Table 1). Theobtained results confirme the possibility that preventionactivities can help to decrease the levels of triglycerides,total and LDL cholesterol, atherosclerosis index, andelevate the HDL cholesterol level. There is clear indica-tion for the secondary prevention of cardiac diseases inthe Romani population of In|ija.

Table II. Mean concentration of lipid status and AI in theexamined and control group.

D49THE ASSOCIATION BETWEEN

HDL PARTICLE SIZE AND PARAOXONASE ACTIVITY IN TYPE 2 DIABETES

MELLITUS PATIENTS

A. Stefanovi}, J. Kotur-Stevuljevi}, S. Spasi}, J. Veki}, A. Zeljkovi},

V. Spasojevi}-Kalimanovska, Z. Jeli}-Ivanovi}

Institute of Medical Biochemistry,Faculty of Pharmacy, Belgrade, Serbia

HDL-cholesterol is recognized as a factor that pro-tects against the development of cardiovascular dis-eases, and low HDL-cholesterol is associated withincreased risk of cardiovascular diseases in subjects withand without diabetes. Many studies showed that HDLcomposition is abnormal in type 2 diabetic patients.Paraoxonase (PON1) is a serum esterase associatedwith HDL which protects LDL particles from oxidativemodifications. Our study examined the association ofHDL2 and HDL3 particles distribution in type 2 diabetesmellitus patients and controls with PON1 status. PON1activities were measured spectrophotometrically in thesera according to the method described by Richter andFurlong in 91 control subjects and 85 type 2 diabetesmellitus patients. HDL particle size was determined by

standardne enzimske metode. U radu su kori{}enikomercijalni reagensi za odre|ivanje lipidnog statusa,firme »Roche Diagnostics GmbH«, na diskretnom bio-hemijskom analizatoru »Roche/Hitachi 912«, a indeksateroskleroze je odre|en ra~unski. Rezultati pokazujuda postoji statisti~ki zna~ajna razlika (P<0,01) u kon-centracijama triglicerida, holesterola, LDL holesterola iindeksa ateroskleroze izme|u ispitivane i kontrolnegrupe, dok (P>0,05) u koncentracijama HDL holes-terola ne postoji statisti~ki zna~ajna razlika (Tabela I). Toukazuje na mogu}nost preventivnog delovanja naspre~avanje razvitka kardiovaskularnih bolesti i daljegporasta koncentracije triglicerida, holesterola, LDLholesterola kao i na pove}anje HDL holesterola. Postojijasna indikacija za sekundarnu prevenciju kardiovasku-larnih bolesti me|u pripadnicima romske populacije nateritoriji op{tine In|ija.

Tabela I. Prose~na koncentracija parametara lipidnog statusapo grupama.

D49VEZA IZME\U VELIÎNE HDL ÊSTICA

I PARAOKSONAZNE AKTIVNOSTI KOD PACIJENATA SA

DIABETES MELLITUS-OM TIPA 2

A. Stefanovi}, J. Kotur-Stevuljevi}, S. Spasi}, J. Veki}, A. Zeljkovi},

V. Spasojevi}-Kalimanovska, Z. Jeli}-Ivanovi}

Institut za medicinsku biohemiju, Farmaceutski fakultet, Beograd, Srbija

Dobro je poznato da su niske koncentracije HDLholesterola udruène sa pove}anim rizikom za razvojkardiovaskularnih bolesti. Mnoge studije su pokazaleda su HDL ~estice strukturno izmenjene kod pacijena-ta sa dijabetes melitusom tipa 2. Paraoksonaza (PON1)enzim je na HDL-u koji {titi LDL ~estice od oksidativnemodifikacije. Ova studija se bavi ispitivanjem veze iz-me|u zastupljenosti HDL2 i HDL3 lipoproteinskihfenotipova i statusa PON1. U serumu 91 zdrave osobei 85 pacijenata sa dijabetes melitusom tipa 2 odre-|ivana je aktivnost PON1 metodom po Richteru i Furl-ongu. Veli~ina HDL ~estica je odre|ivana elektrofore-zom na poliakrilamidnom gelu. Aktivnost PON1 je zna-~ajno manja kod pacijenata u pore|enju sa kontrol-nom grupom (P<0,001), ali nije prona|ena razlika u

Holesterol LDL-H HDL-H Trigliceridi IA

Kontrolna

grupa

4,44 +/–

0,50

2,38 +/–

0,50

1,57 +/–0,28

1,14+/–0,35

1,61 +/–0,62

Ref.

vrednosti

Do 5,2 Do 3,4 Preko1,6

Do 1,7 Do 3,0

Ispitivana

grupa

5,92

+/–

1,24

3,45

+/–1,22

1,48 +/–0,27

1,74 +/–0,86

2,46 +/–1,11

TotalCholesterol

LDL-C HDL-C Triglyceride AI

Control

group

4.44 +/–

0.50

2.38 +/–

0.50

1.57 +/–0.28

1.14+/–0.35

1.61 +/–0.62

Reference

values

Do 5.2 Do 3.4 Preko1.6

Do 1.7 Do 3.0

Examined

group

5.92

+/–

1.24

3,45

+/–1.22

1.48 +/–0.27

1.74 +/–0.86

2.46 +/–1.11

JMB 2008; 27 (2) 273

polyacrylamide gradient gel electrophoresis. PON1activity was significantly reduced in patients comparedwith controls (P<0.001). Furthermore, we determinedPON1 Q192R phenotypes in both groups, but we failedto find any differences in their distribution. Also, wecompared PON1 activity between type 2 diabetes mel-litus patients and controls, both with a predominance oflarger HDL2 particles. As we expected, PON1 activitywas significantly higher in controls (P<0.001). Signifi-cant difference was not observed between the twogroups with a predominance of smaller HDL 3 sub-classes. In the controls with predominant HDL2 parti-cles we found significantly higher PON1 activity com-paring with predominant HDL3 particles (P<0.05), butthe difference in PON1 activity between the subjectswith HDL2 and HDL3 particles was not seen in diabe-tic patients. It suggests that HDL2 subclasses are morepotent inhibitors of LDL oxidation but in diabetics maylose their antioxidant efficiency. Therefore, we conclu-ded that reduced PON1 activity and altered HDL com-position might contribute to the acceleration of athero-sclerosis in type 2 diabetes mellitus patients.

D50TESTING THE ACCURACY OF

TWO NEW FORMULAS FOR THE DETERMINATION OF LOW-DENSITY

LIPOPROTEIN CHOLESTEROL CONCENTRATION COMPARED

WITH A DIRECT ASSAY

A. Vujovi}1, J. Kotur-Stevuljevi}1, N. Buji{i}2, S. Spasi}1

1Institute of Medical Biochemistry, Faculty of Pharmacy, Belgrade

2Biochemical Laboratory »Belladona«, Belgrade-Zemun, Serbia

Reliable and accurate determination of low-densi-ty lipoprotein cholesterol (LDL-C) concentration is veryimportant for the classification of patients into properrisk categories based on cut-off points given by NCEPATP-III. The Friedewald formula for the estimation ofLDL-C concentrations is the most often used formula inclinical practice. In the present study, we attempted toderive and validate more accurate formulas to deter-mine LDL-C levels. Our study included 1640 patients(mean age 54 ± 15, 52.4% women) with triglycerideconcentration 4.5 mmol/L. LDL-C was determined bythe direct method and also calculated with two new for-mulas. The first formula represents a modified Friede-wald formula and the second was derived using multi-ple linear regression analysis. In the studied population,the mean LDL-C levels was 3.86±1.09 mmol/L meas-ured by the direct assay (D-LDL-C), 3.86 ± 1.1 mmol/Lcalculated using the first formula (F1-LDL-C) and3.83±1.06 mmol/L using the second formula (F2-LDL-C). Linear regression analysis shows good andequal correlations between D-LDL-C and LDL-C calcu-

distribuciji PON1 Q192R fenotipa u ispitivanim grupa-ma. Pore|enjem aktivnosti PON1 izme|u kontrole ipacijenata sa predominantno zastupljenim ve}imHDL2 ~esticama, kao {to se i o~ekivalo, dobijene suzna~ajno ve}e aktivnosti PON1 u kontrolnoj grupi(P<0.001). Me|utim, statisti~ki zna~ajna razlika uaktivnosti PON1 izme|u kontrole i pacijenata nije do-kazana kada su upore|ene osobe sa predominantnozastupljenim HDL3 ~esticama. Aktivnost PON1 kodosoba sa predominantnim HDL2 ~esticama bila jezna~ajno ve}a u kontrolnoj grupi (P<0,05) u odnosuna osobe sa predominantnim HDL3, dok kod pacije-nata te razlike nije bilo. Ovi rezultati nas navode nazaklju~ak da HDL2 ~estice imaju ja~i antioksidativniefekat, ali je on zna~ajno umanjen zbog izmenjenestrukture HDL ~estica kod dijabetesa. Smanjena aktiv-nost PON1 udruèna sa strukturno izmenjenim HDL~esticama predstavlja zna~ajan faktor za progresivnirazvoj kardiovaskularnih bolesti kod pacijenata sa dija-betes melitusom tipa 2.

D50PROVERA TA^NOSTI DVE NOVE JEDNAÎNE

ZA IZRAÛNAVANJE KONCENTRACIJE LDL HOLESTEROLA PORE\ENJEM

SA VREDNOSTIMA DOBIJENIM DIREKTNIM ODRE\IVANJEM

A. Vujovi}1, J. Kotur-Stevuljevi}1, N. Buji{i}2, S. Spasi}1

1Institut za medicinsku biohemiju, Farmaceutski fakultet, Beograd, Srbija2Biohemijska laboratorija »Belladona«,

Beograd-Zemun, Srbija

Pouzdano i ta~no odre|ivanje koncentracije LDLholesterola (LDL-H) od velikog je zna~aja zbog pravil-nog razvrstavanja pacijenata u rizi~ne kategorije pred-loène od strane NCEP ATP-III. U klini~koj praksi kon-centracija LDL-H se naj~e{}e odre|uje ra~unanjem po-mo}u Friedewaldove formule. Cilj ove studije je da pro-veri ta~nost dve nove formule za izra~unavanje LDL-H uodnosu na vrednosti dobijene direktnim odre|ivanjem.U studiju je uklju~eno 1640 pacijenata (srednja vred-nost godina 54 ± 15, 52,4% èna) koji su imali kon-centraciju triglicerida 4,5 mmol/L. LDL-H je odre|endirektnom metodom (D-LDL-H) i izra~unat pomo}udve nove formule. Prva formula predstavlja modifiko-vanu Friedewaldovu jedna~inu a druga je izvedena pri-menom multiple regresione analize. Iz dobijenih rezul-tata izra~unata je srednja vrednost LDL-H i to: 3,86 ±1,09 mmol/L za direktnu metodu, 3,86 ± 1,1 mmol/Lza prvu formulu (F1-LDL-H) i 3,83±1,06 mmol/L zadrugu formulu (F2-LDL-H). Linearna regresiona anali-za je pokazala dobre i istovetne korelacije izme|u di-rektnog LDL-H i LDL-H izra~unatog iz obe jedna~ine (R

274

=0,97). Razlika izme|u srednjih vrednosti D-LDL-H iF1-LDL-H nije statisti~ki zna~ajna, dok je razlika srednjihvrednosti D-LDL-H i F2-LDL-H od 0,02±0,25 mmol/Lpokazala statisti~ku zna~ajnost (P<0,01). Slaganje re-zultata po NCEP ATP-III kategorijama rizika izme|udirektno odre|enog i izra~unatog holesterola je 81,1%za prvu formulu i 80% za drugu formulu. Ta~nost rezul-tata u odnosu na direktno odre|en LDL-H bila je zado-voljavaju}a: 90% rezultata za prvu i 93% rezultata zadrugu formulu bilo je u opsegu ±10% razlike. Nove je-dna~ine za izra~unavanje LDL-H treba dodatno testiratina ve}oj populaciji pre nego {to se po~ne sa njihovomimplementacijom u biohemijskim laboratorijama.

D51ZNAÂJ ODRE\IVANJA LDH

I DRUGIH BIOHEMIJSKIH MARKERA KAO PREDIKTORA PREVREMENOG

PORO\AJA I POBAÂJA

M. Milivojevi}1, Lj. Nikoli}2, S. Tomanovi}2, M. Dedi}2, N. Markovi}1

1Institut za medicinsku biohemiju KCS, Beograd, Srbija2Institut za ginekologiju i aku{erstvo KCS,

Beograd, Srbija

Incidenca prevremenog poro|aja se kre}e od 3do 5% i naj~e{}e je posledica infekcije. Intraamnionskeinfekcije, i predstavljaju naj~e{}i uzrok morbiditeta imortaliteta. Ova prospektivna studija sprovedena je saciljem da se uporede vrednosti biohemijskih, hemato-lo{kih i mikrobiolo{kih parametara u amnionskoj te~-nosti (glukoze, laktat dehidrogenaze, leukocita i bojenjapo Gramu) sa pozitivnom kulturom amnionske te~nostiu predikciji intraamnionske infekcije i preterminskogporo|aja. Na Institutu za ginekologiju i aku{erstvo uperiodu od novembra 2003. do aprila 2004. ispitano je125 trudnica. Njima je pri rutinskim ranim amniocen-tezama (izme|u 16. i 18. gestacione nedelje) uzimanaplodova voda za mikrobiolo{ku, biohemijsku i hemato-lo{ku obradu. Mikrobiolo{ka obrada je podrazumevalazasejavanje na standardnim podlogama za aerobnebakterije i bojenje po Gramu. Senzitivnost, specifi~nost,pozitivna i negativna prediktivna vrednost za LDH,glukozu, Gram bojenje, izra~unati su u odnosu na poz-itivnu kulturu amnionske te~nosti. Za statisti~ku obradurezultata kori{}eni su neparametarski Mann-Whitney U-test, ROC-kriva i t-test. Prevalenca pozitivnih kulturaamnionskih te~nosti bila je 13% (15 od 125). Srednjavrednost aktivnosti enzima LDH (414 U/L) u grupi sapozitivnom kulturom bila je zna~ajno ve}a od one ugrupi ~ija je kultura bila negativna (LDH 143 U/L;p<0,01). Granice odlu~ivanja (»cut-off« vrednosti) zasve parametre su izra~unate u odnosu na predikciju po-zitivne kulture amnionske te~nosti i iznosile su za LDH

205 U/L, za broj leukocita 10 cells/mm3

lated by both equations (R=0.97). The mean differ-ence between D-LDL-C and F1-LDL-C showed no sta-tistic significance, while the mean difference betweenD-LDL-C and F2-LDL-C of 0.02±0.25 mmol/L wassignificant (P<0.01). Concordant results for NCEP ATP-III risk categories were present in 81.1% of results cal-culated by the first formula and directly measured LDL-C, and in 80% of the results calculated by the secondformula and directly measured LDL-C. The results cal-culated by both formulas revealed a satisfactory level ofaccuracy, 90% of results for the first formula and 93%of results for the second formula were within the differ-ence range of ±10%. New equations for the determi-nation of LDL-C levels need to be tested on a biggerpopulation before we can start with their implementa-tion into biochemical laboratories.

D51HIGH LEVEL OF LDH AND OTHER BIOCHEMICAL MARKERS IN THE

PREDICTION OF PRETERM LABOR AND ABORTION

M. Milivojevi}1, Lj. Nikoli}2, S. Tomanovi}2, M. Dedi}2, N. Markovi}1

1Institute of Medical Biochemistry2Institute of Gynecology and Obstetrics,


The incidence of preterm labor is 3–5 % and it ismost frequently caused by infection. Bacterial infec-tions of the amniotic cavity (chorioamnionitis) is a sig-nificant cause of perinatal morbidity and mortality. Ourobjective was to analyze the value of an amniotic fluidtest for the detection of microbial invasion of the uter-ine cavity. Amniotic fluid was obtained by amniocente-sis from one hundred and fifteen consecutive patients,from 16 to 18 gestational weeks. Fluid was analyzedfor leukocyte count, glucose level, lactate dehydroge-nase level (LDH), and Gram stain. Cultures for aerobesand anaerobes were performed. Sensitivity, specificity,and positive and negative predictive value were calcu-lated for LDH, leukocyte count, glucose, and Gramstain in the prediction of positive amniotic fluid culture.Receiver-operator characteristic curve analysis, t-test,and nonparametric test (Mann Whitney U-test) wereused. The prevalence of positive amniotic fluid cultureswas 13% (14 of 115). The median LDH level (414U/L) was significantly greater for women with a positiveamniotic fluid culture than for those with a negativeculture (median LDH 143 U/L; p<0.01). Critical val-ues of LDH 205 U/L, leukocyte count 10cells/mm3 (10×106/L) and glucose 1.15 mmol/Lwere selected for optimal performance in the predic-tion of a positive amniotic fluid culture. LDH level hadthe best sensitivity (71%) in contrast to leukocyte count(51%), glucose (65%) and positive Gram stain (36%).Amniotic fluid LDH level has diagnostic value in the

(10×106/L), a za glukozu 1,15 mmol/L. Najsenzitiv-niji test za detekciju mikrobiolo{ke infekcije u amnion-skoj te~nosti je aktivnost LDH (71%), za razliku od ostal-ih dijagnosti~kih markera: glukoze (65%), broja leukoci-ta (51%) i pozitivnog Gram bojenja (36%). Nivo LDH uamnionskoj te~nosti ima dijagnosti~ku vrednost upredikciji pozitivnog nalaza kulture amnionske te~nosti.Odre|ivanje nivoa LDH u amnionskoj te~nosti, kao ra-nog prediktora infekcije, jednostavna je, brza i pouz-dana metoda, lako dostupna svim bolni~kim laboratori-jama.

D52ISPITIVANJE UTICAJA POVI[ENE

FIZI^KE AKTIVNOSTI NA NEKE LIPIDSKEPARAMETRE ZDRAVIH MLADIH OSOBA


Klini~ki centar Vojvodine, Centar za laboratorijsku medicinu,

Novi Sad, Srbija

Brojne studije pokazale su povoljan efekat fizi~keaktivnosti na parametre lipidskog statusa. Ipak, nedo-voljno je ispitano kakav efekat ima intenzivirani fizi~kitrening. Cilj ovog istraìvanja bio je da se ispita uticajpovi{ene fizi~ke aktivnosti na serumske nivoe ukupnogholesterola (TC), triglicerida (TG), HDL holesterola(HDL-C), LDL holesterola (LDL-C), apolipoproteina(apo) A1 i B i lipoproteina (a) (Lp(a)). Ispitivanjem jeobuhva}eno 28 aktivnih ~lanova Sportske sekcije Me-dicinskog fakulteta Univerziteta u Novom Sadu (19mu{karaca i 9 èna), i 25 zdravih studenata sli~ne ì-votne dobi (12 mu{karaca i 13 èna) koji se ne baveaktivno sportom. TC, TG, HDL-C i LDL-C odre|ivani sustandardnim biohemijskim metodama, a apo A1 i B(»Olympus«, Lismehan, Irska) i Lp(a) (»Sentinel«, Mi-lano, Italija) imunoturbidimetrijski. Uzorci ispitanikauzeti su pre i posle osam meseci aktivnog sportskogtreninga (septembar–maj). Dobijeni su slede}i rezultati:

JMB 2008; 27 (2) 275

prediction of a positive amniotic fluid culture. Consi-dering that amniotic fluid infection in a majority ofpregnant women may cause preterm labor or abortionthe determination of amniotic fluid LDH might be ofgreat importance. Lactate dehydrogenase is a readilyavailable, inexpensive, rapid amniotic fluid marker thatcan be measured in any hospital laboratory.

D52EXAMINATION OF THE INFLUENCEOF ENHANCED PHYSICAL ACTIVITY

ON SOME LIPID PARAMETERS IN HEALTHY YOUNG ADULTS


Clinical Centre of Vojvodina, Dept. of Laboratory Medicine, Novi Sad, Serbia

Numerous studies have demonstrated the posi-tive effect of physical activity on some lipid parameters.However, what kind of effect intensified physical prac-tice has is insufficiently examined. The aim of this studywas to examine the influence of enhanced physicalactivity on the serum levels of total cholesterol (TC),triglycerides (TG), HDL cholesterol (HDL-C), LDL cho-lesterol (LDL), apolipoproteins (apo) A1 and B andlipoprotein (a) (Lp(a)). In this study 28 active membersof the Sports Section of the Medical Faculty, Universityof Novi Sad (19 males and 9 females), and 25 healthystudents matched for age (12 males and 13 females)who did not practise sport actively were included. TC,TG, HDL-C and LDL-C were determined by standardbiochemical methods, and apo A1 and B (»Olympus«,Lismeehan, Ireland) and Lp(a) (»Sentinel«, Milano, Ita-ly) by immunoturbidimetry. Samples were taken beforeand after eight months of active sport practise (Septem-ber to May). The following results were obtained:

Ispitivaniparametri

Sportisti (n = 28) Zdrave osobe (n = 25)

bazalnoXsr. SD

posle 8 m.Xsr. SD

% promena

bazalnoXsr. SD

posle 8 m.Xsr. SD

% promena

mmol/L

TC 4,05 0,56 4,13 0,63 +2,03 4,19 0,50 4,43 0,61 +5,63

TG 0,93 0,36 1,09 0,41 +18,28 0,82 0,43 1,07 0,52 +29,71

HDL-C 1,51 0,38 1,42 0,30 –5,88 1,56 0,43 1,47 0,39 –5,90

LDL-C 2,12 0,69 2,15 0,70 +1,38 2,22 0,62 2,42 0,67 +8,83

g/L

Apo A1 1,45 0,19 1,45 0,16 +0,15 1,48 0,19 1,48 0,16 –0,35

Apo B 0,65 0,17 0,75 0,22 +15,41 0,71 0,14 0,79 0,19 +11,99

Lp(a) 0,08 0,11 0,10 0,13 +35,24 0,09 0,11 0,11 0,16 +22,27

Examinedparameters

Sportsmen (n = 28) Healthy subjects (n = 25)

basalXsr. SD

after 8 m.Xsr. SD

%

difference

basalXsr. SD

after 8 m.Xsr. SD

%

difference

mmol/L

TC 4.05 0.56 4.13 0.63 +2.03 4.19 0.50 4.43 0.61 +5.63

TG 0.93 0.36 1.09 0.41 +18.28 0.82 0.43 1.07 0.52 +29.71

HDL-C 1.51 0.38 1.42 0.30 –5.88 1.56 0.43 1.47 0.39 –5.90

LDL-C 2.12 0.69 2.15 0.70 +1.38 2.22 0.62 2.42 0.67 +8.83

g/L

Apo A1 1.45 0.19 1.45 0.16 +0.15 1.48 0.19 1.48 0.16 –0.35

Apo B 0.65 0.17 0.75 0.22 +15.41 0.71 0.14 0.79 0.19 +11.99

Lp(a) 0.08 0.11 0.10 0.13 +35.24 0.09 0.11 0.11 0.16 +22.27

In the examined group, there were no significantdifferences in lipid parameter levels after the period ofeight months of active sport practise. Surprisingly, HDL-C levels in both subgroups as well as apo A1 in the sub-jects who do not practise sport actively were reduced,while the other examined parameter values increased.

