Kniga na apstrakti book of abstrcts

Urinarni proteini kaj febrilni sostojbi od nerenalno poteklo
Svetlana Cekovska
1
, Sne`ana Pal~evska
2
, Velibor Tasi}
2
, Petar Korneti
1
1
Institut za medicinska i eksperimentalna biohemija, 
Medicinski fakultet, 50 Divizija 6, Skopje, R. Makedonija
2
Klinika za detski bolesti, Medicinski fakultet, Vodwanska 17, Skopje, R. Makedonija
Ispituvani se nekolku osnovni parametri povrzani so urinarnata proteinska ekskrecija kaj ispi-
tanici so febrilni sostojbi od nerenalno poteklo.
Analizirani se primeroci na vtora utrinska urina od 30 deca so febrilni sostojbi od nerenalno
poteklo i 24 zdravi deca, site na vozrast od 2-7 godini.
Za sekoj ispitanik se opredeleni: koncentracijata na vkupnite urinarni proteini, P/C soodnosot,
koncentracijata na kataliti~kata aktivnost na enzimot N-acetil-
β-D-glukozaminidaza (β-NAG), a izrabote-
na e i elektroforetska separacija na urinarnite proteini so tenkoslojnata gradientna natrium dodecil
sulfat poliakrilamid gel elektroforeza (SDS-PAGE). Vkupnite urinarni proteini se opredeluvani so
turbidimetriskiot metod spored Meulemans. Koncentracijata na kreatininot vo urinata e odreduvana so
spektrofotometriskiot metod na Jaffe. Aktivnosta na enzimot 
β-NAG e odreduvana so spektrofo-
tometriskiot metod na Horak. Za elektroforetskata separacija se koristeni 4-22% gradientni gelovi (za
Multiphor II sistemot), oboeni so Coomassie Blue R-250.
Kolmogorov-Smirnov testot poka`a deka postoi visoko statisti~ki signifikantna razlika vo
ispituvanite parametri me|u kontrolnata grupa i decata so febrilni sostojbi od nerenalno poteklo (r
< 0,002 za vkupnite urinarni proteini; p < 0,001 za P/C soodnosot i p < 0,001 za 
β-NAG). Vrednostite za vkup-
nite urinarni proteini, P/C soodnosot i aktivnosta na enzimot 
β-NAG pretstavuvaat sredna vrednost od
ispituvaweto vo tek na dva posledovatelni denovi vo koi kaj ispituvanite deca e zabele`ana zgolemena
telesna temperatura (> od 38 
o
C). Najvisoki vrednosti za P/C soodnosot i za koncentracijata na katali-
ti~kata aktivnost na enzimot 
β-NAG se zabele`ani kaj decata so virusna infekcija na gornite respirator-
ni pati{ta. Po primaweto na terapijata i namaluvaweto na telesnata temperatura utvrdeno e brzo nama-
luvawe na vrednostite na site ispituvani parametri, koi po 3 dena gi dostignuvaat vrednostite
karakteristi~ni za zdravi deca.
Kaj ispitanicite so zgolemena telesna temperatura od nerenalno poteklo zabele`an e karakte-
risti~en elektroforetski profil, so izrazena albuminska frakcija i izrazeni frakcii na niskomoleku-
larni proteini. Po primaweto na terapijata i namaluvaweto na telesnata temperatura, elektroforet-
skite profili gi dobivaat karakteristikite na zdravi lica.
Rezultatite govorat vo prilog deka proteinurijata koja se javuva kaj febrilni sostojbi od nere-
nalno poteklo e funkcionalna i tranzitorna.
