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Testicular nitric oxide and thiobarbituric acid reactive substances levels in obstructive azoospermia: a possible role in pathophysiology of infertility.

OBJECTIVE: The aim of the study is to evaluate the levels of nitrite/nitrate and thiobarbituric acid reactive substances (TBARS) and their relationship with seminal parameters in experimental obstructive azoospermic rats to explain the possible mechanism of impaired sperm quality in obstructive azoospermia. METHODS: A total of 10 male Spraque-Dawley rats underwent bilateral vas resection and ligation (Group-1 = vasectomy group). The findings were compared with control group (Group-2 = sham group, n = 10). Animals were sacrificed 8 weeks after surgery. Testes were removed and used for the evaluation of nitrate/nitrite and TBARS levels and for histology. Epididymal-aspirated seminal plasma was used for semen count and morphological analysis according to the Kruger criteria. RESULTS: Testicular tissue nitrate/nitrite and TBARS levels were 35.7 +/- 3.1 micromol/g protein and 3.7 +/- 0.1 nmol/g protein in Group-1, and 19.3 +/- 0.7 micromol/g protein and 3.1 +/- 0.1 nmol/g protein in Group-2, respectively. Both parameters showed statistical differences between the two groups. Testicular tissue nitrate/nitrite and TBARS levels showed negative and statistically significant correlations with sperm motility and morphology. CONCLUSIONS: The present study showed that testicular nitrate/nitrite and TBARS levels were increased in obstructive azoospermia. For that reason, we concluded that antioxidant treatment can be recommended to patients before sperm extraction for artificial reproduction due to obstructive infertility after vasectomy reversal.

Animals↗

Amino acids and thiobarbituric acid reactive substances in cerebrospinal fluid and plasma of patients with septic encephalopathy.

Amino acids and thiobarbituric acid reactive substances (TBARs) in the cerebrospinal fluid (CSF) and plasma were identified and assayed in 5 patients with septic encephalopathy. Levels of all the high molecular weight neutral amino acids (LNAAs) appeared to increase in the CSF. CSF levels of phenylalanine (PHE) and methionine (MET) increased significantly by factors of 20.9 and 9.5, respectively, and the plasma PHE level increased 7.5-fold. No significant changes in branched-chain amino acids were observed in either the CSF or plasma. The CSF/plasma ratios of valine (VAL), tyrosine (TYR), PHE, and MET significantly increased to 0.21, 0.46, 0.52, and 0.52, respectively. TBAR levels increased 4-fold in the CSF and also were slightly increased in the plasma, suggesting that lipid peroxidation in the central nervous tissues is markedly increased. We conclude that increases in LNAA levels and in lipid peroxidation in the central nervous tissues may play important roles in the development of septic encephalopathy.

Aged↗

Activity of antioxidant enzymes and concentrations of thiobarbituric acid reactive substances (TBARS) in melanotic and amelanotic Bomirski melanoma tissues in the golden hamster (Mesocricetus auratus, Waterhouse).

The activity of superoxide dismutase (SOD) and glutathione peroxidase (GSHPx), as well as the concentration of thiobarbituric acid reactive substances (TBARS) in tissues of transplantable melanoma in the golden hamster were measured and compared. Ten inbred male hamsters were used for the experiment. They were divided into two groups and were given Bomirski melanoma cells subcutaneously. The first group was given melanotic (Ma) melanoma cells. The second group was given amelanotic (Ab) melanoma cells. Thirty days after the transplantation the hamsters were dissected and the tumor tissues were taken and homogenized. A statistically significantly higher activity of the measured antioxidant enzymes was found in homogenates of Ma tumor than in homogenates of the Ab tumor. Activity of SOD is 8% higher in melanotic melanoma, 24% higher in CAT, and 45% higher in GSHPx. Statistically significant differences between TBARS concentrations were not confirmed. The higher activity of antioxidant enzymes in the melanotic tumor is a result of increased generation of oxygen-derived free radicals. It is presumed that it is strictly connected with intensified production of quinone and semiquinone radicals in the process of melanogenesis.

