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Oxytocin achieves its effects after binding the oxytocin receptor (OXTR). Oxytocin plays an important role in empathy. The aim of this study was to examine the influence of two single nucleotide polymorphisms (SNPs) of the OXTR gene (rs53576 and rs2254298) on empathy measures and plasma oxytocin levels. Seventy-four university students were screened for the OXTR rs53576 and rs2254298 SNPs using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The level of oxytocin in the plasma was measured by the enzyme linked immunosorbent assay (ELISA). All subjects were assessed with the empathy quotient (EQ) questionnaire and Reading the Mind in the Eyes Test (RMET). Subjects carrying the rs53576 GG genotype had a higher EQ score, compared to GA/AA genotype carriers. By gender analysis, significance was reached only in females. Considering the influence of both examined polymorphisms on empathy, participants with the GGGG diplotype had a higher EQ in comparison with GAGG/AAGG carriers. These hormone effects were not correlated with plasma oxytocin levels. This is the first study implying that higher empathy in rs53576 GG genotype carriers may not depend on plasma oxytocin levels, but possibly on the number and function of OXTRs in the brain. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. III41018]

Introduction: Two most common pharmaceutical formulation of tacrolimus (Tac) used after kidney transplantation (Tx) are immediate-release one, administered twice-daily (TacTD), and prolonged-release one, administered once-daily (Tac-OD). Aim: The aim of this study was to compare daily doses, trough concentrations (C0 ) and dose-adjusted trough concentrations (C0 /D) of Tac between patients who administered different drug formulations, Tac-TD or Tac-OD, during the first year after Tx. Additionally, the aim of the study was to compare the distribution of C0 within and beyond the target therapeutic range (8-12 ng/mL for the first 90 days and 6-10 ng/mL afterwards) after the administration of different drug formulations. Subjects and Methods: A retrospective pharmacokinetic study included 84 patients (56 on Tac-TD and 28 on Tac-OD), with a follow-up period between the first and twelfth month post-transplantation. Following pharmacokinetic data were used: daily dose, daily dose according to patient's body weight, concentration C0 and C0 /D of Tac. Results: The results of the study showed that patients on Tac-OD formulation had higher daily doses and higher C0 during 4-6 months (p

Introduction/Objective. Intensive oxidative stress is proven in patients with diabetes mellitus and important in the development of a microvascular complication of type 2 diabetes mellitus. The aim of the study was to investigate the relationship between morphometric parameters of retinal blood vessels in patients with diabetic retinopathy (DR) and the levels of parameters of oxidative stress: advanced oxidation protein product (AOPP), thiobarbituric acid reactive substances (TBARS), and total sulfhydryl (SH) groups in blood samples. Methods. The patients (the group with DR and controls) were sex- and age-matched. Glycaemia, hemoglobin A1C HbA1C, total cholesterol and its fractions, and triglycerides were measured in blood samples. AOPP and total SH groups were determined in the plasma by specific methods. Modification of the thiobarbituric acid method was used for the determination of TBARS. The number and diameter of retinal blood vessels, as morphometric parameters on digital retinal photography, was determined by using the ImageJ software. Student’s t-test was used as the statistical method for the evaluation of differences between the morphometric and blood test parameters. The significance of differences in morphometric parameters of retinal blood was establish by one-way ANOVA. Results. Significantly higher levels of parameters of oxidative stress (AOPP and TBARS) were in the group of patients with DR than in the controls. This difference was also present among the patients with mild and severe forms of DR (AOPP F 77.03, p < 0.001) (TBARS F 63.28, p < 0.001). The diameter of retinal blood vessels correlated with levels of AOPP, but only in patients with mild DR. Conclusion. Parameters of oxidative stress, AOPP and TBARS, may be important for the follow-up of DR. In early stages in diabetic retinopathy, AOPP can be a valuable biomarker.