Macedonian pharmaceutical bulletin 53 (1,2) 257-258 (2007)
PP - 125
258
^ETVRTI KONGRES NA FARMACIJATA NA MAKEDONIJA SO ME\UNARODNO U^ESTVO
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Serum Cystatin C concentracion in transplanted patients treated 
with glucocorticoid immunosuppression
T. Gruev
1
, M. Bogdanova
1
, K. Chakalarovski
2
, L. Petrushevska-Tozi
3
,
1
Institute for Clinical Biochemistry, University Clinical centre, Skopje, Macedonia
2
Department of Nephrology, University Clinical centre, Skopje, Macedonia
3
Faculty of Pharmacy, Skopje, Macedonia
Cystatin C has been described as meeting many of the characteristics of an ideal GFR marker and has been
reported to be at least as accurate as the commonly used serum creatinine to detect impaired renal function in vari-
ous patients groups, including renal transplant patients.The present study aimed to elucidate the influence of glucor-
ticoid immunosuppression on Cystatin C concentration in serum from renal transplant patients.To evaluate the influ-
ence of immunosuppressive regimens, especially glucocorticoids, on serum Cystatin C, 38 clinically stable patients
on immunosuppression therapy with low-dose glucocorticoids were matched with 30 clinically stable patients receiv-
ing cyclosporin A alone and 18 clinically stable patients receiving cyclosporin A together with azathioprine .The
three groups were matched for estimated creatinine clearance (CrCl) and had comparable gender, age, and time since
transplantation. To reduce the influence of the known biologic variation of Cystatin C,all patients had six measure-
ments during subsequent visits that demonstrated stable clinical condition. Means from cystatin C reciprocals, as
well as from CrCl estimates and CysCGFR were calculated and used for data analysis. Furthermore, 10 patients
receiving a short course of high-dose methylprednisolone (500 mg intravenously per day for 3 days) for deteriorat-
ing renal function were analyzed to observe a possible dose-dependent effect of glucocorticoid administration. The
group receiving short-course, high-dose methylprednisolone had results from four time points available: a) before
methylprednisolone commencement (median, 15 days); b) the day methylprednisolone was started (before medica-
tion); c) after 3 days of methylprednisolone therapy; and d) on a follow-up 9-10 days after last dose. Serum Cystatin
C was measured by a particle-enhanced turbidimetric immunoassay (PETIA; Dako) on a Cobas Mira (Roche) . Serum
creatinine was measured with a modified kinetic Jaffe method. Creatinine clearance was estimated by the formula of
Cockroft and Gault,and the Cystatin C GFR was measured by the formula of Grubb (CysCGFR=84.69.CysC 
-1.680
).
Patients receiving long-term, low-dose glucocorticoid therapy showed higher cystatin C concentrations than con-
trols(2,25;1.9-2.9,P<0.05). High-dose methylprednisolone given intravenously led to significant differences in Cystatin
C values at different time points (before administration, after three doses, and 8 days after discontinuation; P <0.001).
After three daily doses of 500 mg, cystatin C concentrations increased from 2.13 mg/L (1.72–2.80,p<0.05) to 3.69 mg/L
(2.84–4.35; P <0.001). Eight days after discontinuation, cystatin C concentrations significantly decreased to 1.96 mg/L
(1.63–2.4; P <0.05. At these time points, neither the CrCl estimate (54 ± 13 mL · min
-1
· 1.73 m
-2
, 51 ± 15 mL · min
-1
· 1.73 m
-2
, and 56 ± 14 mL · min
-1
· 1.73 m
-2
; P = 0.05) The serum creatinine concentrations (165 µmol/L, 158 µmol/L, and
162 µmol/L, P = 0.19) underwent significant changes. This study demonstrates that renal transplant patients receiv-
ing glucocorticoid medication have higher cystatin C than two comparable groups with glucocorticoid-free immuno-
suppression. Because patients receiving 500 mg of methylprednisolone had significantly higher cystatin C values
than patients receiving 10 mg of prednisone, a dose-dependent influence of the administered glucocorticoid dose is
suggested. Thus, glucocorticoid medication leads to systematic underestimation of GFR in renal transplant patients.