D53THE INFLUENCE OF CHRONIC

HELICOBACTER PYLORI INFECTIONON SERUM LIPID PROFILE PARAMETERS,

APOLIPOPROTEINS A1 AND B AND Lp(a) LIPOPROTEIN

S. Koji}-Damjanov, N. Eremi}, M. \eri}

Clinical Centre of Vojvodina, Centre for Laboratory Medicine, Novi Sad

Recent studies unequivocally show that chronicHelicobacter pylori (HP) infection, as a low-grade per-sistent inflammatory stimulus, has an influence on ath-erosclerosis development. Simultaneously, data on pro-atherogenic alterations in the serum lipid profile due tothis chronic infection are contradictory. The aim of thisstudy was to look for differences in the lipid status,apolipoproteins (apo) A1 and B and Lp(a) lipoprotein ofthe subjects with and without chronic HP infection. Fiftyfive subjects (20 male and 30 female) with chronic HPinfection (IgG>50 U/mL and IgA <20 U/mL) and 55(20 male and 30 female) HP negative subjects (IgG andIgA <20 U/mL) were examined. IgG and IgA antibod-ies were determined from serum samples by the ELISAmethod (Virion/Serion, Würzburg, Germany). Lipid pro-file parameters were determined by standard biochemi-cal methods, apo A1 and B (Olympus, Lismeehan,Ireland) and Lp(a) (Sentinel, Milano, Italy) by immuno-turbidimetry. Levels of total cholesterol (TC), triglycerides(TG), LDL cholesterol (LDL), non-HDL cholesterol (non-HDL) as well as apo B and Lp(a) were higher in HP pos-itive in comparison to HP negative subjects. Differenceswere significant for TC, LDL and non-HDL in all subjects(6.01 1.04 vs. 5.18 0.97 mmol/L, p<0.001;3.69 1.14 vs. 3.23 0.78 mmol/L, p<0.02 i4.62 0.98 vs. 3.87 0.97 mmol/L, p<0.001) and infemale group (5.98 1.07 vs. 5.10 0.82 mmol/L,p<0,001; 3.79 1.11 vs. 3.14 0.66 mmol/L,p<0.01 i 4.58 1.06 vs. 3.75 0.86 mmol/l,p<0.001) and for TG in whole subject group (1.861.16 vs. 1.42 0.78 mmol/L, p<0.05), in male group(2.28 1.29 vs. 1.57 0.75 mmol/L, p<0.05) and forapo B in all subjects (1.10 0.26 vs. 0.90 0.23mmol/L, p<0.001), in males (1.13 0.22 vs. 0.930.23 mmol/L, p<0.01) and the female (1.08 0.28 vs.0.89 0.23 mmol/L, p<0.005) group. The increase inLp(a) levels were not significant, while there were nodiferences in HDL-cholesterol and apo A1 levelsbetween HP positive and negative subjects. Our resultsconfirm the hypothesis that chronic HP infection couldmodify lipid profile in a way that would promote the for-mation of a proatherogenic lipid profile.

U na{oj grupi ispitanika nisu ustanovljene zna-~ajne promene u nivoima ispitivanih lipidskih parame-tara posle perioda od 8 meseci aktivnog treninga. Neo-~ekivano, nivoi HDL-C u obe podgrupe i apo A1 kodosoba koje se ne bave aktivno sportom su se snizili, doksu se svi ostali ispitivani parametri bili povi{eni.

D53UTICAJ HRONI^NE HELICOBACTERPYLORI INFEKCIJE NA PARAMETRE

LIPIDSKOG STATUSA, APOLIPOPROTEINE A1 I B I Lp(a) LIPOPROTEIN

S. Koji}-Damjanov, N. Eremi}, M. \eri}

Klini~ki centar Vojvodine, Centar za laboratorijsku medicinu, Novi Sad

Dosada{nja saznanja pokazuju da hroni~na infek-cija s Helicobacter pylori (HP), kao niskostepeni perzis-tentni inflamatorni stimulus, ima odre|eni uticaj narazvoj ateroskleroze. Istovremeno, podaci o nastankuproaterogenih promena u lipidskom statusu u sklopu tehroni~ne infekcije su kontradiktorni. Cilj istraìvanja bioje da se ispitaju razlike u lipidskom statusu, apolipopro-teinima (apo) A1 i B i Lp(a) lipoproteinu (Lp(a)) kodosoba s hroni~nom HP infekcijom i bez nje. Ispitiva-njem je obuhva}eno 55 osoba (20 mu{karaca i 30èna) s hroni~nom HP infekcijom (IgG >50 U/mL iIgA <20 U/mL) i 55 (20 mu{karaca i 30 èna) HPnegativnih osoba (IgG i IgA <20 U/mL). IgG i IgAantitela protiv HP odre|ivana su ELISA metodom(»Virion/Serion«, Vircburg, Nema~ka). Parametri lipid-skog statusa odre|ivani su standardnim biohemijskimmetodama, a apo A1 i B (»Olympus«, Lismehan, Irska)i Lp(a) (Sentinel, Milano, Italija) imunoturbidimetrijski.Vrednosti ukupnog holesterola (TC), triglicerida (TG),LDL-holesterola (LDL), non-HDL holesterola (non-HDL), kao i apo B i Lp(a) bile su povi{ene kod HP pozi-tivnih u odnosu na HP negativne osobe. Promene subile zna~ajne za TC, LDL i non-HDL u ukupnoj grupi ispi-tanika (6,01 1,04 vs. 5,18 0,97 mmol/L, p<0,001;3,69 1,14 vs. 3,23 0,78 mmol/L, p<0,02 i 4,62

0,98 vs. 3,87 0,97 mmol/L, p<0,001) i kod èna(5,98 1,07 vs. 5,10 0,82 mmol/L, p<0,001; 3,79

1,11 vs. 3,14 0,66 mmol/L, p<0,01 i 4,58 1,06vs. 3,75 0,86 mmol/L, p<0,001), za TG u ukupnojgrupi (1,86 1,16 vs. 1,42 0,78 mmol/L, p<0,05) ikod mu{karaca (2,28 1,29 vs. 1,57 0,75 mmol/L,p<0,05), a za apo B u ukupnoj grupi (1,10 0,26 vs.0,90 0,23 mmol/L, p<0,001), kod mu{karaca (1,13

0,22 vs. 0,93 0,23 mmol/L, p<0,01) i kod èna(1,08 0,28 vs. 0,89 0,23 mmol/L, p<0,005). Povi-{enje Lp(a) nije bilo zna~ajno, dok se nivoi HDL holes-terola i apo A1 nisu zna~ajno razlikovali izme|u HPpozitivnih i HP negativnih osoba. Na{i rezultati po-dràvaju hipotezu da bi hroni~na infekcija s HP moglamodifikovati metabolizam lipida u pravcu formiranjaproaterogenog lipidskog statusa.

276

D54EFEKTI SINTETSKE @U^NE SOLI KAO

MODULATORA LIPIDSKOG METABOLIZMAZDRAVIH I DIJABETI^NIH PACOVA

S. Koji}-Damjanov1, M. \eri}1, M. Mikov2

1Klini~ki centar Vojvodine, Centar za laboratorijsku medicinu

2Medicinski fakultet, Zavod za farmakologiju, toksikologiju i klini~ku farmakologiju, Novi Sad

Otkri}e da su ̀ u~ne kiseline va`ni signalni molekuliuklju~eni i u regulaciju metabolizma lipida dalo je osnovza ispitivanje njihovih sintetskih derivata kao potencijal-no novih hipolipidemijskih agenasa. U ovom istraìvanjuispitivan je uticaj sintetske Na-monoketoholne kiseline(Na-MKHA), aplikovane peroralno (p.o.), subkutano(s.c.) i intravenski (i.v.), 2 mg/kg TT, jednokratno i tokom7 dana, na lipidski status zdravih (n=30) i dijabeti~nih(n=30; dvokratna intraperitonealna aplikacija 100mg/kg TT aloksana) »Wistar« pacova. Odgovaraju}ekontrolne grupe (KG, n=5) zdravih i dijabeti~nih paco-va primale su 10 mL/kg TT fiziolo{kog rastvora p.o, jed-nokratno ili tokom 7 dana. Parametri lipidskog statusaodre|ivani su standardnim biohemijskim metodama.Jednokratna primena Na-MKHA dovela je do neznatnihpromena ukupnog i HDL holesterola, dok je sedmod-nevna primena izazvala zna~ajno sniènje ukupnog (KG1,93 0,45 mmol/L; p.o. 1,21 0,14 mmol/L, p<0,01;s.c. 1,38 0,05 mmol/L, p<0,05; i.v. 1,13 0,08mmol/L, p<0,01) i LDL holesterola (KG 0,49 0,31mmol/L; p.o. 0,03 0,04 mmol/L; s.c. 0,04 0,04mmol/L; i.v. 0,03 0,07 mmol/L; p<0,02 svi) kod dija-beti~nih, i sniènje HDL holesterola i kod zdravih (KG0,91 0,16 mmol/L; i.v. 0,63 0,12 mmol/L, p<0,02)i kod dijabeti~nih (KG 1,08 0,13 mmol/L; s.c.0,87 0,06 mmol/L, p<0,02; i.v. 0,77 0,06 mmol/L,p<0,01) ̀ ivotinja. Trigliceridi su se snizili posle jednokra-tne primene Na-MKHA, zna~ajno samo kod dijabeti~nihpacova (KG 1,38 0,50 mmol/L; s.c. 0,66 0,28mmol/L, p<0,05; i.v. 0,58 0,44 mmol/L, p<0,05),dok je sedmodnevna primena dovela do porastatriglicerida kod zdravih (KG 0,53 0,09 mmol/L; s.c.1,63 0,39 mmol/L, p<0,001; i.v. 0,87 0,11 mmol/L,p<0,001) i nezna~ajnog sniènja kod dijabeti~nih paco-va. Dobijeni rezultati upu}uju na potrebu daljih ispiti-vanja modulacijskog efekta sintetskih ù~nih soli nametabolizam lipida, pre svega putem nuklearnih farne-soid X receptora.

JMB 2008; 27 (2) 277

D54THE EFFECTS OF SYNTHETIC BILE SALT ASA MODULATOR OF THE LIPID METABOLISM

OF HEALTHY AND DIABETIC RATS

S. Koji}-Damjanov1, M. \eri}1, M. Mikov2

1Clinical Centre of Vojvodina, Centre for Laboratory Medicine,

2Medical Faculty, Institute for Pharmacology,Toxicology and Clinical Pharmacology, Novi Sad

The finding that bile acids as important signalmolecules are also involved in lipid metabolism regula-tion give rise to an examination of their synthetic de-rivates as potentially new hypolipidemic agents. In thisstudy, the influence of synthetic sodium-monoketocho-late (Na-MKHA), perorally (p.o.), subcutaneously (s.c.)and intravenously (i.v.) applied, 2 mg/kg b.w., in one-dose and during 7 days, on the lipid status of healthy(n=30) and diabetic (n=30; two-time intraperitonealapplication of 100 mg/kg b.w. of Aloxane) »Wistar« ratswas examined. Corresponding control groups (CG,n=5) of healthy and diabetic rats received 10 mL/kgb.w. of physiological saline p.o., in one-dose or during7 days. Lipid status parameters were determined usingstandard biochemical methods. One-dose applicationof Na-MKHA provoked insignificant changes in totaland HDL cholesterol, while a seven-day applicationprovoked a significant decrease in total (CG 1.930.45 mmol/L; p.o. 1.21 0.14 mmol/L, p<0.01; s.c.1.38 0.05 mmol/L, p<0.05; i.v. 1.13 0.08 mmol/L,p<0.01) and LDL cholesterol (CG 0.49 0.31 mmol/L;p.o. 0.03 0.04 mmol/L; s.c. 0.04 0.04 mmol/L; i.v.0.03 0.07 mmol/L; p<0.02 respectively) in diabetic,and a decrease in HDL cholesterol in healthy (CG0.91 0.16 mmol/L; i.v. 0.63 0.12 mmol/L,p<0.02) and diabetic (CG 1.08 0.13 mmol/L; s.c.0.87 0.06 mmol/L, p<0.02; i.v. 0.77 0.06 mmol/L,p<0.01) animals. Triglycerides significantly decreasedafter the one-dose application of Na-MKHA only in dia-betic rats (CG 1.38 0.50 mmol/L; s.c. 0.66 0.28mmol/L, p<0.05; i.v. 0.58 0.44 mmol/L, p<0.05),while the seven-day application provoked an increase intriglycerides in the healthy (CG 0.53 0.09 mmol/L;s.c. 1.63 0.39 mmol/L, p<0.001; i.v. 0.87 0.11mmol/L, p<0.001) and an insignificant decrease inthe diabetic rats. The obtained results point to the needfor further examination of the modulation effect of syn-thetic bile salts on lipid metabolism, especially throughnuclear farnesoid X receptors.

D55LIPOPROTEIN (a) IN CHRONIC

RENAL FAILURE

V. âbarkapa, M. \eri}, Z. Sto{i},V. Saka~, S. Koji}-Damjanov, N. Eremi}

Department of Laboratory Medicine, Clinic of Immunology and Nephrology,

Clinical center of Vojvodina, Novi Sad, Serbia

In patients with chronic renal failure (CRF) cardio-vascular events are a major cause of mortality. Lp (a)lipoprotein as an independent risk factor for the devel-opment of premature atherosclerosis could have animportant contributing role. The aim of this study was todetermine the relationship between the functional kid-ney reserve and serum Lp(a) level in different CRF sta-diums. This study included 74 patients, 42 (27 males,15 females) of them with CRF and 32 (9 males, 23females) on a chronic hemodialysis program, as well as53 healthy individuals (31 males, 22 females) matchedfor age as a control group. To evaluate the functionalkidney reserve serum urea, creatinine and endogenouscreatinine clearance (except in patients on a chronichemodialysis program) were determined by standardbiochemical methods. Blood was taken directly beforedialysis treatment in patients on chronic hemodialysisprogram on the first day of the week. Moreover, in allsubjects Lp(a) serum levels were determined by immu-noturbidimetry, analyzer Olympus AU 400 (Sentinel,Milano, Italy). Elevated Lp(a) levels (above 0.25 g/L)were found in 31% of hemodialysed patients, 14% ofnon-hemodialysed patients with CRF as well as in 11%of healthy subjects. Extremely elevated Lp(a) levels

0.50 g/L were determined in 16% of hemodialysedpatients, 9% of non-hemodialysed patients with CRFand 0% of healthy subjects. Significant differences werefound between mean Lp(a) levels in hemodialysedpatients and control group (0.22 0.25 vs. 0.08 0.1g/L, p<0.01) as well as in the non-hemodialysed pa-tients with CRF and the control group (0.18 0.21 vs.0.08 0.1 g/L, p<0.05). There were no statistical diffe-rences between mean Lp(a) levels in hemodialysedpatients and non-hemodialysed patients with CRF andno statistical correlation between creatinine clearanceand Lp(a) levels. From our results it can be concludedthat patients with CRF, especially ones on a chronichemodialysis program, have significantly higher Lp(a)levels in comparison with the healthy control group.

D55LIPOPROTEIN (a) U HRONI^NOJBUBRE@NOJ INSUFICIJENCIJI

V. âbarkapa, M. \eri}, Z. Sto{i}, V. Saka~, S. Koji}-Damjanov, N. Eremi}

Centar za laboratorijsku medicinu, Klinika za imunologiju i nefrologiju,

Klini~ki centar Vojvodine, Novi Sad, Srbija

Kardiovaskularne komplikacije predstavljaju vode-}i uzrok mortaliteta kod pacijenata sa hroni~nombubre`nom insuficijencijom (HBI). Lp(a) lipoprotein kaonezavisni faktor rizika za razvoj prevremene ateroskle-roskleroze mogao bi imati zna~ajnu doprinosnu ulogu.Ova studija izvedena je s ciljem ispitivanja odnosa izme-|u funkcione rezerve bubrega i serumskih koncentraci-ja Lp (a) kod bolesnika u razli~itim stadijumima HBI. Ustudiju je uklju~eno 74 ispitanika, 42 (27 mu{karaca, 15èna) sa HBI i 32 (9 mu{karaca, 23 ̀ ena) na hroni~nomprogramu hemodijalize, kao i 53 (31 mu{karaca, 22èna) zdrave osobe, odgovaraju}e ìvotne dobi (kontrol-na grupa). Standardizovanim biohemijskim metodama,a u cilju procene funkcionalne rezerve bubrega, kodbolesnika su odre|ivani klirens endogenog kreatinina(osim kod bolesnika na hemodijalizi), serumski kreatinini urea. Bolesnicima na hemodijalizi krv je va|ena ne-posredno pre samog tretmana dijalize, prvog dana unedelji. Osim toga, kod svih ispitanika, odre|ivan jeserumski nivo Lp(a) imunoturbidimetrijskom metodomna analizatoru »Olympus AU 400« (Sentinel, Milano,Italija). Povi{ene vrednosti Lp(a) (iznad 0,25 g/l) usta-novljene su kod 31% hemodijaliziranih pacijenata, 14%bolesnika sa HBI koji nisu na hemodijalizi, kao i kod 11%zdravih osoba. Izuzetno povi{ene Lp(a) koncentracije,

0,50 g/L, postojale su kod 16% bolesnika na hemod-ijalizi, 9% HBI pacijenata, ali ne i kod zdravih osoba.Ustanovljena je statisti~ki zna~ajna razlika izme|u sred-njih vrednosti Lp (a) ispitanika na hemodijalizi i kon-trolne grupe (0,22 0,25 vs. 0,08 0,1 g/L, p<0,01),kao i nedijaliziranih HBI pacijenata i kontrolne grupe(0,18 0,21 vs. 0,08 0,1 g/L, p<0,05). Izme|u sred-njih vrednosti Lp(a) kod bolesnika na hemodijalizi i onihsa HBI koji nisu na dijalizi ne postoji statisti~ki zna~ajnarazlika. Tako|e, nije postignuta statisti~ki zna~ajnakorelacija izme|u klirensa kreatinina i nivoa Lp (a). Naosnovu dobijenih rezultata moè se zaklju~iti da bolesni-ci sa hroni~nom bubre`nom insuficijencijom, naro~itona hemodijalizi, imaju zna~ajno vi{e vrednosti lipopro-teina (a) u pore|enju sa zdravom kontrolnom grupom.

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D56LIPIDNI STATUS DAVALACA NA PROGRAMU PLAZMAFEREZE

M. Mitrovi}, M. Sin|i}

Institut za transfuziju krvi Srbije, Beograd, Srbija

Plazmafereza je skup postupaka koji omogu}a-vaju odvajanje plazme tj. te~nog dela krvi od ostalihelemenata krvi. Davaocu se reinfuzijom vra}aju svi }eli-jski elementi kao autologna krv. Cilj plazmafereze je dase dobije plazma koja se koristi za proizvodnju stabilnihlekova iz krvi (albumina i imunoglobulina za intraven-sku i intramuskularnu primenu). Radi za{tite zdravljadavalaca plazme primenjuje se redovna kontrola u kojuspada i pra}enje biohemijskih parametara. Prati sefunkcionalno stanje jetre odre|ivanjem sadràja ukup-nih proteina, albumina, ALT, AST,GGT, kao i lipidnogstatusa. Ovim ispitivanjem obuhva}eno je 100 davala-ca plazme kojima je odre|ivan lipidni status. Ispitivanesu vrednosti ukupnog holesterola (UH), triglicerida(TG), HDL i LDL holesterola, indeks ateroskleroze (IA)i utvr|eni su faktori rizika (FR). Odre|ivanja UH i TGsu vr{ena standardnim enzimskim metodama, HDL-holodre|en je u supernatantu nakon precipitacije plazmemagnezijum-hloridom i fosfovolframovom kiselinom.LDL-hol je izra~unat preko Friedwaldove formule.Vrednosti IA i FR su dobijene ra~unskim putem. Dobi-jene su slede}e vrednosti: ukupni holesterol x =5,47 1,089 mmol/L, trigliceridi x = 1,587 0,957mmol/L, HDL-holesterol x=1,276 0,304 mmol/L iLDL – holesterol 3,45 1,144 mmol/L. Dobijene vred-nosti ukazuju na prisustvo aterogenog rizika lipidnogporekla kod davalaca na programu plazmafereze, iakooni pripadaju najzdravijem delu populacije. Neopho-dno je primeniti higijensko-dijetetski reìm ishrane iodgovaraju}u fizi~ku aktivnost. Redovnim davanjemplazme obezbe|ena im je stalna lekarska kontrola ibriga o zdravlju.