Introduction Cytokine network plays an important role in pathogenesis of chronic periapical lesions. The aim of this study was to determine the concentration of interleukin-6 (IL-6) in tissue homogenates of human periapical lesions and correlate its levels with symptomatology and size of the lesions. Materials and Methods 93 samples of chronic periapical lesions were obtained after extraction of teeth. Samples were divided according to the clinical presentation as symptomatic and asymptomatic, and according to the size as large and small. The concentration of IL-6 was analyzed using ELISA. Results Statistically significant difference in IL-6 concentration was observed in symptomatic lesions compared to asymptomatic (p

The influence of ultraviolet (UV) radiation on the leaching of di(2- -ethylhexyl) phthalate (DEHP) from 8 different parts of plastic medical devices made of poly(vinyl chloride) (PVC) that are used in two important medical procedures (peritoneal dialysis and transfusion) was investigated. The investigation was performed for three different extraction times (6, 15 and 30 days). DEHP determination was realized by gas chromatography–mass spectrometry (GC-EI– –MS). All the investigated samples contained a significant amount of DEHP. The results showed that some of the set for peritoneal dialysis contained DEHP in higher amounts than samples from the transfusion set. All samples of tubing material showed higher concentration levels of DEHP than the coupled bags. Results obtained after UV treatment showed that UV radiation has a certain influence on DEHP leaching from samples of PVC medical devices. The smallest difference was in the case of the quadruple blood bag from the transfusion set (about 73 % remained), while the biggest difference was obtained for the SAG-M transfer bag, also from the transfusion set, where just 25 % of total content of DEHP remained. The results obtained for DEHP leaching from investigated samples by time showed that most of the samples showed significant differences in the amounts of DEHP leached after 6 and 30 days. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. III 41018]

Introduction/Objective. An unfavorable prognosis of membranous nephropathy (MN) is determined by the presence of persistent proteinuria and extensive tubulointerstitial lesions at initial biopsy. Our study investigated the value of markers of renal cell dysfunction (glomerular filtration rate, urinary excretion of protein, ectoenzyme proximal tubular epithelial cells, and oxidative stress) in patients with MN, and points to the use of these markers in a possible therapeutic modification. Methods. The study included 28 patients with MN and 30 healthy individuals as control. In addition to the basic laboratory studies, enzyme [aminopeptidase N (APN), plasma cell glycoprotein-1 (PC-1), N-acetyl- β-D-glucosaminidase (NAGA), and dipeptidyl peptidase-4] activity was determined in serum and urine, as well as parameters of oxidative damage [thiobarbituric acid concentration of substance-responders (TBARS), malondialdehyde, and the concentration of the total sulfhydryl (SH) group]. Results. In patients with MN, serum activity of PC-1 and APN and urinary excretion of NAGA were significantly higher than in the control group. Also, significant correlation between daily proteinuria and serum PC-1 activity and urinary excretion of NAGA was found in patients with MN. Serum and urine levels of TBARS as well as total SH group levels were significantly lower in patients with MN than in healthy controls. Conclusion. Kidney damage in MN is accompanied by the release of several tubular enzymes, with potential diagnostic and prognostic significance. The study suggests a possible role of oxidative stress in pathogenesis of MN and the use of antioxidants in preventing impairment as part of future therapy. [Project of the Ministry of Science and Environmental Protection of Serbia, Grant No. 175092]

Immunosuppressive drugs play a crucial role in the inhibition of immune reaction and prevention of graft rejection as well as in the pharmacotherapy of autoimmune disorders. Effective immunosuppression should provide an adequate safety profile and improve treatment outcomes and the patients' quality of life. High-risk transplant recipients may be identified, but a definitive prediction model has still not been recognized. Therapeutic drug monitoring (TDM) for immunosuppressive drugs is an essential, but at the same time insufficient tool due to low predictability of drug exposition and marked pharmacokinetic variability. Parallel therapeutic, biochemical and clinical monitoring may successfully optimize and individualize therapy for transplanted recipients, providing optimal medical outcomes. Modern pharmacotherapy management should include new biomarkers with better sensitivity and specificity that can identify early cell damage. The aim of this study was to point out the importance of finding new biomarkers that would enable early detection of adverse drug events and cell damage in organ transplant recipients. We wanted to confirm the importance of routine biochemical monitoring in improving the safety of immunosuppressive treatment.

