Effects of short- and long-term physical activity on DNA stability and oxidative stress status in young soccer players.

AIM: The aim of the study was to determine whether single soccer training is able to cause oxidative stress and DNA instability. We have also sought to investigate if adaptive response will be developed during 45 days training period and to what extent. METHODS: This study was conducted on 16 soccer players aged 18.13±0.35 years. We used single cells gel electrophoresis (comet assay) to investigate leukocyte DNA stability. The results were presented as DNA score and percent of cells with medium and high damage. Oxidative status of our subjects was estimated through blood levels of superoxide anion, the thiobarbituric acid-reacting substances (TBARs), total antioxidant status (TAS), total oxidant status (TOS), prooxidant-antioxidant balance (PAB) and sulfhydryl-groups (SH-groups). RESULTS: During single soccer training, DNA score and percent of the cells with medium and high damage were increased after training but without significance. Sulphydryl-groups (P=0.033), TOS (P=0.002) and PAB (P=0.045) were significantly lower after training. After 45 days training period DNA score was decreased but with no significance. However, percent of cells with medium and high damage was significantly lower (P=0.01). TOS (P=0.001) and MDA (P=0.038) levels were also significantly lower, while sulphydryl-groups levels were significantly higher (P=0.006). CONCLUSION: This study demonstrated that single soccer training had not compromised DNA stability. Possible development of oxidative stress was effectively neutralized by very well preserved antioxidative mechanisms. It was also shown that during 45 days adaptive response was induced. All measured parameters should be considered as useful information on oxidative status of trainees.

Oxidative stress and anti-oxidative defense in schoolchildren residing in a petrochemical industry environment.

OBJECTIVE: To evaluate the possible relationship between industrial air pollution and oxidative stress in schoolchildren by comparing parameters from children residing in two nearby localities with contrasting environmental conditions. PARTICIPANTS: 42 schoolchildren (12-15 years) from Pancevo (site of Serbias largest petrochemical installation) formed the exposed group. 82 schoolchildren from Kovacica village, located 30 km north of Pancevo, formed the non-exposed group. METHODS: Oxidative stress status, anti-oxidative defense parameters, paraoxonase-1 status, lipid status, glucose concentration and leukocyte counts were compared in two groups. RESULTS: The children from Pancevo showed higher level of oxidative stress demonstrated by an elevated malondialdehyde concentration (P <0.001) and decreased superoxide dismutase activity (P<0.01) in comparison to the non-exposed group. CONCLUSIONS: The results suggested a relationship between the presence of air pollutants and increased oxidative stress in schoolchildren residing in an industrial environment.

Association of oxidative stress and PON1 with LDL and HDL particle size in middle-aged subjects.

BACKGROUND: Alterations in plasma lipoprotein subclass distributions affect atherosclerosis risk. Smaller, denser low-density lipoprotein (LDL) particles (sdLDL) are more susceptible to oxidation. In contrast, most of the protective effects of high-density lipoproteins (HDL) are attributable to larger particles. This study investigates the connection between LDL and HDL particle heterogeneity and oxidative stress, antioxidative defence (AOD) and paraoxonase (PON1) status in a healthy middle-aged Serbian population. MATERIALS AND METHODS: LDL and HDL particle sizes and subclass distributions were measured by gradient gel electrophoresis in 104 men and 103 women, aged 53 +/- 9.4 years. PON1 activities and PON1(Q192R) phenotypes were determined with paraoxon and diazoxon as substrates. The oxidative stress/AOD status was estimated by measuring malondialdehyde (MDA) and superoxide-anion (O2*(-)) levels and superoxide-dismutase (SOD) activity. RESULTS: Subjects with sdLDL had significantly higher MDA (P < 0.001) and O2*(-)(P < 0.05) levels and greater diazoxonase (DZOase) activity (P < 0.05) compared to subjects with larger LDL particles. A high MDA concentration was a significant predictor of the sdLDL phenotype (P < 0.005). Increased levels of and MDA were associated with smaller HDL(3) subclass abundance. Reduced HDL particle size was associated with lower DZOase activity (P < 0.01). CONCLUSIONS: Even in the absence of symptoms of atherosclerosis, sdLDL particles are associated with increased oxidative stress, which may stimulate a compensatory rise in PON1 DZOase activity. Elevated oxidative stress may significantly affect HDL subclass distribution, resulting in the accumulation of smaller, denser HDL particles with diminished antioxidative capacity.