Animals↗

Catalase activity and thiobarbituric acid reactive substances (TBARS) production in a rat model of diffuse axonal injury. Effect of gadolinium and amiloride.

This study evaluated the effect of mechanogated membrane ion channel blockers on brain catalase (CAT) activity and thiobarbituric acid reactive substances (TBARS) production after traumatic brain injury (TBI). A weight drop trauma model was used. Controls were sham-operated and received no weight drop. Gadolinium (GAD) or amiloride (AMI) were administered to control and experimental rats (30 min after TBI). Brain CAT activity and TBARS production were measured. When blood vessels were washed out with saline perfusion brain CAT activity significantly increased up to 6 h after trauma, decreasing significantly by 24 h; GAD or AMI administration preserved CAT activity 24 h after TBI. TBARS production increased after trauma, this effect being significantly reversed by GAD or AMI administration. GAD significantly decreased TBARS production in control animals. Mechanogated membrane ion channels may be involved in the genesis of the ionic disruption leading to oxidative stress and other secondary injury processes in head trauma.

Amiloride↗

Thiobarbituric acid reactive substances and vitamin E in serum and cerebrospinal fluid from patients with cerebral apoplexy.

Thiobarbituric acid (TBA) reactive substances in serum and cerebrospinal fluid from patients with cerebral apoplexy were determined and their relationship to this disease was studied. TBA reactants in serum were elevated in patients with cerebral apoplexy, but there was no difference between patients with brain tumor or other neurological disease and healthy persons. TBA reactants in cerebrospinal fluid in patients with cerebral aneurysm, cerebral arterial and venous anomalies or intracerebral hematoma, and its levels in patients with cerebral infarction or brain tumor were also higher than in healthy persons. Serum alpha-tocopherol levels were elevated in patients with cerebral infarction, but were relatively low in patients with cerebral aneurysm, cerebral arterial and venous anomalies or intracerebral hematoma. The reverse was the case with cerebrospinal fluid levels of alpha-tocopherol. That is, the sequence of decreasing level of alpha-tocopherol in cerebrospinal fluid was intracerebral hematoma, cerebral aneurysm, cerebral arterial and venous anomalies, and cerebral infarction. The levels of TBA reactive substances in patients with cerebral apoplexy showed a positive correlation between serum and cerebrospinal fluid, and the levels of alpha-tocopherol showed a similar correlation.

Aged↗

Plasma thiobarbituric acid reactive substances a predictive marker of cerebral ischemia in patients at high risk?

The clinical meaning of high values of blood lipid peroxides, assessed as thiobarbituric acid reactive substances (TBA-RS), was investigated in 19 selected high risk patients with transient ischemic attacks (TIA). Patients were checked every 3-6 months and followed-up for 3 years. 8 patients experienced further vascular episodes, 4 having minor stroke and 4 TIA; one of the latter died from myocardial infarction. Unlike blood cholesterol and glucose. TBA-RS values discriminated patients with vascular episodes: they, indeed, showed significant higher values of TBA-RS. Discriminant analysis further indicated that TBA-RS levels differentiate patients with and without vascular accidents, suggesting that high blood values of lipid peroxides could represent a predictive sign of vascular ischemia.

Aged↗

Vitamin E, thiobarbituric acid reactive substance concentrations and superoxide dismutase activity in the blood of children with juvenile rheumatoid arthritis.

OBJECTIVE: To study the role of active oxygen species in tissue injury in rheumatoid arthritis. METHODS: We examined the levels of thiobarbituric acid reactive substances (TBARS) and antioxidants of the first line antioxidative defence of the organism, i.e. vitamin E (VE) and superoxide dismutase (SOD) in the blood of 74 young patients with juvenile rheumatoid arthritis (JRA) and in 138 healthy children, all aged 3-15. RESULTS: A statistically significant increase of TBARS was found in the blood plasma of the children with JRA compared with the control group. In the whole group of patients and in the patients over 6 years of age, the VE concentration was in the red blood cells (RBC) was significantly lower in children who had suffered from JRA for more than one year and in those with the systemic form of the disease. The type of treatment also affected the values for the plasma VE and SOD in the RBC. CONCLUSION: Our results seem to confirm the supposition of increased oxidative stress in children with JRA and low antioxidant levels in terms of SOD activity and vitamin E concentrations.

