Antioxidant-associated chemoprevention by sodium selenite in N-nitrosodiethylamine-induced and phenobarbital-promoted hepatocarcinogenesis in rats

The anticarcinogenic/antioxidant potential of sodium selenite (Se), a micronutrient, was evaluated on liver tumourigenesis induced by N-nitrosodiethylamine (DEN) and promoted by phenobarbital (PB; 0.05% in diet). Male, albino rats of the Wistar strain were exposed intravenously to a single dose of DEN (200 mg x kg(-1) body weight). Se (4 ppm in drinking water) was supplemented before initiation, or during initiation and/or during the promotion period of carcinogenesis. At the end of 16 weeks (after DEN administration) nodular incidence, the total number of nodules and non-enzymic antioxidants such as vitamin E, vitamin C, total thiol, protein thiol and non-protein thiol contents were measured in hepatoma, surrounding tissue and kidney tissue of control and experimental groups. In hepatoma-bearing animals the above biochemical changes were decreased when compared with normal control animals. On Se treatment throughout the study, (20 weeks) the above biochemical changes reverted to normal levels. Pre- and post-treatment with Se also shows a tendency to reverse the above changes. The results indicate that prior application of Se significantly reverses the adverse changes produced during the tumourigenesis. Furthermore, prior applications of Se significantly reduced the cumulative number of tumours per tumour-bearing animals. The present study reveals the antitumour potential of Se against DEN-induced liver carcinogenesis.     

Antioxidant effect of diphenyl diselenide on oxidative stress caused by acute physical exercise in skeletal muscle and lungs of mice

This study was designed to examine if diphenyl diselenide (PhSe)₂, an organoselenium compound, attenuates oxidative stress caused by acute physical exercise in skeletal muscle and lungs of mice. Swiss mice were pre‐treated with (PhSe)₂ (5 mg kg^‐1 day^‐1) for 7 days. At the 7th day, the animals were submitted to acute physical exercise which consisted of continuous swimming for 20 min. The animals were euthanized 1 and 24 h after the exercise test. The levels of thiobarbituric acid reactive species (TBARS), non‐protein thiols (NPSH) and ascorbic acid and the activity of catalase (CAT) were measured in the lungs and skeletal muscle of mice. Glycogen content was determined in the skeletal muscle of mice. Parameters in plasma (urea and creatinine) were determined. The results demonstrated an increase in TBARS levels induced by acute physical exercise in the skeletal muscle and lungs of mice. Animals submitted to exercise showed an increase in non‐enzymatic antioxidant defenses (NPSH and ascorbic acid) in the skeletal muscle. In lungs of mice, activity of CAT was increased. (PhSe)₂ protected against the increase in TBARS levels and ameliorated antioxidant defenses in the skeletal muscle and lungs of mice submitted to physical exercise. These results indicate that acute physical exercise caused a tissue‐specific oxidative stress in the skeletal muscle and lungs of mice. (PhSe)₂ protected against oxidative damage induced by acute physical exercise in mice. Copyright © 2009 John Wiley & Sons, Ltd.     

Exogenous Selenium Mitigates Salt Stress in Soybean by Improving Growth,Physiology, Glutathione Homeostasis and Antioxidant Defense

The mechanism of selenium (Se)-induced salt tolerance was studied in moderately sensitive soybean (Glycine max L.) plants. To execute this view, soybean plants were imposed with salt stress (EC 6 dS m⁻¹ ) applying NaCl. In other treatments, Se (0, 25, 50 and 75 µM Na₂SeO₄) was sprayed as co-application with that level of salt stress. Plant height, stem diameter, leaf area, SPAD value decreased noticeably under salt stress. Altered proline (Pro) level, together with decreased leaf relative water content (RWC) was observed in salt-affected plants. Salt stress resulted in brutal oxidative damage and increased the content of H₂O₂, MDA level and electrolyte leakage. Exogenous Se spray alleviated oxidative damage through boosting up the antioxidant defense system by increasing the activity of antioxidant enzymes such as catalase (CAT), peroxidase (POD) and glutathione reductase (GR), as well as by improving non-enzymatic antioxidants like glutathione (GSH) and GSH/glutathione disulfide (GSSG). The upregulated antioxidant defense system, restored Pro and leaf RWC, higher SPAD value conferred better growth and development in Se-sprayed salt-affected soybean plants which altogether put forth for the progressive yield contributing parameters and finally, seed yield. Among different doses of Se, soybean plants sprayed with 50 µM Na₂SeO₄ showed better salt tolerance.     