D57LIPIDNI PROFIL GOJAZNE DECE I ADOLESCENATA PRE I POSLE

ZNAÂJNOG GUBITKA TELESNE MASE

R. Golubovi}1, M. Golubovi}2, M. Nikoli}3

1Institut za javno zdravlje Ni{ 2Medicinski fakultet u Kragujevcu 3Medicinski fakultet u Ni{u, Srbija

Kod gojazne dece mogu se javiti povi{eni nivoiukupnog holesterola, triglicerida i lipoproteina malegustine (LDL) u serumu. Cilj ovog rada bio je pra}e-nje lipidnog profila kod gojazne dece pre i posle zna-~ajnog gubitka telesne mase. Ispitano je 60 gojaznedece (37 devoj~ica i 23 de~aka), prose~nog uzrasta

JMB 2008; 27 (2) 279

D56PLASMAPHERESIS DONORS’

LIPID STATUS

M. Mitrovi}, M. Sin|i}

Blood Transfusion Institute of Serbia, Belgrade, Serbia

Plasmapheresis is a set of procedures that ena-bles separation of plasma, i.e. the liquid part of bloodfrom other blood elements. Donors are then reinfusedwith all blood cell elements as the autologous blood.Goal of plasmapheresis is to obtain plasma used for thepreparation of stable blood products (albumins andimmunoglobulins for intravenous and intramuscularuse). In order to protect plasma donors’ health, regularmedical check-ups are carried, including biochemicalparameters follow-up. Functional liver condition ismonitored by the determination of the total proteincontent, albumin, ALT, AST, GGT, as well as the lipidstatus. This investigation included 100 plasma donorsin which the lipid status had been determined. Totalcholesterol (TCH) values, triglycerides (TG), HDL andLDL cholesterol, index of atherosclerosis (IA) wereinvestigated and risk factors were determined. Deter-mination of TCH and TG was performed using stan-dard enzyme methods, HDL cholesterol was determi-ned in the supernatant following plasma precipitationby MgCl

2and phosphotungstic acid. LDL cholesterol

was calculated using Friedwald formula. IA and FR val-ues had been calculated. The following values wereobtained: total cholesterol x = 5.47 1.089 mmol/L,triglyceride x =1.587 0.957 mmol/L, HDL choles-terol x = 1.276 0.304 mmol/L and LDL – choles-terol 3.45 1.144 mmol/L. The obtained values pointto the presence of the lipid originating an atheroge-nous risk in the plasmapheresis donors, despite the factthat they belong to the healthiest part of the popula-tion. A dietary nutrition regime and adequate physicalactivities are necessary. Regular plasma donation pro-vides them with continuous medical check-up andhealth care.

D57LIPID PROFILE CONCERNING OBESE

CHILDREN AND ADOLESCENTS BEFOREAND AFTER SIGNIFICANT WEIGHT LOSS

R. Golubovi}1, M. Golubovi}2, M. Nikoli}3

1Institut for Public Health Ni{2Medical Faculty Kragujevac3Medical Faculty Ni{, Serbia

The obesity in children can be associated withthe higher level of total cholesterol, triglycerides andlow-density lipoproteins (LDL) in serum. The arm ofthe study was to monitored the lipid profile of obesechildren before and after significant weight loss. Sixtyobese children (37 girls and 23 boys), average age

11,37±2,48 year, who took part in the weight reduc-tion program in The Counseling Service for well-ba-lanced diet of the Institute for Public Health in Nis,were examined. The measurements were taken atthe beginning of the research and 10 weeks later.Anthropometric changes and the changes of thelevel of lipids in serum (total cholesterol, triglyceridesand LDL-cholesterol) were recorded for all partici-pants in the study. Carrying out of the reduced dietduring the period of 10 weeks showed that bodyweight in the examined children had been greatlyreduced (p<0,01). Statistical significant reduction ofall the examined parametres of the lipid profile wasdetected (p<0,01). Results show that the combina-tion of the diet-therapy, increased physician activityand behavior modification is efficient in reducing ofthe total cholesterol, triglycerides and LDL-choles-terol as far as obese children are concerned.

11,37± 2,48 god. koja su bila uklju~ena u programza redukciju telesne mase u Savetovali{tu za una-pre|enje ishrane Instituta za javno zdravlje u Ni{u.Merenja su ra|ena na po~etku ispitivanja i nakon 10nedelja. Za sve ispitanike beleène su antropometri-jske promene, kao i promene nivoa lipida u serumu(ukupnog holesterola, triglicerida i LDL holesterola).Tokom 10 nedelja sprovo|enja redukcione dijete,telesna masa kod ispitane dece je zna~ajno smanje-na (p<0,01). Na|ena je statisti~ki zna~ajna redukci-ja svih ispitivanih parametara lipidnog statusa(p<0,01). Dobijeni rezultati pokazuju da je kombi-nacija dijeto-terapije, pove}ane fizi~ke aktivnosti ipromene ìvotnih navika efikasna u sniàvanjuukupnog holesterola, triglicerida i LDL holesterolakod gojazne dece.

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SLOBODNE TEME

FREE COMMUNICATIONS

E58PRENATALNI BIOHEMIJSKI SKRINING

U II TRIMESTRU TRUDNICA STARIJIH OD 30 GODINA

N. Kosti}, V. Markovi}, S. Ra{ovi}

Biohemijska laboratorija, Zdravstveni centar Kru{evac

Trizomija 18 i trizomija 21 (Dounov sindrom –DS) naj~e{}e su hromozomopatije. Danas se u labora-torijama izvodi test za rano otkrivanje tih anomalija, tzv.tripel test, koji obuhvata odre|ivanje tri markera: alfa-fetoproteina (AFP), nekonjugovanog estriola i horionigonadotropnog hormona (b HCG). Pomo}u softverase izra~unava individualni koeficijent MOM premakarakteristikama trudno}e i ultrazvu~nim parametrimafetusa. Cilj je bio da se ispita da li postoji uve}an rizikda plod ima hromozomsku anomaliju (trizomija 21, iliDS) kod trudnica starijih od 30 g. Odre|ivani subiohemijski markeri: AFP, bHCG i slobodni estriol uvenskoj krvi trudnica od 16 do 20 nedelje gestacije,principom iluminescencije na imunoanalizatoru»Immulit1000«. Ispitivana grupa je brojala 89 trudni-ca starijih od 30 godina {to se jo{ uvek ne smatra dob-nim rizikom. Dobijene su vrednosti: AFP 13,6–93,7IU/mL (MoM 0,48–2,57); estriol 0,412–9,79 ng/mL,(MoM 0,23–3,39); bHCG 8270–128700 mlU/mL(MoM 0,44–4,46). Pove}an rizik za DS dokazan je u 9slu~ajeva (1<50–1<257), 10%. Podgrupu sa visokimdobnim rizikom prose~ne starosti 37 godina (35–44,9), ~inilo je 12 trudnica, tri su imale pove}an rizikod DS-a (1:50–1:96), 25%. Visokodobni rizik (1:33) inajve}i rizik za DS (1:50) i trizomiju 18 (1<102) po-kazao je tripel test kod jedne trudnice, {to je potvr|enoi amniocentezom, dok je u 8 slu~ajeva nalaz bio ure-dan. Rano testiranje trudnica biohemijskim markeri-ma, ultrazvu~ni pregled i amniocenteza, kada je toindikovano, umnogome smanjuju procenat ra|anjadece sa hromozomskim defektom.

E58SECOND-TRIMESTER PRENATAL

BIOCHEMICAL SCREENING IN PREGNANTWOMEN ABOVE THE AGE OF 30

N. Kosti}, V. Markovi}, S. Ra{ovi}

Biochemical Laboratory, Health Centre Kru{evac

Trisomy 18 and trisomy 21 are the most com-mon chromosome anomalies. Nowadays a test for theearly detection of these anomalies, called the tripletest, is performed, that involves three markers: alpha-fetoprotein (AFP), unconjugated estriol and chorionicgonadotropin hormone (b HCG). The software calcu-lates the individual coefficient MOM, related to thecharacteristics of pregnancy and ultrasound parame-ters of the fetus. The aim was to find out if there wasan increased risk of the fetus developing a chromo-some anomaly (trisomy 21, DS or neural tube defect)in pregnant women above the age of 30. Biochemicalmarkers were determined: AFP, bHCG and free estriolin the venal blood of pregnant women in the periodfrom 16–20th week of gestation, using a method ofluminescence, »Immulit 1000«. The studied groupincluded 89 pregnant women above the age of 30,which is still considered an age risk. None of them haddiabetes mellitus, 28 were active smokers, and all ofthem had conceived naturally. The following valueswere acquired: AFP 13.6–93.7 IU/mL (MoM 0.48–2.57); estriol 0.412–9.79 ng/mL (MoM 0.23–3.39)and bHCG 8270–128700 mlU/mL (MoM 0.44–4.46). An increased risk of trisomy 21 was proved in 9cases (1<50–1<257), 10%. The subgroup of highage risk involved 12 pregnant women whose averageage was 37 (35– 44.9). The risk of DS in this groupwas heightened in 3 cases (1:50 – 1:96), 25%. Thetriple test showed high age risk (1:33) and the highestrisk of DS (1:50) and trisomy 18 (1<102) in 1 case.All the pregnants who with an increased risk of chro-mosome anomaly for the fetus were tested were sentto amniocentesis. In 8 cases the findings were normal.Early testing of pregnant women using biochemicalmarkers, an ultrasound check and amniocentesis,when indicated, largely reduces the percentage ofchildren born with chromosomal defects.

JMB 2008; 27 (2) 283

UDK 577.1 : 61 ISSN 1452-8258


Poster sessions

E59PRETERM DELIVERIES –

PROINFLAMMATORY CYTOKINES ASPOSSIBLE MARKERS OF INFECTION

A. Nikoli}1, Lj. Risti}1, J. Oros1, I. Cvetkovi}-Hrnjakovi}1, M. Bogavac1, L. Doì}1

1Clinical Centre of Vojvodina,Department of Laboratory Diagnostics

2Department of Obstetrics and Gynecology3Institute of Public Health of Vojvodina, Novi Sad

Preterm deliveries are the most significant causeof perinatal mortality and morbidity. Bacterial invasionof the choriodecidual space stimulates the deciduaand the fetal membrane to produce cytokines, includ-ing interleukin-1b (IL-1b), interleukin-6 (IL-6), inter-leukin-8 (IL-8), and interferon-g (INT-g). Interleukin 6and interleukin 8 are proinflammatory cytokines whoselevels in the cervical mucus have been reported to beassociated with bacterial vaginosis and intrauterineinfections, while interferon-g is reported in viral infec-tions. Inflammatory cytokines may induce the synthe-sis and release of prostaglandins that stimulate uterinecontractions and metalloproteinases that soften thecervix and weaken the chorioamniotic membranesleading to their rupture. The aim of the study was toinvestigate the possibility of the application of the levelof proinflammatory cytokine IL-8, and interferon-g asbiochemical markers of local infections in patients withclinical symptoms of preterm delivery. The investiga-tion comprised 74 pregnant women at 24–36 weeks'gestation with symptoms of preterm delivery and con-firmed infections. Cytokine levels were determined inthe patients serum using an ELISA method (BecmanCoulter, R&D Quantikine). The values of cytokine IL-8in the investigated group were 18.13 +/–30.87 vs.5.02 +/–4.91 (p< 0.08) in the control group. Thevalues of g-interferon in the group of patients with con-firmed infection were 14.22 +/–28.26 versus 4.66+/–6.45 (p<0.172) in the control group. Cytokine IL-8 and interferon-g are diagnostic markers which couldpoint to the presence of infection in pregnancy, as wellas to an increased risk of preterm deliveries in thesepatients. A combination of these proinflammatorycytokines could be added to the group of predictors ofpreterm delivery.

E60SERUM LEPTIN CONCENTRATION

IN DIALYSIS PATIENTS

S. \or|evi}-Cvetkovi}, S. Ra{ovi}, N. Kosti}, V. Markovi}, S. Kova~evi}

Health Care Center Kru{evac

A large number of patients in the progressivephase of renal insufficiency (HBI) show biochemicalmanifestations of a chronic inflammation condition,

E59PREVREMENI PORO\AJI –

PROINFLAMATORNI CITOKINI KAO POTENCIJALNI MARKERI INFEKCIJE

A. Nikoli}1, Lj. Risti}1, J. Oros1, I. Cvetkovi}-Hrnjakovi}1, M. Bogavac1, L. Doì}1

1Klini~ki centar Vojvodine, Centar za laboratorijsku medicinu

2Klinika za ginekologiju i aku{erstvo3Institut za zdravstvenu za{titu Vojvodine, Novi Sad

Prevremeni poro|aji su najzna~ajniji uzrok peri-natalnog mortaliteta i morbiditeta. Bakterijska invazijau horiodecidualni prostor podsti~e deciduu i fetalnemembrane da proizvode citokine, i to u prvom redu:interleukin-1b (IL-1b), interleukin-6 (IL-6), interleukin-8 (IL-8) i interferon-g (INT-g). Interleukin-6 i inter-leukin-8 su proinflamatorni citokini, ~ije je pove}anje ucervikalnom brisu povezano sa bakterijskom vagino-zom i bakterijskim intrauterinim infekcijama, dok jeskok interferona-g opisan kod virusnih vaginalnih infek-cija. Proinflamatorni citokini mogu indukovati sintezu iosloba|anje prostaglandina i stimulisati uterusne kon-trakcije i metaloproteinaze, koje dovode do omek{a-vanja cerviksa i slabljenja amnionskih membrana, tenjihove prevremene rupture. Cilj studije je bio da seispita mogu}a primena serumskog nivoa IL-8 i g-inter-ferona kao markera lokalne infekcije kod pacijentkinjasa klini~kim simptomima prevremenog poro|aja iinfekcije. Ispitivanje je obuhvatilo 74 gravidne pacijen-tkinje sa simptomima prevremenog poro|aja i po-tvr|enom infekcijom. Citokini su odre|eni u serumupacijentkinja ELISA metodom (Beckman Coulter, R&DQuantikine). Vrednost citokina IL-8 u ispitivanoj grupije bila 18,13 +/–30,87, u odnosu na 5,02 +/– 4,91(p< 0,08) u kontrolnoj grupi. Vrednost g-interferona ugrupi pacijenata sa potvr|enom infekcijom i simptomi-ma prevremenog poro|aja je 14,22 +/–28,26, uodnosu na 4,66 +/–6,45 (p< 0,172) u kontrolnojgrupi. IL-8 i g-interferon predstavljaju neinvazivne mar-kere koji mogu da ukaù na prisustvo infekcije u trud-no}i, kao i na pove}anje rizika od prevremenog poro-|aja kod pacijentkinja sa takvim rizikom.

E60KONCENTRACIJA SERUMSKOG LEPTINA

KOD PACIJENATA NA DIJALIZI

S. \or|evi}-Cvetkovi}, S. Ra{ovi}, N. Kosti}, V. Markovi}, S. Kova~evi}

Zdravstveni centar Kru{evac

Veliki broj bolesnika u uznapredovaloj fazi hro-ni~ne bubre`ne insuficijencije (HBI) pokazuje biohemi-jske manifestacije hroni~nog inflamatornog stanja,

284

koje se odlikuje porastom cirkuli{u}eg nivoa reaktana-ta akutne faze kao {to je C-reaktivni protein (CRP).Anoreksija je dobro poznati efekat inflamacije. Jedanod mehanizama koji indikuje anoreksiju u uslovimainflamacije moè biti povi{eni nivo serumskog leptina.Leptin, produkt ob gena, peptidni je hormon, koji sesmatra klju~nim regulatorom unosa hrane i telesneteìne. Cilj rada je da se utvrdi razlika u nivou leptinakod zdravih ljudi i bolesnika na dijalizi, kao i da seustanovi da li duìna dijaliziranja uti~e na koncentracijuleptina u krvi. Ispitano je 80 bolesnika (47 mu{karacai 33 ène) sa terminalnim stadijumom HBI, prose~neìvotne starosti 56 godina, koji se u proseku 66 mese-ci nalaze na hroni~nom programu dijalize. Od bio-hemijskih parametara odre|ivani su: urea, CRP i leptin(ELISA metod). Rezultati pokazuju da su vrednostiuree, CRP i leptina znatno vi{e u ispitivanoj grupi uodnosu na kontrolnu grupu (p<0,001). Analizomvrednosti CRP i leptina razvrstanih po duìni le~enjadijalizom (do 1 godine, do 5 godina i vi{e od 5 godi-na), moè se re}i da se rizik od hroni~ne inflamacije(CRP) pove}ava sa duìnom le~enja dijalizom (p<0,05),{to nije slu~aj sa leptinom (p>0,05). Ta~ni uzroci povi-{enog nivoa leptina u uremiji nisu sa sigurno{}u obja{-njeni, iako dosada{nji podaci upu}uju na mogu}nostpove}ane produkcije zbog inflamacije ili hiperinsuline-mije.

E61EFEKTI KONZUMIRANJA GAZIRANIH

NAPITAKA NA SERUMSKE LIPIDE I LIPOPROTEINE

M. Popovi}, S. Stojilkovi}

Zdravstveni centar Vranje, Laboratorija »Bela«, Vranje

Promene u lipidnom profilu mogu se smatratiaterogenim faktorima rizika i voditi razvoju kardiovasku-larnih bolesti. Prate}i sve ve}u u~estalost poreme}ajalipidnog profila, do{li smo na ideju da potraìmo vezusa konzumiranjem gaziranih napitaka. Cilj ovog rada jebio da se utvrdi uticaj konzumiranja velike koli~ine gazi-ranih napitaka na lipidni profil. Izabrana je grupa od 55osoba (30 èna i 25 mu{karaca), starosti izme|u 20 i45 godina, koji su konzumirali gazirana pi}a u koli~inive}oj od 1 litra dnevno duè od mesec dana. Kontrolnagrupa od 46 osoba (26 èna i 20 mu{karaca) uop{tene konzumira gazirana pi}a. Svima su ukupni holes-terol i trigliceridi odre|ivani enzimski, HDL holesteroldirektno enzimski, LDLholesterol je odre|ivan pomo}uFriedewaldove jedna~ine, a APO A-1 I APO B imuno-turbidimetrijski. Statisti~ke razlike izme|u grupa sutestirane Studentovim t-testom i p-vrednosti od 0,05 iniè su izabrane za nivo statisti~ke zna~ajnosti. Kodosoba koje konzumiraju gazirana pi}a ukupni holes-

followed by an increase in the circulation level of acutephase reactants such as C-reactive protein (CRP).Anorexia is a well known effect of inflammation. Oneof the mechanisms indicating anorexia in conditions ofinflammation could be an increased level of serum lep-tin. Leptin, an ob gene product, is a peptide hormonethat is considered the basic regulator of food intakeand body weight. The aim of the study was to deter-mine difference in the leptin level between healthysubjects and dialysis treated patients, as well as todetermine if the duration of dialysis treatment has anyinfluence on the leptin concentration in blood. A cross-sectional study included 80 patients, average age 56(47 men and 33 women) with end-stage HBI, mostlyundergoing a chronic dialysis program for 66 months.The biochemical parameters determined were: urea,CRP and leptin (by an ELISA method). The resultsshow that the values of urea, CRP and leptin are sig-nificantly higher in the examined group than in thecontrol group (p<0.001). Having analyzed CRP andleptin values classified according to the duration ofdialysis treatment (up to 1 year, up to 5 years andabove 5 years), we can say that the risk of chronicinflammation (CRP) increases with the duration of dial-ysis treatment (p<0.05), which is not the case withleptin (p>0.05). Precise causes of increased leptin lev-els in uremia have not been explained thoroughly,although actual data point at the possibility of incre-ased production due to inflammation or due to hiper-insulinemia.

E61THE EFFECTS OF A CONSUMPTION OF CARBONATED BEVERAGES ON SERUM

LIPIDS AND LIPOPROTEINS

M. Popovi}, S. Stojilkovi}

Zdravstveni centar Vranje, Laboratory »Bela«, Vranje

Changes in the lipid profile can be considered asatherogenic risk factors and can lead to the develop-ment of cardiovascular diseases. Monitoring theincreased frequency of disturbances in the lipid profile,we came to the idea to look for a connection with theconsumption of carbonated beverages. The purpose ofthis study is to evaluate the effects of a consumption ofcarbonated beverages on the lipid profile. A group of55 individuals (30 females and 25 males) was formed,aged between 20 and 45 years, who consumed largeamounts of carbonated beverages, more than one literper day, longer than one month. The control groupcomprised 46 individuals (26 females and 20 males),who do not consume carbonated beverages at all. Forall individuals we determined total cholesterol andtriglyceride concentrations enzymatically, HDL choles-terol by a direct enzymatic method, LDL cholesterolwas derived by the Frieldewald equation and APO A-1and APO B imunoturbidimetrically. Statistical differ-

JMB 2008; 27 (2) 285

ences between the groups were tested by a Student`st-test and a p-value of 0.05 or less was selected as thelevel of statistical significance. In the group that consu-med carbonated beverages, total cholesterol increasedin comparison with the control group (p<0.05), tri-glycerides also increased (p<0.05), LDL cholesterolincreased in relation to the control group statisticallysignificantly (p<0.05), and HDL cholesterol decreasedstatisticaly significantly (p<0.05). Changes in the va-lues of apolipoproteins were not statistically significant(p>0.05). On the basis of the results of our work it canbe possible to associate increased consumption of car-bonated beverages with changes in the concentrationof serum lipids and lipoproteins as one of the risk fac-tors for cardio-vascular diseases. Further studies areneeded to confirm these results.

TE62HE EXAMINATION OF ANEMIA IN HEMODIALYSIS PATIENTS

Lj. Ba~vanski, R. Jela~i}, G. Stepanov

Clinical Biochemistry Laboratory, Internal Medicine Ward – Hemodialysis Section,

General Hospital, Zrenjanin

Insufficient erythropoietin synthesis and ironshortage are the main causes of normocytic normo-chrome anemia in chronic renal insufficiency (CRI). Toprevent numerous consequences caused by anemia ofrenal failure, an adequate therapy with erythropoietin,iron preparations and folic acid is recommended asnecessary. The examination of anemia and estimationof therapy effectiveness involve the determination ofhematological parameters (erythrocyte, hemoglobin,hematocrit, index erythrocyte: MCV, MCH, MCHC),serum Fe, ferritin, total iron-binding capacity (T.I.B.C.),percentage of transferrin saturation (TSAT) and C-reac-tive protein. The study examines the effect of IV irondosage correction phase in hemodialysis patients treat-ed with erythropoietin. Sixty eight patients were exam-ined over the period of 18 months. Hematological pa-rameters were determined by using a Beckman Coulterautomated analyzer and biochemical ones using aBeckman Synchron analyzer. TSAT was calculated.During the period of examination the results of hema-tological parameters and T.I.B.C. values did not revealany statistical significance. Fe values were considerablylower (p<0.001) as a result of the change (decrease)in therapy. There was a considerable decrease in TSATresults (p<0.01) and ferritin results (p<0.001) andthey were now within the range of recommended CRIvalues, which was the aim of the therapy correction.The obtained results confirmed the necessity of moni-toring the cited parameters, especially TSAT and fer-ritin, in order to estimate the adequate substitutiontherapy in renal anemia.

terol je povi{en u odnosu na kontrolnu grupu(p<0,05), trigliceridi su tako|e povi{eni (p<0,05),LDL holesterol je statisti~ki zna~ajno povi{en u odnosuna kontrolnu grupu (p<0,05), dok je HDL holesterolstatisti~ki zna~ajno snièn (p<0,05). Kod apolipopro-teina promene vrednosti nisu statisti~ki zna~ajne(p>0.05). Na osnovu rezultata na{eg rada mogu}e jepovezati pove}ano konzumiranje gaziranih napitaka sapromenama u koncentraciji serumskih lipida kaojednog od faktora rizika za nastanak kardiovaskularnihbolesti. Dalje i obimnije studije su potrebne da bi sepotvrdili ovi rezultati.

