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AIM: To observe the protective effect of astaxanthin (AST) against hydroquinone (HQ) mediated cell death in the apoptotic cascade and evaluate intracellular Ca2+ release, caspase-3, and -9 activation, reactive oxygen species (ROS) production in ARPE-19 cells. METHODS: We cultured ARPE-19 cells in special mediums and performed MTT tests to determine protective effect of AST, before exposing the cells to HQ in an incubator. We analyzed intracellular Ca2+ release experiments, mitochondrial membrane depolarization, glutathione (GSH), glutathione peroxidase (GSH-Px) and ROS experiments, and apoptosis assay. RESULTS: ROS production ranges depend on the amount of cell death. We computed the correlation between ROS ranges and cell death by 20,70-dichlorofluorescein fluorescence, and Ca2+ levels by Fura-2-AM. HQ-induced cell death found out to rise ranges of caspase-3 and -9, and mitochondrial depolarization. These three steps were delayed by AST management. CONCLUSION: ARPE-19 cells are avoided from HQ-induced ROS production and caspase-3 and -9 activation by AST. AST may limit the range of caspase synthesis, Ca2+ release and excess production of ROS with antiapoptotic effect. This study proposes a new therapeutic approach for the treatment of age-related macular degeneration.

Purpose: Under conditions of oxidative stress, cell apoptosis is triggered through the mitochondrial intrinsic pathway. Increased levels of reactive oxygen species (ROS) are linked to excess cell loss and mediate the initiation of apoptosis in a diverse range of cell types. The aims of this study were to assess intracellular Ca(2+) release, ROS production, and caspase-3, and -9 activation in ARPE-19 cells during the blue light-mediated cell death, and to examine a potential protective effect of melatonin and amfenac, in the apoptotic cascade.Methods: ARPE-19 cells were cultured in their medium. First, MTT tests were performed to determine the protective effects of amfenac and melatonin. Cells were then exposed to blue light irradiation in an incubator. Intracellular Ca(2+) release experiments, mitochondrial membrane depolarization, apoptosis assay, glutathione (GSH), glutathione peroxidase (GSH-Px), and ROS experiments were done according to the method stated in the Materials and methods section.Results: Cell death was clearly associated with increased levels of ROS production, as measured by 2',7'-dichlorofluorescein fluorescence, and associated increase in Ca(2+) levels, as measured by Fura-2-AM. Blue light-induced cell death was associated with an increased level of caspase-3 and 9, suggesting mediation via the apoptotic pathway. Cell death was also associated with mitochondrial depolarization. Melatonin was shown to delay these three steps.Conclusion: Melatonin, amfenac, and their combination protect ARPE-19 cells against blue light-triggered ROS accumulation and caspase-3 and -9 activation. The antiapoptotic effect of melatonin and amfenac at doses inhibiting caspase synthesis modified Ca(2+) release and prevented excessive ROS production, suggesting a new therapeutic approach to age-related macular degeneration. [ABSTRACT FROM AUTHOR]

Contrast media (CM)-induced nephropathy is a common cause of iatrogenic acute renal failure. The aim of the present review was to discuss the mechanisms and risk factors of CM, to summarize the controlled studies evaluating measures for prevention and to conclude with evidence-based strategies for prevention. A review of the relevant literature and results from recent clinical studies as well as critical analyses of published systematic reviews used MEDLINE and the Science Citation Index. The cytotoxicity induced by CM leads to apoptosis and death of endothelial and tubular cells and may be initiated by cell membrane damage together with reactive oxygen species (ROS) and inflammation. Cell damage may be aggravated by factors such as tissue hypoxia, properties of individual CM such as ionic strength, high osmolarity and/or viscosity. Clinical studies indeed support this possibility, suggesting a protective effect of ROS scavenging with the administration of N-acetylcysteine, ascorbic acid erdosteine, glutathione and bicarbonate infusion. The interaction between extracellular Ca(2+), which plays a central role in intercellular contacts and production of ROS, and the in vitro toxicity of CM was also reviewed. The current review addresses the role of oxidative stress in the pathogenesis of CM in the kidney as well as current and potential novel treatment modalities for the prevention of neutrophil activation and CM-induced kidney degeneration in patients. ROS production through CM-induced renal hypoxia may exert direct tubular and vascular endothelial injury. Preventive strategies via antioxidant supplementation include inhibition of ROS generation or scavenging. [ABSTRACT FROM AUTHOR]

Daily moderate exercise and supplementation of vitamins C and E (VCE) can be beneficial in diabetes by ameliorating the effects of free radical production. The present study sought to analyze the effect of moderate exercise accompanying VCE supplementation on lipid peroxidation (LP) and antioxidative systems in the blood of streptozotocin-induced diabetic rats. Forty female Wistar rats were randomly divided 4 groups. The 1st and 2nd groups served as the control and diabetic groups, respectively. The 3rd group was the diabetic-exercise group. The 4th group, also diabetic-exercise rats, received VCE-supplemented feed. Animals in the exercised groups were moderately exercised on a treadmill 5 days a week for 3 weeks. Diabetes was induced on Day 0 of the exercise. Plasma and red blood cell (RBC) samples were taken from all animals on Day 20. Glutathione peroxidase, catalase, and reduced glutathione levels in plasma and RBCs, and vitamins A, E, and beta-carotene in plasma were lower in diabetic rats than in control animals, whereas there was a significant increase in platelet counts in both plasma and RBC LP levels. The decreased antioxidant enzymes and vitamins, and the increased LP levels and WBC counts, did improve through exercise only, although their levels were mostly increased by exercise + VCE supplementation. There were no significant changes in the hemoglobin and hematocrit values in the 4 groups. In conclusion, these data demonstrate an increase in LP in the blood of diabetic animals whereas there was a decrease in the antioxidant vitamins and enzymes. However, dietary VCE with moderate exercise may strengthen the antioxidant defense system by decreasing reactive oxygen species. [ABSTRACT FROM AUTHOR]