Effects of excess selenomethionine on selenium status indicators in pregnant long-tailed macaques (Macaca fascicularis)

Forty pregnant long-tailed macaques were treated daily for 30 d with 0, 25, 150 or 300 μg selenium as L-selenomethionine/kg body weight. Erythrocyte and plasma selenium and glutathione peroxidase specific activities, hair and fecal selenium, and urinary selenium excretion were increased by and were linearly related to L-selenomethionine dose. Hair selenium was most sensitive to L-selenomethionine dose, with an 84-fold increase in the 300 μg selenium/(kg-d) group relative to controls (r=0.917). Daily urinary selenium excretion (80-fold,r=0.958), plasma selenium (22-fold,r=0.885), erythrocyte selenium (24-fold,r=0.920), and fecal selenium (18-fold,r=0.911) also responded strongly to L-selenomethionine. Erythrocyte and plasma glutathione peroxidase specific activities increased 154% and 69% over controls, respectively. Toxicity was associated with erythrocyte selenium >2.3 μg/mL, plasma selenium >2.8 μg/mL, and hair selenium >27 μg/g. Plasma, erythrocyte, and hair selenium concentrations may be useful for monitoring and preventing the toxicity of L-selenomethionine administered to humans in cancer chemoprevention trials.     

Antioxidant activity of fluoxetine: Studies in mice melanoma model

In vivo effects of the antidepressant fluoxetine on spleen antioxidant status of C57BL/6 mice were studied using a melanoma experimental model. After a 14‐day treatment with fluoxetine (10 mg kg^?1 day^?1, i.p.), the endogenous antioxidant non‐enzyme (glutathione) and enzyme (superoxide dismutase (SOD) and glutathione peroxidase (GPx)) defense systems in spleen of healthy animals were not changed; the lipid peroxidation (LP) was also unchanged. When B16F10 melanoma cells were introduced in C57BL/6 mice 2 h before fluoxetine treatment, a drug‐protective effect against the melanoma‐induced oxidative changes (increased LP and decreased total glutathione (GSH)‐level, as well as antioxidant enzyme activities) in spleen was observed. Fluoxetine dose‐dependently reduced the amounts of free oxygen radicals (hydroxyl and superoxide anion radicals), generated in chemical systems. Taken together, the present results suggest that fluoxetine, acting as antioxidant, prevents from melanoma‐induced oxidative changes in mice spleen. Copyright © 2010 John Wiley & Sons, Ltd.     

Cyclophosphamide pretreatment in tumor cryotherapy: A murine model

Following the protocol of Turk and Lagrange for selective potentiation of cell-mediated immunity, a single high dose of cyclophosphamide was given to inbred mice bearing subcutaneous tumors 3 days before cryosurgery. Cyclophosphamide pretreatment caused a rise in the rate of total tumor ablation and local recurrence was delayed. The dose response to tumor size, cyclophosphamide, and freeze parameters indicated that these effects were the result of synergistic rather than additive processes. Instances of metastatic tumor growth and immunologically mediated complications of cryosurgery were also lower when the cryosurgery was accompanied by cyclophosphamide pretreatment.     