Macedonian pharmaceutical bulletin 53 (1,2) 259-260 (2007)
PP - 126
259
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Koncentracija na Cistatin C kaj transplantirani pacienti 
tretirani so glukokortikoidnata imunosupresivna terapija
T. Gruev
1
, M. Bogdanova
1
, K. ^akalarovski
2
, L. Petru{evska-Tozi
3
,
1
Institut za klini~ka biohemija, UKC, Skopje, Makedonija
2
Klinika za Nefrologija, UKC, Skopje, Makedonija
3
Farmacevtstki fakultet, Skopje, Makedonija
Cystatin C e opi{an prema svoite karakteristiki kako idealen GFR marker i prema dosega{nite
soznanija mo`e da se koristi vo sledewe na bubre`nata funkcija kaj razli~ni zaboluvawa, vklu~uvajki
gi i transplantiranite pacienti. Ova studija ima za cel da uka`e na vlijanieto na glukokortikoidnata
imunosupresija vrz koncentracijata na serumskiot CysC kaj transplantirani pacienti. Vlijanieto e sle-
deno kaj 38 klini~ki stabilni transplantirani pacienti so imunosupresivna terapija so niski dozi na
glukokortikosteroidi sporedeniso 30 klini~ki stabilni pacienti so Cyclosporin A terapija i 18 klini~ki
stabilni pacienti na terapija so Cyclosporin A i Azathioprine. Trite grupi se sporeduvani preku odreduva-
we na kreatinin klirensot(CrCL), starosta i vremetraeweto po transplantacijata. Za ostranuvawe na
vlijanieto na poznati biolo{ki varijabli vrz CysC kaj site pacienti se vr{eni {est odreduvawa za
uka`uvawe na stabilna klini~ka sostojba. Kaj drugi 10 pacienti e primenuvana kratkotrajna (tri dnev-
na) terapija so 500 mg na metilprednizalon intravenski. Efektot e sleden 9-10 dena pred terapijata, vo
tekot na terapijata i 9-10 dena po zavr{uvaweto na terapijata. Koncentracijata na CysC e merena imuno-
turbidimetriski (DAKO) testovi, a taa na kreatininot so Jaffe metodata. Klirensot na kreatininot
e odreduvan prema Cockroft-Gault formulata, a na CysC prema Grubb (CysCGFR=84.96.CysC.-
1.680)
. Paci-
entite koi primat dolgotrajna terapija so prednizolon poka`uvaat signifikantno zgolemuvawe na CysC
(2.34-3.5 mg/L,p< 0.05). Dozi na metilprednizolonot(500 mg) predizvikuvaat visoko signifikantno
zgolemuvawe na CysC koncentracijata vo zavisnost od vremeto na odreduvaweto (najizrazeno vo deno-
vite na terapijata,vo po~etakot 2.13mg/L (1.72-2.80, p<0.05) do 3.69 mg/L (2.84-4.35, p<0.001). Osum dena po
terapijata signifikantno se namaluva koncentracijata na 1.96 mg/L (1.63-2.4, p<0.05). Vo isto vreme e
odreduvan i klirensot na kreatininot koj ne poka`a signifikantni otstapuvawa za razlika od ovoj
na cistatinot (54 ± 13 mL · min
-1
· 1.73
m-2
, 51 ± 15 mL · min
-1
· 1.73 
m-2
, and 56 ± 14 mL · min
-1
· 1.73 m
-2
; P = 0.05).
Koncentracijata na serumskiot kreatinin be{e 165 mmol/L,158 mmol/L I 162 mmol/L,P=0.19). Ova studija
uka`uva deka transplaniranite pacienti podlo`eni na terapija so gkukortikoidi imaat povisoki
vrednosti na CysC sporedeni si kontrolnata grupa, pogotovo ako se primenuvaat visoki dozi. Nivnata
primena bara kontinuirano sledewe na sostojbata na glomerulranata filtraciona brzina kaj site
bubre`no transplantirani pacienti.