Low serum alpha-1-antitrypsin specific activity in monoclonal gammopathies.

BACKGROUND: Serum alpha 1-antitrypsin (alpha 1AT) antigen concentration is elevated in malignancies as the result of acute phase reaction. In the present study, we examined whether the alpha 1AT elevation in monoclonal gammopathies was accompanied by an adequate increase of its functional activity. METHODS: In this case-control study, serum alpha 1AT concentration was measured in 187 ambulatory patients with monoclonal gammopathies and 320 healthy blood donors matched according to sex and age. The alpha 1AT antigen concentration was assayed by immunonephelometry, whereas its functional activity was measured as trypsin inhibitory capacity (TIC). The specific alpha 1AT inhibitory activity (SIA) was calculated, defined as the TIC/antigen concentration ratio. RESULTS: The alpha 1AT antigen concentrations obtained in the patients' samples were very significantly higher as compared with the corresponding values in the control group (mean +/- SD = 134 +/- 41.9% of normal, p < 0.001). However, the TIC values were higher in the patients than in the healthy controls only by 4% (104 +/- 23.8%, p < 0.05). The specific alpha 1AT activity was very significantly lower in the patients, as compared with the controls (p < 0.001), indicating that serum alpha 1AT in monoclonal gammopathies was partially inactive. CONCLUSIONS: As poor correlation between the TIC values and the antigen concentrations was obtained in the patient group as well as the decreased specific alpha 1AT activities, the TIC values in patients with monoclonal gammopathies should be interpreted with caution.

Simple and rapid method for the measurement of nitrite and nitrate in human plasma and cerebrospinal fluid by capillary electrophoresis.

Nitrite and nitrate levels in physiological fluids are commonly used as an index of nitric oxide production. We developed simple and rapid method for the determination of these anions by capillary zone electrophoresis employing borate buffer (pH 10, 100 mmol/l) as running electrolyte. The anions were analyzed in plasma and cerebrospinal fluid (CSF) without deproteinization of the samples. Electrophoresis was carried out in a capillary (36.5 cm x 75 microm) at a potential of 15 kV, with on-column UV detection at 214 nm. Mean retention times for nitrite and nitrates were 4.631 and 5.152 min, respectively. The method was linear (r=0.999) within a 1-500 micromol/l concentration range. Physiological levels of nitrate in plasma (40.2 micromol/l) and CSF (15.3 micromol/l) could be determined with good precision (coefficients of variation <6%) and accuracy (recoveries of added nitrate to plasma and CSF were 97.4 and 104.5%, respectively). Measurements of the physiological levels of nitrite in plasma (6.1 micromol/l) and CSF (0.9 micromol/l) were less precise and accurate.

Low frequency of PIM3 gene in patients with monoclonal gammopathies.

The distribution of PI (protease inhibitor) phenotypes and PI M subtypes was studied in 200 patients with monoclonal gammopathies and 320 healthy blood donors by isoelectric focusing in thin-layer polyacrylamide gels, pH range 4-5. The distribution of PI phenotypes and gene frequencies in the patients differed significantly from the corresponding values found in the blood donors, as shown by the chi 2 test (chi 2 = 16.5, p = 0.02 and chi 2 = 17.6, p = 0.0014, respectively). A lower frequency of the M3 variant was observed in patients, both in homozygous (PI M3) and heterozygous forms (PI M1M3 and PI M2M3). No significant difference between PIz allele frequencies in patients and healthy controls was found.

alpha-1-Antitrypsin (Pi) polymorphism in Serbia: deviation of Pi M subtype distribution from the Hardy-Weinberg equilibrium.