Stromelysin-3 suppresses tumor cell apoptosis in a murine model

Stromelysin-3 (STR-3) is a matrix metalloproteinase with a unique pattern of expression and substrate specificity. During embryogenesis and remodeling of normal adult tissues, STR-3 is produced by stromal cells in direct contact with epithelial cells undergoing regional apoptosis and selective cell survival. STR-3 is also overexpressed by interdigitating stromal cells in primary epithelial malignancies. Although STR-3 does not degrade classic extracellular matrix components, the enzyme promotes the establishment of local tumors in nude mice by as yet undefined mechanisms. STR-3 is induced when malignant epithelial cells come into contact with surrounding stromal elements; the active stromal cell-derived 45 kDa enzyme is subsequently processed to a 35 kDa protein without enzymatic activity. We have generated MCF-7 transfectants expressing wild type or catalytically inactive 45 kDa STR-3 (STR-3wt and STR-3cat-) or secreted 35 kDa STR-3 (35 kDa STR-3sec) and evaluated their implantation and survival in nude mice. Tumors developed significantly more rapidly in animals receiving STR-3wt, rather than vector-only, STR-3cat- or 35 kDa STR-3sec transfectants. Most importantly, STR-3wt tumors had a significantly lower percentage of apoptotic cells than tumors derived from vector-only, STR-3cat- or 35 kDa STR-3sec transfectants. Taken together, these studies suggest that the active STR-3 enzyme may increase tumor take by suppressing tumor cell apoptosis and that 45 kDa to 35 kDa STR-3 processing limits STR-3 activity at the tumor/stromal interface. Because STR-3 is secreted as an active enzyme rather than a proform, subsequent 45 kDa to 35 kDa STR-3 processing may represent a novel mechanism for regulating enzymatic activity.     

硫硒配施对衰老期大蒜生长、品质及抗氧化能力的影响

本文研究了衰老过程中硫硒配施对大蒜生长、品质及抗氧化能力的影响。结果显示,硒的施用显著提高大蒜干鲜重、鳞茎鲜重和横径,降低了MDA含量,提高了处理200 d的Fv/Fm值与叶绿素含量,促进了大蒜的生长。硒处理均显著提高了谷胱甘肽过氧化物酶(GSH-Px)活性;降低了超氧化物歧化酶(SOD)活性;低硫浓度下低硒处理提高了过氧化氢酶(CAT)活性,高硫浓度下, CAT活性随硒浓度升高而升高;处理160 d的过氧化物酶(POD)活性变化与CAT相同,硒处理200 d时,低硫浓度下POD活性降低,高硫浓度下POD活性呈先上升后下降趋势。硒处理提高了大蒜鳞茎Vc和大蒜素含量。可溶性蛋白含量在低硫浓度下随硒浓度升高而降低,而在高硫浓度随之升高;可溶性糖含量变化与可溶性蛋白恰好相反。综合各指标以低硫低硒处理(2 mmol·L-1 S+3μmol·L-1 Se)为最优。     

TNF-alpha-treated DC exacerbates disease in a murine tumor metastasis model

Due to the pivotal role that dendritic cells (DC) play in eliciting functional anti-tumor T cell responses, immunotherapeutic approaches utilizing DC-based vaccines have readily been exploited. It has been argued that, in the setting of immunotherapy, mature DC will be more efficient at T cell priming and, therefore, required for effective vaccination. As TNF-alpha is commonly used as a DC maturation factor, we have examined the efficacy of treatment with DC matured with TNF-alpha (DC-TNF) in a murine model of melanoma. We have now shown that treatment with DC-TNF leads to an increase in the number of lung metastases as compared to mice treated with immature DC. No differences in the number of CD4⁺CD25⁺ T-regulatory cells were measured in the lungs of DC-TNF-treated mice. On examination of the infiltrating lymphocytes, an enhanced secretion of IL-10 and a higher percentage of CD4⁺IL-10⁺ T cells were measured in the lungs of DC-TNF-treated mice. However, treatment with DC-TNF did not enhance the number of melanoma lesions in the lungs of IL-10 knockout mice or in mice depleted of CD4⁺ T cells. Together, these studies indicate that treatment of melanoma-bearing mice with DC treated with TNF-alpha can induce IL-10 production by resident cells at the tumor site, leading to immune tolerance and exacerbation of disease.     