E62ISPITIVANJE ANEMIJE KOD PACIJENATA

NA HEMODIJALIZI

Lj. Ba~vanski, R. Jela~i}, G. Stepanov

Klini~ko-biohemijska laboratorija, Interno odeljenje – odsek hemodijalize,

Op{ta bolnica, Zrenjanin

Nedovoljna sinteza eritropoetina i nedostatakgvo`|a su osnovni uzroci razvoja normocitne nor-mohromne anemije kod hroni~ne bubre`ne insuficijen-cije (HBI). U cilju prevencije brojnih posledica anemijebubre`nog porekla, prema preporukama, neophodnaje adekvatna terapija primenom eritropoetina, prepara-ta gvo`|a, folne kiseline. Ispitivanje anemije i procenaterapije obuhvataju odre|ivanje hematolo{kih para-metara (eritrocita, hemoglobina, hematokrita, indeksaeritrocita: MCV, MCH, MCHC), serumskog Fe, feritina,kapaciteta vezivanja gvo`|a (T.I.B.C.), procenta satu-racije transferina (TSAT), C-reaktivnog proteina. U na-{em radu ispitivan je efekat korekcije doze iv. prepara-ta Fe kod pacijenata na hemodijalizi koji su primalieritropoetin. Pra}eno je 68 pacijenata u periodu od 18meseci. Hematolo{ki parametri odre|ivani su na bro-ja~u Coulter Beckman, a biohemijski na SynchronBeckman automatskom analizatoru. TSAT je izra~unat.U ispitivanom periodu, rezultati hematolo{kih parame-tara i vrednosti T.I.B.C. nisu pokazali statisti~ku zna~aj-nost. Vrednosti za Fe su bile zna~ajno niè (p<0,001)posle promene (smanjenja) terapije. Rezultati za TSAT(p<0,01) i feritin (p<0,001) zna}ajno su se snizili i u{liu opseg preporu~enih vrednosti za HBI, {to je bio ciljkorekcije terapije. Dobijeni rezultati su potvrdili ne-ophodnost pra}enja navedenih parametara, posebnoTSAT i feritina, za procenu primene adekvatne supsti-tucione terapije u bubre`noj anemiji.

286

E63SERUMSKI BIOMARKERI

U BOLESTIMA JETRE

D. ]uji}, S. Golubovi}, @. Boji}-Trbojevi}, N. Ili},I. Bari~evi}-Jones, O. Nedi}, Lj. Vi}ovac

Institut za primenu nuklearne energije – INEP, Zemun, Srbija

Mnogi proteini krvi sinteti{u se u jetri. Nivoi tihproteina u serumu mogu usled poreme}aja u jetri bitisnièni ili povi{eni, stoga neki od njih mogu posluìtikao korisni biomarkeri. Odre|ivanje koncentracije razli-~itih biomarkera u serumu ima va`nu ulogu u dijag-nostici oboljenja jetre i u pra}enju toka bolesti. U ovomradu izvr{eno je pore|enje tri serumska markera koji supovezani sa oboljenjima jetre: a-fetoproteina (AFP),feritina i insulinu-sli~nog faktora rasta I (IGF-I). Visokekoncentracije AFP u serumu ukazuju na primarni he-patocelularni karcinom (HCC), a vrednosti feritina userumu ~esto su pove}ane kod neoplazija u jetri. Kom-ponente IGF sistema su tako|e uklju~ene u hepa-tokarcinogenezu. Cilj ovog rada je bio da se proceni dali pomenuti biomarkeri, sintetisani u jetri, ispoljavajuspecifi~nu distribuciju u razli~itim oboljenjima jetre: vi-rusnom hepatitisu, parazitarnoj infekciji izazvanoj ehi-nokokusom i primarnom HCC. Kontrolnu grupu ~inilesu zdrave osobe. Koncentracije AFP i IGF-I u serumuodre|ene su IRMA, odnosno RIA testom (INEP, Beo-grad, Srbija), a koncentracija feritina odre|ena je po-mo}u ELISA testa »Ferritin« (»DIMA Diagnostika«,Getingen, Nema~ka). Statisti~ka analiza izvr{ena je pri-menom programa »Primer of Biostatistics«. VrednostiAFP i feritina u serumu ispoljile su sli~an obrazac kodpacijenata sa virusnim hepatitisom ili primarnim HCC,a koncentracije ta dva markera bile su zna~ajno vi{e uodnosu na kontrolnu grupu (p<0,05). S druge strane,koncentracija IGF-I bila je zna~ajno snièna kod paci-jenata sa primarnim HCC ili ehinokokozom u odnosuna kontrolnu grupu (p<0,01 odnosno p<0,05). Kodpacijenata sa primarnim HCC sva tri analizirana se-rumska biomarkera pokazala su bilo povi{ene, bilosniène koncentracije u odnosu na kontrolnu grupu.Rezultati su pokazali da odre|ivanje IGF-I diferenciravirusni hepatitis od primarnog HCC i da moè pomo}iu proceni o{te}enja jetre kod pacijenata sa ehinokoko-zom i primarnim HCC.

JMB 2008; 27 (2) 287

E63SERUM BIOMARKERS

IN LIVER DISEASES

D. ]uji}, S. Golubovi}, @. Boji}-Trbojevi}, N. Ili}, I. Bari~evi}-Jones, O. Nedi}, Lj. Vi}ovac

Institute for the Application of Nuclear Energy – INEP,Zemun, Serbia

Many human blood proteins are synthesized inthe liver. Their serum levels may decrease or increasedue to liver disorders, therefore some of them couldserve as useful biomarkers. The determination of theserum concentration of different biomarkers has animportant role in the diagnosis of liver diseases and inmonitoring the course of disease. In this work threeserum markers, linked to liver disorders, were compa-red: a-fetoprotein (AFP), ferritin and insulin-like growthfactor I (IGF-I). High serum concentrations of AFP arestrongly predictive of primary hepatocellular carcinoma(HCC) and serum ferritin levels are often increased inliver neoplasia. Components of the IGF axis are alsoinvolved in hepatocarcinogenesis. The aim of this workwas to assess whether the above-mentioned liver bio-markers exhibit a specific distribution pattern in differ-ent types of liver disease: viral hepatitis, parasitic infec-tion echinococcosis and primary HCC. The controlgroup consisted of healthy subjects. Serum concentra-tions of AFP and IGF-I were determined using IRMAand RIA assays, respectively (INEP, Belgrade, Serbia),while the concentration of ferritin was determinedusing a Ferritin ELISA test (»DIMA Diagnostika«,Goettingen, Germany). Statistical analysis was per-formed using program »Primer of Biostatistics«. SerumAFP and ferritin levels exhibited similar patterns inpatients with viral hepatitis or primary HCC, and con-centrations of these two markers were significantlyincreased compared to the control group (p<0.05).On the other hand, the concentration of IGF-I was sig-nificantly decreased in patients with HCC or echinococ-cosis compared to the control group (p<0.01 andp<0.05, respectively). In patients with HCC, all threeanalysed serum biomarkers exhibited either elevated orreduced concentrations compared to the controlgroup. Our results showed that the determination ofIGF-I differentiates between viral hepatitis and primaryHCC and may aid in assessing hepatic damage inpatients with echinococcosis and primary HCC.

E64TPOAb AND TgAb DURING THE TREATMENT

OF GRAVES DISEASE WITH RADIOIODINETHERAPY AND MEDICAMENTOSE THYROSUPPRESSIVE THERAPY

R. Mijovi}1, N. ]uri}1, R. Avramovi}2

1Department of Diagnostic Endocrinology2Department of Nuclear Medicine, Clinical Center of Vojvodina, Serbia

The aim of the study was to compare TPOAb andTgAb values in patients (pts) with Graves disease (GD)treated with radioiodine therapy (RAITh) and thoseonce who were treated with medicamentose thyrosup-pressive therapy (MTh), and to evaluate the connec-tions between TPOAb and TgAb levels and indicatorsof thyroid gland function. We investigated 15 femaleswith GD treated with RAITh and 15 females newlydiagnosed with GD treated with MTh. Fifteen healthysex and age-matched persons made up the controlgroup. All pts treated with RAITh received only onedose, average 202 MBq. Levels of TPOAb, TgAb, TT3,TT4 and TSH were measured by an electrochemilumi-nescence immunoassay, using an »Elecsys 2010«.These parameters were estimated before treatment, 3and 6 months after RAITh or MTh. Before treatment,93.3% of pts on MTh had increased levels of TPOAbcompared to the controls, the average was 332.2IU/mL. During MTh, TPOAb constantly decreased and6 months after MTh their average was 217.4 IU/mL.Before RAI application, 86.6% of pts had increased le-vels of TPOAb, average was 814.3 IU/mL, 3 monthsafter, they increased twice, average 1579.3 IU/mL,and 6 months after the treatment we noticed somedecrement, average was 1352 IU/mL. In both groups46.6% pts had increased basal levels of TgAb. We didnot notice any significant change between the levels ofTgAb at the beginning of MTh (x=193 IU/mL) andafter six monts of MTh (x=199 IU/mL). Basal levels ofTgAb in the group treated with RAITh were average260.7 IU/mL and they increased to an average level of533 IU/mL after 6 months of RAITh. Basal levels ofTPOAb and TgAb in GD in the analysed groups aresignificantly higher compared to the controls. Therewas a significant difference between the basal levels ofTPOAb and TgAb in the group treated with RAITh andwith MTh. MTh decreased levels of TPOAb. TPOAblevels rose after RAITh, but showed some decrementafter six months of RAITh. TgAb levels rose constantlyafter RAITh, but there was no change in TgAb duringMTh. There was no direct correlation of TPOAb andTgAb levels with thyroid hormone levels and TSH ineither group of patients.

E64KRETANJE TPOAt I TgAt TOKOM LEÊNJA

GRAVESOVE BOLESTI RADIOJODNOM I MEDIKAMENTOZNOM

TIREOSUPRESIVNOM TERAPIJOM

R. Mijovi}1, N. ]uri}1, R. Avramovi}2

1Odeljenje za endokrinolo{ku dijagnostiku, 2Odeljenje za nuklearnu medicinu,Centar za laboratorijsku medicinu,

Klini~ki centar Vojvodine, Srbija

Cilj ovog istraìvanja je bio da se uporede vred-nosti TPOAt i TgAt u grupi pacijenata obolelih od Mb.Graves-Basedowi (MGB) koji su le~eni radiojodnom ter-apijom (RAJTh) sa vrednostima tih antitela kod pacije-nata le~enih medikamentoznom tireosupresivnom tera-pijom (MTh), te da se utvrdi eventualna povezanostizme|u nivoa TPOAt i TgAt i funkcijskog stanja {titaste`lezde u tim grupama pacijenata. U istraìvanju jeu~estvovalo 15 osoba ènskog pola obolelih od MGBle~enih RAJTh. Sve pacijentkinje su primile jednu dozuRAJ, srednje veli~ine 202 MBq. U drugoj grupi je bilo15 èna kojima je tek dijagnostikovana MGB i uvedenaMTh. Kontrolnu grupu sa~injavalo je 15 zdravih ènasli~ne ìvotne dobi. Kod obe grupe bolesnica odre|iva-ne su vrednosti TT3, TT4, TSH, TPOAt i TgAt viso-koosetljivom elektrohemiluminiscentnom metodom naautomatizovanom aparatu »Elecsys 2010«. Svi navede-ni parametri odre|ivani su bazalno, pre uvo|enja MTh iRAJTh, te nakon 3 i 6 meseci. Pre MTh, 93,3% pacije-nata imalo je povi{en nivo TPOAt, x=332,2 IU/mL kojikonstantno opada u toku MTh dostiù}i x=217,4 IU/mLposle {est meseci terapije. Pre aplikacije RAJTh, 86,6%pacijenata je imalo povi{ene vrednosti TPOAt, x=814,3 IU/mL, za 3 meseca vrednosti antitela dvostrukorastu na 1579,3 IU/mL, a 6 meseci od primene RAJThsniàvaju se na x=1352 IU/mL. U obe ispitivane grupe46,6% pacijenata imalo je povi{ene bazalne vrednostiTgAt. Nema zna~ajne razlike izme|u prose~nih bazal-nih vrednosti TgAt (193 IU/mL) pre MTh i vrednostinakon {est meseci po zapo~etoj MTh (x=199 IU/mL).Bazalna vrednost TgAt u grupi le~enoj RAJTh iznosiprose~no 260,7 IU/mL i raste konstantno u toku {estmeseci po primenjenoj RAJTh dostiù}i prose~ne vred-nosti od 533 IU/mL. Bazalne vrednosti TPOAt i TgAt uosoba obolelih od MGB zna~ajno su vi{e u odnosu nanivo antitela u kontrolnoj grupi zdravih. MTh dovodi dosmanjenja titra TPOAt, a RAJTh do zna~ajnog povi{enjavrednosti TPOAt nakon tri meseca, sa tendencijomsmanjenja nivoa antitela nakon {est meseci posleRAJTh. Nivo TgAt se ne menja u toku MTh, a kon-stantno raste nakon primene RAJTh. Nema korelacijevrednosti nivoa tireoidnih hormona i TSH sa nivoimaTPOAt i TgAt u obe grupe pacijenata.

288

E65PROMENE REDOKS STATUSA VITAMINA C

U TOKU ASEPTI^NOG MENINGITISA

D. Miri}, Z. Milanovi}, R. Katani}, B. Kisi}, I. Dragojevi}

Institut za biohemiju, Institut za fiziologiju, Klinika za infektivne bolesti,

Medicinski fakultet Pri{tina – Kosovska Mitrovica, Srbija

Mo`dano tkivo je zbog svog sastava i fiziolo{kihosobenosti izuzetno osetljivo na dejstvo slobodnih ra-dikala. U inflamatornoj reakciji koja prethodi i prati me-ningealnu lokalizaciju bolesti, aktivirani fagociti i drugiu~esnici inflamacije stvaraju veliku koli~inu slobodnihradikala kiseonika. U cerebrospinalnom likvoru (CSF)aktivnost enzima antioksidativne za{tite je niska, zbog~ega se odbrana subarahnoidnog prostora zasnivauglavnom na antioksidantnom dejstvu vitamina C. Uovoj studiji su odre|ivane koncentracije redoks para vi-tamina C (vit C), askorbata (Asc) i dehidroaskorbata(DHAsc) u cerebrospinalnom likvoru (CSF) i serumukod 53 pacijenata sa asepti~nim meningitisom, na pri-jemu, zatim 2–3. dana i 7–8. dana hospitalizacije.Kontrolnu grupu ~inilo je 15 pacijenata sli~nog uzrastai istog pola sa dijagnozom meningizma. Osim toga,odre|ivane su koncentracije ukupnih proteina u likvo-ru, albumina, albuminski koeficijent, kao i koncentraci-ja stabilnog produkta lipidne peroksidacije malondi-aldehida (MDA). Rezultati pokazuju da u toku asep-ti~nog meningitisa ne dolazi do promene koncentracijeukupnog vit C, ve} do zna~ajnog pove}anja odnosaredoks para DHAsc/Asc (p<0,01). Normalizacija togodnosa zapaèna je na kraju hospitalizacije, izuzev kodpacijenata sa po~etnom proteinorahijom ve}om od1,01 g/L, kod kojih tako|e postoji i stalni pad odnosaCSF/serum za askorbat (p<0,05). Dobijeni rezultatisugeri{u da pacijenti sa virusnim meningitisom imajurazli~ite brzine intratekalne akumulacije askorbata kojazavisi ne samo od aktuelnih koncentracija Asc u seru-mu, ve} i od brzine cerebralnog protoka krvi i o~u-vanosti mehanizama aktivnog transporta tog vitamina.Ne{to vi{e vrednosti odnosa askorbata CSF/serum naotpustu kod grupe sa proteinorahijom do 1,00 g/L uodnosu na kontrolne vrednosti (p<0,05) verovatnopredstavljaju kompenzatornu akciju horioidnog plek-susa, koja kod pacijenata sa zna~ajnim o{te}enjemkrvno-likvorske i krvno-mo`dane barijere izostaje.

JMB 2008; 27 (2) 289

E65CHANGES IN THE VITAMIN C REDOX

STATUS DURING ASEPTIC MENINGITIS

D. Miri}, Z. Milanovi}, R. Katani}, B. Kisi}, I. Dragojevi}

Institute of Biochemistry, Institute of Phisiology, Clinic for Infective Diseases,

Medical Faculty Pri{tina – Kosovska Mitrovica, Serbia

Due to its specific composition and physiologicalfunctions, cerebral tissue is extremely sensitive to thefree radical injury. In inflammatory reactions that pro-ceed and accompany the meningeal localization of dis-ease, activated phagocytes and other inflammatoryresponse effectors produce huge amounts of oxygen-derived free radicals. In cerebrospinal fluid (CSF) theactivities of antioxidative enzymes are low, therefore theantioxidative defense of the subarachnoid space ismostly based on protective actions of vitamin C. In thisstudy we have followed up the concentration changesof vitamin C redox pair, ascorbate (Asc) and dehy-droascorbate (DHAsc) in CSF and serum of 53 patientswith aseptic meningitis on admission, between the 2ndand 3rd day, and between the 7th and 8th day of hos-pitalization. The results were compared with a group of15 sex- and age-matched meningismus patients. Theconcentrations of CSF total protein, as well as concen-trations of albumin, malondialdehyde (MDA) and albu-min quotient in CSF and serum were also measured.Results show that during the course of viral meningitisno significant changes occur in total vitamin C concen-trations in CSF and serum, but a significant increase inDHAsc/Asc ratio was found on admission (p<0.01).During hospitalization that increase reversed towardscontrol values, except in cases with initial proteino-rachia above 1.01 g/L where, besides a permanentincrease in DHAsc/Asc ratio, a permanent decrease inAsc CSF/serum ratio also occurred (p<0.05). Theobtained results suggest that patients with asepticmeningitis have different swiftness of intrathecal ascor-bate accumulation that depends not only on the actu-al ascorbate serum concentration, but also on the cere-bral blood-flow and conserved active transport mecha-nisms of this vitamin. Somewhat higher values of ascor-bate CSF/serum ratio were observed on dismissal with-in the group with proteinorachia up to 1.00 g/L(p<0.05 vs. control) which probably represents a com-pensatory action of the chorioid plexus, absent inpatients with severe blood-CSF and blood-brain barrierdysfunction.

E66THE INFLUENCE OF THYROID DYSFUNCTION

ON THE HOMOCYSTEINEMIA LEVEL

D. Mirkovi}1, N. Antonijevi}2, A. Beleti}1, S. Popovi}3,M. Petakov3, V. Sre}kovi}3, M. Petrovi}3, S. Savi}3,

J. ]iri}3, M. Stojanovi}3, B. Jakovljevi}4, B. Trbojevi}3,N. Majki}-Singh1


2Institute of Cardiovascular Diseases, Clinical Center of Serbia, Belgrade, Serbia

3Institute of Endocrinology, Diabetes and Diseases ofMetabolism, Clinical Center of Serbia, Belgrade, Serbia

4Institute of Hygiene and Medical Ecology, School of Medicine, Belgrade, Serbia

Hyperhomocysteinemia (HHcy) is a risk factor forthe development of premature atherothrombosis andthrombotic complications. Hypothyroidism is one ofthe acquired causes of HHcy. Our study comprised 68patients with hypothyroidism, 64 patients with hyper-thyroidism and 68 euthyroid persons, matched for age.Homocysteine (Hcy) was measured in serum, using anHPLC method with fluorescent detection. Statisticalanalyses included ANOVA, Post Hoc, Chi-square,Mann-Whitney U and Student’s t tests. The Hcy level inpatients with hypothyroidism (12.75±7.18 mmol/L)was higher compared to that in controls (11.26±4.39mmol/L) and patients with hyperthyroidism (9.49±3.34 mmol/L), but the difference was significant only inthe comparison between hypothyroid and euthyroidpersons (p=0.001). The incidence of HHcy (Hcy>12mmol/L) was significantly higher (p=0.008) in patientswith hypothyroidism (46.4%), than in euthyroid con-trols (27%) and hyperthyroid patients (21%). Thegroup of patients with severe hypothyroidism (TSH>50 mU/L) had a significantly higher (p=0.008) Hcylevel (14.23±5.6 mmol/L) compared with the Hcylevel in euthyroids. A comparison of Hcy levels in pa-tients with moderate hypothyroidism (TSH 50 mU/L)(11.36 mmol/L) and euthyroids revealed no significantdifference. The Hcy levels in euthyroid patients showedno significant difference compared to the Hcy levels inpatients with subclinical and clinical forms of hypothy-roidism. We can conclude that the increase in the Hcylevel is influenced by the decrease of thyroid function,as well as the degree of reduction of thyroid activity inpatients with hypothyroidism. The probability thatHHcy will express its deleterious effects on the induc-tion of atherothrombotic events and venous throm-boembolism is rather high in patients with hypothy-roidism, especially if it has a severe form. In the afore-said risk groups it is necessary to perform all the ade-quate diagnostic and therapeutic procedures in orderto prevent the development of diseases in whose pato-genesis HHcy has a important role.