The presence of DEHP in dialysis and infusion sets for peritoneal dialysis and parenteral nutrition, which are made of PVC and other plastic polymeric materials, were investigated. Phthalate determination was carried out by gas chromatography - mass spectrometry method (GC-MS). The results showed that the peritoneal dialysis set (bag and tubing) made of PVC contains DEHP in significant amount, about 31 - 34%. Solution for peritoneal dialysis which was stored in the investigated PVC bag, contains low amount of DEHP, about 3.72 μg dm-3. Infusion bottles which are made of LDPE, also contain DEHP but in lower amount than PVC bags. LDPE bottle for packaging physiological saline solution (0.9% NaCl) showed higher amount of DEHP than LDPE bottle for packaging Ringer’s solution. In contrast, solution stored in bottle with lower DEHP level, i.e. Ringer’s solution contained about three times higher concentration of DEHP than physiological saline solution stored in bottle with higher DEHP level. Concentrations of DEHP in Ringer’s solution and physiological saline solution are 17.30 and 5.83 μg dm-3, respectively. The obtained values are under estimated upper-bound dose of DEHP received by adult patients undergoing procedures of peritoneal dialysis and parenteral nutrition. [Projekat Ministarstva nauke Republike Srbije, III 41018]

An interaction of Co(II), Ni(II) and Zn(II) with humic acid and O-donor humic-like ligands was studied by ion-exchange and electrospray ionization - mass spectrometry (ESI-MS) method. Interactions were confirmed by differences between values of chromatogram peak areas for monocomponent (ligand) and binary systems (ligand with metal ion) by newly developed ESI-MS 5 μL Loop injection technique. Schubert’s ion exchange method was used for determination of stability constants of formed complexes at pH 4.0 and I = 0.01. Comparing the values of log Kmn for complexes formed by dipositive metal ions with humic, benzoic and salicylic acids, it can be concluded that the interaction strengths of transitional series metals follow the Irving-Williams order: Co(II) < Ni(II) < Cu(II) > Zn(II). The obtained values of log Kmn indicate that Pb(II) ions effect the strongest interactions with all investigated ligands. Complexation of humic acids macromolecules possessing O-donor binding sites as major binding sites can be predicted and modelled following the same order of interaction strength like Irving-Williams order. [Projekat Ministarstva nauke Republike Srbije, br. III41018]

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Background: The primary goal of this study was to evaluate the influence of cytochrome P450 (CYP) 3A5 (6986A>G) and ABCB1 (3435C>T) polymorphisms on tacrolimus (TAC) dosage regimen and exposure. Second, we evaluated the influence of TAC dosage regimen and the tested polymorphisms on renal oxidative injury, as well as the urinary activities of tubular ectoenzymes in a long-term period after transplantation. Also, we aimed to determine the association between renal oxidative stress and tubular damage markers in the renal transplant patients. Methods: The study included 72 patients who were on TAC based immunosuppression. Allele-specific PCR was used for polymorphism determination. We measured the urinary thiobarbituric acid reactive substances (TBARS) and reactive carbonyl derivates (RCD) in order to evaluate oxidative injury, as well as the urinary activities of ectoenzymes (N-acetyl-b-Dglucosaminidase, aminopeptidase N and dipeptidyl peptidase IV) to evaluate tubular damage. Results: The carriers of CYP 3A5*1 allele required statistically higher daily doses of TAC than CYP *3/*3 carriers, as well as the carriers of C allele of ABCB1 gene compared to those with T T genotype. Also, there were no differences in TBARS, RCD and the activities of ectoenzymes between the patients' genotypes. Our results showed significant correlations between urinary TBARS and RCD and the ectoenzymes' activities. Conclusions: Our findings suggest that CYP 3A5 and ABCB1 3435 polymorphism may affect TAC daily doses, but not the drug's tubular toxicity. Furthermore, tubular damage may be associated with increased renal oxidative stress.

Background: The aim of the study was to evaluate parameters of oxidative and nitrosative stress as well as antioxidative parameters in a group of renal transplant recipients with stable graft function and no clinical signs of cardiovascular disease. We also aimed to determine the correlations among these parameters and to evaluate potential differences in all the biomarkers with regard to the immunosuppression protocol. Methods: We enrolled 57 renal transplant recipients and 31 controls who were age and sex matched with the renal transplant recipients. All of the patients included in this study had post-renal transplant surgery at least 12 months earlier and were on standard immunosuppressive therapy. In this study, we determined thiobarbituric acid-reactive substances in plasma and red blood cells and advanced oxidation protein products, nitrosative stress parameters (asymmetric and symmetric dimethylarginine - ADMA and SDMA), and antioxidative parameters (total SH groups and catalase activity). Results: The results of our study demonstrated that the levels of oxidative and nitrosative stress were significantly increased compared to the healthy population (p < 0.01 except for plasma catalase activity p < 0.05). Correlation analysis showed significant positive correlations between: ADM A and SDMA (p < 0.01); ADM A and nitrates (p< 0.05); SDMA and nitrates (p < 0.05); between OS parameters in the experimental group; AOPP and SH groups (p < 0.05) and TBARS in plasma and SH groups (p < 0.01), SDMA and AOPP (p< 0.05); SDMA and TBARS in plasma (p < 0.05); SDMA and SH groups (p < 0.01); nitrates and SH groups (p < 0.05). Conclusion: There was no significant difference in oxidative and nitrosative stress parameters with respect to the immunosuppressive protocol.