Concerted Action Of Reduced Glutathione And Superoxide Dismutase In Preventing Redox Cycling Of Dihydroxypyrimidines, And Their Role In Antioxidant Defence

Dialuric Acid, the reduced form of the β-cell toxin alloxan, and the related fava bean derivatives divicine and isouramil, autoxidize rapidly in neutral solution by a radical mechanism. GSH promotes redox cycling of each compound, with concomitant GSH oxidation and H₂O₂ production. With superoxide dismutase present, there is a lag period in which little oxidation occurs, followed by rapid oxidation. GSH extends this lag and decreases the subsequent rate of oxidation, so that with superoxide dismutase and a sufficient excess of GSH. coupled oxidation of GSH and each pyrimidine is almost completely suppressed. This mechanism may be a means whereby GSH in combination with superoxide dismutase protects against the cytotoxic effects of these reactive pyrimidines. Superoxide dismutase may also protect cells against oxida-tive stress in other situations where GSH acts as a radical scavenger. and we propose that the concerted action of GSH and superoxide dismutase constitutes an important antioxidant defence     

Glutathione peroxidase activity in insects: A reassessment

In vertebrate species, cytotoxic H₂O₂ and other lipid or organic hydroperoxides (ROOH) formed in aerobic metabolism are removed by a selenoprotein, glutathione peroxidase (GPOX). The GPOX activity in most rat tissues ranges from 100 to 1,000 units (1 unit = 1 nmol NADPH oxidized·mg protein^?1·min^?1), except for muscles (20–30 units). In contrast, GPOX activities of two strains of the housefly (Musca domestica), cabbage looper (Trichoplusia ni), southern armyworm (Spodoptera eridania), and black swallowtail butterfly (Papilio polyxenes), were found to be in the range 2–12 units. Trivial GPOX activity was detected in the confused flour beetle (Tribolium confusum). In the earthworm (Lumbricus terrestris), banana slug (Ariolimax columbianus), and market squid (Loligo opalescens), the GPOX activity ranged from 1 to 5 units. Tissue selenium concentrations were about 500–1,000 ppb for adult M. domestica, 600 ppb in T. confusum, 32 ppb in T. ni, 17 ppb in S. eridania, and 31 ppb in P. polyxenes larvae. The form of selenium incorporated at such high levels in tissues of invertebrates such as M. domestica remains an unresolved issue. Peroxidase activity of non-selenium glutathione-S-transferase (GT) against ROOH may compensate for the low GPOX activity. Catalase (CAT) has high activity and wide subcellular distribution in insects. This may be an evolutionary adaptation to GT's inability to catalyze the reduction of H₂O₂. The GT's peroxidase and CAT activities were not assessed for other invertebrate species, and warrants an investigation due to their reported low GPOX levels.     

Physiological and Proteomic Responses of Different Willow Clones (Salix fragilis X alba) Exposed to Dredged Sediment Contaminated by Heavy Metals

High biomass producing species are considered as tools for remediation of contaminated soils. Willows (Salix spp.) are prominent study subjects in this regard. In this study, different willow clones (Salix fragilis x alba) were planted on heavy-metal polluted dredging sludge. A first objective was assessment of the biomass production for these clones. Using a Gupta statistic, four clones were identified as high biomass producers (HBP). For comparison, a group of four clones with lowest biomass production were selected (LBP). A second objective was to compare metal uptake as well as the physiological and proteomic responses of these two groups. All these complementary data's allow us to have a better picture of the health of the clones that would be used in phytoremediation programs. Cd, Zn, and Ni total uptake was higher in the HBPs but Pb total uptake was higher in LBPs. Our proteomic and physiological results showed that the LBPs were able to maintain cellular activity as much as the HBPs although the oxidative stress response was more pronounced in the LBPs. This could be due to the high Pb content found in this group although a combined effect of the other metals cannot be excluded.     

