Daria Pašalić
Department of Medical Chemistry, Biochemistry and Clinical Chemistry
Zagreb University School of Medicine
Šalata ul 2.
10 000 Zagreb, Croatia
Phone +385 (1) 4590 205; +385 (1) 4566 940
E-mail: dariapasalic [at] gmail [dot] com

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Zec I, Bukovec Megla Ž, Krpan R, Marout J, Petek-Tarnik I, Krilić D. PO6-1: Quantitative automated measurement of gonadotropins and ovarian steroid hormones during the menstrual cycles on the Roche-Ellecsys analyzer. Biochemia Medica 2009;19(Suppl 1):S130.
Laboratory of Endocrinology, Clinics of oncology and nuclear medicine, Sestre milosrdnice University Hospital, Zagreb, Croatia
Corresponding author:ikulas [at] kbsm [dot] hr
Objective: To determine concentrations of hormones relevant for the evaluation of function of pituitary-ovarian axes in fertile women during the menstrual cycle.
Subjects and methods: A total of 35 women aged 32 ± 6 years, with regular menstrual cycles (30 ± 2 days) were selected for the study. Normal hormone concentrations were confirmed by measurement of lutropin (LH), folitropin (FSH), estradiol (E2), total testosterone (T) and sex-hormone binding globulin (SHBG) in follicular phase (cycle day 3-5), LH, FSH and E2 in ovulation (cycle day 13-15) and progesterone (P) in lutheal phase (cycle day 20-23). Measurements of hormones were performed by electrochemiluminiscence immunoassay (ECLIA) on Roche-Ellecsys analyzer. Concentrations of free testosterone (FT) were calculated on the basis of total T and SHBG concentrations.
Results: Normal distributions of measured hormones were confirmed by Kolmogorov-Smirnov test. Mean concentrations of hormones in follicular phase were: LH 5.6 ± 3.0 IJ/L, FSH 7.1 ± 2.0 IJ/L, E2 174 ± 65 pmol/L, total T 1.03 ± 0.5 nmol/L, free T 11.4 ± 5.5 pmol/L and SHBG 73.9 ± 28.8 nmol/L, respectively. Average concentrations of hormones in ovulatory phase were: LH 11.9 ± 7.0 IJ/L, FSH 6.0 ± 2.1 IJ/L, odnosno E2 734 ± 477 pmol/L. Mean concentration of P in luteal phase was 48.4 ± 19.5 nmol/L.
Conclusion: Determination of hormone levels on automated Roche-Ellecsys platform enables simple and fast measurements of relevant parameters for women’s menstrual cycles. This is particularly important when assessment of hormones is needed for appropriate classification and treatment of hormone disorders. Also, this is an appropriate method for monitoring of ovarian function and ovarian stimulation in the procedures of assisted reproduction techniques.
Petlevski R1, Frank S2. PO6-2: Myricetin, a naturally occurring flavonoid, prevents D-glucose induced oxidative damage in Hep G2 cells. Biochemia Medica 2009;19(Suppl 1):S131.
1University of Zagreb, Faculty of Pharmacy and Biochemistry, Zagreb, Croatia
2Institut für Medizinische Biochemie und Medizinische Molekularbiologie-Karl-Franzens-Universität, Graz, Austria
Corresponding author: rpetlevski [at] pharma [dot] hr
Introduction: Hyperglicemia in diabetes can induce oxidative stress in Hep G2 cells via several mechanisms. These include glucose autoxidation, the formation of advanced glycation end-products (AGE), and activation of the polyol pathway. Myricetin is a flavonoid compound found in many fruits and vegetables, including red wine, that acts as a powerful antioxidant.
Materials and methods: The aim of the present study was to investigate effect of the flavonoid myricetin (Sigma, Co) within a low concentration range (5 x 10-7 and 1 x 10-6 M) on the concentration of total glutathione (GSH) in Hep G2 cells. The major roles of GSH (gama-glutamylcysteinylglycine) are to maintain the intracellular redox balance and to eliminate ROS in cells.
Rezultati: Compared with control incubation, D-glucose (20 mM) significantly lowering GSH content in Hep G2 cells (N = 9). Exposure the Hep G2 cells plus glucose to low dose myricetin (5 x 10-7 M) for 4 hours at 37 degrees C resulted in significant increased of the GSH level when compared with control cells.