Introduction. The balance between proinflammatory and anti-inflammatory cytokines plays an important role in the pathogenesis of chronic periapical lesions. The aim of this study was to determine the concentration of TNF-α in tissue homogenates of periapical lesions and analyze its levels in relation to the symptomatology and the size of lesions. Materials and Methods. 93 samples of chronic periapical lesions were obtained after extraction of teeth. Samples were classified according to the clinical presentation as symptomatic and asymptomatic, and according to the size as large and small. The concentration of TNF-α was analyzed using ELISA. Results. The results showed increased production of TNF-α in symptomatic lesions compared to asymptomatic. Higher concentration of TNF-α was demonstrated in large lesions compared to small. Large symptomatic lesions showed greater concentration of TNF-α compared to small symptomatic lesions, while bigger asymptomatic lesions demonstrated higher amount of the cytokines compared to small asymptomatic lesions. Conclusion. Higher concentration of TNF-α in large symptomatic lesions indicates that TNF-α is an important factor responsible for the progression of lesions.

Introduction. In spite of the growing number of reports on the study of anti-nucleosome and anti-C1q antibodies, there are still controversies on their significance as disease activity markers in patients with systemic lupus erythematosus (SLE) and their use in everyday clinical practice. Objective. Our aim was to assess the presence of anti-dsDNA, anti-nucleosome and anti-C1q antibodies in SLE patients, as well as to establish their sensitivity, specificity, positive and negative predictive value, and their correlation with SLE and lupus nephritis clinical activity. Methods. The study enrolled 85 patients aged 45.3±9.7 years on the average, with SLE of average duration 10.37±7.99 years, hospitalized at the Institute „Niška Banja“ during 2011, and 30 healthy individuals as controls. Disease activity was assessed using Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). In all examinees the levels of anti-dsDNA, anti-nucleosome and anti-C1q antibodies were measured using the ELISA method with Alegria Test Strips Orgentec (Germany). Results. Patients with active lupus nephritis had a higher presence of anti-C1q antibodies and higher co-positivity of anti-dsDNA, anti-nucleosome, and anti-C1q antibodies compared to those with inactive lupus nephritis (77.77% vs. 21.74%; p

The complexation of humic acid with certain heavy metal ions (Co(II), Ni(II), Cu(II), Zn(II) and Pb(II)) was investigated. The stability constants of humate complexes were determined by method which is based on distribution of metal ions between solution and resin in the presence and the absence of ligand, known as Schubert’s ion exchange method. Experiments were performed at 25 ºC, at pH 4.0 and ionic strength of 0.01 mol dm-3. It was found that the 1:1 complexes were formed between metal ions and humic acid. Obtained results of the stability constants, log βmn, of complexes formed between the metal ions and humic acid follow the order Co(II) < Ni(II) < Cu(II) > Zn(II) which is the same like in the Irving-Williams series for the binding strength of divalent metal ion complexes. Stability constant of complex between Pb(II) ions and humic acid is greater than stability constants of other investigated metal-humate complexes. The investigation of interaction between heavy metal ions and humics is important for the prediction of the distribution and control of the migration of heavy metals in natural environment. [Projekat Ministarstva nauke Republike Srbije, br. III 41018]