Adolescent↗

GM1 ganglioside attenuates convulsions and thiobarbituric acid reactive substances production induced by the intrastriatal injection of methylmalonic acid.

The effects of the administration of monosialoganglioside (GM1) on methylmalonic acid (MMA)-induced convulsions, production of thiobarbituric acid reactive substances (TBARS) and on the striatal content of ascorbic acid and total non-protein thiol (SH) groups were evaluated in adult male rats. Animals received two intraperitoneal injections of GM1 (50 mg/kg) or saline (0.85% NaCl) spaced 24h apart. Thirty minutes after the second GM1 or saline injection, L-MMA (6 micromol) or NaCl (9 micromol) was injected into the right striatum and the animals were observed for the appearance of convulsions for 15 min. The animals were sacrificed and their striatal content of ascorbic acid, SH groups and TBARS was measured. The effect of GM1 on MMA-induced TBARS production in striatal homogenates was also evaluated in vitro.MMA injection caused convulsions (Sal-MMA: 9.8+/-1.4 episodes, which lasted 271+/-48 s) and increased the striatal content of TBARS (Sal-MMA: 149.0+/-11.5 nmol MDA/g tissue), but did not alter total striatal SH or ascorbic acid contents. GM1 pretreatment decreased MMA-induced convulsions (GM1-MMA: 6.3+/-2.0 episodes, which lasted 115.1+/-42.2s) and TBARS increase (GM1-MMA: 102.4+/-19.5 nmol MDA/g tissue). GM1 pretreatment increased ascorbic acid content of the striata (saline-pretreated: 1514+/-75.9; GM1-pretreated: 1878.6+/-102.8 microg ascorbic acid/mg tissue). MMA increased TBARS production in vitro, and GM1 had no effect on such MMA-induced effect. This study provides evidence that GM1 increases striatal ascorbic acid content and decreases MMA-induced neurotoxicity assessed by behavioral and neurochemical parameters.

Animals↗

A phospholipase A2 inhibitor decreases generation of thiobarbituric acid reactive substance during lung ischemia-reperfusion.

A novel active-site directed specific inhibitor of phospholipase A2 (PLA2), 1-hexadecyl-3-trifluoroethylglycero-sn-2-phosphomethanol (MJ33), administered endotracheally co-dispersed in liposomes, significantly reduced the formation of thiobarbituric acid reactive substances (TBARS) in isolated rat lungs subjected to ischemia-reperfusion. Elevated conjugated dienes were unaffected. This contrasts with the effects of the cyclo-/lipoxygenase inhibitor 5,8,11,14-eicosatetraynoic acid (ETYA), which decreased formation of both TBARS and conjugated dienes (CD). The effects of MJ33 plus ETYA were additive for TBARS but results for CD were similar to ETYA alone. A similar dissociation of inhibition of TBARS and CD formation by MJ33 was observed with t-butyl hydroperoxide induced lipid peroxidation of isolated lung microsomes. Assay of lung homogenate with phosphatidylcholine as substrate showed that MJ33 selectively inhibited the Ca(2+)-independent acidic PLA2. MJ33 had no effect on thromboxane B2 release by the isolated lung, indicating the effects of acidic PLA2 inhibition do not involve the arachidonate cascade. MJ33 also partially prevented lung edema and lactate dehydrogenase release associated with ischemia-reperfusion. The observations show that this PLA2 inhibitor can be delivered to oxidant-sensitive lung sites by its co-dispersal in liposomes, and that oxidant-induced lipid peroxidation in this model of lung injury occurs in a complex lipid prior to PLA2 activity.

5,8,11,14-Eicosatetraynoic Acid↗

Chlorpyrifos induced alterations in levels of thiobarbituric acid reactive substances and glutathione in rat brain.