Summary The aim of the current study was to determine whether a rumen protected palm oil based diet affect malondialdehyde (MDA), glutathione peroxidase (GSH-Px), reduced glutathione (rGSH) and vitamin A levels in the tissues of cornu uteri , corpus uteri and corpus luteum over the barley based isoenergetic and isonitrogenous diet, and whether the response is different between ewes and ewe-lambs. During the breeding season, half of Morkaraman ewes (2–4-year-old, n =10) and ewe-lambs (7–8-months-old, n =10) was offered a barley based diet and the other half was offered a protected palm oil based diet for 42 ± 0.7 days. At the end of the experiment all animals were slaughtered and measurements carried out in the tissues collected. In all animals tested, cornu uteri had the highest MDA levels followed by corpus uteri and corpus luteum (P <0.01) but no differences were between the tissues observed in GSH-Px and rGSH levels (P >0.05). Vitamin A levels were, however, higher in corpus luteum than in cornu uteri and corpus uteri (P <0.05). Corpus uteri MDA levels were not different (P >0.05) but rGSH levels were higher for the palm oil fed groups (P <0.05). GSH-Px and rGSH levels were higher for ewe-lambs than ewes (P <0.05). In conclusion, it appears that MDA, rGSH, GSH-Px, and vitamin A work in a different fashion for corpus uteri , cornu uteri and corpus luteum , and for ewes and ewe-lambs. Dietary palm oil did not significantly affect the parameters studied except higher rGSH levels in corpus uteri . Levels of anti-oxidatively active substances, such as rGSH and GSH-Px were lower in ewes compared with those in ewe-lambs. [ABSTRACT FROM AUTHOR]

Hypoxia induces cell death through excessive production of reactive oxygen species (ROS) and calcium (Ca 2+ ) influx in cells and TRPM2 cation channel is activated by oxidative stress. Zinc (Zn), selenium (Se), and glutathione (GSH) have antioxidant properties in several cells and hypoxia-induced TRPM2 channel activity, ROS and cell death may be inhibited by the Zn, Se, and GSH treatments. We investigated effects of Zn, Se, and GSH on lipid peroxidation (LPO), cell cytotoxicity and death through inhibition of TRPM2 channel activity in transfected HEK293 cells exposed to hypoxia defined as oxygen deficiency.We induced four groups as normoxia 30 and 60 min evaluated as control groups, hypoxia 30 and 60 min in the HEK293 cells. The cells were separately pre-incubated with extracellular Zn (100 µM), Se (150 nM) and GSH (5 mM). Cytotoxicity was evaluated by lactate dehydrogenase (LDH) release and the LDH and LPO levels were significantly higher in the hypoxia-30 and 60 min-exposed cells according to normoxia 30 and 60 min groups. Furthermore, we found that the LPO and LDH were decreased in the hypoxia-exposed cells after being treated with Zn, Se, and GSH according to the hypoxia groups. Compared to the normoxia groups, the current densities of TRPM2 channel were increased in the hypoxia-exposed cells by the hypoxia applications, while the same values were decreased in the treatment of Zn, Se, and GSH according to hypoxia group. In conclusion, hypoxia-induced TRPM2 channel activity, ROS and cell death were recovered by the Se, Zn and GSH treatments.

Background: Cisplatin (CDDP) is one of the most active cytotoxic agents in the treatment of cancer. We investigated the effect of selenium (Se) with high dose vitamin E (VE) administration to prevent CDDP-induced nephrotoxicity in rats., Materials and Methods: In this study, 40 female Wistar rats were randomly divided into five equal groups. The first group, which served as the control, was administered physiological saline (2.5 cc/day, 5 days) intraperitoneally (IP), while group A was administered cisplatin (6 mg/kg BW/ single dose) plus physiological saline IP. Groups B, C, D received IP five doses of Se (1.5 mg/kg BW), and a high dose of VE (1000 mg/kg BW) (Se-VE) in combination before, simultaneously, and after CDDP, respectively. The rats were sacrificed five days after CDDP administration. Plasma malondialdehide (MDA), glutathione peroxidase (GSH-Px), reduced glutathione (GSH), catalase, urea, creatinine levels, renal histopathological changes were measured., Results: The histopathological injury score, plasma levels of MDA, urea, creatinine were found to increase in group A compared to the control (p<0.05), while plasma levels of GSH-Px, GSH and catalase decreased (p<0.05). In contrast, plasma levels of MDA decreased (p<0.05) in groups B, C, D, which were treated with Se- VE, whereas levels of GSH-Px, GSH were found to increase only for group D (p<0.05). Plasma urea, creatinine levels improved in the treatment groups compared to group A (p<0.001). Histopathological changes caused by CDDP were also significantly improved after Se-VE treatment (p<0.05)., Conclusions: Oxidative stress increases with CDDP-induced nephrotoxicity in rats. Se-VE supplementation might thus play a role in the prevention of CDDP-induced nephrotoxicity in patients.

Objectives: Hypericum perfortarum (HP, St John's wort) is a modulator of Ca(2+) entry in neutrophils and it may modulate intracellular free Ca(2+) ([Ca(2+)]i) entry in leukocytes of patients with multiple sclerosis (MS). We investigated effects of HP on oxidative stress, apoptosis, and [Ca(2+)]i concentrations in serum and leukocytes of patients with MS., Methods: Neutrophils of nine newly diagnosed MS patients and nine healthy subjects within four subgroups were used in the study. The first group was a control; the second group was patients with MS. The neutrophils from patient group were incubated non-specific TRPM2 channel blocker (2-APB), voltage-gated calcium channel blockers, verapamil and diltiazem (V + D) with HP before N-formyl-L-methionyl-L-leucyl-L-phenylalanine stimulation, respectively., Results: Neutrophil and serum lipid peroxidation, neutrophil apoptosis and [Ca(2+)]i levels in patients with MS were higher than in control although their levels were decreased by HP, 2-APB, and V + D incubations. The modulator role of V + D in MS and MS + HP groups was higher than in the 2-APB group. Neutrophilic glutathione peroxidase (GSH-Px) and serum vitamin A and E concentrations were lower in the MS group than in control. However, the neutrophil GSH-Px activity was increased by HP incubation. The neutrophil reduced glutathione, serum vitamin C and β-carotene concentrations did not change in control and patients., Discussion: We observed that HP-induced protective effects on oxidative stress and [Ca(2+)]i concentrations by modulating transient receptor potential and voltage gated calcium channel in the patients with MS. Thus, it may provide useful treatment of neutrophil activity in the patients.