Bacground/Aim. Hepcidin may play a pathogenetic role in iron metobolism disorders. The aim of this study was to determine the correlation between hepcidin concentration and parameters of iron metabolism in patients with different stage of chronic kidney disease (CKD). Methods. The study involved 104 patients with CKD: 64 on hemodialysis (HD) and 40 patients in pre-dialysis stadium (pre-HD) with adequate erythropoetin therapy and iron supplementation. The HD group was divided in four subgroups according to the level of serum ferritin (up to 100; 100-199; 200-499 and over 500 ng/mL). Parameters of anemia, iron status, inflamation and hepcidin level were evaluated. Results. The HD patients had a significantly lower eritrocyte count, erythrocytes indexes, hemoglobin and transferrin saturation and significantly higher iron, ferritin, hepcidin and total iron binding capacity (TIBC). The HD subgroups up to 199 ng/mL of serum feritin had lower high-sensitivity Creactive protein (hsCRP), iron and higher unbuffered iron binding capacity (UIBC), transferrin saturation and TIBC compared to the HD subgroups over 200 ng/mL. The lowest and the highest ferritin subgroups had the highest hepcidin level and it showed significant correlation with ferritin. Conclusion. Hepcidin may serve as a marker for better diagnosing and monitoring anemia and iron metabolism disorders in CKD.

Nitric oxide (NO) is produced by many cells in the body; however, its production by vascular endothelium is particularly important in the regulation of blood flow. Vascular actions of NO include the following: direct vasodilation, indirect vasodilation by inhibiting the vasoconstrictor influences, anti-thrombotic, anti-inflammatory and anti-proliferative effects. Due to its importance in vascular function, abnormal production of NO, occurring in different diseases can adversely affect blood flow and other vascular functions. It has been suggested that alterations in NO generation are a critical cause of injury in the ischemic heart. A biologic link between the endothelial damage and atherosclerotic coronary arterial disease has been presumably related to decreased arterial bioavailability of NO through the increased leucocyte and platelet adhesions, vasoconstriction and smooth muscle cell proliferation. However, the precise mechanism of the impaired NO generation is not known, and there is a considerable controversy regarding whether myocardial ischemia results in increased or decreased NO formation. Asymmetric dimethylarginine (ADMA) is a natural, competitive inhibitor, and one of the primary factors controlling the nitric oxide production. ADMA was found to be elevated and closely correlated with the impaired vasodilator function in conditions associated with the endothelial dysfunction, such as hypercholesterolemia, hypertension, insulin resistance and type 2 diabetes, and renal failure. But ADMA also seems to be involved in myocardial ischemia, since its plasma levels predict future coronary events in patients with the elevated cardiovascular risk. It has been recently reported that the elevated plasma ADMA concentrations in the acute coronary events are an independent cardiovascular risk factor.

Decreased nitric oxide (NO) production and/or impaired NO bioavailability may occur in patients with the chronic kidney disease (CKD), and could contribute to elevation of blood pressure, cardiovascular disease (CVD) and progression of renal injury in these patients. Free guanidinomethylated arginine residues occur endogenously as a result of proteolysis of post-translational methylated tissue proteins. The asymmetric dimethyl arginine (ADMA) is a competitive inhibitor of the nitric oxide synthase (NOS) enzymes. The kidney has a predominant role in ADMA elimination by combining two mechanisms; urinary excretion and metabolization of ADMA The degradation of ADMA is accomplished intracellularly by the enzyme dimethylarginine dimethylaminohydrolase (DDAH). ADMA is not only a uremic toxin, but also a strong marker of the endothelial dysfunction and atherosclerosis and a stronger independent predictor of all-cause mortality and cardiovascular outcome in patients with the chronic renal failure. There are at least four mechanisms that may explain the accumulation of ADMA in CKD: increased methylation of proteins, increased protein turnover, decreased metabolism by DDAH and impaired renal excretion. A strong positive correlation between symmetric dimethyl arginine (SDMA) and creatinine suggests that SDMA might be of value as a marker of the renal function. Reduced NO elaboration secondary to accumulation of ADMA and elevated inflammation may be important pathogenic factors for endothelial dysfunction in patients with the renal disease. Elevation of ADMA may be a missing link between CVD and CKD.

Nitric oxide (NO) is an important signaling molecule in a variety of physiological processes. NO, a gas, is produced from L-arginine by different isoforms of the nitric oxide synthase and serves as mediator in important physiological functions, such as promoting vasodilation of blood vessels and mediating communication between nervous system cells. Contradictory to its physiologic actions, free radical activity of NO can cause cellular damage by the induction of nitrosative stress with significant implications on nervous system diseases. Although the mechanism of NO-mediated neurodegeneration still remains unclear, numerous studies suggest its crucial role in modification of protein functions by nitrosylation and nitro-tyrosination. NO contributes to glutamate excitotoxicity, participates in organelle fragmentation, inhibits mitochondrial respiratory complexes and mobilizes zinc from the internal stores. Recently, NO has been emerged as a mediator of epigenetic gene expression and chromatin changes. Besides, NO is a key mediator in the regulation of inflammatory and immune response of the central nervous system. It is involved in down regulation of several aspects of CNS inflammation, but also has a dual role in that it is required for inflammation in some situations.