E66UTICAJ TIREOIDNE DISFUNKCIJE

NA NIVO HOMOCISTEINEMIJE

D. Mirkovi}1, N. Antonijevi}2, A. Beleti}1, S. Popovi}3,M. Petakov3, V. Sre}kovi}3, M. Petrovi}3, S. Savi}3,

J. ]iri}3, M. Stojanovi}3, B. Jakovljevi}4, B. Trbojevi}3,N. Majki}-Singh1

1Institut za medicinsku biohemiju, KCS, Beograd, Srbija

2Institut za kardiovaskularne bolesti,KCS, Beograd, Srbija

3Institut za endokrinologiju, dijabetes i bolesti metabolizma, KCS, Beograd, Srbija4Institut za higijenu i medicinsku ekologiju,

Medicinski fakultet, Beograd, Srbija

Hiperhomocisteinemija (HHcy) faktor je rizika zanastanak ubrzane ateroskleroze i trombotskih komp-likacija. Hipotireoidizam je jedan od ste~enih uzrokaHHcy. U radu je ispitivano 68 bolesnika sa hipotireo-idizmom, 64 bolesnika sa hipertireoidizmom i 68 euti-roidnih ispitanika ekvivalentne starosne dobi. Homo-cistein (Hcy) je odre|ivan u serumu, HPLC metodomuz fluorescentnu detekciju. U statistickoj analizi kori{-}eni su slede}i testovi: ANOVA, Post Hoc, Chi-square,Mann-Whitney U i Student’s t. Nivo Hcy kod bolesnikasa hipotireoidizmom (12,75±7,18 mmol/L) vi{i je ne-go kod kontrola (11,26±4,39 mmol/L) i kod bolesnikasa hipertireoidizmom (9,49±3,34 mmol/L), pri ~emuje razlika bila zna~ajna samo izme|u hipotireoidnih ieutireoidnih osoba (p=0,001). Incidencija HHcy(Hcy>12 mmol/L) bila je zna~ajno ve}a (p=0,008)kod bolesnika sa hipotireoidizmom (46,4%) nego kodu eutireoidnih kontrola (27%) i hipertireoidnih bolesni-ka (21%). Grupa hipotireoidnih bolesnika te{kog stepe-na (TSH>50 mU/L) imala je zna~ajno vi{u (p=0,008)koncentraciju homocisteina (14,23±5,6 mmol/L) uodnosu na koncentraciju homocisteina u eutireoidnihosoba. Pore|enjem vrednosti Hcy izme|u hipotireoid-nih bolesnika umerenog stepena (TSH 50mU/L)(11,36 mmol/L) i eutiroidnih osoba nije dobijena sta-tisti~ki zna~ajna razlika. Pore|enjem vrednosti Hcy ubolesnika sa subklini~kim hipotireoidizmom sa euti-reoidnim kontrolama, kao i bolesnika sa klini~ki ispolje-nim hipotireoidizmom i eutireoidnih kontrola, nisuuo~ene statisti~ki zna~ajne razlike. Smanjenje tireoidnefunkcije, kao i stepen smanjenja tireoidne funkcije ugrupi bolesnika sa hipotireoidizmom uti~e na povi{enjenivoa homocisteinemije. Kod bolesnika sa hipotireoidi-zmom, naro~ito ako je prisutan u vi{em stepenu, ve}aje verovatno}a ispoljavanja negativnih uticaja HHcy naindukciju aterotrombotskih doga|aja i nastanak ven-skog tromboembolizma. U navedenim rizi~nim grupa-ma potrebno je primeniti odgovaraju}e dijagnosti~ke iterapijske procedure radi spre~avanja nastanka obo-ljenja u ~ijoj patogenezi HHcy ima definisano mesto.

290

JMB 2008; 27 (2) 291

E67ANAEMIA DIFFERENTIATION IN CHRONICDISEASES (ACD) CAUSED BY ANAEMIA

AS RESULT OF Fe DEFICIENCY (IDA)

V. Kneèvi}

Department for Laboratory Diagnostics, »Health Center Kragujevac«, Kragujevac

Problems in the diagnostic of anaemia occurwhen the iron reserves are depleted and no clinicsymptoms found. This is actually iron deficiency with-out any clinic symptoms (also known as iron deficiencywithout anaemia). A second problem is connected tochronic inflammations, infections and neoplasm. Inthese situations it is often difficult to make an ironinsufficiency diagnosis which is important for determin-ing proper therapy. A quantitative analysis of iron in theserum without TIBC and transferrin in the serum is oflittle importance. This is the reason why quantitativeanalysis of Fe and TIBC, namely transferrin, must occursimultaneously. The amount of TIBC correlates withthe concentration of transferrin, wherewith it serves fordirect quantitative analysis of transferrin in the blood.Them main problem is that bout iron and TIBC are re-latively senseless, as a result of descending concentra-tion of iron and ascending concentration of TIBC whenthe hypochronic anaemia appears as a clinic manifes-tation of iron deficit. When the ratio of Fe/TIBC is lessthan 15%, we have a certain diagnostic indicator ofiron deficiency. This is why quantitative analysis of fer-ritin is important. The amount of ferritin descends inthe early stages of iron deficiency, before changes inthe concentration of haemoglobin, size of erythrocytesor amount of Fe and TIBC occur. Ferritin is a maindepot of iron formation in the organism and is imma-nent in every cell. In this research the values of iron,TIBC, Fe/TIBC ratio and ferritin in serum were measu-red in 30 patients with iron deficiency (IDA) and 20patients with anaemia in chronic diseases (ACD) in thisresearch. Measuring was performed using Biosystemtests on an automatic analyzer BTI 2000. Fe and TIBCare expressed in mmol/L and ferritin in μg/L. Theresults shows that iron is significantly lower in patientswith ACD anaemia and patients with IDA anaemia(p<0.05). Ferritin is considerably higher in patientswith ACD anaemia (p<0.05) and lower among pa-tients with IDA anaemia (p<0.05). TIBC is noticeablyhigher among patients with IDA anaemia but loweramong patients with ACD anaemia (p<0.05). Diffe-rential diagnosis of these two anaemia formations ishighly important because of the treatment which is sig-nificantly different depending of what kind of anaemiawe are dealing with. The existence of an acute reactionphase must be excluded during the result examinationof the quantitative analysis of ferritin in anaemia, as aresult of iron deficiency, by determining inflammationmarkers (CRP, sedimentation and fibrinogen).

E67DIFERENCIJACIJA ANEMIJE U HRONI^NIM

BOLESTIMA (ACD) OD ANEMIJE USLEDDEFICIJENCIJE Fe (IDA)

V. Kneèvi}

Slu`ba za laboratorijsku dijagnostiku, Dom zdravlja Kragujevac, Kragujevac

Problemi u dijagnostici anemije nastaju kada jerezerva gvo`|a ispra`njena, a nema klini~kih simpto-ma. To je, u stvari, deficijencija gvo`|a bez ikakvih kli-ni~kih simptoma (tzv. deficijencija gvo`|a bez anemije).Drugi problem javlja se kod hroni~nih inflamacija,infekcija i neoplazmi. Tu je ~esto te{ko postaviti dijag-nozu nedostatka gvo`|a, i time se mo`da propu{ta pri-mena adekvatne terapije. Odre|ivanje koncentracijegvo`|a u serumu bez TIBC i transferina u serumu je odmalog klini~kog zna~aja. Zato se istovremeno morajuodre|ivati koncentracije Fe i TIBC, odnosno transferi-na. Vrednost TIBC korelira sa koncentracijom transferi-na, pa sluì za indirektnu procenu koli~ine transferina ukrvi. Glavni problem je {to su Fe i TIBC relativno ne-osetljivi, jer koncentracija Fe pada a TIBC raste ve}kada se javi hipohromna anemija, kao kasna klini~kamanifestacija deficita gvo`|a. Samo kad je odnosFe/TIBC manji od 15% pouzdan je dijagnosti~ki po-kazatelj deficijencije Fe. Zato je veoma bitno odre|iva-nje feritina. Koncentracija feritina opada vrlo rano koddeficijencije gvo`|a pre nego {to do|e do promene ukoncentraciji hemoglobina, veli~ini eritrocita ili nivou Fei TIBC. Feritin je glavni depo oblik gvo`|a u organizmui prisutan je u svim }elijama. U ovoj studiji odre|ivanesu vrednosti Fe, TIBC, odnos Fe/TIBC i feritin u seru-mu kod 30 pacijenata sa deficijencijom Fe (IDA) i 20pacijenata sa anemijom u hroni~nim bolestima (ACD).Odre|ivanje je vr{eno Biosystem-ovim testovima naautomatskom analizatoru BTI 2000. Fe i TIBC suizraàvani u mmol/L a feritin u mg/L. Rezultati pokazu-ju da je Fe zna~ajno snièno i kod pacijenata sa IDAanemijom i kod pacijenata sa ACD anemijom(p<0,05). Feritin je zna~ajno vi{i kod pacijenata saACD anemijom (p<0,05) a niì kod pacijenata sa IDAanemijom (p<0,05). TIBC je zna~ajno vi{i kod pacije-nata sa IDA anemijom (p<0,05) a niì kod pacijenatasa ACD anemijom (p<0,05). Diferencijalna dijagnozate dve anemije je veoma bitna jer se njihova terapijaznatno razlikuje. Zato se pri tuma~enju rezultata kon-centracije feritina u anemiji usled deficijencije Fe moraisklju~iti postojanje reakcije akutne faze odre|ivanjemmarkera inflamacije (CRP, sedimentacije i fibrinogena).

E68THE APPLICATION OF THE LAW

ON MEDICINES AND MEDICAL DEVICES IN »SPEKTAR« LTD., ÂÂK

B. Mihailovi}

»Spektar« Ltd., 144/1 Dr Dragi{e Mi{ovi}a Street,32000 â~ak

The purpose of this thesis is to point out the sig-nificance of the Law on Medicines and Medical Devi-ces (»Official Herald« No. 84/04) for medical devicemanufacturers. The material used was obtained from»Spektar« Ltd., as well as from the Serbian Chamber ofCommerce and the Medicines and Medical DevicesAgency of Serbia. A method of descriptive epidemiolo-gy was used. By monitoring the harmonization of thework process with valid standards and directives of theEU in »Spektar« Ltd., the entire course of adjusting themedical devices production procedure according tocurrent legislation and up to obtaining the ISO 9001,ISO 13485 certificates and the CE sign for medicaldevices, was analyzed. The above procedure made theregistration of our products at the Medicines and Me-dical Devices Agency of Serbia much easier. Accordingto the Serbian Chamber of Commerce and ALIMSdata, only 15% of medical device manufacturers inSerbia have the Permit to distribute a medical devicedespite the necessity of such a permit when distributinga product in the Serbian market. Having analyzed thematerial used, we concluded that manufacturing con-ditions and the quality of medical devices have to beharmonized with the current legislation. We expect thatthe obtained permit for distributing our medical de-vices, as well as the CE sign they carry, will enlarge ourdistribution in both the domestic and foreign market,provide us with a competitive advantage in the domes-tic market and improve the work process in the healthfacilities that use our products.

E69BIOCHEMICAL ALTERATIONS

OF THE FOLATE AND METHYL-TRANSFERPATHWAY IN THE CSF OF A PATIENT WITH METHOTREXATE ASSOCIATED

CHRONIC NEUROTOXICITY

S. Vezmar1, U. Bode2, U. Jaehde3

1Institute of Pharmacokinetics, Faculty of Pharmacy,University of Belgrade

2Dept. of Pediatric Hematology/Oncology, University Children's Hospital, University of Bonn

3Dept. of Clinical Pharmacy, Institute of Pharmacy, University of Bonn

Methotrexate (MTX) is a widely used antifolate,administered in a high-dose and/or intrathecally in thetreatment of malignancies such as acute lymphoblastic

E68PRIMENA ZAKONA O LEKOVIMA I

MEDICINSKIM SREDSTVIMA U PRIVREDNOMDRU[TVU »SPEKTAR« D.O.O. ÂÂK

B. Mihailovi}

»Spektar« d.o.o, Dr Dragi{e Mi{ovi}a 144/1, 32000 â~ak

Cilj rada je da ukaè na zna~aj Zakona o lekovi-ma i medicinskim sredstvima (»Sl. glasnik RS« 84/04)za proizvo|a~e medicinskih sredstava. Kao materijal jekori{}ena dokumentacija »Spektra« d.o.o. kao i podacidobijeni od Privredne komore Srbije i Agencije za le-kove i medicinska sredstva Srbije. Kori{}ena je deskrip-tivna epidemiolo{ka metoda. Pra}enjem uskla|ivanjana~ina rada sa vaè}im standardima i direktivama EUu »Spektru« d.o.o., analiziran je kompletan tok prilago-|avanja proizvodnje medicinskih sredstava zakonskimodredbama, sve do dobijanja sertifikata ISO 9001, ISO13485 kao i CE znaka za medicinska sredstva. Pred-hodna procedura je olak{ala registraciju na{ih proizvo-da kod ALIMS-a. Na osnovu podataka dobijenih iz PKSi ALIMS-a, samo 15% proizvo|a~a medicinskih sred-stava u Srbiji ima Re{enje za pu{tanje u promet medi-cinskog sredstva, neophodno da bi se proizvod plasiraona trì{tu Srbije. Analizom kori{}enog materijala utvrdilismo da je neophodno usagla{avanje uslova proizvod-nje i kvaliteta medicinskih sredstava sa zakonskom reg-ulativom. O~ekujemo da }e nam dobijena Re{enja zapu{tanje u promet medicinskih sredstava i posedovan-je CE znaka za proizvode omogu}iti bolji plasman pro-izvoda na doma}em i stranom trì{tu, ostvariti konku-rentsku prednost na doma}em trì{tu i unaprediti rad uzdravstvenim ustanovama koje koriste na{e proizvode.

E69BIOHEMIJSKE PROMENE FOLATNOG

I METIL-TRANSFERNOG PUTA U CEREBROSPINALNOJ TE^NOSTI

PACIJENTA SA HRONI^NOM NEUROTOKSI^NOSTI ASOCIRANOM

SA PRIMENOM METOTREKSATA

S. Vezmar1, U. Bode2, U. Jaehde3

1Institut za farmakokinetiku, Farmaceutski fakultet, Univerzitet u Beogradu

2Pedijatrijska hematologija/onkologija,Univerzitetska de~ja bolnica, Univerzitet u Bonu

3Odeljenje za klini~ku farmaciju, Farmaceutski institut, Univerzitet u Bonu

Metotreksat (MTX) antifolat je koji se ~esto pri-menjuje u visokim dozama i/ili intratekalno u terapijimaligniteta poput akutne limfoblasti~ne leukemije

292

(ALL). Slu~ajevi neurotoksi~nosti koja se klasifikuje kaoakutna, subakutna i hroni~na dovedeni su u vezu s pri-menom MTX kod malignih oboljenja. Patofiziolo{kimehanizam nastanka neurotoksi~nosti jo{ nije rasve-tljen, mogu}e je da u njemu va`nu ulogu igraju biohe-mijske promene folatnog i metil-transfernog metaboli-~kog puta za koje se smatra da su uzrokovane dejstvomMTX. Opisan je slu~aj odrasle pacijentkinje na terapijirecidiva ALL sa simptomima hroni~ne leukoencefalo-patije za koju se smatra da je uzrokovana primenomMTX. U cilju procene folatnog i metil-transfernog putau cerebrospinalnoj te~nosti (CST) pacijentkinje merenasu smo koncentracije 5-metiltetrahidrofolata (5-metil-THF), MTX, S-adenozilmetionina (SAM) i S-adenozil-homocisteina (SAH). Tri uzorka CST su dobijena lum-balnom punkcijom u periodu od ~etiri meseca.Koncentracije su merene pomo}u validiranih bioanali-ti~kih metoda zasnovanih na primeni HPLC sa ultralju-bi~astom i fluorescentnom detekcijom. Rezultati supokazali dvostruko sniènje koncentracije 5-metil-THF(29,3–31,8 nmol/L) u pore|enju sa referentnim vred-nostima u svim uzorcima, dok su koncentracije SAMbile vi{e od pet puta manje u dva uzorka (5–34,2nmol/L). Koncentracije SAH su bile u rasponu 7,5–14,3 nmol/L. Pacijentkinja je imala izraène promeneu folatnom i metil-transfernom metaboli~kom putukoje ukazuju na to da biohemijske promene ovih meta-boli~kih puteva za koje se smatra da su uzrokovane dej-stvom MTX mogu imati va`nu ulogu u razvoju leuko-encefalopatije.

E70HIPERKALEMIJA KOD PACIJENATA

NA HEMODIJALIZI

N. Kosti}, S. Ra{ovi}, S. Kova~evi}

Biohemijska laboratorija, Zdravstveni centar Kru{evac,Kru{evac, Srbija

Bubreg ima klju~nu ulogu u homeostatskommehanizmu i odràva eksterni balans kalijuma (K+)konstantnim pode{avanjem urinarne ekskrecije kaliju-ma (bubreg izlu~uje 92% K+). Organizam je veomaosetljiv ~ak i na male promene K+ u serumu. Uzrocihiperkalemije su mnogi i veoma zna~ajni po posledica-ma te se zbog toga le~enju pristupa ozbiljno. Dijalizadovodi do brzog odstranjivanja K+ iz organizma. Ciljovoga rada je bio da se ispita hiperkalemija kod pacije-nata u terminalnoj fazi bubre`ne insuficijencije koji sele~e hemodijalizom, i povezanost tog katjona sa kon-centracijama kreatinina. Ispitivana grupa je imala 122bolesnika (@=46; M=76). Prose~na ìvotna dob bo-lesnika je bila 59,2±12,0 godina, minimalno 20 god,maksimalno 81 god. (M=58,7±12,6, @=59,6±11,0).Duìna dijalize kod mu{karaca iznosila je 4,6±4,1 god,kod èna 4,5±4,3 god. Odre|ivani su: urea, kreatinini kalijum u serumu (pre uklju~enja na aparat za dijalizu)na analizatoru »Bayer Advia« standardnim metodamaa kalijum jonselektivnom elektrodom. Prema nivou K+

leukaemia (ALL). Cases of neurotoxicity classified asacute, subacute and chronic were associated withMTX-treatment of malignant diseases. The pathogenicmechanism of neurotoxicity is not yet clear, and possi-bly the MTX associated biochemical alterations of thefolate and methyl-transfer metabolic pathway may playan important role. A case is reported of an adult patienttreated for ALL recidive with signs of chronic leuko-encephalopathy associated with MTX administration.In order to asses the folate and methyl-transfer pathwayin the cerebrospinal fluid (CSF) of the patient, 5 met-hyltetrahydrofolate (5-methyl-THF), MTX, S-adenosyl-methionine (SAM) and S adenosylhomocysteine (SAH)were determined. Three CSF samples were obtainedby lumbar punction within four month period. Themeasurement was performed using validated bioana-lytical methods based on HPLC with UV and fluores-cence detection. The results showed a two-fold de-crease in 5-methyl-THF levels (29.3–31.8 nmol/L) inall obtained samples compared to reference values,whereas SAM concentrations were more than five-foldlower in two samples (5–34.2 nmol/L). SAH was in therange 7.5–14.3 nmol/L. The patient had pronouncedalterations in the folate and methyl-transfer pathwaywhich indicate that MTX-associated biochemical alter-ations of these pathways may play an important role inthe development of leukoencephalopathy.

E70HYPERPOTASSEMIA IN THE SERA

OF PATIENTS ON DIALYSIS

N. Kosti}, S. Ra{ovi}, S. Kova~evi}

Biochemical Laboratory, Health Centre Kru{evac,Kru{evac, Serbia

The kidney has a vital role in the homeostaticmechanism and it regulates the external balance ofpotassium (K+) by the constant adjustment of urinaryexcretion of potassium. The causes of hyperpotassemiaare many and very significant concerning their conse-quences. This is the reason why we must seriously dealwith the treatment. Dialysis leads to a very fast removalof K+ from the organism. The aim of this work is toconfirm the presence of hyperpotassemia in the serumof patients in the terminal phase of kidney insufficien-cy who are on a hemodialysis treatment, and to checkthe connection of this cation with the concentration ofcreatinine. The examined group comprised 122patients (W=46, M=76). Average age of the patientswas 59.2±12.0 years old. The duration of dialysis inmen was 4.6±4.1 years and in women 4.5±4.3 years.We determined urea, nitrogen, creatinine and potas-sium in the serum (before starting dialysis) on a »BayerAdvia« analyzer using standard methods, and for po-tassium the ionselective electrode was used. According

JMB 2008; 27 (2) 293

to the level of K+ in the serum all the tested patientswere divided into two groups: the first group (N=78)with a concentration of potassium K+ of up to 5.5mmol/l, the second group (N=42) with a concentra-tion of K+>5.6 mmol/L. The patients in the secondgroup (xpotas= 6.261 mmol/L, median 5.93) hadhigher levels of urea (x=27.84 mmol/L, median 26.8)and creatinine (901.7 mmol/L mediane, 878.2) in rela-tion to the group of tested patients who had K withinthe reference range (xpotas=4.832 mmol/L, xurea=26.0 mmol/L, xcreat=857.8 mmol/L). The given re-sults show that the increase of urea and creatinine inthe serum is followed by hyperpotassemia in patients inthe terminal phase of CRF who are on a chronic he-modialysis programme (at the level of P=0.05).

E71CALCIUM, PHOSPHORUS, MAGNESIUM AND INTACT PARATHYROID HORMONE

IN HAEMODIALYSED PATIENTS

S. Ra{ovi}, N. Kosti}, S. \or|evi}-Cvetkovi}, S. Kova~evi}, V. Markovi}, D. Nikoli}

Health Center Kru{evac, Pharmacological Institution Kru{evac, Kru{evac, Serbia

In chronic renal failure patients secondary hyper-parathyroidism occurrs as a result of stimulation of theparathyroid gland by low calcium levels. The purposeof the study was the follow-up of laboratory parame-ters: calcium (Ca), phosphorus (P), magnesium (Mg)and intact parathyroid hormone (iPTH) in haemodial-ysed patients before dialysis. The group included 74patients of both sexes – 28 females and 46 men, theduration of haemodialysis was 1–24 years. Parameterswere determined in the serum: Ca, P and Mg concen-trations were measured using a biochemical analyzer»Advia 1650« and »Advia Chemistry« tests. IPTH wasdefined on an »Immulite 1000« analyzer (Immulite1000 iPTH is a solid phase, to-site chemiluminescentenzyme labeled immunometric assay). The results ofpatients were compared with those obtained from thecontrol group. Mean serum levels of Ca and Mg inpatients were within the normal reference range, but ofP were higher than the normal reference range. Themedian of iPHT was 269.5 pg/mL. Mean serum levelsof calcium were significantly lower in patients (p<0.001). Mean serum levels of phosphorus were alsosignificantly lower in patients (p<0.001). Mean serumlevels of phosphorus were significantly higher inpatients (p<0.001). Magnesium levels were not signif-icantly different. Mean serum levels of iPTH were in thecontrol group 41.61±11.15 pg/mL, the median ofiPTH in patients was 296.5 mg/mL. IPTH is a poten-tial diagnostic tool in the prediction of renal bone dis-eases in haemodialysed patients, as is known in litera-ture. Although levels of iPTH were many times higher

u serumu svi ispitanici su razvrstani u dve grupe: Igrupa (N=78) sa konc. K+ do 5,5 mmol/L, II grupa(N=42) konc. K+>5,6 mmol/L. Pacijenti u II grupi(xkalijum=6,261 mmol/L, medijana 5,93) imali suvi{e vrednosti uree (x=27,84 mmol/L, medijana26,80) i kreatinina (901,7 mmol/L, medijana 878,2) uodnosu na grupu ispitanika koja je imala K+ u referen-tnim granicama (xkalijum=4,832 mmol/L, xurea=26,0 mmol/L, x kreat=857,8 mmol/L). Dobijeni rezul-tati pokazuju da porast vrednosti uree i kreatinina userumu pacijenata u HBI prati hiperkalemija (na nivouP=0,05) i stoga je neophodno njihovo redovno labo-ratorijsko kontrolisanje.