To improve skin flap healing, one promising strategy in reconstructive surgery might be to optimize platelet rich plasma (PRP) bioactivity and the ischemia-altered expression of genes. We studied both the effect of PRP on ischemic flaps, and whether in vivo bioluminescence imaging (BLI) is a suitable method for the longitudinal monitoring of angiogenesis in surgical wounds. Axial murine skin flaps were created in four experimental groups. In vivo measurements of VEGFR2 expression levels were made every other day until the 14th day. The local VEGF level and microvessel density were quantified on the 14th day via ELISA and immunohistochemistry, and flap survival rates were measured. We demonstrated that PRP and induced ischemia have a beneficial influence on angiogenesis and flap healing. Combining the two resulted in a significantly robust increase in angiogenesis and flap survival rate that was corroborated by bioluminescence imaging of VEGFR2 activity. This study shows that angiogenic effects of PRP may be potentialized by the stimulus of induced ischemia during free flap harvesting, and thus the two procedures appear to have a synergistic effect on flap healing. This study further demonstrates that BLI of modulated genes in reconstructive surgery is a valuable model for longitudinal in vivo evaluation of angiogenesis., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Melatonin has antitumor activity via several mechanisms including its antiproliferative and proapoptotic effects in addition to its potent antioxidant action. Thus, melatonin has proven useful in the treatment of tumors in association with chemotherapeutic drugs. This study was performed to evaluate the effect of melatonin on the cytotoxicity and apoptosis induced by three different chemotherapeutic agents, namely 5-fluorouracil (5-FU), cisplatin, and doxorubicin in the rat pancreatic tumor cell line AR42J. We found that both melatonin and the three chemotherapeutic drugs induce a time-dependent decrease in AR42J cell viability, reaching the highest cytotoxic effect after 48 hr of incubation. Furthermore, melatonin significantly augmented the cytotoxicity of the chemotherapeutic agents. Consistently, cotreatment of AR42J cells with each of the chemotherapeutic agents in the presence of melatonin increased the population of apoptotic cells, elevated mitochondrial membrane depolarization, and augmented intracellular reactive oxygen species (ROS) production compared to treatment with each chemotherapeutic agent alone. These results provide evidence that in vitro melatonin enhances chemotherapy-induced cytotoxicity and apoptosis in rat pancreatic tumor AR42J cells and, therefore, melatonin may be potentially applied to pancreatic tumor treatment as a powerful synergistic agent in combination with chemotherapeutic drugs., (© 2012 John Wiley & Sons A/S.)

Diabetes mellitus is a well-recognized cause of male sexual dysfunction and impairments of male fertility. Streptozotocin (STZ) is used for medical treatment of neoplastic islet β-cells of pancreas and producing of animal model of diabetes mellitus type 1 that is characterized by suppression of reproductive activity due to the hyperglycaemia-induced oxidative stress and histopathological alterations in testes. Seeking for the agents that could alleviate diabetes-induced damage to reproductive system is yet the important area of inquiry. The present study was designed to evaluate whether hydrated C(60) fullerene (C(60)HyFn), which is known to be powerful bioantioxidant, eliminate testicular dysfunction induced by STZ-diabetes in rats. Wistar strain male albino rats were divided into four groups of six animals each: (1) control group, (2) C(60)HyFn-treated nondiabetic group, (3) STZ-diabetic group and (4) C(60)HyFn-treated diabetic group. Once hyperglycaemia was induced by STZ, rats in the second and fourth groups were treated with C(60)HyFn (in the form of drinking water) at the dose of 4μg/kg daily for 5 weeks. In diabetic rats, relative weights of right cauda epididymis, seminal vesicles, prostate, sperm motility and epididymal sperm concentration were significantly less than those of control group, but which were restored in the fourth group treated with C(60)HyFn (p<0.001). In hematoxylin and eosin staining, marked histopathological changes including degeneration, desquamation, disorganisation and reduction in germinal cells, interstitial oedema and congestion were evident in the testis of diabetic rats, but C(60)HyFn treatment resulted in recovery of histopathological changes and an increase in Johnsen's testicular score significantly (p<0.001). C(60)HyFn treatment restores the increased apoptosis induced by STZ-diabetes. In diabetic rats, levels of serum testosterone, testicular reduced glutathione (GSH) and alpha-tocopherol were significantly reduced and testicular lipid peroxidation level was increased (p<0.001). Nevertheless, treatment of diabetic rats with C(60)HyFn resulted in significant corrective effects on these parameters towards the control levels. C(60)HyFn, applied alone, did not exert any toxic effects in testicular tissues. Furthermore, C(60)HyFn treatment in diabetic and nondiabetic rats resulted in considerable elevations of some important polyunsaturated fatty acids. In conclusion, we have presented for the first time substantial evidence that administration of C(60)HyFn significantly reduces diabetes-induced oxidative stress and associated complications such as testicular dysfunction and spermatogenic disruption., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

The aim of this study was to investigate possible effects of 50 Hz-1 mT magnetic field (MF) on action potential in isolated rat sciatic nerve. We used 16 Wistar rats in the study. They were divided into control (n = 10) and MF (n = 6) groups. The sciatic nerve of left legs in the MF group was exposed to 50 Hz-1 mT MF for 30 min by using a Helmholtz applicator and then action potentials in control and experimental groups were recorded extracellularly. Maximum amplitude and hyperpolarization time and action potential were significantly (p ≤ 0.025) lower in the MF group than in control. However, conduction time, minimum amplitude, depolarization and repolarization times of the action potential was not different between control and MF groups evaluated. In conclusion, 50-1 mT MF caused to decrease amplitude value and hyperpolarization time of action potential in the rat nerve.

The levels of liver lipid peroxidation, glutathione peroxidase, reduced glutathione, and vitamins A and E were used to follow the level of oxidative damage caused by ionizing radiation in pregnant rats. The possible protective effects of selenium and vitamin E supplemented to rats housed in concrete-protected cages using hematite and colemanite were tested and compared to untreated controls. Ninety-six rats were randomly divided into four main equal groups namely control (A), normal concrete (B), concrete containing colemanite (C), and concrete containing hematite (D). Except group A, all groups exposed to 7 Gy radiation. The four main groups were divided into four subgroups each as follows: subgroups 1 (n = 6): nonpregnant control rats. Subgroups 2 (n = 6): selenium and vitamin E combination was intraperitoneally (i.p.) given to the nonpregnant rats for 20 days. Subgroups 3 (n = 6): pregnant control rats. Subgroups 4 (n = 6): selenium and vitamin E combination was i.p. given to the pregnant rats for concessive 20 days. Lactate dehydrogenate, alkaline phosphates, and lipid peroxidation values were higher in subgroups 1 and 3 than in no radiation group although glutathione peroxidase and vitamin E levels in liver were lower in radiation group than in no radiation group. Lactate dehydrogenate activity and lipid peroxidation levels were found to be decreased in subgroups 2 and 4 protected with concrete containing hematite and colemanite when compared to subgroup 1 and 3 with normal concrete. The radiation doses in rats housed by concrete without colemanite and hematite exposed radiation clearly showed liver degeneration. In conclusion, selenium and vitamin E supplementations and housing by concrete with colemanite was found to offer protection against gamma-irradiation-induced liver damage and oxidative stress in rats, probably by exerting a protective effect against liver necrosis via its free radical scavenging and membrane stabilizing. Protective effects of colemanite in the liver seem to be more important than in hematite.

