Association of the Brain-derived Neurotrophic Factor Val66Met Polymorphism with Body Mass Index, Fasting Glucose Levels and Lipid Status in Adolescents.

Brain-derived neurotrophic factor (BDNF) has an important role in energy balance. It suppresses food intake, reduces hepatic glucose production and converts white fat into brown fat in adipose tissue, leading to energy dissipation, lowered blood glucose and a lean phenotype. Studies have shown that the single nucleotide polymorphism (SNP) Val66Met within BDNF may be associated with obesity, insulin sensitivity, type 2 diabetes mellitus (T2DM) and dyslipidemia. The objective of the study was to investigate the association of the Val66Met polymorphism with body mass index (BMI), fasting glucose levels and lipid profile in Serbian adolescents. The study included 308 randomly selected healthy adolescents, 153 (49.68%) boys and 155 girls (50.32%), 15 years of age. Data including age, gender, height, weight, lipid profile and fasting glucose were recorded. Genotyping was performed by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. No association of this polymorphism was found with BMI and lipid profile. However, significant association was observed between this polymorphism and fasting blood glucose (FBG). Carriers of a Val/Val genotype had significantly higher mean values of fasting glucose level compared to carriers of Val/ Met and Met/Met genotypes (p = 0.01). To confirm these results multiple linear regression analysis was performed. Body mass index and gender were taken as covariates. Carriers of the Val/Val genotype had significantly higher levels of FBG (ß = -0.152, p = 0.02). A statistically significant association between BMI and glucose level was also observed (ß = 0.124,p = 0.033). This polymorphism could be associated with fasting glucose level in Serbian adolescents, thus further research would be of great interest to validate these results.

Serum cystatin C levels in normal pregnancy.

AIM: The aim of this study was to determine the levels of cystatin C, creatinine and creatinine clearance in different trimesters of uncomplicated pregnancy in women with normal kidney function. SUBJECTS AND METHODS: A total of 109 pregnant women were included: group 1 - 38 women (average age 29.63 ± 4.3 y) in the first trimester, Group 2 - 32 women (average age 33.56 ± 5.95 y) in the second trimester and Group 3 - 39 pregnant women (average age 30.1 ± 6.95 y) in the third trimester. Serum cystatin C was determined by the PENIA method (Particle-Enhanced Nephelometric Immuno-Assay), using Behring tests (Behring Diagnostics GmbH, Marburg, Germany). Results were statistically analyzed using the ANOVA. RESULTS: A statistically significant increase in serum cystatin C level was found in the third trimester of pregnancy (0.69 ± 0.16 mg/l vs. 0.78 ± 0.26 mg/l vs. 1.21 ± 0.30 mg/l). CONCLUSION: It appears that cystatin C is not a reliable marker of kidney function in pregnancy and that its increase is connected with a combination of several factors, including endotheliasis, hormonal influence and glomerular filtration rate (GFR) alterations.

Reticulocyte haemoglobin content vs. soluble transferrin receptor and ferritin index in iron deficiency anaemia accompanied with inflammation.

Ferritin concentration, as a parameter of iron status that is commonly used in the diagnosis of iron deficiency anaemia (IDA), often has limited values if the iron deficiency is accompanied by inflammatory disease. This study evaluated the value of reticulocyte haemoglobin content (CHr) and soluble transferrin receptor-ferritin index (sTfR/F) in the diagnosis of IDA and differential diagnosis of IDA and anaemia of chronic disease. The study included 66 nonanaemic individuals as controls, 86 patients with IDA divided into noninflammatory and inflammatory subgroups, and 32 patients with anaemia of chronic disease. Blood count, iron, transferrin saturation, total iron binding capacity, ferritin, C-reactive protein, sTfR and CHr were determined. Receiver operator characteristic curve analysis showed very high discriminating power for CHr, soluble transferrin receptor (sTfR) and sTfR/F in the diagnosis of IDA. In patients with anaemia of chronic disease these parameters showed no significant difference from the control. CHr and sTfR enabled recognition of iron deficiency and were not affected by acute phase reaction. They are sensitive markers of body iron status with additional value to conventional tests for the detection of iron deficiency.

Serum lipids and anti-oxidized LDL antibodies in primary antiphospholipid syndrome.