The distribution of the alpha 1-antitrypsin (Pi) phenotypes and subtypes was investigated in a population sample of 1060 unrelated individuals from Serbia (Yugoslavia). The allele frequencies estimates were: Pi*M1: 0.702; Pi*M2: 0.183; Pi*M3: 0.088; Pi*Z: 0.013, Pi*S: 0.007; Pi*P: 0.004; Pi*F: 0.003. The observed phenotype frequencies differed very significantly from those expected assuming H.W. equilibrium (chi 2 = 49.51, p < 0.0005). The deviation from equilibrium involved the three Pi*M subtypes: an excess of Pi*M1, Pi*M2 and Pi*M3 homozygotes was found, with the corresponding decreased number of M1M2 and M1M3 heterozygotes. The possible significance of this finding is discussed.

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.

Plasma C1 inhibitor in malignant diseases: functional activity versus concentration.

Plasma C1 inhibitor (C1-INH) was determined in 64 patients with different malignant diseases and in 58 healthy persons. C1-INH antigen concentration was measured by radial immunodiffusion (RID), whereas its functional activity was assayed with chromogenic substrate. Within-run and day-to-day precision of both methods were good, with CVs ranging from 3.6 to 5.4%. Plasma C1-INH antigen concentrations were significantly higher in the patients than in healthy controls (P = 4.0 x 10(-3)), as were their C1-INH functional activities (P = 3.5 x 10(-3)). C1-INH activities obtained in the patient plasma samples were in correlation with their antigen concentrations (r = 0.914), showing that C1-INH synthesized in malignant disease was functional. However, the specific activity of the C1-INH (functional activity/antigen concentration ratio) was significantly lower in the patient group as compared with the controls (P = 2.1 x 10(-6)), indicating partial inactivation of plasma C1-INH in malignant diseases. The C1-INH specific activity in patients was inversely proportional to its antigen concentration.

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.

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.

Polymorphism of human red cell galactose-1-phosphate uridyl transferase in Serbia, Yugoslavia.

Phenotypes of human red cell galactose-1-phosphate uridyl transferase (GALT) were determined in 283 unrelated adults from Serbia (Yugoslavia). The gene frequencies were 0.959 for GALT N, 0.018 for GALT D and 0.023 for GALT N.

Effects of some anti-inflammatory drugs on 12 blood constituents: protocol for the study of in vivo effects of drugs.

We investigated the in vivo effects of acetylsalicylic acid, diclofenac, indomethacin, ibuprofen, and ketoprofen on the concentrations of various blood constituents. Total protein, glucose, calcium, and inorganic phosphate were not significantly affected by any of these drugs. Ketoprofen had no definite influence on any constituent. Acetylsalicylic acid induced an increase in cholesterol, triglyceride, and iron; albumin, uric acid, and creatinine decreased with ibuprofen therapy. Urea nitrogen increased in patients treated with diclofenac or indomethacin. Our protocol for the study of in vivo drug effects is discussed.

Interference by analgesic and antirheumatic drugs in 25 common laboratory assays.

Twenty five different analytical procedures, commonly used in clinical laboratories, were investigated for interference by eight analgesic and antirheumatic drugs. Ten of the investigated assays showed no statistically significant interference. Acetylsalicylic acid interfered in six assays (for glucose, uric acid, protein and cholesterol). Aminophenazone significantly decreased glucose, bilirubin and protein values, whereas caffeine affected four methods (for glucose, protein and iron). No definite influence of phenobarbital could be detected on any assay. Glucose, uric acid and iron values were altered in the presence of diclofenac. Indomethacin interfered in glucose, urea, uric acid and protein assays. Samples containing ibuprofen had altered creatinine, bilirubin and iron values, whereas ketoprofen interfered in glucose and iron determination.

Polymorphism of red cell glyoxalase I in Serbia, Yugoslavia.

Red cell glyoxalase I (GLO) phenotypes were determined in 258 unrelated adults from the population of Serbia (Yugoslavia). The GLO1 gene frequency was estimated to be 0.384.