Imidacloprid (IMI) is widely used systemic insecticide that acts as an agonist on nicotinic acetylcholine receptors (nAChRs). IMI has been reported to be more active against insect nAChRs (EC(50) 0.86-1 microM) than it is against mammalian nAChRs (EC(50) 70 microM). The objective of this study was to determine to what extent IMI affects the nAChRs of the stellate cells of mouse cochlear nucleus (CN), using whole-cell patch-clamp recording. Puff application of 1 microM IMI had no significant effect on the membrane properties of the neurons tested, while a concentration of 10 microM caused a significant depolarizing shift in the membrane potential and resulted in increases in the fluctuation of the membrane potential and in the frequency of miniature postsynaptic potentials (mpps) within less than a minute of exposure. IMI at concentrations >or=50 microM caused a significant depolarizing shift in the membrane potential, accompanied by a marked increase in the frequency of action potential. IMI decreased the membrane input resistance and the membrane time constants. Bath application of 50 microM d-tubocurarine (d-TC) reversibly blocked the depolarizing shift of the resting membrane potential and the spontaneous firing induced by IMI application in current clamp and blocked the inward currents through nicotinic receptors induced by IMI application in voltage clamp. Similarly, 100 nM alpha-bungarotoxin (alpha-BgTx) blocked the spontaneous firing induced by IMI (n=3). The amplitude of the 100 microM IMI-induced inward current at -60 mV holding potential was 115.0+/-16.2 pA (n=7). IMI at a concentration of 10 microM produced 11.3+/-3.4 pA inward current (n=4). We conclude that exposure to IMI at concentrations >or=10 microM for <1 min can change the membrane properties of neurons that have nAChRs and, as a consequence, their function., (2009 Elsevier Inc. All rights reserved.)

The levels of blood lipid peroxidation, glutathione peroxidase, reduced glutathione, and vitamin C were used to follow the level of oxidative damage caused by 2.45 GHz electromagnetic radiation in rats. The possible protective effects of selenium and L-carnitine were also tested and compared to untreated controls. Thirty male Wistar Albino rats were equally divided into five groups, namely Groups A1 and A2: controls and sham controls, respectively; Group B: EMR; Group C: EMR + selenium, Group D: EMR + L-carnitine. Groups B–D were exposed to 2.45 GHz electromagnetic radiation during 60 min/ day for 28 days. The lipid peroxidation levels in plasma and erythrocytes were significantly higher in group B than in groups A1 and A2 (p<0.05), although the reduced glutathione and glutathione peroxidase values were slightly lower in erythrocytes of group B compared to groups A1 and A2. The plasma lipid peroxidation level in group A2 was significantly lower than in group B (p<0.05). Erythrocyte reduced glutathione levels (p<0.01) in group B; erythrocyte glutathione peroxidase activity in group A2 (p<0.05), group B (p<0.001), and group C (p<0.05) were found to be lower than in group D. In conclusion, 2.45 GHz electromagnetic radiation caused oxidative stress in blood of rat. L-carnitine seems to have protective effects on the 2.45-GHz-induced blood toxicity by inhibiting free radical supporting antioxidant redox system although selenium has no effect on the investigated values.

Neurons in the cochlear nucleus (CN) have distinct anatomical and biophysical specializations and extract various facets of auditory information which are transmitted to the higher auditory centres. The aim of the present study was to determine if the principal neurons (stellate, bushy and octopus cells) of the ventral cochlear nucleus (VCN) in 2-week-old dog brain slices share common electrophysiological properties with the principal neurons of mouse VCN. Stellate cells (n=21, of which three were anatomically identified), fired large, regular trains of action potentials in response to depolarizing current pulses. Input resistance and membrane time constant were 176+/-35.9 MOmega (n=21) and 8.8+/-1.4 ms (n=21), respectively. Bushy cells, (n=6, of which three were anatomically identified) responded with a single action potential at the onset of depolarizing current steps and showed large hyperpolarizing voltage changes that sag back toward rest to hyperpolarizing current pulses. Input resistance and membrane time constant were 120.4+/-56.1 MOmega (n=5) and 7.6+/-2.3 ms (n=5), respectively. Octopus cells (n=17, of which seven were anatomically identified) fired a single action potential at the start of a depolarizing current step and exhibited a pronounced depolarizing sag of the membrane potential towards the resting value to hyperpolarizing current steps. Input resistance and membrane time constant were 17.58+/-1.3 MOmega (n=15) and 1.34+/-0.13 ms (n=15), respectively. While stellate cells did not have a threshold rate of depolarization (dV/dt(thresh)), bushy and octopus had a dV/dt(thresh) of 5.06+/-1.04 mV/ms (n=4) and 10.6+/-2.0 mV/ms (n=6), respectively. In octopus cells, the single action potential was abolished by tetrodotoxin (TTX). An alpha-dendrotoxin (alpha-DTX)-sensitive, low-voltage-activated potassium conductance (g(KL)) together with a ZD7288-sensitive, mixed-cation conductance (g(h)) were responsible for the low input resistance, and as a consequence for the brief time constant of the octopus cells. We conclude that the principal neurons of the dog VCN are, as in mouse and cat, distinguishable on the basis of whole-cell patch-clamp recordings., (2009 Elsevier B.V.)