Heavy metals as pollutants in the working and living environment are a serious health and environmental problem because they are toxic, non-biodegradable, accumulate in living systems and have a long half-life in soil. Sources of lead contamination are combustion products in the chemical industry and metallurgy, industrial waste water, landfills, traffic etc. Lead enters into the body via the food chain and drinking water. In the body lead is deposited in the liver, kidneys, brain and mineral tissues. Excretion of lead causes damage to the epithelial cells of certain organs. High level exposure to cadmium is usually the result of environmental pollution by human activities. Exposure to cadmium can lead to acute and chronic tissue damage of various organs, including liver and kidneys in humans and in animals. In this paper we analyzed the effects of lead and cadmium exposure, in working and living environment, on the model system of experimental animals, particularly the activity of certain liver enzymes, acid and alkaline DNase, and standard biochemical blood parameters. The study showed that lead and cadmium significantly affect the protein content, red blood cells, hemoglobin and hematocrit, and the activity of liver enzymes. This harmful effect of this toxic metal can be reduced by the supplements.

Background/Aim. Reperfusion of previously ischemic tissue leads to injuries mediated by reactive oxygen species. The aim of the study was to investigate the effects of different anesthesia techniques on oxidative stress caused by tourniquetinduced ischemia-reperfusion (IR) injury during extremity operations at children's age. Methods. The study included 45 patients American Society of Anesthesiologists (ASA) classification I or II, 8 to 17 years of age, undergoing orthopedic procedures that required bloodless limb surgery. The children were randomized into three groups of 15 patients each: general inhalational anesthesia with sevoflurane (group S), total intravenous anesthesia with propofol (group T) and regional anesthesia (group R). Venous blood samples were obtained at four time points: before peripheral nerve block and induction of general anesthesia (baseline), 1 min before tourniquet release (BTR), 5 and 20 min after tourniquet release (ATR). Postischemic reperfusion injury was estimated by measurement of concentration of malondialdehyde (MDA) in plasma and erythrocytes as well as catalase (CAT) activity. Results. Plasma MDA concentration in the group S was significantly higher at 20 min ATR in comparison with the groups T and R (6.78 ± 0.33 μmolL-1-1 vs 4.07 ± 1.53 and 3.22 ± 0.9. μmolL-1-1, respectively). There was a significant difference in MDA concentration in erytrocythes between the groups S and T after 5 min of reperfusion (5.88 ± 0.88 vs 4.27 ± 1.04 nmol/mlEr, p < 0.05). Although not statistically significant, CAT activity was slightly increased as compared to baseline in both groups S and R. In the group T, CAT activity decreased at all time points when compared with baseline, but the observed decrease was only statistically significant at BTR (34.70 ± 9.27 vs 39.69 ± 12.91 UL-1, p < 0.05). Conclusion. Continuous propofol infusion and regional anesthesia techniques attenuate lipid peroxidation and IR injury connected with tourniquet application in pediatric extremity surgery.