Age-related changes in the glutathione redox system

The effect of aging on the glutathione redox system was evaluated in this study. For this purpose, we determined reduced glutathione (GSH) and oxidized glutathione (GSSG) in whole blood, glutathione peroxidase (GPx) and glutathione reductase (GSSGR) in erythrocytes and selenium (Se) in plasma in 176 healthy individuals. We also calculated GSH/GSSG molar ratios. These subjects were divided into five groups: group 1 (n=25; 0.2-1 years old); group 2 (n=28; 2-11 years old); group 3 (n=23; 12-24 years old); group 4 (n=40; 25-40 years old); group 5 (n=60; 41-69 years old). GSH levels in groups 1 and 5 were significantly lower than the other groups (p<0.001). Conversely, GSSG levels were significantly high in these periods (p<0.001). The GSH/GSSG molar ratio was found to be low both in the first year of life and in the oldest group (p<0.001, respectively). GPx activity in group 5 was increased as compared to the other groups (p<0.001). GSSGR activity was significantly lower in the oldest groups than in the other groups (p<0.001). Se levels were found to be low in the oldest group (p<0.001). Selenium levels of women in group 5 were significantly high as compared to the men (p<0.01). We found negative correlations between age and GSH levels (r=0.402; p<0.001), selenium levels (r=0.454; p<0.001), GSH/GSSG molar ratio (r=0.557; p<0.001) and GSSGR activity (r=0.556; p<0.001). There were positive correlations between age and GPx (r=0.538; p<0.001) and GSSG level (r=0.551; p<0.001). In conclusion, our findings show that the glutathione redox system is affected by age. Oxidative stress increases during the aging process. There is no effect of aging on the glutathione redox system according to sex except for the Se level.     

Concerted Action Of Reduced Glutathione And Superoxide Dismutase In Preventing Redox Cycling Of Dihydroxypyrimidines,And Their Role In Antioxidant Defence

Dialuric Acid, the reduced form of the β-cell toxin alloxan, and the related fava bean derivatives divicine and isouramil, autoxidize rapidly in neutral solution by a radical mechanism. GSH promotes redox cycling of each compound, with concomitant GSH oxidation and H₂O₂ production. With superoxide dismutase present, there is a lag period in which little oxidation occurs, followed by rapid oxidation. GSH extends this lag and decreases the subsequent rate of oxidation, so that with superoxide dismutase and a sufficient excess of GSH. coupled oxidation of GSH and each pyrimidine is almost completely suppressed. This mechanism may be a means whereby GSH in combination with superoxide dismutase protects against the cytotoxic effects of these reactive pyrimidines. Superoxide dismutase may also protect cells against oxida-tive stress in other situations where GSH acts as a radical scavenger. and we propose that the concerted action of GSH and superoxide dismutase constitutes an important antioxidant defence     

镉胁迫下3种藓类植物抗氧化酶活性变化的比较研究

采用水培实验研究了重金属Cd对3种藓类植物尖叶拟船叶藓(Dolichomitriopsis diversiformis)、湿地匍灯藓(Plagiomnium acutum)和匍枝青藓(Brachythecium procumbens)的抗氧化酶系统(SOD、POD和CAT)的影响.结果表明,在1-100 mg·L-1的镉胁迫浓度范围内,Cd对3种藓类植物的SOD活性都具有显著的促进作用,SOD对Cd胁迫的种间敏感性顺序为尖叶拟船叶藓>湿地匍灯藓>匍枝青藓;3种藓类植物的POD和CAT活性对Cd胁迫均表现为先升后降,POD对Cd胁迫的种间敏感性顺序为匍枝青藓>湿地匍灯藓>尖叶拟船叶藓;高浓度Cd对CAT具有明显的抑制效应,CAT对Cd胁迫的种间敏感性顺序为湿地匍灯藓>尖叶拟船叶藓>匍枝青藓.在Cd长期胁迫下,SOD对3种藓类植物体内活性氧的清除起到关键作用,可作为藓类植物抗镉能力强弱的敏感生理指标;3种鲜类植物中匍枝青藓的抗镉能力最强,湿地匍灯藓次之,尖叶拟船叶藓最弱,并且这种抗性与cd在植物体内的积累成反相关.     

Antioxidant properties of mesenchymal stem cells against oxidative stress in a murine model of colitis

Objective

To investigate the effects of oxidative stress injury in dextran sulfate sodium (DSS)-induced colitis in mice treated with mesenchymal stem cells (MSC).

Results

Mice exposed to oral administration of 2% DSS over 7 days presented a high disease activity index and an intense colonic inflammation. Systemic infusion of MSC protected from severe colitis, reducing weight loss and diarrhea while lowering the infiltration of inflammatory cells. Moreover, toxic colitis injury increased oxidative stress. Administration of DSS decreased reduced glutathione (GSH) and superoxide dismutase (SOD) activity, and increased thiobarbituric acid-reactive substances levels in the colon. No alteration was found in catalase (CAT) and glutathione peroxidase (GPx) activity. Otherwise, MSC transplantation was able to prevent the decrease of GSH levels and SOD activity suggestive of an antioxidant property of MSC.