An imbalance between oxidative stress and antioxidative capacity may play an important role in the development and progression of bronchial asthma (BA) and chronic obstructive pulmonary disease (COPD). We carried out a study to assess the systemic oxidant-antioxidant status during the exacerbation and the stable period in patients with BA and COPD. A total of 33 patients, 16 with BA and 17 with COPD were included in the study. During the exacerbation and the stable periods, levels of malondialdehyde (MDA), activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione reductase (GRd), and catalase (CAT) in erythrocytes and serum melatonin concentrations were investigated. Blood counts, respiratory functions, and blood gases of the patients were also performed. During an exacerbation period of BA, despite the decreases in GSH-Px, GRd and melatonin levels, MDA and CAT levels, and the white blood cell count, the percentage of eosinophils were significantly higher than in the stable period. Also, it was found that FEV(1)/L (where FEV(1) is the forced expiratory volume in 1 s), FVC/L (where FVC is forced vital capacity), PEF/L/s (where PEF is peak expiratory flow), pO(2) (where pO(2) is oxygen pressure) levels increased during the stable period in patients with BA. MDA and SOD values were higher in the exacerbation period than in the stable period although GSH-Px, GRd, melatonin, pH, and pO(2) values were lower in the exacerbation period than in the stable period. The blood counts and the respiratory function tests did not change between the exacerbation and the stable period of patients with COPD significantly. In conclusion, we observed that oxidative stress in the exacerbation period of patients with BA and COPD increased whereas the antioxidant enzymes and melatonin values reduced. The episodes of BA or COPD might be associated with elevated levels of oxidative stress., ((c) 2009 John Wiley & Sons, Ltd.)

In the dysfunctional splice variant TRPM2-DeltaN, a stretch of 20 amino acids (aa 537-556) is missing within the N-terminal cytosolic tail of the cation channel TRPM2. The DeltaN-stretch overlaps with two IQ-like calmodulin-binding domains. Moreover, it contains two PxxP motifs implicated in protein-protein interactions. Here, we constructed variants to test whether any of these motifs may explain why TRPM2-DeltaN does not respond to stimulation with either ADP ribose or hydrogen peroxide. Each of the two IQ-motifs could be removed without loss of channel function. Similarly, deletion of either one or both PxxP motifs had no effect. Moreover, the single point mutation D543E associated with bipolar disorder does not change the activation of TRPM2. We conclude that no functional role can be attributed to any of the structural motifs within the DeltaN-stretch that may be a spacer segment for other functional sites in the N terminus.

Previous studies have shown that generation of free radicals is increased following pentylenetetrazol kindling, due to increased cytosolic Ca2+ concentrations. Topiramate, a voltage-gated calcium channel inhibitor, has an evident effect in the treatment of childhood epilepsy; however, topiramate may cause nephrotoxicity. We investigated the effects of topiramate and vitamin E administration on pentylenetetrazol-induced nephrotoxicity in rats by evaluation of lipid peroxidation, nitric oxide, glutathione peroxidase, catalase and superoxide dismutase values. Forty male Wistar rats were randomly divided into five equal groups. Group 1 was used as control and group II received a single dose of pentylenetetrazol. Fifty and 100 mg/kg topiramate daily were intragastrically administered to rats in groups III and IV for 7 days, respectively. Intragastric 100 mg topiramate (daily for 7 days) and intraperitoneal vitamin E (150 mg/kg, daily for 3 days) combination were given to animals in group V before a single-dose pentylenetetrazol administration. Serum and kidney samples were taken after 3 hr of pentylenetetrazol administration. Pentylenetetrazol resulted in a significant increase in nitric oxide levels of serum and kidney, and lipid peroxidation levels of kidney although superoxide dismutase and catalase activities in the kidney was reduced by pentylenetetrazol administration. The lipid peroxidation levels in serum and kidneys and the nitric oxide levels in kidneys of groups III, IV and V were decreased by topiramate although the superoxide dismutase and catalase activities in the kidneys were increased. Lipid peroxidation and nitric oxide levels were reduced by the topiramate and vitamin E combination compared to only topiramate. Glutathione peroxidase activity was not affect by pentylenetetrazol, topiramate and vitamin E administrations. In conclusion, topiramate and vitamin E have protective effects on pentylenetetrazol-induced nephrotoxicity by inhibition of free radicals and by support of the antioxidant redox system.

In this study, we evaluated the hypothesis that long term dietary restriction would have beneficial effects on the oxidative stress and antioxidant enzyme systems in liver, heart and kidney in adult male rats undergoing different intensities of swimming exercise. Sixty male, Sprague-Dawley rats were assigned as either dietary restricted on every other week day (DR) or fed ad libitum (AL) groups, and each group was further subdivided into sedentary, endurance swimming exercise training (submaximal exercise) and exhaustive swimming exercise (maximal exercise) groups. Animals in the submaximal exercise group swam 5 days/week for 8 weeks, while maximal exercise was performed as an acute bout of exercise. In parallel with the increase in the intensity of the exercise, the degree of lipid peroxidation and protein oxidation were increased in both the DR and AL groups; however the rate of increase was lower in the DR group. Reduced glutathione (GSH), glutathione peroxidase (GSH-Px) and glutathione reductase (GR) enzyme activities were lower in the DR group than in the AL group. In parallel with the increase in exercise intensity, GSH and GR enzyme activities decreased, whereas an increase was observed in GSH-Px enzyme activity. In conclusion, the comparison between the DR and AL groups with the three swimming exercise conditions shows that the DR group is greatly protected against different swimming exercise-induced oxidative stress compared with the AL group., (Copyright 2005 John Wiley & Sons, Ltd.)

Behcet's disease (BD) has been known for many years, yet the etiology of the systemic vasculitis remains unknown. Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide (NO) synthase. ADMA is involved in endothelial dysfunction in various vascular diseases and its level in BD is unclear. This study was performed to evaluate the relationship between ADMA and NO levels in plasma of patients with BD. There were 3 groups of 30 subjects: (a) controls, (b) BD patients with mucocutaneous involvement, and (c) BD patients with vascular involvement. Plasma NO levels were assayed by spectrophotometry and plasma ADMA levels were assayed by an ELISA test. Plasma ADMA levels were higher in both groups of BD patients than in the controls; the ADMA levels were higher in the BD patients with vascular involvement than in the mucocutaneous group. Plasma NO levels were lower in both groups of BD patients than in controls; plasma NO levels were lower in the BD patients with vascular involvement than in mucocutaneous group. In the combined groups of 60 BD patients, there was significant inverse correlation between the plasma concentrations of ADMA and NO (r = -0.570, p <0.001). Plasma lipid profiles did not differ significantly between the BD patients and the controls. These results are evidence for increased plasma ADMA levels and decreased plasma NO levels as risk factors for cardiovascular events in BD patients. Inhibition of NO synthesis by ADMA may contribute to vascular involvement in BD.