Chlorpyrifos, O,O'-diethyl-O-3,5,6-trichloro-2-pyridyl phosphorothionate, exposure in rats caused significant inhibition of acetylcholinesterae activity in different regions of brain and generated oxidative stress as evidenced by increase in the level of thiobarbituric acid reactive substances and decrease in the ratio of reduced to oxidized glutathione in all the three regions of brain. Malondialdehyde level was increased significantly in all regions of brain and the increase was dose dependent. Mid brain showed highest level of lipid peroxidation.

Animals↗

The effect of ascorbic acid and ethanol on the level of thiobarbituric acid reactive substances (TBARS) in selected tissues of albino BALB/c mice.

The aim of the paper was to study the effect of ascorbic acid (vitamin C) and ethanol on the level of thiobarbituric acid reactive substances (TBARS) in the brain, liver, lungs and blood serum of inbred albino BALB/c mice. Sixty male mice divided into 4 groups were used for the study; the mice in each group were injected intraperitoneally (i.p.): Group 1--control, saline; Group 2--H2O2 and FeSO4; Group 3--ascorbic acid (AA); Group 4--AA and ethanol. The level of TBARS in the investigated tissues of the mice in all of the groups was statistically significant higher than in the control group. The highest content of TBARS in blood serum, brain, liver and lung tissue was observed in mice which were given ascorbic acid and ethanol.

Animals↗

Broad-band ultraviolet B phototherapy is associated with elevated serum thiobarbituric acid reactive substance and nitrite-nitrate levels in psoriatic patients.

BACKGROUND: Although the local anti-inflammatory, immunosuppressive and oxidative activity of UVB is known, the systemic effect of UVB phototherapy in dermatological patients has not been investigated. OBJECTIVE: We aimed to investigate the lipid peroxidation status (as represented by thiobarbituric acid reactive substance, TBARS) and nitrite-nitrate levels in psoriatic patients under broad-band ultraviolet B (BB-UVB) phototherapy in order to determine the systemic effects of UVB. SUBJECTS AND METHODS: Thirty-two psoriatic patients and 20 healthy controls were included. Blood samples were obtained at the beginning, after 6-10 exposures to BB UVB phototherapy (mean 5 weeks) and at the end of the therapy period (mean 21 weeks). Serum TBARS and nitrite-nitrate levels were evaluated. RESULTS: There was no statistically significant difference in serum TBARS and nitrite-nitrate levels between psoriatic patients (basal) and healthy volunteers. There was no statistically significant correlation between disease duration, disease severity, or the total cumulative dose of UVB and serum levels of TBARS and nitrite-nitrate in psoriatic patients. Total nitrite levels in samples obtained during and at the end of therapy were significantly higher than basal levels (P=0.033 and P=0.005, respectively). TBARS levels in samples obtained during and at the end of therapy were significantly higher than basal levels (P=0 and P=0.026, respectively). There was a negative correlation (r=-0.576, P=0.039) between the total nitrite and TBARS levels in psoriatic patients at the end of therapy. CONCLUSION: Our study showed that chronic exposure to UV irradiation may lead to a systemic effect on lipid peroxidation and NO levels, which are shown by a significant elevation in TBARS and nitrite-nitrate levels in serum.

Adult↗

Thiobarbituric acid reactive substances, Fe3+ reduction and enzymatic activities in cultures of Ganoderma australe growing on Drimys winteri wood.

Ganoderma australe is a basidiomycete responsible for a natural process of selective and extensive lignin degradation. Fatty acids, thiobarbituric acid reactive substances (TBARS), Fe3+-reduction and enzymatic activities were monitored in cultures of G. australe growing on Drimys winteri wood chips. Linoleic acid was de novo synthesized, and steadily increased during 12 weeks of cultivation. Part of the unsaturated fatty acids underwent peroxidation as TBARS accumulated with biodegradation time. TBARS accumulation was proportional to the wood weight and component losses. Manganese-dependent peroxidase and lignin peroxidase were not detected in the culture extracts, whereas laccase-induced oxidation of syringaldazine peaked after 2 weeks (104+/-9 micromol oxidized min(-1) kg(-1) of dry wood), subsequently decreasing. On the other hand, nonenzymatic Fe3+-reducing activity increased as a function of cultivation time and could be involved in the initiation of lipid peroxidation.