Introduction/Aim. By analyzing activity of some of the enzymes normally present in the saliva and the level of malondialdehyde in gingivitis, it is possible to estimate the functional condition of parodontium, and the examined parameters can be considered as biochemical markers of its functional condition. The aim of this paper was to examine activity of alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, lactate dehydrogenase and the level of malondialdehyde in the saliva of children affected with gingivitis, as well as the values of the mentioned parameters in relation to the level of the inflammation of gingiva. Methods. The research included 120 children at the age of 12.2 with permanent dentition. Löe and Silness gingival index was used to estimate the condition of gingiva, based on which the children were classified into four groups: the children with healthy gingiva (the control groups), the children with mild, moderate and severe inflammation of gingiva (the study group). Enzymes of the saliva were determined by the use of original tests and measured by the autoanalyser (Bio Systems A25, Spain). A modified method with tiobarbituric acid was used to determine malondialdehyde in nonstimulated mixed saliva. Results. The results of the examined enzyme activity and the level of malondialdehyde in the saliva of the study groups showed statistically considerably higher values for the level of malondialdehyde (p < 0.001), for the activity of aspartate aminotransferase and gamma glutamyl transferase (p < 0.01), as well as for alanine aminotransferase (p < 0.05) in comparison with the control group, whereas the activity of lactate dehydrogenase did not show a statistically significant increase. In relation to the level of the inflammation of gingiva, the results of the examination of the enzyme activity in the study groups showed statistically significantly higher values in the group with severe inflammation in comparison with those with mild, as well as the moderate inflammatory, except for the gamma glutamyl transferase, and in the group with moderate inflammation compared to that with the mild one, except for alanine aminotransferase. The results of the examination of the level of malondialdehyde in the saliva of the study groups did not show a statistically significantly increase in relation to the level of the inflammation of gingiva. Conclusion. There is a higher level of alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase and lactate dehydrogenase enzyme activity together with the higher level of malondialdehyde in the saliva of children with gingivitis in comparison with the activity of the same enzymes and the level of malondialdehyde in the saliva of children without gingivitis. The activity of the examined enzymes in the saliva of children with gingivitis increases in relation to the intensity of the pathological process, whereas the level of malondialdehyde shows no significant difference in relation to the level of the inflammation of gingiva.

Oxidative stress is considered to be a unifying link between diabetes mellitus (DM) and its complications, including nephropathy (DN). The aim of this study was to determine the parameters of oxidative injury of lipids and proteins as well as the activity of ectoenzymes in the urine of DN patients. The study included 40 individuals: 10 patients with type 2 diabetes mellitus and microalbuminuria (DMT2-MIA), 10 type 2 diabetic patients with macroalbuminuria (DMT2- MAA), 10 patients with type 1 diabetes and microalbuminuria (DMT1-MIA) and 10 age- and sex-matched healthy subjects (control). In the urine we determined TBA reactive substances (TBARS), reactive carbonyl groups (RCG), and the activity of ectoenzymes N-acetyl-b-d-glucosaminidase (NAG), plasma cell differentiation antigen (PC-1), aminopeptidase N (APN) and dipeptidyl peptidase IV (DPP IV). A higher concentration of TBARS in the urine was found in DMT2-MIA and DMT1-MIA, compared to the control group (p

Numerous factors can influence the onset of SLE and development of some clinical disease manifestations with various organ involvements and occurrence of characteristic symptoms and disease signs. This paper studies the balance between proinflammatory and antiinflammatory cytokines investigates the presence of oxidative stress measuring certain prooxidative factors and determines the activation of antioxidative protection pathways aiming to establish possible correlations between the studied parameters. ELISA, enzymatic spectrophotometry and colorimetric methods were used to determine the above-mentioned parameters. The results obtained indicate that disturbed pro/antioxidative status is associated with the change of antioxidative factors, with the fall od SOD activity and increase of GPx and CAT activity in the erythrocytes of all studied groups of patients. At the same time, the cytokine production was altered, not only compared to the healthy control samples, but also in various clinical disease manifestations. Altered relationships of pro and antiinflammatory cytokines and the consequential disorders of other studied systems provide us with useful strategic targets for diagnostic monitoring and possible therapeutic interventions in SLE patients.

Wine is an alcoholic beverage created as a product of alcoholic fermentation. It is a complex compound consisting of a large number of components (water, alcohols, acids, sugars, mineral substances, phenols, esters, glycerol, etc.). The chemical composition and quality of wine largely depend on the variety, wine type, production, and storage methods. Maceration is an essential part of the red wine production process. During maceration, numerous compounds are separated from the solid parts of the grape and berry and they are extracted into wine, and thanks to this process, red wine acquires most of its characteristics. It is considered that temperature and duration are the most important maceration factors that affect the quality of red wines. This paper aims to determine the influence of different maceration regimes on the quality of red wines of the Vranac and Merlot varieties. Vinification was carried out in 2021 in the Microbiology Laboratory of the Faculty of Technology in Banja Luka, where maceration regimes for 6 and 12 days were applied, at temperatures of 16 and 25°C. Wine quality analyses were done in the Laboratory for Ampelography and Winemaking at the Faculty of Agriculture in Banja Luka. Differences in wine quality were observed depending on the maceration regime. The highest alcohol content was observed after 6 days of maceration at 25°C, while prolonged maceration led to reduction in the alcohol content. The increased temperature had a positive effect on the total extract, as well as on the wine chromatic characteristics of both varieties. The wines of the Merlot variety had lower total acidity at longer maceration, while the wines of the Vranac variety had lower total acidity at shorter maceration. All analyzed wines had a satisfactorily low volatile acidity, as well as pH. The Merlot wines had the highest ash content after 12 days of maceration at 25°C, and the Vranac wines after 6 days of maceration at 25°C.