Cisplatin is one of the most active cytotoxic agents in the treatment of cancer. High doses of cisplatin have also been known to produce hepatotoxicity. Several studies suggest that supplementation with an antioxidant can influence cisplatin-induced hepatotoxicity. The present study was designed to determine the effects of cisplatin on the liver oxidant/antioxidant system, and the possible protective effects of caffeic acid phenethyl ester (CAPE) on liver toxicity induced by cisplatin. Twenty-four adult female Wistar albino rats were divided into four groups of six rats each: control, cisplatin, CAPE, and cisplatin+CAPE. Cisplatin and CAPE were injected intraperitoneally. Liver tissue was removed to study the activities of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), myeloperoxidase (MPO), xanthine oxidase (XO), adenosine deaminase (ADA), and the levels of malondialdehyde and nitric oxide (NO). The activities of SOD and GSH-Px increased in the cisplatin+CAPE and CAPE groups compared with the cisplatin group. CAT activity was higher in the cisplatin +CAPE group than the other three groups. XO activity was lower in the cisplatin group than the control group. MPO activity was also increased in the cisplatin group compared to the control and CAPE groups. It can be concluded that CAPE may prevent cisplatin-induced oxidative changes in liver by strengthening the antioxidant defence system by reducing reactive oxygen species and increasing antioxidant enzyme activities., (Copyright (c) 2006 John Wiley & Sons, Ltd.)

Microwaves (MW) from cellular phones may affect biological systems by increasing free radicals, which may enhance lipid peroxidation levels of the brain, thus leading to oxidative damage. Melatonin is synthesized in and secreted by the pineal gland at night and exhibits anti-oxidant properties. Several studies suggest that supplementation with anti-oxidant can influence MW-induced brain damage. The present study was designed to determine the effects of MW on the brain lipid peroxidation system, and the possible protective effects of melatonin on brain degeneration induced by MW. Twenty-eight Sprague-Dawley male rats were randomly divided into three groups as follows: (1) sham-operated control group (N = 8); (2) study 900-MHz MW-exposed group (N = 8); and (3) 900-MHz MW-exposed+melatonin (100 microg/kg sc before daily MW exposure treated group) (N = 10). Cortex brain and hippocampus tissues were removed to study the levels of lipid peroxidation as malonyl dialdehyde. The levels of lipid peroxidation in the brain cortex and hippocampus increased in the MW group compared with the control group, although the levels in the hippocampus were decreased by MW+melatonin administration. The brain cortex lipid peroxidation levels were unaffected by melatonin treatment. We conclude that melatonin may prevent MW-induced oxidative changes in the hippocampus by strengthening the anti-oxidant defense system, by reducing oxidative stress products.

There is a possible role of reactive oxygen species (SROS) in the complication of implants although there is presently little information. The aim of this study was to investigate the alterations in lipid peroxidation (LP) and antioxidant enzyme activities in tissues surrounding implants in rabbits. Thirty New Zealand albino male rabbits were used. They were randomly divided into five groups. The first group (I) was used as control. Groups II, III, IV and V were implanted with stainless steel, ceramic, titanium and polyethylene, respectively. One month after the administration of implant, the tissues surrounding the implant were carefully removed for antioxidant enzyme analysis. Glucose-6-phosphate dehydrogenase (G6PD), glutathione reductase (GR), superoxide dismutases (SOD), glutathione peroxidase (GPx), catalase (CAT) in tissues surrounding the implants in the groups II, III and IV were significantly (p<0.05-p<0.001) lower than in the control group although glutathione S-transferase (GST) activities and LP values were increased. CAT activity and LP level did not decrease in group V. In conclusion, these data demonstrate that there is an increase in lipid peroxidation in the tissues surrounding ceramic and titanium implants of animals whereas there is a decrease in antioxidant enzymes. Oxidative stress plays a very important role in the complications of ceramic and titanium implants. The polyethylene implant seems to be the best of the four implant materials tested., (Copyright (c) 2005 John Wiley & Sons, Ltd.)

Fatty acids containing stearic acid, which are found in hydrogenated fat, may have a detrimental effect on the cholesterol and triacylglycerol (TAG) content of plasma lipoproteins, and on the absorption of fatty acids and fat-soluble vitamins. The aim of our study was to examine the tissue concentration of lipids and vitamins A and E after feeding a hydrogenated soybean oil (HSO) diet to rats. Twenty male Sprague-Dawley rats were randomly divided into two groups, fed on coconut oil (control) and HSO, respectively in amounts corresponding to 15% of the total feed. Plasma total cholesterol, VLDL- and LDL-cholesterol, lipid peroxidation and daily excretion of the TAG and cholesterol in feces were higher in the HSO than in the control group. TAG values in plasma and liver, and HDL-cholesterol levels in plasma were lower in the HSO than in the control group. The same was true for phospholipids in plasma and for saturated fatty acids, mono- and polyunsaturated fatty acids levels in the liver and vitamin E in plasma, LDL and adipose tissue. The results of this study provide new evidence concerning the effect of dietary hydrogenated fat on lipid, TAG and vitamin E status, which are important for maintenance of good health. Consumption of dietary HSO may be associated with cardiovascular disease.

Cisplatin is one of the most active cytotoxic agents in the treatment of cancer but its clinical use is associated with nephrotoxicity. Several studies suggest that supplementation with antioxidant can influence cisplatin induced nephrotoxicity. In the present study, we investigated the effect of selenium with high dose vitamin E administration on lipid peroxidation (MDA) and scavenging enzyme activity in kidneys, liver and lens of cisplatin-induced toxicity in rats. Forty female Wistar rats were used. They were randomly divided into five groups. The first and second groups were used as control and cisplatin (6 mg/kg BW) intraperitoneally administrated groups. Groups III, IV and V received intraperitoneally five doses of selenium (1.5 mg/kg BW) and a high dose of vitamin E (1000 mg/kg BW) combination before, simultaneously and after with cisplatin, respectively. Glutathione peroxidase (GSH-Px), vitamin E and beta-carotene levels in the kidney, lens and liver, vitamin A and reduced glutathione (GSH) levels in the kidney were significantly (P<0.05 to <0.001) lower in the cisplatin group than in the control whereas there was a significant increase in kidney, liver and lens MDA levels in rats treated with cisplatin. The decreased antioxidant enzymes and vitamins and increased MDA levels in the kidney, lens and liver of animals administered with cisplatin were significantly (P<0.05 to <0.001) improved with selenium and a high dose vitamin E injection. In conclusion, this data demonstrates that there is an increase in lipid peroxidation in the kidney, liver and lens of animals administered with cisplatin whereas there is a decrease in antioxidant vitamins and enzymes. However, intraperitoneally injected selenium combined with a high dose of vitamin E seem to produce a significant improvement on antioxidants concentrations in rats treated before, simultaneously and after with cisplatin. The selenium with high dose vitamin E injection may play a role in preventing cisplatin-induced nephropathy and cataract formation in cancer patient.