Conclusion

The oxidative stress is a pathomechanism underlying the pathophysiology of colitis and MSC play an important role in preventing the impairment of antioxidants defenses in inflamed colon.

     

Combination cytokine therapy inhibits tumor growth by generation of tumor-specific T-cell responses in a murine melanoma model

Various cytokines, including interferon α (IFNα), tumor necrosis factor α (TNFα), and granulocyte–macrophage colony-stimulating factor (GM-CSF), have been used as adjuvant therapy for advanced-stage melanoma with some success but with marked toxicity, which appears to be related to higher doses. We investigated the efficacy of IFNα, GM-CSF, and TNFα in various combinations to induce antitumor and immune responses in a B16F10 murine melanoma model. These studies showed that GM-CSF, IFNα, and TNFα, when injected together intratumorally, mediated significant inhibition of tumor growth. Tumor regression correlated with local tumor necrosis and significant infiltration of T cells. In addition, this injected intralesional cytokine cocktail also induced lymphadenopathy, with an increase in both CD4⁺ and CD8⁺ T cells in the draining lymph nodes. Furthermore, tumor-specific CD8⁺ T cells were identified from draining lymph nodes. These investigations identify the combined effects of IFNα, GM-CSF, and TNFα in induction of the adaptive immune response and generation of antigen-specific T-cell reactivity. These results support potential clinical trials of the low-dose cytokine combination as adjuvant therapy for melanoma.     

Antioxidant status in patients with psoriasis

Psoriasis is a chronic inflammatory skin disease with an unknown aetiology that has been associated with abnormal plasma lipid metabolism and oxidative stress. There are controversial results in the previous studies investigating oxidant/antioxidant systems in psoriasis. The aim of this work was to evaluate the plasma levels of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), total bilirubin (T. Bili), direct bilirubin (D. Bili), uric acid (UA), apolipoproteins (ApoA1 and ApoB), Lp(a) and activities of paraxonase 1 (PON1) in 100 patients with psoriasis and 100 controls, and to look for a correlation between these parameters in psoriasis. PON1, bilirubin and UA were measured spectrophotometrically, MDA by the high‐performance liquid chromatography method, apolipoproteins and Lp(a) by immunoprecipitation assays, and lipid and other biochemical parameters were determined by routine laboratory methods. In patients with psoriasis, there was a significant decrease in PON1, SOD and CAT activities (P < 0.05) and an increase in MDA levels (P < 0.01). Also, the levels of bilirubin (total and direct) and UA were decreased in patients with psoriasis but were not significant (P > 0.05). These results suggest that psoriasis was in a state of oxidative stress and that the protective effects of high‐density lipoprotein against atherosclerosis may be dependent on PON1 activity. Moreover, there is a negative correlation between antioxidant with Lp(a), apoB and MDA levels, suggesting that subjects with higher levels of Lp(a) and apoB and lower levels of antioxidant are more exposed to oxidative damage. These findings may explain in part the reported increase in cardiovascular mortality in psoriasis. Copyright © 2013 John Wiley & Sons, Ltd.     

Distinguishing effects on tumor multiplicity and growth rate in chemoprevention experiments

In some types of cancer chemoprevention experiments and short-term carcinogenicity bioassays, the data consist of the number of observed tumors per animal and the times at which these tumors were first detected. In such studies, there is interest in distinguishing between treatment effects on the number of tumors induced by a known carcinogen and treatment effects on the tumor growth rate. Since animals may die before all induced tumors reach a detectable size, separation of these effects can be difficult. This paper describes a flexible parametric model for data of this type. Under our model, the tumor detection times are realizations of a delayed Poisson process that is characterized by the age-specific tumor induction rate and a random latency interval between tumor induction and detection. The model accommodates distinct treatment and animal-specific effects on the number of induced tumors (multiplicity) and the time to tumor detection (growth rate). A Gibbs sampler is developed for estimation of the posterior distributions of the parameters. The methods are illustrated through application to data from a breast cancer chemoprevention experiment.