Conclusion: These results demonstrate that myricetin in low concentration attenuates D-glucose induced damage in Hep G2 cells.
ZecI1, Tišlarić-MedenjakD1, PosavecLj1, BukovecMegla Ž1, Šimundić AM2, KralikOguić S3. PO6-3: Importance of measurement of Anti-Mullerian hormone serum concentrations in women of reproductive age with regular menstrual cycles. Biochemia Medica 2009;19(Suppl 1):S131-S132.
1Sestre milosrdnice University Hospital, Laboratory of Endocrinology,
2Clinics of oncology and nuclear medicine, Zagreb, Croatia
3University Department of Chemistry, Sestre Milosrdnice University Hospital, Zagreb, Croatia
4Clinical Institute for Laboratory Diagnostics, Clinical Hospital Centre Zagreb, Zagreb, Croatia
Corresponding author: ikulas [at] kbsm [dot] hr
Objective: To estimate the changes of Anti-Mullerian hormone concentrations in sera of fertile women with regular menstrual cycles.
Subjects and methods: The study included 35 women from 19 to 44 years of age with regular menstrual cycles (30 ± 2 days) and normal hormonal profile. Women were separated in two groups, according to their age. First group consisted of women younger than 30 years and second, of women of 30 years or older, respectively. Hormone concentrations were examined by measurement of lutropin (LH), folitropin (FSH), estradiol (E2), total testosterone (T) and sex-hormone binding globulin (SHBG) in follicular phase (cycle day 3-5), LH, FSH and E2 in ovulation (cycle day 13-15) and progesterone (P) in lutheal phase (cycle day 20-23). Measurements of hormones were performed by electrochemiluminiscence immunoassay (ECLIA) on Roche-Ellecsys analyzer. Concentrations of AMH were determined with enzyme immunoassay (ELISA, DSL Inc.) in follicular and ovulatory phase, respectively.
Results: The first group comprised 15 women at age 26 ± 3 and the second one included 20 women at age 36 ± 4. Normal distributions of all measured parameters were confirmed by Kolmogorov-Smirnov test. Mann-Whitney test didn’t prove significantly different AMH concentrations between groups (P = 0.0693). The measurements of AMH concentrations in both, follicular and ovulatory phase were performed in 19 women. Wilcoxon test did not confirmed significant difference of AMH concentrations between follicular phase and ovulation (P = 0.3124).
Conclusion: Preliminary results didn’t show significant difference of AMH levels in relation to woman’s age who still having regular menstrual cycles and normal hormonal profile. In order to show usefulness of AMH determination as better indicator than other hormones which are usually measured as parameters of women’s fertility, further studies on larger number of patients are necessary.
Čepić K, Dujmov I, Roje D, Mladina B, Šupe Domić D, Stanišić L. PO6-4:Human placental growth factor, pregnancy-associated plasma protein A and human chorionic gonadotropin-beta in idiopathic intrauterine growth restriction. Biochemia Medica 2009;19(Suppl 1):S132-S133.
Split Universty Hospital Center, Split, Croatia
Corresponding author:katarina [dot] cepic [at] kbsplit [dot] hr
Introduction: Complex genetic and environmental mechanisms of maternal, fetal and placental origin regulate fetal growth and may contribute to intrauterine growth restriction (IUGR). Placental growth factor (PGF), a member of the vascular endothelial growth factor family, is produced chiefly by the placenta and is a potent angiogenic factor. Pregnancy-associated plasma protein A (PAPP-A) is a large zinc-binding protein produced by the developing placenta. Human chorionic gonadotropin-beta (β-hCG) is a sialoglycoprotein and is initially secreted by the trophoblastic cell of the placenta. The aim of this study (as a part of a large, comprehensive study) was to evaluate PGF, PAPP-A and beta-hCG levels in groups of pregnant women with IUGR and control group as a indicator for diagnosis of IUGR.
Subjects and methods: 32 women with IUGR were compared with 48 normal controls at the beginning of labour. PGF and PAPP-A serum level was assayed with an enzyme-linked imunosorbent assay. Serum level of Total beta-hCG was measured by chemiluminiscent microparticle imunoassay. Comparisons were made by using the Mann-Whitney U test for nonparametric data. Also, the Pearsons coefficient of correlations was used for correlation analysis.