OBJECTIVE: The link between specific antibodies and atherogenesis in primary antiphospholipid syndrome (PAPS) is less strong than for thrombosis, although clearly the two processes are related and thrombosis is the main complication of atherosclerosis, a process known as atherothrombosis. The aim of this study was to investigate the influence of serum lipid levels and anti-oxidized LDL (oxLDL) antibodies on the clinical features of 42 patients with PAPS (mean age 40.45+/-13.37; 32 women and 10 men), and to compare them with 47 control subjects (mean age 39.68+/-13.93; 33 women and 14 men). METHODS: Total cholesterol, HDL and triglyceride concentrations were determined by enzymatic methods. LDL was calculated according to the Friedwald formula. Anticardiolipin, anti-oxidized LDL and anti-Beta2glycoprotein I antibodies were detected by ELISA. RESULTS: A significant association was found between arterial events and triglyceride, LDL and cholesterol concentrations, but multivariate analysis showed that cholesterol concentrations were the most important predictor of arterial events (p=0.012). Cerebrovascular insults were the most significantly associated with cholesterol concentrations (p=0.011). Myocardial infarctions were more frequently present in patients more than 40 years of age (p=0.032). No significant association of the investigated parameters with venous thromboses was found. Recurrent abortions were not associated with the presence or concentrations of the investigated parameters. Although patients had increased concentrations of anti-oxLDL antibodies, no significant association was found between the titres of anti-oxLDL antibodies and clinical features of APS. CONCLUSION: In patients with PAPS, lipid concentrations are a better predictor for arterial events than anti-oxLDL antibodies.

Usefulness of soluble transferrin receptor and ferritin in iron deficiency and chronic disease.

Soluble transferrin receptor (sTfR) is a biochemical parameter used for the detection of iron deficiency in situations where ferritin has limited diagnostic value owing to the present chronic disease. The sTfR concentration was determined in 118 patients divided according to their inflammatory status and underlying disease into groups of patients with iron-deficiency anemia (IDA), anemia of chronic disease (ACD) and patients with a coexisting state of iron deficiency and anemia of chronic disease (ID+ACD). All patients with iron deficiency had elevated sTfR levels, but ferritin concentrations were normal or increased in patients with inflammatory characteristics. Diagnostic efficiencies of sTfR, sTfR/log ferritin index (sTfR/F) and ferritin were evaluated by receiver operating characteristic curve (ROC) analysis. According to the results obtained, the best diagnostic efficiency for differential diagnosis of anemic patients with iron deficiency compared to the control group had a sTfR concentration (0.884) that was significantly higher than ferritin (0.638), but not higher than the calculated ratio sTfR/F (0.820). The cut-off value of the sTfR/F index differentiating the best control group from the IDA and ID+ACD groups was 1.30, and for differentiation of ACD from IDA and ID+ACD, the value was 0.90. Soluble transferrin receptor is an additional parameter to ferritin for the diagnosis of IDA and differential diagnosis of ID+ACD, but calculation of the sTfR/F index did not improve the diagnostic value of determining sTfR alone.

Changes in the plasma levels of protein C system parameters in pregnancy.

Resistance to activated protein C (APC) is a condition that leads to a hypercoagulative state with an increased risk for venous thrombosis. The aim of this study was to test the functionality of the protein C system in normal and complicated pregnancies and APC resistance. A total of 131 patients were tested, including pregnant women with normal and complicated pregnancies at different periods, e.g. from weeks 1 to 20, 21 to 30 and 31 to 38 of gestation. The following hemostatic parameters were determined: protein S, protein C, protein C Global and protein C APC sensitivity. Commercial "Behring" tests were used to determine the parameters of hemostasis. The values for protein C activity were within normal limits. Protein S values were below the lower limits. Significantly lower PC-NR (protein C normalized ratio) and APC-NR (activated protein C normalized ratio) values were found in all three of the gestation periods in pregnant women with a history of repeated miscarriages and hypertension in relation to pregnant women with normal pregnancies. In order to assess the diagnostic accuracy of investigated hemostatic parameters as markers of prethrombotic changes in pregnant women, the obtained values of investigated hemostatic parameters were evaluated by ROC analysis. PC-NR and APC-NR showed satisfactory diagnostic accuracy as markers of prethrombotic changes in pregnant women: more precisely, they were found to be good indicators of resistance to activated protein C in pregnancy.

Opposite effects of dexamethasone and ACTH on the adrenal cortex response to ethane dimethanesulphonate (EDS).