Macedonian pharmaceutical bulletin 53 (1,2) 259-260 (2007)
PP - 126
260
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Fluoride Levels in Enamel Samples of Children 
from Fluorotic Region of Vranje
Z. Mandinic
1
, M. Curcic
2
, B. Antonijevic
2
, M. Nedeljkovic
2
, M. Carevic
1
1
Clinic of Preventive and Pediatric Dentistry, Faculty of Dentistry, University of Belgrade, Dr Subotica 11, Belgrade, Serbia
2
Institute of Toxicology, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, Belgrade, Serbia
During a process of amelogenesys in early childhood, fluorides are of particular importance for the integrity of
teeth. However, long term intake of water containing large amounts of fluorides may induce development of fluorosis.
Aim of this work was to examine fluoride content in enamel samples taken from children who live in a fluo-
rotic region (Vranje city) that is known of high fluoride concentration in drinking water. Children from non-florinat-
ed region (Valjevo city) were included as a control group. This study was approved by Ethical Committee of Faculty
of Dentistry, Belgrade University (No.727/1; 2006). Fluoride content was determined potentiometrically by means of
ion-selective electrode.
Average fluoride concentrations in enamel samples and drinking water samples from Vranje region were
182.2 
µg F-/g (182.2 ppm), and 10 mg/L (10 ppm), respectively. Obtained results indicated that fluoride levels in
drinking water of Vranje were significantly higher than levels recommended by WHO (0.7-1.2 ppm). Consequently,
fluoride levels in enamel samples were higher than in samples taken from children who live in region where fluo-
rides content in water is in the range of recommended values. 
Results of this pilot study indicated some quantitative correlation between fluoride content in enamel and
water concentration of fluoride. However further studies are needed to assess exposure model in relation to water
consumption, and consequent fluoride intake.
Macedonian pharmaceutical bulletin 53 (1,2) 261 (2007)
PP - 127
261
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Involvement of NMDA receptors in diquat neurotoxicity 
Marijana Curcic Jovanovic, Mirjana Djukic, Milica Ninkovic, Ivana Maksimovic, Marina Jovanovic
Institute of Toxicological – academician Danilo Soldatovic, Faculty of Pharmacy, Belgrade, 
Vojvode Stepe 450, 11221 Belgrade, Serbia
Institute for Medical Research, Military Medical Academy, Crnotravska 17, 11000 Belgrade, Serbia 
Nitrate, longlasting endproducts of nitric oxide (NO) metabolism, in brain regions can be used as a marker
of diquat (DQ) neurotoxicity. In order to reveal mechanisms of DQ neurotoxicity throughout NMDA receptors, 
2-amino-5-phosphonovaleric acid (APV), an NMDA receptor antagonist, was used. Influence of APV pretreatment
in DQ poisoning was estimated by measuring nitrate levels in striatum homogenates of Wistar rats. Diquat was intras-
triatally (i.s.) applied in single dose of 50 mg/kg, and APV was administrated, in the same manner, in single dose of
100 mg/kg, 30 minutes before DQ. It is known that APV exerts its effects via NMDA receptors and Ca
2+
signaliza-
tion pathways, while trigger NO production. Moreover, i.s. DQ administration promotes NOS activity, and on the
top of the results obtained in the experiment, anticipated high nitrate concentration occurred in brain regions.
Achieved nitrate levels (NO
3
- nmol/mg of proteins) in Wistar rats’striatum homogenates depend on treatment
and time. Group pretreated with APV did not show statistically significances in both, ipsilateral and contralateral
striatum comparing to control group (16.35 ± 4.09 nmol/mg of proteins), 30 min, 24h and 7 days after the treatment.
Contrary, nitrate levels in the striatum of Wistar rats poisoned with DQ were significantly higher after the treatment.
Efficacy of the APV was confirmed in all time intervals. Based on obtained results, this pilot study cofirms NMDA
receptors involvement in DQ neurotoxicity and NO is probabdly one of the messengers involved in this mechanism.