Melatonin, the main secretory product of the pineal gland, has been shown to be potentially effective in prevention of numerous types of neurodegenerative disorders in which free radical processes are involved. Homocysteine (Hcy), an independent risk factor for atherosclerosis, undergoes auto-oxidation and generates reactive oxygen species. The purpose of this study was to test whether intracerebroventricular (ICV) injection of Hcy leads to neural lipid peroxidation and also to investigate the protective effects of melatonin on the brain tissue from oxidative stress of Hcy. Adult male Wistar rats under anaesthesia were injected ICV with Hcy at a dose of 143 microg/kg. Melatonin was administered intraperitoneally to a group of rats for three consecutive days before Hcy injection. The rats were decapitated and brain tissues were removed and hippocampus, cortex and cerebellum were dissected. There was a significant development of oxidative stress as indicated by an increase in malondialdehyde in hippocampus, cortex and cerebellum of rats injected with Hcy, whereas melatonin prevented the elevation of lipid peroxidation. Furthermore, melatonin significantly increased glutathione levels and glutathione peroxidase activity in all brain regions. The present study demonstrates that Hcy, in high levels, may be a causal factor in generation of free radicals in the brain and it may be one of the mechanisms which cause neurodegeneration in elderly people. It also shows that melatonin could potentially be beneficial in prevention of neurodegeneration caused by hyperhomocysteinemia.

Behçet's disease (BD) is known for many years, yet its etiology remains unknown. In BD, the increased production of reactive oxygen species from activated neutrophils may reduce concentrations of antioxidant vitamins and enzymes in plasma and red blood cells (RBC). Vitamin E is an important fat soluble antioxidant and its role on antioxidant parameters of BD is unclear. The study was undertaken to evaluate the role of vitamin E on lipid peroxidation (MDA) levels, antioxidant vitamin and enzyme concentrations in plasma and RBC in patients with BD. There were three groups i.e., control, patient and treatment groups with twenty-five subjects in each. Nonsmoking patients with BD, patient group, was compared with an equal number of healthy control subjects (control group). Blood samples were taken from both control and patient groups and then oral vitamin E was daily supplemented to the patients with BD for six weeks (treated group). At the end of six weeks, blood was taken from the treated group once more. RBC and plasma MDA levels, serum neopterin, complement system (C(3) and C(4)), ASO, CRP, rheumatoid factor, plasma lipoprotein (a), total cholesterol concentrations and erythrocyte sedimentation rate were higher in the patient group than in the control group, but they were lower in the treatment group than in the patient group. While vitamins A, E and beta-carotene concentrations in plasma, catalase, glutathione peroxidase activities and reduced glutathione levels in RBC and plasma were lower in patient group than in the controls, they were found to be higher in the treatment group than in the patient group. These results provide some evidence for a potential role of increased lipid peroxidation and decreased enzymatic and non enzymatic antioxidants in BD by its inflammatory character and vitamin E which may strengthen the antioxidant defense system, and may contribute to the treatment of BD., (Copyright 2002 The Canadian Society of Clinical Chemists)

Reactive oxygen species can be generated by daily exposure of the skin to ultraviolet light and may cause some subchronic and chronic skin disorders. The aim of this study was to investigate a possible preventive role of alpha-tocopherol acetate (ATA) on ultraviolet B (UVB) induced peroxidation by assessing lipid peroxide (LPO) levels and activity of reactive oxygen scavenging enzymes including glutathione peroxidase and superoxide dismutase (SOD) in guinea pigs. ATA was topically applied to the skin for three weeks before a single dose of 0.9 J/cm2 UVB irradiation on the skin and lipid peroxide levels and antioxidants in plasma, skin and liver and erythrocytes were determined after decapitation. Topical application of ATA prevented the UVB irradiation-induced reduction of scavenging enzyme activities in skin and erythrocytes. In conclusion, we suggest that topical applications of ATA before UVB irradiation is effective in protecting the skin from unwanted effects of UVB irradiation.

The aim of this research was to determine levels in blood of vitamin E, beta-carotene, lipid peroxidation as thiobarbituric-acid reactive substances (TBARS) and reduced glutathione (GSH) and activity of glutathione peroxidase (GSH-Px) in patients with alopecia. Studies were carried out on 37 patients with alopecia and 34 healthy age-matched controls. Red blood cell (RBC) and plasma samples from healthy and patient subjects were taken. Beta-cartotene levels (P<0. 001) in plasma and levels of GSH (P>0.05) and the activity of GSH-Px (P<0.05, P<0.01) in both plasma and RBC samples were significantly lower in patients with alopecia than in controls, whereas TBARS levels in plasma (P<0.05) and RBC (P<0.001) samples were significantly higher in patients with alopecia than in controls. However, vitamin E levels in plasma did not differ statistically. Although being far from conclusive, these results provide some evidence for a potential role of increased lipid peroxidation and decreased antioxidants in alopecia., (Copyright 2000 John Wiley & Sons, Ltd.)

The aim of this study was to investigate the effects of feed withdrawal and darkening on the antioxidative status of laying hens in high ambient temperatures between 30-40 degrees C. We determined vitamins A and E, lipid peroxidation as thiobarbituric-acid reactive substances (TBARS), glutathione peroxidase (GSH-Px) and reduced glutathione (GSH) in the serum, liver and muscle. Sixteen week old hens (Ross Brown) were divided into 3 groups of 30 hens each. The first group was used as a control. Hens in the second (feed withdrawal) group were subjected to feed removal between 14-18 h during the summer. Hens in the third (darkening) group were subjected to darkening of the hens house with black curtains between 14-18 h. At the end of 16 weeks, blood serum, liver and breast muscle samples were taken from all animals and analysed. Vitamins A and E levels in serum and liver were significantly (P < 0.05, P < 0.01) higher in feed withdrawal and darkened groups than in control group whereas the serum levels of the darkening group were highest. GSH-Px activity and GSH level in liver and muscle were also significantly (P < 0.05, P < 0.01) higher in feed withdrawal and the darkening group than in control. However, TBARS levels in liver and muscle were significantly (P < 0.05, P < 0.01) lower in feed withdrawal and darkening groups compared with control group, whereas the level of the darkening group was the lowest. In conclusion, we observed that feed withdrawal and darkening of hens in high temperatures during summertime resulted in an increase of antioxidant enzymes and vitamins levels and in a decrease of lipid peroxidation.