Recently we have reported that ethane dimethane-sulphonate (EDS), the Leydig cell cytotoxin, caused marked atrophy of the adrenal cortex of adult male rats. The aim of this work was to examine whether a 9-day treatment with dexamethasone (0.25 mg/kg/d) or ACTH (40 IU/kg/d), which started 4 days prior to administration of a single dose of EDS (75 mg/kg), influenced the response of the inner adrenocortical zones to the toxin. On day 15 after administration of EDS, adrenal weight was significantly decreased in saline treated rats, but glandular and serum corticosterone levels were not altered. In dexamethasone-suppressed rats, the effect of EDS was augmented; an additional decrease in adrenal weight was accompanied by reduced adrenal and serum corticosterone levels. In ACTH-treated animals EDS was ineffective. These results demonstrate that the deleterious effects of EDS on rat adrenal cortex can be prevented by ACTH and potentiated by dexamethasone.

The importance of antioxidant enzyme and total antioxidant status of patients with acute myocardial infarction on thrombolytic therapy.

It is believed that intensive production of free radicals occurs immediately after reperfusion, and that it explains a series of adverse effects of reperfusion. The aim of the study was to establish the importance of the antioxidative enzymes superoxide dismutase (SOD), glutathione peroxidase (GPX) and the total antioxidant status (TAS) in patients with acute myocardial infarction according to success or failure of reperfusion. Enzyme activities in erythrocyte hemolysate and concentration of TAS in plasma were determined by Randox kits in 50 patients with acute myocardial infarction. No significant differences in TAS between the mean values against either clinical status or the 10 time intervals were recorded. The SOD and GPX values were significantly influenced by the presence or absence of reperfusion (p<0.05). Monitoring of SOD and GPX at ten time points: before administration of streptokinase, 1, 3, 6, 12, 18, 24 hours, 2, 3 and 5 days after administration of streptokinase revealed the lowest SOD levels up to one hour before administration of streptokinase, supporting the fact that the maximum production of free radicals is achieved at that time. GPX values differed for 24 hours according to whether reperfusion was achieved or not. The obtained results suggest that free radical production is increased after reperfusion, and monitoring of antioxidant enzymes may help to evaluate the success of thrombolytic therapy.

Biochemical evaluation of patients with acute pancreatitis.

In this study we determined the clinical accuracy of alpha2-macroglobulin, alpha-amylase, C-reactive protein, lipase, non-esterified fatty acids, pancreatic alpha-amylase and phospholipase A in the diagnosis and prognosis of acute pancreatitis in a group of patients with acute abdominal pain using receiver operator characteristic curve analysis. We investigated 59 patients with acute pancreatitis and 72 patients with extrapancreatic diseases of gastrointestinal origin. On the basis of initial enzyme activities, at cut-offs of 245 U/l for amylase, 656 U/l for lipase, and 182 U/l for pancreatic alpha-amylase, the diagnostic efficiencies were 0.993, 0.980, and 0.975, respectively. Receiver operator characteristic curve analysis showed the same diagnostic accuracies. We evaluated the accuracy of serum alpha2-macroglobulin, C-reactive protein, non-esterified fatty acids and phospholipase A for differentiation between acute necrotizing pancreatitis and acute oedematous pancreatitis. C-reactive protein had the highest prognostic accuracy of the parameters studied (the area under curve = 0.9082) and at a cut-off value of 126 mg/l, sensitivity and specificity were 0.759 and 0.912, respectively. The role of the clinical laboratory in the investigation of patients with acute pancreatitis continues to evolve and biochemical parameters are a good diagnostic and prognostic option.

Serum beta-endorphin level in patients with depression on fluvoxamine.

The main interest of the present study was to determine possible alternations in beta-endorphin serum levels in healthy volunteers and in patients with depression, as well as changes in beta-endorphin serum levels caused by fluvoxamine treatment. Fluvoxamine maleate (Fevarin) was administered orally at a dose of 200 mg/day for 4 weeks. The serum levels of beta-endorphin were lower in patients with 'nonendogenous' depression (104.68 +/- 5.29 pg/ml) and those with 'endogenous' depression (36.34 +/- 2.23 pg/ml) than in healthy volunteers (125.19 +/- 1.64 pg/ml). The endogenously depressed patients had significantly lower beta-endorphin levels than the nonendogenous patients. A 4-week treatment of fluvoxamine (200 mg/day) caused a statistically significant increase in beta-endorphin serum levels in all patients (nonendogenous depression 132.10 +/- 2.38 pg/ml and endogenous depression 50.09 +/- 2.45 pg/ml) in comparison to values found before the onset of the therapy. The efficacy of fluvoxamine was 11.0 (+/- 9.0) evaluated by the Hamilton Rating Scale for Depression (HAMD) in the patients with a diagnosis of depression. These results indicate that determination of beta-endorphin serum levels could be a valuable laboratory test in the diagnosis of depression.