Macedonian pharmaceutical bulletin 53 (1,2) 262 (2007)
PP - 128
262
^ETVRTI KONGRES NA FARMACIJATA NA MAKEDONIJA SO ME\UNARODNO U^ESTVO
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Determination of diazepam in serum by GC- ECD
Suzana Angelova
1
, Desa Jakimova
1
, Lidija Petrusevska-Tozi
2
1
Institute of Public Health, 11 Oktomvri bb, 1300 Kumanovo, R. Macedonia
2
Faculty of Pharmacy “Ss Cyril and Methodius” University, Vodnjanska 17, 1000 Skopje, R. Macedonia
Rapid proven of the benzodiazepines in blood/ plasma/ serum or/and urine in the clinical practice is done
by immunochemical techniques (FPIA, RIA, EIA…) due to improve the rapidness and efficiency of the appropriate
therapy. These techniques are simple and sensitive which make them applicable to routine use.
The presence of the conjugation metabolite and lack of the specification lead to false negative and/or false
positive results. Due to these facts there is a need of another more sophisticated method which would provide a big-
ger benzodiazepines specifics. 
Due to this a procedure of simultaneous separation, identification and quantification of some benzodiazepines
in serum is developed, by gas chromatography with ECD detection. Optimal conditions for simultaneous separation
of benzodiazepines from mixture are made with choose of the HP-5 columns, with 5% phenyl methyl silicon sta-
tionary phase as well as choose of a gradient temperature program (which starts from 140
0
C for 1 min; 30
0
C/min to
260
0
C for 5 min; 50
0
C/min to 300
0
C). To extract the benzodiazepines from serum Extrelut columns with capacity
of 3 ml are used. Their usage excludes the protein denaturation procedure, time reduce, high analytic income improve-
ment and results reproducibility.
This method is used to identify and quantify the diazepam in serum after administration of its single thera-
peutic dose.
Upon the surface of the peaks and with usage of the equivalency of the linear function the value of the
diazepam in serum expressed in ng./ml is received.
This method is taken on 9 patients and the received results are presented in the following table. 
Table 1: Results received from diazepam determination in serum.
Presented results are in correlation with the literature dates for a therapeutic concentration. 
Macedonian pharmaceutical bulletin 53 (1,2) 263-264 (2007)
PP - 129
263
^ETVRTI KONGRES NA FARMACIJATA NA MAKEDONIJA SO ME\UNARODNO U^ESTVO
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Patient No.  
1 
2 
3 
4 
5 
6 
7 
8 
9 
C
diazepam
 (ng/ml) 
30.9 
56.4 
82.7 
191.8 
58.3 
194.1 
36.7 
97.9 
134.9 

Primena na gasna hromatografija so ECD detekcija 
vo opredeluvawe na diazepam vo serum
Suzana Angelova
1
, Desa Jakimova
1
, Lidija Petru{evska-Tozi
2
1
JZU Zavod za zdravstvena za{tita, 11 Oktomvri bb, 1300Kumanovo, R. Makedonija
2
Farmacevtski Fakultet, Univerzitet “Sv. Kiril i Metodij”, 
Vodwanska 17, 1000 Skopje, R. Makedonija 
Voobi~aeno, vo klini~kata praksa, brzo doka`uvawe na prisustvo na benzodiazepini vo krv/plaz-
ma /serum i/ili urina se vr{i so imunohemiskite tehniki (FPIA, RIA, EIA...), so cel brzo i efikasno da se
reagira so soodvetna terapija. Se raboti za ednostavni, dovolno osetlivi i ne mnogu skapi analizi, povol-
ni za rutinska rabota.