Results: We found that maternal levels of PGF, PAPP-A and Total beta-hCG were no significantly different in women with IUGR compared to controls (median; min-max; 84.72, 26.90-626.59 vs 74.83, 8.17-474.04 pg/mL, P = 0.885; 410.8, 66.7-2039.3 vs 324.9, 38.9-1469.9 ug/mL, P = 0.105; and 16182, 846-42391 vs 16938, 1527-105672 mIU/mL, P = 0.960; respectively). Between Total beta-hCG and PAPP-A we found positive coefficient of correlation (r = 0.3642, P = 0.001).
Conclusion: We conclude that there was no significant difference in the serum level of PGF, PAPP-A and Total beta-hCG between IUGR and control group. But, statistically significant correlations were found between levels of PAPP-A and Total beta-hCG.
Tandara L, Tandara M. PO6-5:Glucose and pyruvate metabolism and morphological structure in preimplantation human embryos. Biochemia Medica 2009;19(Suppl 1):S133-S134.
Split Universty Hospital Center, Split, Croatia
Corresponding author:leida [dot] tandara [at] gmail [dot] com
Aim: The aim of the study was to establish whether the measurement of consumed glucose and pyruvate can be helpful in foreseeing development of preimplantation embryos and increasing the success rate of in-vitro fertilisation.
Materials and methods: Extracorporeal fertilization was applied in 80 subjects. The study subjects were divided into groups according to the quality of the embryos and the level of parameters measured. Day 3 embryos were classified into five groups according to their cellularity, the identity of blastomeres and fragmentation. The quantity of consumed glucose and pyruvate was measured in 580 preimplantation embryos. The level of pyruvate and glucose were determined in 15 µl of the medium in which the embryos were incubated and in the control medium by microfluorometric method.
Results: The consumption of pyruvate was much higher than the consumption of glucose in early preimplantation embryos before glucose became the main substrate at the blastocyst stage. Deterioration of the quality of day 3 embryos was related to a decrease in the consumption of pyruvate and glucose. The day 3 embryos rated 2 and 3 showed a statistically significant difference in the consumption of pyruvate (P < 0.001). A statistically significant difference in the consumption of glucose (P = 0.002) was found to exist between the day 3 embryos rated 2 and 3.
Conclusion: An appropriate relationship between morphology quality of preimplantation embryos and consumption of glucose and pyruvate in cultivation medium can be helpful in choosing the quality day 3 embryos and ensuring a higher success rate of in-vitro fertilization.
Galetović A, Salamunić I, Bilopavlović N, Tandara L. PO6-6:Catalitic concentrations of superoxide dismutase, glututhione peroxidase and catalase in diabetic patients with and without diabetic retinopathy. Biochemia Medica 2009;19(Suppl 1):S134-S135.
Department of Medical Laboratory Diagnosis, Split Universty Hospital Center, Split, Croatia
Corresponding author:agalet [at] krizine [dot] kbsplit [dot] hr
Introduction: Superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase are the enzymes of antioxidative system. The role of antioxidative system is elimination of free radicals and oxidants. SOD converts O2 to H2O2, GPX H2O2 and lipide peroxide to less reactive molecules and catalase removes H2O2 produced by activity of oxidases in peroxisomes. Autooxidation of glucose, glycosilated proteins and setting up of sorbitole pathway during hyperglycemia raise the amount of reactive oxygen species (ROS) and free radicals. The increased level of ROS and compromised antioxidative defence of diabetic could possibly interact with the development of diabetic retinopathy (DR).
This study aims to evaluate whether there is a difference in catalitic concentrations of antioxidative enzymes between two diabetic groups (Group A with DR and Group B without DR N = 30) and to compare them with the control group (N = 30).
Materials and methods: Catalitic concentrations of SOD and GPX were determined in RBC lysate and catalase in plasma.
Rezultati: The results for all three enzymes were significantly lower in both diabetic groups (Group A: SOD, P < 0.001; GPX, P = 0.030; catalase, P = 0.001; Group B: SOD, P < 0.001; GPX, P = 0.005; catalase, P < 0.001) compared to the control group, and confirm the compromised antioxidative status of diabetic patients. There was not statistically significant difference of enzymes catalitic activity between the two diabetic group in our study (SOD, P = 0.451; GPX, P = 0.375; catalase, P = 0.546).
Conclusion: Inconsistent results obtained in a number of similar studies could be explained by different number of participants, different methods used, or differences in patients’ age, type and duration of diabetes.