[Diagnostic importance of fibronectin in chronic liver diseases]. / Dijagnosticki znacaj fibronektina u hronicnim bolestima jetre.

Plasma fibronectin was determined in 29 patients with decompensated cirrhosis (7 patients had bacterial infection) and 23 patients with malignant liver disease. The obtained values were compared with the fibronectin values in 20 healthy subjects belonging to the control group in order to determine the possible diagnostic value of this dimer glycoprotein of high molecular weight whose role in the organism has not been completely explained. Fibronectin was determined on nephelometer with the use of specific antiserum by Behringwerke. The results expressed as mean values and SD were compared with monofactorial variance analysis (method One-way ANOVA). Fibronectin values in patients with liver cirrhosis were statistically significantly lower than in the control group (p < 0.01), which is also the case with correlation with malignant liver disease (p < 0.01). The fibronectin values in patients with malignant diseases were almost the same as the control group values (p < 0.01). In 7 patients with liver cirrhosis and bacterial infection the fibronectin values were statistically significantly higher in relation to those in the remaining 22 patients with cirrhosis but without bacterial infection (p < 0.001). The investigation in this study indicated that the decrease of mean fibronectin values is related to hepatic failure which is of diagnostic value, while normal values in malignant diseases do not favor the opinion on fibronectin as a tumor marker. Higher fibronectin values in infection in patients with liver cirrhosis are not clear, which indicated the total complexity of the relation between fibronectin as a dimer glycoprotein and chronic liver diseases including malignant.

Plasma and urinary serotonin and 5-HIAA in children treated with lamotrigine for intractable epilepsy.

Alteration of monoamine levels by some antiepileptic drugs (AEDs) was elucidated in this study. Lamotrigine (LTG) is a new AED, acting the sodium-channels. LTG was given as add-on therapy to 16 patients aged 4.5-18 yrs with intractable epilepsy and comedicated with carbamazepine or valproate. An equal group of epileptics with comparable clinical characteristics and treatment served as control. Plasma and urinary (24 h-samples) serotonin and 5-HIAA were determined before onset of LTG therapy and after 2-3 months. HPLC and electrochemical detection was used for the determination of serotonin (5-HT) and 5-hydroxy indoleacetic acid (5-HIAA). No significant effect of LTG on both urinary 5-HT and 5-HIAA levels was found, whereas plasma 5-HT concentrations significantly decreased in comparison with levels before LTG starting and relevant values in controls. This findings was noted in 7/16 children with favourable response to LTG. Increased serotonin catabolism may be result of LTG action.

Densitometric determination of urinary 4-hydroxy-3-methoxymandelic acid (vanillylmandelic acid).

We propose a simple and precise densitometric method for measuring vanillylmandelic acid (VMA). The method comprises direct urine application, thin-layer chromatographic (TLC) separation, postchromatographic derivatization, and in situ reading of the formed derivative at 560 nm. Quantitation was done by measuring peak area with a computer-controlled TLC Scanner and applying a five-point calibration function. The calibration function was 7.57-49.23 mumol/L. This method was used to determine urinary concentrations of excreted VMA; concentrations ranged from 4.84 to 56.06 mumol/day.

Reference values for apolipoproteins A-I and B in healthy subjects, by age.

We determined reference values of apolipoproteins A-I (apo A-I) and B (apo B) in serum from a population of 448 healthy subjects (265 men and 183 women, ages 18 to 61 years) by a kinetic immunonephelometric procedure. Frequency distributions of apo A-I were normal, whereas those of apo B were not and yielded asymmetrical curves. Thus, reference intervals for apo A-I were determined as mean +/- 2SD (1.08-1.89 g/L), but a nonparametric method was used for determining reference intervals for apo B (0.60-1.94 g/L). Apo B concentrations were significantly higher (P less than 0.001) in men than in women (0.63-2.01 g/L, mean 1.21 g/L; and 0.54-1.91 g/L, mean 1.08 g/L, respectively). No significant differences for apo A-I between men and women were observed. Concentrations of both proteins increased with age, but apo B increased more than apo A-I. We conclude that not only sex but also the age of the subjects must be considered in interpreting laboratory results for apolipoproteins.