Me|utoa, poradi prisustvo na kowugirani metaboliti i nedovolnata specifi~nost, mo`e da se
dobijat la`no negativni i/ili la`no pozitivni rezultati. Poradi toa, po`elno e da se napravi potvrda
na rezultatite so nekoja posofisticirana metoda, koja manifestira pogolema specifi~nost kon benzo-
diazepinite.
Za taa cel razrabotena e postapka za ednovremeno razdvojuvawe, identifikacija i kvantifikaci-
ja na nekoi benzodiazepini vo serum so gasna hromatografija so ECD detekcija. Optimalnite uslovi za
ednovremeno razdvojuvawe na benzodiazepini od smesa, se vospostaveni so izbor na HP-5 kolona, so 5%
phenyl methyl silicon stacionarna faza, kako i izbor na gradienten temperaturen program ( se startuva od
140
0
C za 1 min; 30
0
C/min do 260
0
C za 5 min; 50
0
C/min do 300
0
C). Za ekstrakcija na benzodiazepini od serum
koristeni se Extrelut koloni so kapacitet od 3 ml. Nivnata primena ovozmo`uva izbegnuvawe na postapka-
ta za denaturacija na proteinite, namaluvawe na vremetraewe na analizata i obezbeduvawe na visok anal-
iti~ki prinos i reproducibilnost na rezultatite.
Vospostavenata metoda, so dobienite vrednosti za statisti~ki parametri, e primeneta za identi-
fikacija i kvantifikacija na diazepam vo serum, po administrirawe na poedine~na doza na istiot, vo ter-
apevtski celi.
Vrz osnova na dobienite povr{ini za pikovite, so primena na prethodno dobienata ravenka na
linearna funkcija, dobieni se vrednostite za koli~estvoto na diazepam izrazeno vo ng/ml serum.
Napraveno e opredeluvawe na diazepam vo serum na devet pacienti, a dobienite rezultati se pret-
staveni vo Tabela 1. 
Tabela 1: Rezultati od opredeluvawe na diazepam vo serum
Dobienite rezultati pretstaveni vo Tabela 1 se vo korelacija so podatocite za terapevtski kon-
centracii dobieni od literatura. 
Macedonian pharmaceutical bulletin 53 (1,2) 263-264 (2007)
PP - 129
264
^ETVRTI KONGRES NA FARMACIJATA NA MAKEDONIJA SO ME\UNARODNO U^ESTVO
FOURTH CONGRESS OF PHARMACY OF MACEDONIA WITH INTERNATIONAL PARTICIPATION
Pacient br.  
1 
2 
3 
4 
5 
6 
7 
8 
9 
C
diazepam
 (ng/ml) 
30.9 
56.4 
82.7 
191.8 
58.3 
194.1 
36.7 
97.9 
134.9 

Relationship between lipoprotein levels and antioxidant capacity 
in patients with coronary heart disease
R. Gosevska, M. Sudzukovic, E. Bagasovska
PZU RUZA-BIOLAB Mladinski Kej br. 2 Berovo Macedonija
PZU Chemist shop – SOFORA Kej Revolucija br. 3a Berovo Macedonija
Lipid abnormality and deficiency in the antioxidant system are major risk factors for coronary heart disease
and myocardial infarction (MI).Raised plasma triglycerides (TG), free radicals levels and low HDL- cholesterol (HDL-
C) concentration increases coronary heart diseases (CHD) risk in a number of ways in both men and women. It was
analyzed the correlation between TOAS, and lipid parameters as risk factors in patients with myocardial infarction.
The serum levels of total cholesterol (TC), TG, HDL-C and total antioxidant status (TOAS) were determi-
nate in 35 males and 12 females age 43-70 years, 6 to 18 moths after MI. The control group consisted of 79 healthy
individuals, 25-49 years old.
It has been established that differences were significant for all the parameters. TG were increased in all
patients (males:2.10
+
0,99 mmol/L, range:0,85-4,42 mmol/L, females:2,16
+
0,844 mmol/L, range:1,24-5,36 mmol/L)
in comparison with the control group (1,34