The study aimed to investigate the protective effects of chokeberry fruit products and by-products against cisplatin-induced acute nephrotoxicity in rats. Potential mechanisms involving oxidative stress and inflammatory responses were examined through biochemical and histopathological analyses of kidney tissue. Chokeberry waste, along with the whole fruit extract and juice, was evaluated as a potential raw material for pharmaceutical use. The chemical composition of chokeberry juice and extracts was analyzed using spectrophotometry and HPLC. Rats were treated with chokeberry preparations via intragastric tube for ten days, with a single intraperitoneal dose of cisplatin (8 mg/kg BW) administered on the third day. Post-sacrifice, plasma samples were analyzed for biochemical nephrotoxicity markers, oxidative stress, and inflammatory markers. Kidneys were removed for histopathological and biochemical analysis. Cisplatin-induced acute nephrotoxicity was confirmed by elevated plasma creatinine and blood urea nitrogen levels. Additionally, lipid peroxidation was significantly elevated, while reduced glutathione and catalase activity were significantly reduced. Pro-inflammatory mediators IL-1β, TNF-α, and IL-6 levels were significantly increased in the cisplatin group. Treatment with chokeberry extracts and juice significantly mitigated these nephrotoxic effects, as confirmed by histopathological examination and biochemical marker analysis. Notably, the waste extract demonstrated greater efficacy than the whole fruit extract, likely due to its higher concentration of polyphenolic compounds, especially anthocyanins. These results highlight the potential of chokeberry as a therapeutic and preventive agent for kidney protection, emphasizing the value of by-products rich in biologically active compounds. [ABSTRACT FROM AUTHOR]

1. Introduction: The study aimed to investigate the influence of interleukin (IL)-6 -174 G/C gene polymorphism on graft function (defined as estimated glomerular filtration rate, eGFR), as well as on the tacrolimus (Tac) pharmacokinetics during the five years after kidney transplantation. 2. Methods: The study included 115 Caucasian kidney transplant recipients on Tac-based immunosuppression. The patients were followed between 6 and 60 post-transplantation months. Interleukin-6 and CYP3A5 genotyping were performed. 3. Results: Patients carrying the IL-6 -174GG genotype had lower eGFR values compared to the patients with the IL-6 -174GC and −174CC genotypes at the 12th, 48th and 60th post-transplantation months. The linear regression analysis indicated that eGFR at the 6th post-transplantation month and IL-6 -174 G/C polymorphism are independent predictors of eGFR values in the late post-transplantation period. The IL-6 -174GG genotype carriers had lower dose-adjusted trough concentration (C0/D) of Tac compared to the IL-6 C allele carriers during the entire observation period (except at the 24th month), while this effect was independent of the CYP3A5 genotype within three years post-transplantation. 4. Conclusion: Interleukin-6 genotyping could be an additional tool to categorise patients towards the risk of graft deterioration in the long-term post-transplantation period. The IL-6 genotyping could be supportive in genotype-guided dosing of Tac. [ABSTRACT FROM AUTHOR]

In July 2015, 179 grapevine plants belonging to 16 grapevine autochthonous cultivars were assessed for sanitary status using DAS ELISA test for the presence of: Grapevine fanleaf virus (GFLV), Grapevine leafroll-associated virus 1 (GLRaV-1), Grapevine leafroll-associated virus 2 (GLRaV-2) and Grapevine leafroll-associated virus 3 (GLRaV-3). Furthermore, survey for the phytoplasma presence and laboratory analyses using nested-PCR/RFLP assay was conducted at the beginning of September 2015 on grapevine cultivars which were not positive in DAS ELISA test for the presence of the four viruses. Out of 179 tested plants with DAS ELISA test, 146 (81%) were positive for the presence of at least one virus. The most widespread viruses were GFLaV-1 and GFLaV-3 with approximately 80 % of grapevines infected. Nested-PCR/RFLP assay showed that out of 33 tested samples 2 were positive for the presence of phytoplasmas from 16SrXII group. Sanitation of infected grapevine cultivars is needed in near future.