A study of several red cell enzyme markers in the Rumanian ethnic group in Yugoslavia.

The present investigation reports the polymorphism of eight red cell enzymes, studied in 308 unrelated voluntary blood donors from the Rumanian ethnic group of Yugoslavia. Only common phenotypes were detected, which are distributed as in European populations. The estimated gene frequencies were: GLO1*1 = 0.401, GPT*1 = 0.533, PGM1*1 = 0.707, ESD*1 = 0.878, AK1*1 = 0.982, PGD*A = 0.974, ADA*1 = 0.939, ACP1*A = 0.384, ACP1*B = 0.550 and ACP1*C = 0.065. The observed gene frequencies are discussed in the context of other European populations and other populations from Yugoslavia.

Red cell enzyme polymorphisms of the Hungarian ethnic group in Yugoslavia.

A genetic study was carried out on phenotype and gene frequencies of the genetic markers in eight red cell enzymes: glyoxalase I (GLO1), glutamic pyruvate transaminase (GPT), phosphoglucomutase (PGM1), esterase D (ESD), adenylate kinase (AK1), 6-phosphogluconate dehydrogenase (6-PGD), adenosine deaminase (ADA), acid phosphatase (ACP1), in the Hungarian ethnic group living in Yugoslavia. The gene frequencies obtained were: GPT*1 = 0.542, PGM1*1 = 0.760, ESD*1 = 0.909, AK*1 = 0.971, PGD*A = 0.971, ADA*1 = 0.939, GLO1*1 = 0.417, ACP1*A = 0.329, ACP1*B = 0.591 and ACP1*C = 0.080. The distribution of these phenotype and gene frequencies was examined and compared with the phenotype and gene frequencies found for the Hungarian population living in Hungary and for other populations living in the northeast of Yugoslavia.

Eight red cell enzymes polymorphisms in the Slovakian ethnic group of Yugoslavia.

The polymorphisms of eight red cell enzymes were studied in 211 unrelated voluntary blood donors from the ethnic group of Slovakians (Yugoslavia). Only common phenotypes were detected, which are usually present in European populations. The allele frequencies found were: GLO1*1 = 0.410, ESD*1 = 0.887, AK1*1 = 0.962, PGM1*1 = 0.780, ACP1*A = 0.315, ACP1*B = 0.637, ACP1*C = 0.047, GPT*1 = 0.535, ADA*1 = 0.952 and PGD*A = 0.940. These findings are discussed in the context of other European populations and the population of Serbia, Yugoslavia.

Determination of platelet monoamine oxidase by new continuous spectrophotometric method.

A simple, continuous spectrophotometric method for the determination of tissue monoamine oxidase based on the oxidation of 2,2'-azino-di-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) using peroxidase has already been described (Ivanovic, I. & Majkic-Singh, N. (1986) Fresenius Z. Anal. Chem. 324, 307). In the present study the method is optimised for platelet monoamine oxidase assay and applied to healthy persons and schizophrenic patients. The obtained data were statistically analysed. The continuous ABTS method is sensitive, precise (CV below 6.9%) and linear up to 83 U/g protein. Comparison with the end-point method of Szutowicz et al. (1984) Anal. Biochem. 138, 86-94) gave a good correlation (r = 0.983). The reference values for the activity of human platelet monoamine oxidase by the new continuous ABTS method are 25 to 42 U/g protein (means = 33.2 U/g protein, CV = 15.5%, n = 67). No differences were found between females and males, or between three age groups ranging from 21 to 52 years. The patients with chronic (n = 76) or acute (n = 17) schizophrenia had significantly lower monoamine oxidase activities compared with normal values (p less than 0.005), which indicates that platelet monoamine oxidase can be a possible marker for schizophrenic diseases.

Esterase D polymorphism in Serbia (Yugoslavia).

Phenotypes of human red cell esterase D (EsD) were determined in 351 unrelated adults from Serbia (Yugoslavia). The calculated allele frequencies were 0.911 for EsD1 and 0.089 for EsD2. The phenotype distribution was in good agreement with the Hardy-Weinberg equilibrium.