Objective: The aim of the present study was to investigate the direct influence of cigarette smoke on the bacterial colonization of the lower respiratory tract and the effect of supplementary vitamin E on the colonization of instilled bacteria into the trachea of rats that do not have the chronic airway pathology associated with smoking., Methodology: Thirty male Wistar albino rats, weighing approximately 250 g, were used as experimental animals. A 0.1 mL bacterial suspension containing six bacterial species (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumonia, Proteus mirabilis, Haemophilus influenza, Peptostreptococci spp.), isolated previously from pharynx of rats, were instilled into the trachea of three groups of rat (10 control, 10 cigarette smoke-treated and 10 cigarette smoke- and vitamin E-supplemented rats). The smoke-treated rats were exposed to cigarette smoke for 3 days prior to and after intratracheal instillation. The third group of rats were given vitamin E supplements (100 mg/kg per day). Tracheobronchial lavage samples of all rats were quantitatively cultured after 3 days from the instillation., Results: The colony numbers of isolated bacteria were significantly higher in cigarette smoke-treated rats than in the control group and in the smoke- and vitamin E-supplemented rats (P < 0.05). Only S. aureus and S. epidermidis were isolated from vitamin E-supplemented rats, while instilled all six bacterial species were isolated from the other two groups., Conclusions: It is concluded that smoking impairs the elimination of bacteria or facilitates colonization of bacteria in the lower respiratory tract of rats. Supplementary treatment with vitamin E reduces the effect of cigarette smoke; however, some bacteria may be resistant to this action of vitamin E.

The aim of this investigation was to determine serum levels of vitamin A, E, beta carotene, glutathione peroxidase (GSHPx), lipid peroxidation (MDA) and biochemical and haematological parameters during enflurane anaesthetised dogs. Ten kangal dogs were used and all animals were anaesthetised with enflurane for two hours and blood samples were taken before and 30, 120 minutes, 24 hours and 7 days during the anaesthesia. Vitamin E and beta carotene content were significantly (p<0.05 and p<0.01) higher before anaesthesia than after whereas serum GSHPx activity was not statistically different. However, serum levels of vitamin A and MDA were significantly (p<0.05) increased during the anaesthesia. In general, serum levels of aspartate aminotransferase, alanine aminotransferase, albumin, glucose, urea and creatinine were significantly (p<0.05 and p<0.01) increased during anaesthesia and returned to near normal values after 7 days of anaesthesia, whereas the white blood cell count was significantly (p<0.05 and p<0.01) decreased during the anaesthesia. However, the red blood cell count, haemoglobin and packed cell volume values, and levels of total cholesterol, triglycerides, total protein and globulin were apparently not influenced by the anaesthesia. In conclusion, we observed that the serum level of vitamin E and beta carotene were significantly decreased, whereas serum MDA and vitamin A levels were significantly increased during the enflurane anaesthesia., (Copyright 1999 John Wiley & Sons, Ltd.)

Cataractous lenses have been found to have an altered distribution of the intracellular ionic environment: the concentrations of potassium and magnesium being decreased and the concentrations of sodium and calcium increased. These changes arise as a result of changes to lens membrane characteristics causing an increase in lens membrane permeability. In this study flame atomic absorption spectroscopy (AAS) was used for calcium, magnesium, iron and zinc determination, and flame atomic emission spectroscopy (AES) was used for sodium and potassium contents in normal and cigarette smoke-exposed rat lenses. The methods are sensitive enough to detect quantitatively all six cations in a single rat lenses. In this work, six elements, including Ca2+, K+, Na+, Zn2+, Fe2+ and Mg2+ in experimental rat eye lenses and normal transparent lenses were determined. It was found that the concentrations of Ca2+, Na+, Zn2+, and Fe2+ were increased dramatically while K+ and Mg2+ decreased in smoke-exposed rat lenses when compared to the control rat lenses. There were no significant changes between 'smoked' rats supplied with vitamin C and control groups. A positive correlation was found also in the other two groups of 'cigarette smoked' animals supplemented with selenium plus vitamin E and selenium when compared with 'cigarette smoked' without any supplements. These data provide support for the hypothesis that cigarette smoking increases the risk of cataract formation. We investigated whether vitamin C is the most important antioxidant in the body. The roles of diet with optimum amounts of antioxidant vitamins C and vitamin E and the antioxidant mineral selenium are discussed.

The effects of supplemented selenium and vitamin E on a number of rumen parameters such as the population of rumen protozoa, pH, concentration of volatile fatty acids and ammonia nitrogen in the rumen content were studied. Eight lambs were randomly allocated into two groups: a control group and an experimental group receiving vitamin E (DL-alpha-tocopheryl acetate, 250 mg/kg of feed) and selenium (sodium selenite, 0.3 mg/kg of feed) supplementation. Samples of rumen content were taken from all lambs three times daily once a week (before feeding as well as 3 and 6 h after feeding) over a period of 10 consecutive weeks. In addition, the lambs were weighed at the end of experiment. The total counts and percentage proportions of rumen protozoa, the pH value, and the levels of ammonia nitrogen and volatile fatty acids were determined in the samples of rumen content. The levels of acetic acid, propionic acid, butyric acid and total volatile fatty acids, the total counts of protozoa, and the percentage proportion of Diplodinium were found to be significantly higher in the supplemented than in the control group (P < 0.05, P < 0.01), whereas the pH values and the percentage proportion of Dasytricha ruminantium were significantly lower in the supplemented group than in the control (P < 0.05). However, there was no statistically significant difference between the two groups in ammonia nitrogen levels. The body mass gain of lambs in the supplemented group was significantly higher than that of the control animals (P < 0.01). Combined selenium and vitamin E supplementation of the lambs' ration caused an increase in the levels of volatile fatty acids, total counts of protozoa, and body mass gain while decreasing the pH value of the rumen content.