Biodegradation, Environmental↗

Effects of magnetic fields on the accumulation of thiobarbituric acid reactive substances induced by iron salt and H(2)O(2) in mouse brain homogenates or phosphotidylcholine.

In this study, we examined the effects of magnetic fields (MFs) on the generation of thiobarbituric acid reactive substances (TBARS) in the mouse brain homogenates or phosphotidylcholine (PC) solution, incubated with FeCl(3) and/or H(2)O(2). Active oxygen species were generated and lipid peroxidation was induced in mouse brain homogenates by incubation with iron ions, resulting in the accumulation of TBARS. Lipid peroxidation was induced in PC by incubation with iron ions and H(2)O(2). Exposure to sinusoidal MFs (60 Hz, 0.2-1.2 mT), symmetric sawtooth-waveform MFs (50 Hz, 25-600 mT/s), rectangular MFs (1/0.4-1/16 Hz, 3.3 mT) and static MFs (1, 5-300 mT) had no effect on the accumulation of TBARS in brain homogenates induced by FeCl(3). In contrast, when the homogenates were incubated with FeCl(3) in static MFs (2-4 mT), the accumulation of TBARS was decreased. However, this inhibitory effect disappeared when EDTA was added to the homogenate and incubated with H(2)O(2). The accumulation of TBARS in PC solution incubated with FeCl(3) and H(2)O(2) was also inhibited by the static MF. These results indicate that only static MFs had an inhibitory effect on iron-induced lipid peroxidation and the effectiveness of this magnetic field on iron ion-induced active oxygen species generation is restricted to a so called 'window' of field intensity of 2-4 mT.

Journal Article↗

A method to reduce interference by sucrose in the detection of thiobarbituric acid-reactive substances.

A thiobarbituric acid (TBA) reaction for measuring lipid peroxidation products was evaluated for interference by several ingredients commonly used in solutions to prepare or analyze tissue homogenates or subcellular organelles. These included sucrose (up to 100 mM final concentration in the assay medium), Tris-maleate (up to 40 mM), imidazole (up to 20 mM), inorganic phosphate (up to 10 mM), and 4- morpholinepropanesulfonic acid (up to 20 mM). When the samples were heated at 95 degrees C as recommended in some procedures, only sucrose significantly affected color development. Sucrose concentrations as low as 10 mM significantly increased absorbance at 532 nm of aqueous tetramethoxypropane (TMP) standards, and so the assay could not be applied reliably to tissue samples prepared in sucrose. Sucrose interference was only partially reduced by subsequent organic extraction (n-butanol plus pyridine), with measured absorbances remaining significantly greater (50-100%) than sucrose-free controls at sucrose concentrations of 20 mM or more. Modifying the assay to include sucrose in blanks and TMP standards failed to adequately correct for interference when the absorbance of unextracted (aqueous) solutions was measured. Further modification by adding sucrose to blanks and TMP standards, followed by butanol-pyridine extraction, gave standard curves that were linear, through the origin, and had slopes equivalent to those of sucrose-free standards. This modification enabled almost complete recovery (average 2% error) of known amounts of TMP added to aliquots of tissue homogenates containing amounts of sucrose that otherwise significantly interfered. Also, with the modified method the content of TBA-reactive substances in tissues homogenized in sucrose was found to be not significantly different from that measured in tissues homogenized in a noninterfering substance, KCl.

Animals↗

Probucol scavenged 1,1-diphenyl-2-picrylhydrazyl radicals and inhibited formation of thiobarbituric acid reactive substances.