E71KALCIJUM, FOSFOR, MAGNEZIJUMI INTAKTNI PARAT HORMON KOD PACIJENATA NA HEMODIJALIZI

S. Ra{ovi}, N. Kosti}, S. \or|evi}-Cvetkovi}, S. Kova~evi}, V. Markovi}, D. Nikoli}

Zdravstveni centar Kru{evac, Apotekarska ustanova Kru{evac, Kru{evac, Srbija

Kod pacijenata sa hroni~nom renalnom insufici-jencijom javlja se sekundarni hiperparatireodizam kaorezultat stimulacije paratireoidnih `lezda usled niskognivoa kalcijuma. Predmet rada je pra}enje biohemijskihparametara – kalcijuma (Ca), fosfora (P), magnezijuma(Mg) i intaktnog parat hormona kod hemodijaliziranihpacijenata pre dijalize. Grupa uklju~uje 74 pacijentaoba pola – 28 èna i 46 mu{karaca, podvrgnutih he-modijalizi tokom 1–24 godine. Parametri su odre|ivaniu serumu: Ca, P i Mg odre|ivani su na biohemijskomanalizatoru »Advia 1650« komercijalnim testovima»Advia chemistry«. Intaktni parat hormon je odre|ivanna analizatoru »Immulite 1000« hemiluminiscencijom(»Immulite-Siemens« testovima). Rezultati pacijenatapore|eni su sa rezultatima kontrolne grupe od 20 do-brovoljnih davalaca krvi. Srednje serumske vrednostiCa i Mg pacijenata dobijene su u okviru normalnih re-ferentnih vrednosti, a fosfora iznad gornje referentnegranice. Medijana iPTH bila je 269,5 pg/mL. Srednjevrednosti kalcijuma niè su kod pacijenata (p<0,001).Srednja vrednost fosfora vi{a je kod pacijenata(p<0,001). Vrednosti magnezijuma se statisti~ki zna-~ajno ne razlikuju u ispitivanim grupama. Srednja vred-nost iPTH je u kontrolnoj grupi 41,61±11,15, a medi-jana pacijenata je 296,5 pg/mL. Parat hormon moèbiti potencijalno dijagnosti~ko sredstvo u prevencijiko{tanih oboljenja hemodijaliziranih pacijenata i kaotakav se pominje u literaturi. Mada su vrednosti iPTH uovoj grupi vi{estruko povi{ene u odnosu na normalnevrednosti, klini~ke manifestacije na kostima ih ne prateu istom stepenu. One su u ovoj grupi diskretne i ume-

294

JMB 2008; 27 (2) 295

than the normal reference interval in the examinedgroup, clinical manifestations on the bones do not havethe same gravity. In our patient group renal bone chan-ges were discrete and moderate. It seems that there issome kind of bone resistence to iPTH in chronic renalfailure patients.

E72INFLAMMATION MARKERS AND OXIDATIVE

STRESS IN DIABETES MELLITUS TYPE 2

M. Jeli}, G. Damnjanovi}, B. Jovovi}, N. Le~i}

Clinical Biochemical Laboratory,Internal ward, Military Hospital, Ni{, Serbia

Recent theories of atherosclerosis have revealedthat the inflammatory response of the artery wall is asignificant pathogenesis factor. Not only does thisinflammation process contribute to a cardiovascularchange, but it also initiates and helps atherosclerosisprogress. However, the characteristics of the inflamma-tory process in type 2 diabetes mellitus are still vaguetaking into account that the disease is characterised bya coexistence of myriads of proinflammatory, prooxida-tive and procoagulating risk factors. Besides the lipidstatus and glicoregulation levels in patients with type 2diabetes mellitus, the following aspects have beendetermined: lipid peroxidation – malondialdehyde(MDA), high-sensitivity C-reactive protein – acute pha-se reactant (hsCRP), intracellular adhesive molecule-1(ICAM-1), and vascular cellular adhesive molecule-1(VCAM-1). MDA values have been determined usingthe Andreev et al. method (1988), hsCRP values by acommercial test of the Dade Behring Company with aDimension-Xpand analyzer, and ICAM-1 and VCAM-1values by the ELISA method, commercial test of theBeckman Coulter Company with the BioSystems-ELISAreader. The subjects were divided into the followinggroups: type 2 DM and myocardial revascularizationpatients, type 2 DM without revascularisation, patientswith ischemic cardiac disease, and a healthy controlgroup. The levels of HbA1c, lipid parameters, MDA,hsCRP, ICAM-1 and VCAM-1 were significantly elevat-ed in the diabetic patients compared to the othergroups (p<0.05). There was a negative correlationbetween the values of hsCRP and HDL cholesterol(p<0.05). A positive correlation was found betweenthe hsCRP and ICAM-1 values (p<0.05). Elevatedoxidative stress and proinflammatory marker valueswere found in the type 2 DM patients and have a signi-ficant role in the occurence of cardiovascular diseases.

rene. Izgleda da postoji neka vrsta rezistencije ko{tanogtkiva na parat hormon kod obolelih od hroni~ne bu-bre`ne insuficijencije, pa su promene na kostima malei pored velikog lu~enja paratireoidnih `lezda.

E72INFLAMATORNI MARKERI I OKSIDATIVNISTRES U DIJABETES MELITUSU TIPA-2

M. Jeli}, G. Damnjanovi}, B. Jovovi}, N. Le~i}

Klini~ko-biohemijska laboratorija,Interno odeljenje, Vojna bolnica, Ni{, Srbija

Najnovije teorije o aterosklerozi ukazuju na to daje inflamatorni odgovor arterijskog zida zna~ajan pato-genetski faktor. Inflamacija ne samo da doprinosi akut-nom kardiovaskularnom doga|aju, ve} je klju~ inici-jacije i progresije ateroskleroze. Me|utim, karakteris-tike inflamatornog procesa u dijabetes melitusu tipa 2jo{ uvek su nedovoljno poznate, budu}i da to obolje-nje karakteri{e udruènost brojnih proinflamatornih,prooksidativnih i prokoagulacionih faktora rizika. Kodbolesnika sa dijabetes melitusom tipa 2 (DM tipa 2),pored lipidnog statusa i stepena glikoregulacije, odre-|ivani su: lipidna peroksidacija – malondialdehid(MDA), reaktant akutne faze – visokoosetljivi C-reak-tivni protein (hsCRP), intracelularni adhezivni molekul-1 (ICAM-1) i vaskularni }elijski adhezivni molekul-1(VCAM-1). Vrednosti MDA su odre|ivane po metodiAndreeve i sar. (1988), hsCRP komercijalnim testomfirme »Dade Behring« na analizatoru »Dimension-Xpand«, a ICAM-1 i VCAM-1 ELISA metodom, komer-cijalnim testom firme »Beckman Coulter Company« na»ByoSistems« ELISA rideru. Ispitanici su podeljeni ugrupe bolesnika sa: DM tipa 2 i revaskularizacijommiokarda, DM tipa 2 bez revaskularizacije, bolesnici saishemi~nom sr~anom bole{}u, i grupu zdravih ispitani-ka. Nivoi HbA1c, koncentracija lipidnih parametara,MDA, hsCRP, ICAM-1 i VCAM-1 bili su znatno vi{i kodpacijenata sa dijabetesom u odnosu na ostale ispi-tanike (p<0,05). Kod grupe pacijenata sa dijabetesompostojala je negativna korelacija izme|u vrednostihsCRP i HDL holesterola (p<0,05). Pozitivna korelaci-ja postojala je izme|u vrednosti hsCRP i ICAM-1(p<0,05). Pove}ani nivo oksidativnog stresa i proinfla-matornih markera javlja se u DM tipa 2 i ima zna~ajnuulogu u pojavi kardiovaskularnih oboljenja.

E73INTERDEPENDENCE BETWEEN VALUES

OF BCL-2 AND DIFFERENT AGE

V. ]osi}, L. Zvezdanovi}, T. Risti}, S. Kundali}, V. B. \or|evi}

Department of Medical Biochemistry, Clinical Center, Ni{, Serbia

The Bcl-2 presents a family of genes and proteinswhich are involved in the homeostasis of cell prolifera-tion and death. The Bcl-2 probably realizes its effectsby the regulation of mitochondrial permeability transi-tion pore which is involved in the maintenance of Ca2+,pH and voltage. It is known that decreases in Bcl-2 le-vels lead to cell death by apoptosis while an overex-pression of Bcl-2 protects cells from death. The Bcl-2gene has been implicated in a number of cancers,including melanoma, breast, prostate, and lung carci-nomas, as well as schizophrenia and autoimmunity dis-eases. It is also may be involved in resistance to con-ventional cancer treatment. In this study we investigat-ed the effects of age and sex on anti-apoptotic protein– Bcl-2. The values of Bcl-2 were measured by theELISA method in the serum of two healthy groups ofvolunteers: the first, older group (58 5.6 years) and asecond, younger group (30 6.8 years). We observedstatistically significant lower Bcl-2 values in older per-sons compared with younger persons 0.2500.067ng/mL vs. 0.294 0.67ng/mL; p<0.05). Simul-taneously, we noted unchanged values of Bcl-2 bet-ween different sexes in both investigated groups. Wecan conclude that a low level of Bcl-2 in aged personsmay be one of possible risks in the adequate regulationof apoptosis, including oncogenesis as well as resistan-ce to therapy of cancer in comparison with youngerperson.

E74OXIDATIVE STRESS IN WISTAR RAT LIVER

CAUSED BY CLOFIBRATE

J. Bogdanska, B. Todorova, P. Korneti, D. Labudovi}

Institute of Medical and Experimental Biochemistry,Medical Faculty, Skopje, Republic of Macedonia

It is well known that clofibrate, as well as otherperoxisomal proliferators, can induce peroxisomal pro-liferation and a variety of associated effects in rodents.Long-term treatment of rodents with peroxisomal pro-liferators has been found to cause hepatocarcinogene-sis and there are also reports of tumor formation inother organs. One of the proposed mechanisms under-lying the hepatocarcinogenicity of peroxisomal prolifer-ators is the hypothesis of oxidative stress. The aim ofour study was to determine whether antioxidant enzy-mes that facilitate the removal of reactive oxygen

E73ME\UZAVISNOST VREDNOSTI BCL-2

I RAZLIÎTOG @IVOTNOG DOBA

V. ]osi}, L. Zvezdanovi}, T. Risti}, S. Kundali}, V. B. \or|evi}

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

Bcl-2 predstavlja familiju gena i proteina koji suuklju~eni u homeostatske mehanizme }elijske prolif-eracije i smrti. Bcl-2 verovatno ostvaruje sopstvene efe-kte regulacijom permeabilnosti prolaznih pora mito-hondrija koje su uklju~ene u odràvanje Ca2+, pH ivoltaè. Poznato je da sniènje nivoa Bcl-2 vodi u }elij-sku smrt putem procesa apoptoze dok pove}ana pro-dukcija Bcl-2 {titi }elije od smrti. Bcl-2 gen ima u~e{}au pojavi brojnih kancera, uklju~uju}i melanome, karci-nome dojke, prostate kao i pojavi {izofrenije i razli~itihautoimunih bolesti. On tako|e igra va`nu ulogu usmanjivanju rezistencije na tradicionalnu onkolo{ku te-rapiju. U ovoj studiji pra}en je uticaj godina starosti ipola na anti-apoptoti~ni protein – Bcl-2. Nivo Bcl-2odre|ivan je ELISA metodom, u serumu dve grupezdravih ispitanika: prvoj grupi starijih osoba (58 5,6god.) i drugoj grupi mla|ih osoba (30 6,8 god.).Ustanovljen je statisti~ki zna~ajan niì nivo Bcl-2 vred-nosti kod starijih osoba u odnosu na mla|e osobe(0,250 0,067ng/mL prema 0,294 0,67ng/mL;p<0,05). Istovremeno, zabeleèna je neizmenjenavrednost Bcl-2 izme|u razli~itih polova obe ispitivanegrupe. Moèmo zaklju~iti da nizak Bcl-2 nivo kod star-ijih osoba moè biti jedan od mogu}ih rizika za nead-ekvatnu regulaciju apoptoze, uklju~uju}i proces onko-geneze kao i rezistenciju na onkolo{ku terapiju u odno-su na mla|e osobe.

E74OKSIDATIVNI STRES U JETRI VISTAR

PACOVA IZAZVAN KLOFIBRATOM

J. Bogdanska, B. Todorova, P. Korneti, D. Labudovi}

Institut za medicinsku i eksperimentalnu biohemiju,Medicinski fakultet, Skoplje, Republika Makedonija

Poznato je da klofibrat, kao i drugi peroksizomal-ni proliferatori, mogu izazvati peroksizomalnu prolif-eraciju i u vezi s tim brojne druge efekte kod glodara.Otkriveno je da dugotrajan tretman glodara peroksi-zomalnim proliferatorima izaziva hepatocanceroge-nezu a ima izve{taja i o nastanku tumora u drugimorganima. Jedan od navedenih potencijalnih meha-nizama hepatokancerogenosti peroksizomalnih prolif-eratora jeste hipoteza o oksidativnom stresu. Cilj na{estudije bio je da se utvrdi da li je do{lo do modulacijeantioksidantnih enzima koji olak{avaju uklanjanje reak-

296

JMB 2008; 27 (2) 297

species (ROS) were modulated following short-termadministration of clofibrate and whether the modula-tion had the same pattern as the prooxidant one. Thestudy was performed in male Wistar rats weighting250–350 g, divided into two groups: control one,N=14 (injected i.p. with sterile saline) and experimen-tal one N= 14 (injected i.p. 250 mg/1000 g/24h for12 days). Whole liver homogenates, as well as subcel-lular fractions, were assayed for activity of D-aminoacid oxidase, urate oxidase and palmitoyl CoA oxidaseas prooxidant enzymes, and for the activity of catalase,superoxide dismutase and glutathione peroxidase asthe antioxidant enzymes. Standard spectrophotometermethods were employed for the measurement of theenzyme activities. Specific activity of prooxidantenzymes showed a statistically significant increase, withthe exception of D-aminoxidase. On the contrary, onlycatalase showed a slight increase in activity. Superoxidedismutase and gluthatione peroxidase showed a statis-tically significant decrease of the activity. We may con-clude that clofibrate treatment might lead to oxidativestress in Wistar rat liver.

E75ADHESION MOLECULES AND SYSTEMIC

LUPUS ERYTHEMATODES

L. Zvezdanovi}-êlebi}1, V. \or|evi}1, V. ]osi}1,T. Cvetkovi}2, S. Kundali}1, D. Stankovi}-Ferle`1

1Department of Medical Biochemistry, Clinical Centre, Ni{, Serbia2Institute of Biochemistry,

Faculty of Medicine Ni{, Serbia

Systemic lupus erythematodes (SLE) is a disorderof complex etiology, involving initial factors in the envi-ronment reacting with the background of clear poly-gene susceptibility. One of the most important pre-con-ditions of SLE is the disturbance of immune regulationwith inadequate removal of immune complexes lead-ing to tissue damage and organ function disorders. Sin-ce adhesion molecules are important products of cellactivation with a key role in the intracellular signallingprocess, this paper aims to determine the concentra-tion of intercellular (ICAM) and vascular adhesion mol-ecule (VCAM-1) in patients with different clinical man-ifestations of SLE in the clinical exacerbation phase ofthe disease: skin (S-SLE); neurolupus (N-SLE); joint (J-SLE); and vascular (V-SLE). Healthy blood donor vol-unteers comprised the control group. Adhesion mole-cule values were measured by the ELISA method, witha commercial R&D assay. ICAM, as one of the integralfactors in cell-cell and cell-matrix receptors and an ini-tial marker of inflammatory response, shows only aslight increase, without statistical significance com-pared to control values. As for VCAM, produced byendothelial cells, there was a marked increase(P<0.05) in the group of total SLE compared to con-

tivnih kiseoni~nih vrsta (ROS) posle kratkotrajne pri-mene klofibrata i da li takva modulacija ima isti obrazackao i prooksidantna. Studija je sprovedena na mu`jaci-ma Vistar pacova, teìne 250–350 g, podeljenim u dvegrupe: kontrolnu, N=14 (injekcije i. p. fiziolo{kog ras-tvora) i eksperimentalnu N= 14 (injekcije i. p. 250mg/1000 g/24h 12 dana). U homogenatima jetre,kao i subcelularnim frakcijama, odre|ene su aktivnostioksidaze D-aminokiselina, urat oksidaze i palmitoil CoAoksidaze kao prooksidantnih enzima, kao i aktivnostikatalaze, superoksid dismutaze i glutation peroksidazekao antioksidantnih enzima. Standardne spektrofoto-metrijske metode kori{}ene su za merenje aktivnostienzima. Specifi~na aktivnost prooksidantnih enzimapokazala je statisti~ki zna~ajan porast, sa izuzetkom D-aminooksidaze. Nasuprot tome, samo je katalaza po-kazala blag porast aktivnosti. Aktivnosti superoksid dis-mutaze i glutation peroksidaze bile su statisti~ki zna-~ajno smanjene. Zaklju~ujemo da tretman klofibratommoè izazvati oksidativni stres u jetri Vistar pacova.

E75ADHEZIONI MOLEKULI I SISTEMSKI

LUPUS ERITEMATODES

L. Zvezdanovi}-êlebi}1, V. \or|evi}1, V. ]osi}1,T. Cvetkovi}2, S. Kundali}1, D. Stankovi}-Ferle`1

1Centar za medicinsku biohemiju, Klini~ki centar, Ni{, Srbija

2Institut za biohemiju, Medicinski fakultet, Ni{, Srbija

Sistemski lupus eritematodes (SLE) jeste poreme-}aj kompleksne etiologije koji obuhvata inicijalne fak-tore spolja{nje sredine koji reaguju sa zale|em jasnoprisutne poligene podlo`nosti. Jedan od osnovnih uslo-va za njegov nastanak je poreme}aj imunolo{ke regu-lacije uz neadekvatno odstranjivanje imunih kompleksakoji dovode do o{te}enja tkiva i funkcije organa. Budu-}i da adhezioni molekuli predstavljaju zna~ajne pro-dukte aktivacije }elija sa klju~nom ulogom u procesuinter}elijske signalizacije, cilj ovog rada je bio odreditikoncentracije intercelularnog (ICAM) i vaskularnogadhezionog molekula (VCAM-1) kod bolesnika sarazli~itim klini~kim manifestacijama SLE u fazi klini~kogpogor{anja bolesti: ko`ni (K-SLE); neurolupus (N-SLE);zglobni (Z-SLE) i vaskularni (V-SLE). Kontrolnu grupusu sa~injavali uzorci zdravih osoba, dobrovoljnih davao-ca krvi. Merenje vrednosti adhezionih molekula jevr{eno ELISA metodom, komercijalnim testom R&D.ICAM, kao jedan od integralnih ~inilaca }elija-}elija i}elija-matriks receptora i inicijalni marker inflama-tornog odgovora, pokazuje samo neznatan porast bezstatisti~ki zna~ajne razlike u odnosu na kontrolne vred-nosti. [to se ti~e VCAM, koga stvaraju }elije endoteliju-ma, postoji statisti~ki zna~ajan porast (P<0,05) u grupi»ukupni SLE« u odnosu na kontrolne vrednosti. Do-

trols. The results we obtained suggest that VCAM canbe an appropriate marker for surveillance of SLE pa-tients. VCAM is also one of the NADP-H oxidase acti-vation markers in endothelial cells, necessary in thelymphocyte migration process. VCAM-1 promotertranscription is supported by NF-kB composed of p50and p65 subunits, with p50 subunit having an inhibito-ry role in the suppression of VCAM-1 activation by var-ious cytokines, and with the p65 subunit being a posi-tive, cytokine-inducible specific regulator of VCAM-1gene expression. Future determination of NF-kB valuecan thus be of special importance in SLE patients.

E76THE USE OF HOMA MODELING

IN THE ASSESSMENT OF INSULINRESISTENCE IN WOMEN WITH

POLYCYSTIC OVARY SYNDROME

B. Gli{i}1, \. Macut2, N. Majki}-Singh1

1Institute of Medical Biochemistry, 2Institut of Endocrinology,

Diabetes and Metabolic Diseases, Clinical Center of Serbia, Belgrade, Serbia

Insulin resistance (reduced actions of insulin onglucose homeostasis) is present in many PCOS (poly-cystic ovary syndrome) women and plays a role in thepathogenesis of the reproductive disturbances as wellas PCOS complications (diabetes, hypertension, dyslip-idaemia, atherosclerosis). The »gold standard« for theevaluation of insulin resistance (IR) is euglycemic hy-perinsulinemic clamp, but it is not feasible for the rou-tine screening of patients. The HOMA (homeostasisassessment model) index uses the basal glucose andinsulin levels for assessment of IR. The aim of this studywas to identify PCOS in women who are most insulinresistant using the HOMA index. We investigated 67PCOS women and 46 healthy, age- and BMI-matchedcontrols. Based on the BMI (body mass index), bothPCOS and control groups were divided into two sub-groups: normal weight, BMI 25 kg/m2 (A1-PCOS(n=36) and B1-CONT (n=32)) and overweight, BMI> 25 kg/m2 (A2-PCOS (n=31) and B2-CONT(n=14)). Blood samples were collected in the follicularphase of the cycle for the determination of basal glu-cose (enzymatic method), insulin (ELISA method) andtestosterone (RIA method). HOMA was calculatedusing the formula fasting insulin (mU/L) x fasting glu-cose (mmol/L) /22.5. We have found no significantdifferences in these parameters between control sub-groups (B1 vs. B2). Analyzing the PCOS subgroups (A1vs. A2), we have found significant differences in thelevels of insulin (p<0.005) and HOMA (p<0.005).Normal weight PCOS women and controls (A1 vs. B1)showed statistically significant differences in insulin le-vels (p<0.001) and HOMA index (p<0.001) as well as

bijeni rezultati sugeri{u da VCAM moè biti obe}avaju-}i marker za pra}enje bolesnika sa SLE. Tako|e, VCAMje jedan od markera aktivacije NADP-H oksidaze uendotelnim }elijama koja je neophodna u procesu mi-gracije limfocita. Transkripciju VCAM-1 promoterapodràva NF-kB sastavljen od p50 i p65 subjedinica,pri ~emu p-50 subjedinica ima inhibitornu ulogu usupresiji aktivacije VCAM-1 razli~itim citokinima, dok jep65 subjedinica predstavljena kao pozitivan, citokin-inducibilan specifi~an regulator ekspresije gena zaVCAM-1. Otuda budu}e odre|ivanje vrednosti NF-kBmoè biti od posebnog zna~aja kod bolesnika sa SLE.

E76PRIMENA HOMA MODELA ZA PROCENU

INSULINSKE REZISTENCIJE KOD PACIJENATA SA SINDROMOMPOLICISTI^NIH OVARIJUMA

B. Gli{i}1, \. Macut2, N. Majki}-Singh1

1Institut za medicinsku biohemiju,2Institut za endokrinologiju,

dijabetes i bolesti metabolizma,Klini~ki centar Srbije, Beograd, Srbija

Insulinska rezistencija (smanjen efekat date dozeinsulina na homeostazu glukoze) ~esto je prisutna kodèna sa sindromom policisti~nih ovarijuma (PCOS) ismatra se odgovornom za pojavu reproduktivnih pore-me}aja kao i komplikacija PCOS (diabetes mellitus tip2, hipertenzija, dislipidemija, ateroskleroza). Zlatnistandard za procenu insulinske rezistencije (IR) pred-stavlja euglikemijski hiperinsulinemijski klamp. Zbogzametnog postupka klampa, {iru klini~ku primenu imaHOMA (Homeostasis Model Assessment) koji za pro-cenu IR koristi bazalne vrednosti glukoze i insulina. Popreporuci SZO vrednosti HOMA indeksa >2,5 ukazu-ju na pojavu insulinske rezistencije. Rad je obuhvatio67 pacijentkinja sa PCOS i 46 ispitanica koje su ~inilekontrolnu grupu. Obe grupe su prema vrednosti BMIpodeljene u dve podgrupe: negojazne, BMI 25 kg/m2

(A1 – PCOS (n=36) i B1 – KONT (n=32)) i gojazne,BMI > 25 kg/m2 (A2 – PCOS (n=31) i B2 – KONT(n=14)). Kod svih ispitanica odre|ivani su glukoza(enzimskom metodom), insulin (ELISA metodom) itestosteron (RIA metodom). HOMA je izra~unata pri-menom formule insulin (mU/L) x glukoza (mmol/L) /22,5. Dobijene su slede}e vrednosti HOMA indeksa(izraène kao Xsr±Sd): A1-PCOS 2,62±1,18; A2-PCOS3,93±2,65; B1-KONT 1,68±0,60; B2-KONT 1,97±1,04. Statisti~kom obradom podataka nisu dobijene zna-~ajne razlike odre|ivanih parametara unutar kontrolnegrupe (B1 prema B2). Unutar grupe PCOS pacijenata(A1 prema A2) statisti~ki zna~ajnu razliku su pokazivalevrednosti insulina i HOMA indeksa (p<0,005), dok sevrednosti glukoze i testosterona nisu razlikovale. Me-|usobnim pore|enjem podgrupa pacijenata i odgo-

298

varaju}ih kontrolnih podgrupa nisu dobijene zna~ajnerazlike u vrednostima glukoze u obe podgrupe, dok su sevrednosti HOMA indeksa i insulina zna~ajno razlikovale(p<0,001 u grupi negojaznih i p<0,005 za insulin ip<0,01 za HOMA u grupi gojaznih ispitanica). Na osno-vu dobijenih rezultata moè se zaklju~iti da je IR prisutnai kod velikog broja negojaznih èna sa PCOS, a da seHOMA indeks moè koristiti kao screening test za IR i se-lekciju èna sa PCOS koje bi dale dobar odgovor na pri-menu terapije u cilju pobolj{anja insulinske senzitivnosti.

E77NEFROTOKSI^NI EFEKTI GENTAMICINA KOD

PACIJENATA U NEONATALNOM PERIODU

T. Vuji}1, B. Davidovi}2, S. Uletilovi}2, J. Predojevi}-Samardì}3, D. Mal~i}3, @. Sani~anin2

1Agencija za lekove Republike Srpske, Banjaluka2Medicinski fakultet, Univerzitet u Banjaluci

3Klinika za de~ije bolesti, Klini~ki centar Banjaluke, Republika Srpska, Bosna i Hercegovina

Rezultati na{e ranije studije su pokazali da ~ak inormalne doze gentamicina izazivaju, kod pacijenatastarosti od 3 do 10 godina oba pola, nefrotoksi~ne efe-kte pra}ene pove}anjem aktivnosti enzima dominantnolokalizovanih u proksimalnim tubulama u urinu, alani-naminopeptidaze (AAT, EC 3.4.11.2), gama-glutamil-transferaze (GGT, EC 2.3.2.1) i N-acetil-beta-D-glu-kozaminidaze (NAG, EC 3.2.1.30). Kod pacijenata uneonatalnom periodu manje je izraèna renalna aku-mulacija aminoglikozida {to moè da se objasni morfo-lo{kim i funkcionalnim karakteristikama bubrega u tomperiodu i zbog toga bi se moglo o~ekivati manje nefro-toksi~no delovanje gentamicina, uz iste uslove primene,u odnosu na decu starijeg uzrasta. Mi smo, u uzorku 12-~asovnog urina, pratili aktivnosti enzima proksimalnihtubula, AAP, GGT i NAG, kod 30 pacijenata kojima je uokviru terapije apliciran gentamicin i kod 30 pacijenataiz kontrolne grupe. U sastav ispitanika bila su uklju~enaoba pola, u neonatalnom periodu. Tretman je spro-vo|en do 10 dana uz davanje gentamicina u dozama od2,5 mg/kg telesne mase dnevno. Utvr|ena je zna~ajnarazlika izme|u AAP i GGT, u U/mmol kreatinina, osmogdana sprovo|enja terapije izme|u ispitanika u eksperi-mentalnoj i kontronoj grupi (p<0,01) (kod odrasle deceje do porasta aktivnosti, pri ranijem ispitivanju, do{lo dru-gog dana sprovo|enja terapije) i nije utvr|ena zna~ajnarazlika u aktivnosti NAG izme|u ispitanika u eksperi-mentalnoj i kontrolnoj grupi (u odrasle dece, signifikant-na razlika je ranije utvr|ena osmog dana nakon spro-vo|enja terapije). Dobijeni rezultati pokazuju da je nefro-toksi~no delovanje gentamicina kod pacijenata u neona-talnom periodu manje izraèno, verovatno zbog nepot-pune morfolo{ke razvijenosti bubrega i brè glomeru-larne filtracije, nego kod dece uzrasta od 3 do 10 godi-na. Kod dece tog uzrasta rani i veoma osetljivi indikatorinefrotoksi~nosti gentamicina su AAP i GGT.

JMB 2008; 27 (2) 299

overweight patients and controls (A2 vs. B2): p<0.005for insulin and p<0.01 for the HOMA index. Therewere no differences in glucose levels between groupsand subgroups. Based on these results, we can con-clude that IR is present in a significant number of nor-mal weight PCOS women. The HOMA index can beused as a screening test for IR and for selecting PCOSwomen most likely to respond to therapy for improve-ment of insulin sensitivity.

E77NEPHROTOXIC EFFECTS OF GENTAMICIN IN PATIENTS IN THE NEONATAL PERIOD

T. Vuji}1, B. Davidovi}2, S. Uletilovi}2, J. Predojevi}-Samardì}3, D. Mal~i}3, @. Sani~anin2

1Drug Administration Agency of the Republic Srpska, Banjaluka

2Faculty of Medicine, University of Banjaluka, 3Pediatric Clinic, Clinical Centre of Banjaluka,Republic of Srpska, Bosnia and Herzegovina

From the results obtained throughout our earlierstudy it can be concluded that even normal gentamicindoses in children patients aged 3–10 years old of bothsexes, express nephrotoxic effects followed by elevat-ed enzymatic activities of alaninaminopeptidase (AAT,EC 3.4.11.2), gamma-glutamyltransferase (GGT, EC2.3.2.1) and N-acetyl-beta-D-glucosaminidase (NAG,EC 3.2.1.30) i.e. enzymes dominantly localized withinthe proximal tubules. Patients in the neonatal periodshow lower renal accumulation of aminoglycosides,which can be explained by the morphometric and func-tional characteristics, and there would be less renal tox-icity. We studied the enzyme activities in 12-hour urinesamples of the proximal tubular enzymes, AAP, GGTand NAG, in 30 patients who received gentamicin andin 30 similar control patients. The patients were of bothsexes, all in the neonatal period. The tretment was con-ducted during a period of 10 days using gentamicin indoses of 2.5 mg/kg body weight daily. A significantdifference in AAP and GGT, in units/mmol creatinine,between the experimental and control group wasnoted on the 8th day (p<0.01) (earlier, in adults on the2nd day) and no significant differences in NAG activi-ties were noted between the experimental and controlgroup (in adults we had noted a significant differenceon the 8th day). The date indicate that the renal toxic-ity of aminoglycosides seems to be less frequent innewborn infants compared to adults even though glo-merular filtration mechanisms are subjected to adap-tive processes during the neonatal period, and in pa-tients in the neonatal period only high urinary AAP andGGT levels are among the earliest and extremely sen-sitive indicators of nephrotoxicity.

E78RENAL INSUFFICIENCY BIOMARKERS

IN DIABETES MELLITUS TYPE II PATIENTS

Lj. Bokan, Z. Ra{i}-Milutinovi}, G. Peruni~i}, D. Mili}evi}

Laboratory Service Department of Endocrinology,Department of Nephrology,

Hospital Center Zemun, Belgrade, Serbia

Renal failure (RF) is a chronic manifestation ofdiabetes melitus (DM). Early recognition of factors thatspeed up RF development in DM patients is veryimportant. This cross-sectional study included DM typeII patients with hypertension (25), 13 patients with RF(Gr1), average age 64.67 years, elevated body mass(BMI=28.79 kg/m2) with blood pressure 151.67/91.67mmHg) and 12 patients without RF (Gr2), average age70 years, similar elevated body mass (BMI= 28.50kg/m2) with blood pressure 138.33/78,33 mmHg. Thecontrol group (Gr3) included 12 voluntary blood donorswithout DM presence, average age 51.57 years, with mildto moderate hypertension (TA=153/96 mmHg).Amongst all tested biochemical serum parameters. Wefound significant differences in glucose (p1/3=0.06;p2/3= 0.007), urea (p1/3= 0.009), creatinine(p1/3=0.02), microalbuminuria/ proteinuria (p1/3=0.007) and age (p1/3=0.002; p1/2=0.02). Indepen-dent predictors of RF development, in the tested pop-ulation with DM type II, were (after age and hyperten-sion presence adjustments) glycemia, urea, creatinineand microalbuminuria/proteinuria (R2=0.943).

E79SYNERGISTIC EFECTS OF NUTRITION, PHYSICAL ACTIVITY AND BEHAVIORAL

THERAPY ON GLUCOREGULATION PARAMETERS IN PATIENTS

WITH SYNDROME X

M. Vidin1, K. Go{i}1, O. Vranje{evi}1, M. Bukilica1,Lj. Tushup-Petrovi}2, V. Simovska3

1Institute of Rheumatology, Belgrade, Serbia2Public of Health Center »Savski venac«,

Belgrade, Srbija3Public of Helth Institute of Sports Medicine,

Nutrition Unit, Skopje, Makedonia

Syndrome X (Sy X) is estremely potent athero-genic risk factor for most CVD. With its insulin resist-ance (IR), Syndrom X is a prephase of DM type 2, ie.prediabetes. Gerald Reaven in 1988 made a hypothe-sis that syndrome X is a cluster of insulin resistance (IR)and at least one atherogenic risk factor (ATRF), but in1998 the diagnostic criteria (DG) were changed by theWHO: insulin resistance and at least two ATRF (Alberti

E78BIOMARKERI RENALNE INSUFICIJENCIJE

KOD BOLESNIKA SA DIJABETES MELITUSOM TIPA II

Lj. Bokan, Z. Ra{i}-Milutinovi}, G. Peruni~i}, D. Mili}evi}

Laboratorijska slu`ba, Endokrinolo{ko odeljenje,Nefrolo{ko odeljenje

Klini~ko bolini~ki centar Zemun, Beograd

Renalna insuficijencija (RI) jeste hroni~na mani-festacija dijabetes melitusa (DM). Zna~ajno je pra-vovremeno prepoznavanje faktora koji ubrzavaju nas-tanak RI kod obolelih od DM. Ovom presek studijomobuhvatili smo pacijente sa DM tip II i hipertenzijom(25), od kojih je sa RI bilo 13 pacijenata (Gr1), pro-se~ne starosne dobi 64,67 god, prekomerne telesnemase (BMI=28,79 kg/m2) sa tenzijom 151,67/91,67mmHg, i 12 pacijenata bez RI (Gr2), prose~ne starosti70,0 god, sli~ne, prekomerne telesne mase (BMI=28,50 kg/m2) sa tenzijom 138,33/78,33 mmHg.Kontrolnu grupu (Gr3) ~inili su dobrovoljni davaoci krvi(12) bez DM, prose~ne starosti 51,57 god, sa blagomdo umerenom hipertenzijom (TA=153/96 mmHg). Odsvih odre|ivanih biohemijskih parametara u serumu zna-~ajno su se razlikovali glukoza (p1/3=0,06; p2/3=0,007), urea (p1/3=0,009), kreatinin (p1/3=0,02),mikroalbuminurija/proteinurija (p1/3=0,007) kao i sta-rosno doba (p1/3=0,002; p1/2=0,02). Nezavisni pre-diktori za nastanak RI, u ispitivanoj populaciji DM tipa 2,bili su, posle prilago|avanja za ìvotno doba i prisustvohipertenzije, glikemija, urea, kreatinin i mikroalbumi-nurija/proteinurija (R2=0,943).

E79SINERGISTI^KI EFEKTI ISHRANE, FIZI^KEAKTIVNOSTI I BIHEVIORALNE TERAPIJENA GLUKOREGULACIONE PARAMETRE

KOD BOLESNIKA SA SINDROMOM X

M. Vidin1, K. Go{i}1, O. Vranje{evi}1, M. Bukilica1,Lj. Tushup-Petrovi}2, V. Simovska3

1Institut za reumatologiju, Beograd, Srbija, 2Dom zdravlja »Savski venac«, Beograd, Srbija

3Narodni institut sportske medicine, Odeljenje ishrane, Skopje, Makedonija

Sindrom X (Sy X) jeste maksimalno potentni ate-rogeni faktor rizika za veliki broj KVB. Zbog obavezneinsulinske rezistencije (IR) sindrom X ~ini prefazu DMtipa 2, tj. predijabetesa. Gerald Reaven je 1988. godi-ne postavio hipotezu o sindromu X kao klasteru obave-zne insulinske rezistencije (IR) i najmanje jednog atero-genog faktora rizika (ATFR), ali 1998. godine kriteriju-mi za dijagnozu promenjeni su od strane SZO: obavez-

300

na insulinska rezistencija i najmanje dva ATFR (AlbertiKG, 1998. i SZO, 1999.) Mi smatramo da je Sy Xodgovor tela na egzogene stimuluse: prejedanje, sma-njena fizi~ka aktivnost i stres uz uticaj podignutog pra-ga osetljivosti hipotalami~kih glikoreceptora i pove}anekoli~ine abdominalnog (visceralnog) masnog tkiva. Na{cilj je bio da istraìmo sinergisti~ke efekte fizi~ke aktiv-nosti, ishrane i bihevioralne terapije na glukoregulacio-ne poreme}aje kod bolesnika sa sindromom X. Pa-cijenti: sedamdeset patijenata sa Sy X, starosti 51 14,7godina stari podeljeni su u dve grupe (DIJ i BIH). Svakagrupa je sadràla 35 bolesnika (27 ` i 8 m). Zdravakontrolna grupa je brojala 35 osoba (27 `. i 8 m.), 4711,2 god. Dva kombinovana terapijska programa (DIJand BIH) bili su kori{}eni. Bila su sli~na u dijetoterapijii aerobno acikli~nim ve`bama. Program BIH je imaojednu komponentu vi{e: bihevioralnu terapiju. Istraìlismo dve razli~ite faze terapije: prva je trajala dva mese-ca (intenzivni dijetoterapijski program sa paralelnimprogramom fizi~kih ve`bi), druga faza je trajala dvegodine. Terapijski program u celini trajao je dve godinei dva meseca. Svi parametri su mereni paralelno triputa: inicijalno, onda na prvoj kontroli (posle dva me-seca) i kona~no (posle 26 m. od po~etka terapije).Koncentracije serumskog insulina (I) bile su mereneRIA komercijalnim testom (n. vrednosti od 5 do 25mIU/L). Glukoza (Gly) je bila odre|ivana kolorimetri-jskom GOD-PAP tehnikom. Normalne vrednosti bile su3,6–6,1 mmol/L. Koncentracije (STH), bile su merenespektrofluorimetrijski, donja i gornja kvartila (0,762–6,345 mmol/L) i »Quicki index« senzitivnosti periperal-nih tkiva na delovanje insulina je ra~unat (Katz A,2000). Mediana kontrolne grupe je bila 0,338, sniòm i vi{om kvartilom: 0,307 i 0,348. Pored inicijal-nih vrednosti, klasi~nim Anova i Manova tehnikamaistraìli smo intragrupne i intergrupne promene kon-centracija: glukoze Gly, insulina (I) i hormona rasta(STH) i vrednosti »Quicki indexa«. Prose~na koncen-tracija glucoze bila je inicijalno vi{a u obe grupe(p<0,05). I prose~na koncentracija insulina bila jepovi{ena u obe grupe: medijana i (nià i vi{a kvartila)bile su: u grupi BIH = 26,20 (22–34,3) i u DIJ: 27,92(24,6–31,1); zdrava kontrola (med. 16,20 i (11,8–18,9) – donja i gornja kvartila). U obe grupe prose~navrednost »Quicki indexa« inicijalno bila je nià (u grupiBIH: 0,286±0, 0012, i 0,287±0,008 u DIJ). U nor-malnoj kontroli bila je 0,326±0,018 tj. (0,313 nià i0,335 vi{a 0,335 kvartila). Koncentracija STH bila jesnièna u obe grupe tako|e (medijana; nià i vi{a kvar-tila) bile su: (u grupi BIH: 0,711 (0,264–0,65 i 0,728(0,304–2,09) u DIJ. U zdravoj kontroli bila je 1,55, tj.(0,762–6,35), nià i vi{a kvartila. Napravili smo grup-nu tabelu koja pokazuje statisti~ku zna~ajnost razlikaprose~nih vrednosti izme|u dve eksperimentalne grupeu prvoj fazi terapije (inicijalno-prva kontrola) na nivou(p<0,05) i tokom ~itavog perioda od 26 meseci (po{toprogram terapije se smatra uspe{nim samo akobolesnici o~uvaju dobre rezultate najmanje dve godine(stat. zna~ajnost razlika izme|u inicijalnog i finalnogmerenja na nivou p<0,05). Naàlost nije bilo stat. zna-

KG, 1998 and WHO, 1999). We presume that func-tional Sy X that is the body's answer to exogenous stim-ulances: overeating, lowered physical activity and stresswith the contribution of elevated insensitivity of hypo-thalamic glycoreceptors and elevated amount of abdo-minal (visceral) fat tissue. Our purpose was to investi-gate the sinergistic effects of physical activity, nutritionand behavioral therapy on glucoregulatiory distur-bances in patients with syndrome X. Seventy patientswith Sy X, 51 14.7 y. old were divided in two groups(DIJ and BIH). Each group consisted of 35 patients (27f and 8 m). Healthy control numbered 35 persons (27F and 8 M), 47 11.2 y. old. Two combined therapy pro-grams (DIJ and BIH) used were. The patients had sim-ilar dietotherapy and aerobic acyclic exercises pro-grams. The program BIH had one component more:behavioral therapy. We investigated two different phas-es of therapy: first one last two months (intensive die-totherapy program with parallel physical exercises), se-cond phase lasted two years. The therapy programslasted two years and two months in total. All parame-ters were measured paralelly three times: initialy, thenon the first control (after two months) and finally (after26 m. from the beginning of therapy). Concentrationof serum insulin (I) were mesured with RIA commercialtest (n. values from 5 to 25 mIU/L). Glycose (Gly) wasmeasured with colorimetric GOD-PAP technicque.Normal values were 3.6–6.1 mmol/L. Concentrationof STH was measured spectrofluorimetrically, Lower-upper qu.(0.762–6.345 mmol/L) and Quicki index ofpertipheral tissue sensitivity to insulin action were cal-culated (Katz A, 2000). The median in the healthycontrols was 0.338, lower and upper quart were 0.307and 0.348. Beside the initial values, using classicAnova and Manova technics, we investigated inter- andintragroup changes of the conc.: of glycose (Gly), insu-line (I) and growth hormone (STH) and the value ofQuicki index. Mean conc. of glucose were higher inboth exp. groups initialy (p<0.05). Mean conc of insu-lin were higher in both exp. groups: (medians andlower and upper quartile) were: in group BIH = 26.20(22–34.3) and in DIJ: 27.92 (24.6–31.1), respective-ly; healthy control med. 16.20 (11.8–18.9) up-lowquartile. In both exp. groups, means of Quicki indexinitialy were lowered (in BIH group: 0.286 ± 0, 0012,and 0.287±0.008 in DIJ), respectively. In normal con-trols it was 0.326 ± 0.018 (ie. 0.313 lower and 0.335upper quartile). Conc. of STH were lowered too in bothexp. Groups (med.; lower and upper quartile) in BIHgroup: 0.711 (0.264–1.65), and 0.728 (0.304–2.09)in DIJ, respectively. In normal controls in median was1.55 ie. (0.762–6.35, lower and upper quartile). Wemade a group table which represents the stat. signifi-cance of the differencies of means beetwen two exp. ingroups in the first phase of th. (initial-first control) atlevel p<0.05 and during the whole period of 26months (because the therapy program is efficient onlyif the patient keeps good results for two years (stat. sig-nificances of stat differences beetwen inicial-final mea-surement, at level p<0.05). Unfortunately there is no

JMB 2008; 27 (2) 301

statistically significant difference beetwen the inicialand final value of means of Gly, I, Quicki index andSTH in Group DIJ but the same measurements weresignificantly different in group DIJ (behavioral therapy).At The first Yugoslav Congress on obesity (1992) prof.M. Pecelj-Gec said that all intentions to eliminate glu-coregulation disturbances in patients with Sy X are onlySisyphean work bringing no permanent bihevioralchanges. This examination proved that.