Probucol is suggested to have antioxidant properties. The direct scavenging action of probucol on hydroxyl radicals, superoxide and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals were examined using electron spin resonance (ESR) spectrometry. Probucol scavenged DPPH radicals dose dependently but showed no effect on hydroxyl radicals and superoxide generated by Fenton reaction and by hypoxanthine-xanthine oxidase system, respectively. It inhibited the formation of thiobarbituric acid reactive substances (TBARS) in rat cortex homogenate induced by ascorbic acid and FeCl2 at low dose, but it increased TBARS formation at high doses. Probucol showed no effect on the carbon centered radicals. Iron injection into the rat cortex, which is an experimental model for traumatic epilepsy, increased TBARS level in the cortex, hippocampus, striatum and cerebellum, but pretreatment with probucol inhibited the increase in these brain parts except for the hippocampus. These results suggest that the antioxidant property of probucol is partly due to its free radical scavenging effect.

Animals↗

Effect of oxidative stress on (3H)N-methylscopolamine binding and production of thiobarbituric acid reactive substances in rat cerebral cortex membranes.

We investigated the effect of lipid peroxidation, in vitro induced by H2O2 or FeSO4 and ascorbic acid, on binding properties of muscarinic receptors in rat cerebral cortex membranes. Simultaneously the concentrations of thiobarbituric acid reactive substances (TBARS) were measured to assess the extent of lipid peroxidation. In conditions of increased TBARS levels the density of (3H)N-methylscopolamine [(3H)NMS] binding sites in rat cerebral cortex membranes was not affected. Decreased numbers of (3H)NMS binding sites observed in the presence of high concentrations of H2O2 (100 and 1000 mmol.l-1) accompanied by a decrease of TBARS levels might be related to a nonspecific effect of H2O2 on cellular proteins.

Animals↗

Gingival crevicular fluid prostaglandin E(2) and thiobarbituric acid reactive substance levels in smokers and non-smokers with chronic periodontitis following phase I periodontal therapy and adjunctive use of flurbiprofen.

BACKGROUND: It has been established that smoking is an important risk factor for the initiation and progression of chronic periodontitis (CP). This study investigates the effects of phase I periodontal therapy and adjunctive flurbiprofen administration on prostaglandin E(2) (PGE(2)) and thiobarbituric acid reactive substance (TBARS) levels in gingival crevicular fluid (GCF) samples from smoker and non-smoker patients with CP. METHODS: Twenty-one non-smoker and 21 smoker patients with CP were divided into four groups according to treatment modalities. Group 1 (non-smokers with CP) and group 3 (smokers with CP) patients received daily 100-mg flurbiprofen tablets in a 2 x 1 regimen for 10 days together with scaling and root planing (SRP). Patients in group 2 (non-smokers with CP) and group 4 (smokers with CP) received placebo tablets in a 2 x 1 regimen for 10 days together with SRP. Plaque index (PI), gingival index (GI), probing depth (PD), and clinical attachment level (CAL) measurements were recorded and GCF samples were collected at baseline and on day 10 of drug intake from each sampling area by a single examiner who was unaware of the treatment modality. Assays for GCF PGE(2) and TBARS were carried out by an enzyme-linked immunosorbent assay and fluorometric method, respectively. RESULTS: All groups showed statistically significant reductions in PI and GI scores following the phase I periodontal treatment on day 10 (P <0.05), but no statistical differences were observed in PD and CAL scores after the therapy. In groups 1 and 2, the reduction of GCF PGE(2) and TBARS levels were not significant after the therapy compared to baseline levels. In group 3, GCF PGE(2) and TBARS levels exhibited a statistically significant decrease (P <0.05) after the therapy. Group 4 showed significant reductions (P <0.05) in GCF PGE(2) levels after the therapy. No statistically significant reductions were observed in group 4 with regard to GCF TBARS levels. When groups 1 and 3 were compared according to GCF TBARS levels after the therapy, a more statistically significant reduction was observed in group 3 (P = 0.001). CONCLUSION: These results suggest that additional flurbiprofen administration may have more inhibitory effects on GCF levels of PGE(2) and TBARS in the groups of smokers compared to non-smokers with CP.

Adult↗