E80OBESITY – RISK FACTOR FOR GALLSTONE DEVELOPMENT

J. \or|evi}1, D. Tomi}2, D. Davidovi}3

1Institute of Medical Biochemistry, 2Institute of Gastroenterology,

Clinical Center of Belgrade, Belgrade, Serbia3Institute for Hygiene, Medical University,

Belgrade, Serbia

Cholelitiasis is one of the most common diseasesof the digestive system. Obesity is an important risk fac-tor for gallstone development. The aim of this studywas to evaluate the impact of obesity and standard bio-chemistry parameters on gallstone development.Cholelitiasis patients (19 male and 37 female) wereevaluated as well as control grupu (20 male and 41female), with similar demographic characteristics:stature, education, and marriedge status. The allimen-tation was estimated based on antropometric and bio-chemistry parameters. Level of allimentation wasexpressed through body mass index (BMI). Standardbiochemistry parameters (albumine, proteins, CRP, glu-cose, iron, TIBC, cholesterol, HDL- and LDL- choles-terol, tryglicerides, bilirubin, uric acid, and enzymeactivities AST, ALT, ALP, g-GT, a-amilase) were analyzedon Vitros 350 Ortho Clinical Diagnostics system(Johnson & Johnson company). There were confirmeda statistical difference in BMI values between two eval-uated groups (p 0,01). Patients with cholelitiasis werewith higher levels of BMI compared to control group.Lower HDL level and high tryglicerides were connect-ed with high risk for gallstone development.Hiperuricemia increase risk for gallstone development2.5 times (Exp (B) = 2.519; p=0.046), and hyper-glicemia increase risk for 3 times in cholelitiasis pa-tients compared to control group. CRP level higherthan 3 mg/L increase risk for gallstone development.In conclusion, it is very important to keep from obesityto decrease a risk for gallstone development and keeptotal energy diet in recommended levels.

~ajne razlike izme|u inicialnog i finalnog prose~nihvrednosti Gly, I, »Quicki indeksa« i STH u grupi DIJ, aliista merenja dala su zna~ajne razlike u grupi BIH (bihe-vioralna terapija.) Na prvom jugoslovenskom kongresuo gojaznosti (1992) prof. M. Pecelj-Gec rekla je da svanastojanja da se elimini{u glukoregulacione disturban-cije kod bolesnika sa sindromom X samo su Sizifovposao bez stalne promene pona{anja. Ovo istraìvanjeje to dokazalo.

E80GOJAZNOST KAO FAKTOR RIZIKA ZA

POJAVU @U^NIH KAMENACA

J. \or|evi}1, D. Tomi}2, D. Davidovi}3

1Institut za medicinsku biohemiju,2Institut za gastroenterologiju, KCS, Beograd, Srbija

3Institut za higijenu, Medicinski fakultet, Beograd, Srbija

Holelitijaza je jedna od naj~e{}ih bolesti diges-tivnog trakta. Gojaznost predstavlja zna~ajan faktor urazvoju ù~nih kamenaca. Cilj ovog rada bio je da seproceni povezanost gojaznosti i standardnih biohemij-skih parametara sa pojavom ù~nih kamenaca. Ispi-tivanje je obuhvatilo grupu pacijenata sa holelitijazom(19 mu{karaca/37 ̀ ena) i kontrolnu grupu (20 mu{ka-raca/41 èna) pribli`no istih demografskih karakteristi-ka: starosne dobi, obrazovanja i bra~nog statusa. Pro-cena uhranjenosti ispitanika izvr{ena je na osnovuantropometrijskih i biohemijskih parametara. Stanjeuhranjenosti izraèno je preko indeksa telesne mase(BMI), a standardne biohemijske analize (albumin, pro-tein, CRP, glukoza, gvo`|e, TIBC, ukupan, HDL i LDLholesterol, trigliceridi, bilirubin, mokra}na kiselina, kaoi aktivnosti enzima AST, ALT, ALP, g-GT, a-amilaze)ura|ene su na aparatu »Vitros 350« (»Ortho-ClinicalDiagnostics«, Johnson & Johnson company). Utvr|enaje statisti~ki zna~ajna razlika u vrednostima BMI izme|udve ispitivane grupe (p<0,01), a bolesnici sa holeliti-jazom su imali prose~no ve}i BMI u odnosu na kon-trolnu grupu. Snièn nivo HDL holesterola i visok nivotriglicerida su zna~ajno povezani sa pove}anim rizikomza nastanak ̀ u~nih kamenaca. Hiperurikemija 2,5 putapove}ava {ansu za nastanak te bolesti (Exp (B) =2,519; p=0,046), a hiperglikemija ~ak 3 puta kodosoba sa holelitijazom, u pore|enju sa zdravim osoba-ma. Nivo CRP-a iznad 3 mg/L zna~ajno pove}ava {an-su za nastanak kalkuloze ù~ne kese. Moè se zaklju~itida je u cilju smanjenja rizika va`no spre~iti pojavu go-jaznosti i odràvati ukupan energetski unos u prepo-ru~enim granicama.

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E81NOVA TEHNOLOGIJA U UPRAVLJANJU

MEDICINSKIM OTPADOM U SLU@BI LABORATORIJSKE DIJAGNOSTIKE

DOMA ZDRAVLJA »NOVI SAD«

B. Vukeli}, J. O{ap, I. Skeli}

Slu`ba laboratorijske dijagnostike,Dom zdravlja »Novi Sad«, Novi Sad, Srbija

Brojne analize stanja ìvotne sredine u RepubliciSrbiji pokazale su da neadekvatno postupanje saotpadom predstavlja jedan od najve}ih ekolo{kih prob-lema na ovom prostoru. Po preporuci Evropske agen-cije za rekonstrukciju i Ministarstva zdravlja RS i nakonkarakterizacije, uvedena je trijaà ~vrstog zaraznogzdravstvenog otpada u Domu zdravlja, te tako i u Slu`bilaboratorijske dijagnostike. Kako Slu`ba laboratorijskedijagnostike uzima uzorke biolo{kog materijala za oko1000 pacijenata, nametnuo se problem na koji na~inupravljati zaraznim zdravstvenim otpadom, a ne dozvo-liti {tetan uticaj na zdravlje ljudi i ìvotnu sredinu.Nakon vi{egodi{njih bezuspe{nih poku{aja spaljivanjaigala, Dom zdravlja je nabavio sistem za dezinfekciju isterilizaciju ~vrstog zaraznog zdravstvenog otpada –»SINTION« sa drobilicom »SHREDTION«. Tehnologijarada tog aparata se zasniva na kombinaciji vakuuma,mikrotalasne energije i zasi}ene pare, {to omogu}avaefikasno uni{tavanje mikroorganizama u razli~itimmaterijalima i u najuìm {upljinama. Svi ciklusi rada supotpuno automatizovani. Sistem je zatvoren i operaterna ure|aju nije izloèn nikakvoj opasnosti od emisija,otpadnih voda ili struje. Proces upravljanja otpadom una{oj laboratoriji zapo~inje trijaòm, pakovanjem u spe-cijalne plasti~ne kese i transportom do sistema speci-jalnim namenskim vozilom, gde pomo}u pomenutogaparata na zdravstveno i ekolo{ki najprihvatljiviji na~inre{avamo problem zaga|ivanja ìvotne sredine.Prakti~na iskustva u re{avanju problema adekvatnogupravljanja otpadom govore da se ova pitanja mogure{iti samo stalnom i kontinuiranom saradnjom i edu-kacijom svih subjekata na celom prostoru, a u okvirusistema za{tite ìvotne sredine.

E82TRIJA@A MEDICINSKOG OTPADA U DOMU ZDRAVLJA »NOVI SAD«

K. [kori}, J. O{ap, I. Frankovi}

Slu`ba laboratorijske dijagnostike,Dom zdravlja »Novi Sad«, Novi Sad, Serbija

Dosada{nja praksa upravljanja, rastu}a produk-cija, i prate}i zdravstveni rizici aktuelizovali su i potenci-rali neophodnost re{avanja problema medicinskogotpada. Zabrinutost javnosti zbog odlaganja medicin-skog otpada raste usled straha od {irenja krvno preno-

E81A NEW TECHNOLOGY FOR MANAGEMENT

MEDICAL WASTE IN THE LABORATORY SERVICE OF HEALTH CENTER »NOVI SAD«

B. Vukeli}, J. O{ap, I. Skeli}

Laboratory Diagnostic Service, Health Center »Novi Sad«, Novi Sad, Serbia

Many analyses of evironmental conditions in theRepublic of Serbia have led to the conclusion that ina-dequate dealing with the waste is one of the greatestecological problems in this area. Following the rec-ommendation from the European Agency of Recon-struction and the Ministry of Health of the RepublicSerbia and after caracterisation we made a selection ofhard infectious medical waste both in the »Novi Sad«Health Center and also in the Laboratory Service. Ourservice of laboratory diagnostics prepares the biologi-cal material of about 1000 people and it has becomea problem to remove tall infectious medical waste toaviod its harmful influence on people’s health and onthe environment. After many years of unsuccessfulattempts to burn the needles, our Health Center haspurchased the disinfection and sterilization system forall kinds of infectious medical waste – »SINTION« withthe division part SHREDTION. It is based on a combi-nation of vacuum, microwave energy and saturatedsteam and this combination allows for effectivedestruction of microorganisms in different materialsand in the tightest of hollows. All work cycles are com-pletely automated. The system is closed and the tech-nician operating the apparatus is not exposed to theinfluence of bad emissions, foul water or the current.The waste management in our laboratory begins with-the division, packing in to special plastic bags andtransportation to the system in a specially designedvehicle where this modern new system helps us to solvethe problem. In practice we have learned that the ma-nagement of waste is a big problem for the whole com-munity. The adequate removal of waste can be achie-ved only through permanent cooperation and educa-tion of the all subjects in the community and within thesystem of environmental protection.

E82MEDICAL WASTE DIVISION IN THE

HEALTH CENTER »NOVI SAD«

K. [kori}, J. O{ap, I. Frankovi}

Laboratory Diagnostic Service, Health Center »Novi Sad«, Novi Sad, Serbia

Former inadequate management practice, grow-ing production and health risks have brought the prob-lem of medical waste to the open line. The public isworried about the transmission of blood diseases andabout toxic materials. Medical waste is all waste from

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health institutions, research institutes and laboratories.In the health center »Novi Sad«, we perform medicalwaste division, as advised by the European Agency forReconstruction and the Ministry of Health of theRepublic of Serbia into five categories: communalwaste, communal recyclable waste, infectious healthwaste, harmful waste and waste to be buried. Thecommunal waste is placed into plastic cans with blackbags inside, and then taken to containers for trans-portation to the waste depot. The recyclable commu-nal waste is collected into bags or boxes and is thensent to be recycled. Infectious health waste is placedinto appropriately marked metall pedal cans with blackbags inside, and sharp things (needles) are moved tosuitable carton boxes. All infectious waste is removedinto specific central cans and from there the waste istransported by suitable vehicles to the system for wastemanagement. There waste is transported into commu-nal waste by a special technology. Harmful waste isspecially marked, locked and protected in the pharma-cy. Some European studies have supplied the data thatonly about 10% of all medical waste is potentially infec-tious and dangerous and, because many medical insti-tutions have no sufficient financial means and equip-ment for adequate waste management, it is necessaryto work very seriously on waste division and make it apart of the daily work routine.

E83BIOCHEMICAL MARKERS AS A TRACER IN FOLLOW-UP OF THERAPY EFFECT ONPATIENTS WITH RHEUMATOID ARTHRIT

T. Gruev1, S. Dimitrov3, D. Spasovski2, N. Marina2, N. Gruev1, A. Grueva1

1Institute for Clinical Chemistry, University Clinical centre, Skopje2Department of Rheumatology,University Clinical centre, Skopje

3Health center, Skopje, Macedonia

The biochemical markers (urinary enzymes,albuminuria and serum Cystatin C ) was investigatedin patients with RA to evaluate the eventually toxicityof non-steroidal anti-inflammatory drugs( NSAIDs).Urinary enzyme activity of b-NAG,AAP, g-GT wasdetermined with standardised kinetic methods. Uri-nary albumin concentration and serum Cystatin Cwas measured immunoturbidimetric methods usingDAKO tests. Mean +/– SD values of urinary NAG,AAP and g-GT In seropositive (N=56), in seronega-tive RA(N=32) patients and in normally subjects(N=30) were found to be: b-NAG (2,27+/–0.65,1.26+/– 0.34,0.75+/–0.43 U/mmol creatinine);AAP(2.98+/–0.36,1.30+/–0.74,0.50+/–0.35U/mmol creatinine), g-GT(3.84+/– 0.30,1.36+/–

sivih bolesti, kao i izlaganja toksi~nim materijama.Medicinski otpad se defini{e kao sav otpad koji nastajeu zdravstvenim institucijama, istraìva~kim ustanovamai laboratorijama. U Domu zdravlja »Novi Sad« vr{i serazdvajanje celokupnog otpada, prema preporukamaEvropske agencije za rekonstrukciju i Ministarstvazdravlja RS, u pet kategorija: komunalni, komunalnikoji je mogu}e reciklirati, zarazni zdravstveni otpad,{tetni otpad i otpad koji se zatrpava. Komunalni otpadse odlaè u plasti~ne kante sa crnim kesama, a zatim ukontejnere koje komunalno preduze}e transportuje dodeponije. ^vrsti komunalni otpad koji je mogu}e reci-klirati sakuplja se u odgovaraju}e kese ili kutije i pro-sle|uje na reciklaù. ^vrsti zarazni zdravstveni otpad sepo slu`bama odlaè u adekvatno ozna~ene metalne,pedalne kante sa poklopcem i crnom kesom kaoulo{kom, dok se o{tri predmeti (igle) odlaù u namen-ske, kartonske kutije. Celokupan zarazni zdravstveniotpad se odlaè u centralne tipske kante, odakle setransportuje namenskim vozilom do sistema za uprav-ljanje otpadom, gde se specijalnim tehnolo{kim pos-tupkom prevodi u komunalni. [tetni otpad se ~uvazaklju~an u apoteci i posebno obeleèn. Budu}i daevropske studije pokazuju da zarazni zdravstveni otpad~ini najvi{e 10% ukupne koli~ine medicinskog otpada,neophodno je ozbiljno shvatiti zna~aj pravilne trijaèotpada i uvesti je u svakodnevnu praksu.

E83BIOHEMIJSKI MARKERI KAO

SMERNICA U PRA]ENJU EFEKTA TERAPIJE KOD PACIJENATA

SA REUMATOIDNIM ARTRITISOM

T. Gruev1, S. Dimitrov3, D. Spasovski2, N. Marina2, N. Gruev1, A. Grueva1

1Institut za klini~ku hemiju, Univerzitetski klini~ki centar, Skoplje,

2Odsek za reumatologiju, Univerzitetski klini~ki centar, Skoplje

3Zdravstveni centar, Skoplje, Makedonija

Biohemijski markeri (enzimi u urinu, albuminurijai cistatin C u serumu) ispitivani su kod pacijenata sa RAkako bi se procenila mogu}a toksi~nost nesteroidnihantiinflamatornih lekova (NSAID). Enzimska aktivnostb-NAG, AAP, g-GT u urinu odre|ena je standardizo-vanim kineti~kim metodama. Koncentracije albuminau urinu i cistatina C u serumu merena je imunotur-bidimetrijskim metodama primenom »DAKO« testova.Srednje vrednosti ± Sd NAG, AAP i g-GT u urinu kodseropozitivnih (N=56), seronegativnih RA (N=32)pacijenata i zdravih osoba (N=30) prona|ene su: b-NAG 2,27±0,65; 1,26±0,34; 0,75±0,43 U/mmolkreatinina; AAP 2,98±0,36; 1,30±0,74; 0,50±0,35U/mmol kreatinina; g-GT (3,84±0,30; 1,36±0,28;1,32±0,50 U/mmol kreatinina). Utvr|ene srednje

304

vrednosti NAG i g-GT u urinu kod RA pacijenata su bilezna~ajno vi{e (p<0,01) u pore|enju sa srednjim vred-nostima kod seronegativnih RA pacijenata i normalnihzdravih osoba kada su analizirani one-way ANOVAtestom. Kod pacijenata koji su tretirani metatreksatomi dekortinom pokazano je da su koncentracije cistatinaC u serumu zna~ajno povi{ene u toku terapije (2,85±0,56 vs. 0,70±0,50 mg/L). Koncentracija albumina uurinu je bila pove}ana, ali ne zna~ajno, u toku prvihdana tretmana. Odre|ivanje enzima b-NAG i g-GT uurinu, kao i koncentracije cistatina C u serumu moèprema tome sluìti kao osetljiviji test za o{te}enje bu-brega kod pacijenata sa reumatoidnim artritisom. Ranootkrivanje visokih NAG enzimurija i pove}anja nivoaalbumina u urinu pre primene MTX terapije moè bitikorisno za predvi|anje mogu}e toksi~nosti MTX koja jeverovatno povezana sa smanjenim renalnim klirensomMTX.

E84RAZLIKE U HEMATOLO[KIM PARAMETRIMA

KOD PACIJENATA SA DIJABETESOM

S. Panchevska1, T. Gruev1, S. Shubeska-Stratova2, S. Dimitrov3

1Institut za klini~ku biohemiju, Univerzitetski klini~ki centar, Skoplje, Makedonija

2Odsek za endokrinologiju, Univerzitetski klini~ki centar, Skoplje, Makedonija

3Zdravstveni centar, Skoplje, Makedonija

Ova studija bavi se ispitivanjem razlika u hemato-lo{kim parametrima kod pacijenata sa dijabetes meli-tusom (tipa 1 i tipa 2). Objavljen je mali broj studijakoje se odnose na dobru kontrolu dijabetes melitusa ibroja trombocita ili kontrolu funkcije krvnih }elija.Pove}anje kontrolne glikemije kod pacijenata sa tipom2 dijabetesa prouzrokuje redukciju u trombogenosti.Dobra kontrola glikemije moè pomo}i u korelacijivisokog pove}anja koagulobilnosti. U ovoj studiji ispiti-vani su broj eritrocita, leukocita i trombocita, kao idrugih parametara za razlikovanje dva tipa dijabetesa(tip 1 N=160, tip 2 N=180) kod dijabeti~nih pacije-nata i pore|eni sa kontrolnom populacijom. Uzorci krvi(K3EDTA) pacijenata sa dijabetesom tipa 1 ili 2 i kon-trolne grupe (N=50) sa referentnim vrednostima glu-koze su kori{}ene u ovoj studiji. Koncentracija glukozeje odre|ena pomo}u GOD metode, a hematolo{ki pa-rametri na »Sysmex KX-21« i »PENTRA 120« analiza-torima. Rezultati su pokazali pove}anje vrednosti he-matokrita, broja leukocita, limfocita i monocita kodpacijenata sa dijabetesom, dok su MCV, MCH (osetljivna koncentraciju glukoze) i broj neutrofila bili smanjeni.Pacijenti sa dijabetesom tipa 1 pokazali su pove}anjeeritrocita, Hct, leukocita, limfocita (p<0,05) i koncen-tracije glukoze (p<0,01), u pore|enju sa kontrolnom.

JMB 2008; 27 (2) 305

0.28,1.32+/–0.50 U/mmol creatinine. The meanurinary NAG and g-GT values in RA patients wasfound to be a significant higher (p< 0.01) comparedto the mean values in seronegative RA patients andnormal healthy subjects when analysed by one wayANOVA. In patients who were treated with Metha-trexat and Decortin the serum Cystatin C concentra-tion showed significantly elevated values in the ther-apeutic days (2.85+/– 0.56; v.c.g. 0.70 +/–0.50mg/L). Urinary albumin concentration increased, butnot significantly in the first days of treatment. Dete-rmination of the urinary enzymes b-NAG and g-GT,as well as the serum concentration of Cystatin C metherefore serve a more sensitive test for kidney injuryin patients with Rheumatoid arthrit.Early detection ofhigh NAG enzymuria and elevated albumin levels inurine before the initiation of MTX therapy could behelpful in predicting possible MTX toxicity probablyrelated to impaired renal clearence of MTX.

E84DIFFERENCES IN HEMATOLOGY

PARAMETERS IN DIABETIC PATIENTS

S. Panchevska1, T. Gruev1, S. Shubeska-Stratova2, S. Dimitrov3

1Institute of Clinical Biochemistry,University Clinical Centre, Skopje, Macedonia

2Department of Endocrinology, University Clinical Centre, Skopje, Macedonia

3Health Centre, Skopje, Macedonia

This study is related to the investigation differ-ences in hematology parameters in patients with dia-betes mellitus (type 1 and type 2). Few studies relat-ing good control of diabetes mellitus and thrombo-cyte number, or the control of the function of bloodcells have been reported so far. A rise a glycemic con-trol in patients with type 2 diabetes causes a reduc-tion in thrombogenicity. Good glycemic control canhelp in the correlation of increased coagulability. Inthis study erythrocyte, leukocyte and thrombocytecounts were investigated, as well as other parametersfor differentiating between the two types of diabetes(type 1 N=160, type 2 N=180) in relation to a con-trol population. Blood samples (K

3EDTA) from two

diabetic and one control group (N=50) with referen-ce values for glucose were used in this study. The glu-cose concentration was analyzed with the GOD met-hod and hematologic parameters with the »SysmexKX-21« and »PENTRA 120« analyzers. Our resultsshowed an increase in hematocrit, leukocyte, lym-phocyte and monocyte number in diabetic patients,while MCV, MCH (affected by the glucose concentra-tion) and neutrophils number were decreased. Pa-tients with type 1 diabetes showed an increase in ery-throcytes, Hct, leukocytes, lymphocytes (p<0.05)

and glucose (p<0.01) concentration compared withthe control. Hematocrit and thrombocytes are signi-ficantly increased (p<0.05) while neutrophils andmonocytes are decreased (p<0.05) in patients withtype 2 diabetes in relation to type 1 diabetes.

Hematokrit i trombociti su zna~ajno povi{eni (p<0,05),dok su neutrofili i monociti snièni (p<0,05) kod paci-jenata sa dijabetesom tipa 2 u pore|enju sa dijabete-som tipa 1.

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Documents

POSTERSKE SEKCIJE POSTER SESSIONS razlika ima manju zna~ajnost u odnosu na kontrolnu grupu (p