Effects of methanolic extracts from broad beans on cellular growth and antioxidant enzyme activity

Objective There are several reports of cellular-aging-dependent alterations in the antioxidant capacity of human fibroblasts. Fibroblasts show slower the growth rate at late passages (referred to hereafter as old cells) than at early passages (referred to hereafter as young cells). Antioxidants may control cellular growth by modulating reactive oxygen species (ROS). Methanolic extracts from broad beans (MEBB) contain phenolic compounds and have ROS-scavenging activities. In this study, we investigated the effects of MEBB on cellular growth and antioxidant levels in normal human lung fibroblasts. Methods To determine cytosolic superoxide dismutase (SOD) activities, cytosolic glutathione peroxidase (GSH-Px) activities, catalase activities, reduced glutathione (GSH) concentrations, and growth rate, MEBB treatments were performed on young and old cells. Results In young and old cells treated with 120 μg/ml MEBB, the growth rates increased by 28.1 and 15.2%, respectively, compared with controls. The MEBB treatment of young cells caused a 62.5% increase in SOD activity, but the treatment of old cells caused a 39.5% decrease. The catalase activities of the young and old cells treated with MEBB were equal to those of control cells. Young and old cells treated with MEBB were equal to the control cells in terms of GSH-Px activity. The GSH concentrations in the young and old cells treated with 120 μg/ml MEBB increased by 22.1 and 45.9%, respectively. Conclusion These studies elucidated a new cellular growth mechanism whereby human lung fibroblasts modulate intracellular GSH levels via the action of MEBB.     

Effects of radical scavenger protein from broad beans on glutathione status in human lung fibroblasts

Objective Human diploid cells are more susceptible to oxidative stress at late passage than at early passage, presumably because of the decrease in cellular-reduced glutathione (GSH) concentration. Water-soluble protein (WSP) from broad beans scavenges free radicals. The effects of WSP on the glutathione system were examined in PDL 20 (early passage) and PDL 50 (late passage) human lung fibroblasts (TIG-1). Methods To determine cytosolic glutathione peroxidase (GSH-Px) activities, glutathione reductase (GR) activities, oxidized glutathione (GSSG) concentrations, and GSSG/reduced glutathione (GSH) ratios, WSP and hydrocortisone (HC) treatments of TIG-1 cells (PDL 20→50 and PDL 50→75) were performed for 40 days. We also investigated the GSSG concentrations and GR activities in PDL 20 cells that were continuously treated with WSP until PDL 39 and 55. Results GSSG concentrations decreased in WSP- and HC-treated PDL 50→75 cells. The GSSG/GSH ratios in PDL 50→75 cells became low after the treatments. Increases in GR activities were observed in treated PDL 50→75 cells. The decline in the GSSG concentration of PDL 50→75 cells correlated with the increase in GR activity. The GSSG levels in control cells were higher following cellular age, whereas the levels in treated cells were lower than those in the control. The studies on cellular age-related changes indicated that greater increases in GR activity were found in treated cells than in the control. Conclusion These results indicated that WSP influences the GSSG concentration that is associated with cellular aging, but the mechanism of GSSG reduction by WSP remains unknown. The enhancement of glutathione status following WSP treatment may be related to the delay in the cellular aging.     

人体内自由基的研究进展

自由基是人体代谢的中间产物,广泛存在于人体中。正常情况下,自由基在人体新陈代谢,细胞信号传导和基因调节,诱导细胞增殖、凋亡,维持人体正常生长发育以及在抑制细菌病毒和癌症等方面起着重要作用。然而,当机体发生病变或受外界环境因素影响、体内自由基产生/清除平衡被打破时,自由基又会与人体内脂质、蛋白质和核酸发生反应,从而危及健康。本文从人体内自由基的种类、寿命、来源、检测方法、清除机制及其与人类健康的关系等方面综述了近年来对自由基的研究进展,并分析了多环芳烃等环境污染物介导体内氧化应激及体内自由基清除的可能机制。     

Sub-lethal effects of profenofos on tissue-specific antioxidative responses in a Euryhyaline fish, Oreochromis mossambicus

The euryhaline fish, Oreochromis mossambicus was exposed to sub-lethal concentration (30 μg/L) of profenofos (PF) for 28 days and allowed to recover for 7 days. Responses were evaluated through antioxidant enzyme activities in various tissues of fish. Glutathione-S-transferase (GST) and superoxide dismutase (SOD) activities showed transient increases in gill, viscera and muscle, but decreased in brain. However, catalase (CAT) and glutathione reductase (GR) were inhibited in gill and viscera. Reduced glutathione (GSH) levels were depleted in all the tissues and a significant induction in lipid peroxidation (LPO) was observed. Significant recovery in all the studied parameters was noticed at the end of recuperation period. The enhanced LPO in the present study suggests that reactive oxygen species (ROS)-induced oxidative damage could be one of the main toxic effect of PF. The increased LPO and alterations in the antioxidant defense system can be used as biomarkers in the pesticide-contaminated aquatic streams.     

Responses of environmental Amycolatopsis strains to copper stress

Copper is a redox-active metal, which acts as a catalyst in the formation of Reactive Oxygen Species (ROS) encouraging oxidative stress. Protection against oxidants is intrinsic to every living cell; however, in stress conditions, cells are forced to increase and expand their antioxidative network. In this work, the novel copper-resistant strain Amycolatopsis tucumanensis and the copper-sensitive Amycolatopsis eurytherma were grown under copper increasing concentrations in order to elucidate the dissimilar effects of the metal on the strains viability, mainly on morphology and antioxidant capacity. Although biosorbed copper encouraged ROS production in a dose-dependent manner in both strains, the increase in ROS production from the basal level to the stress conditions in A. tucumanensis is lesser than in the copper-sensitive strain; likewise, in presence of copper A. eurytherma suffered inexorable morphological alteration while A. tucumanensis was not affected. The levels of antioxidant enzymes and metallothioneins (MT) were all greater in A. tucumanensis than in A. eurytherma; in addition MT levels as well as superoxide dismutase and thioredoxin reductase activities in A. tucumanensis, were higher as higher the concentration of copper in the culture medium. This work has given evidence that an efficient antioxidant defense system might aid microorganisms to survive in copper-stress conditions; besides it constitutes the first report of oxidative stress study in the genus Amycolatopsis and contributes to enlarge the knowledge on the copper-resistance mechanisms of A. tucumanensis.     

Lipid-lowering effects of methanolic extract of Vernonia amygdalina leaves in rats fed on high cholesterol diet

We investigated the lipid-lowering effects of methanolic extract of Vernonia amygdalina (VA) leaves in rats fed an high cholesterol diet, and compared with a standard hypolipidemic drug, Questran (Qu). The effects of VA on the lipid profile were assessed by measuring the levels of total cholesterol, triglyceride, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol, lipid peroxidation (LPO), phospholipid, and glutathione (GSH) in the plasma and liver of the rats. Administration of cholesterol at a dose of 30 mg/0.3 ml, five times in a week for nine consecutive weeks resulted in a significant increase (p < 0.05) in plasma and post mitochondrial fraction (PMF) cholesterol levels by 33% and 55%, respectively. However, treatment with extract of VA at doses of 100 and 200 mg/kg caused a dose dependent reduction in the plasma and PMF cholesterol by 20%, 23% and 23%, 29%, respectively. Similar reduction in cholesterol levels was obtained in Qu-treated rats. Furthermore, VA at 200 mg/kg decreased the plasma and PMF LDL-cholesterol levels by 23% and 49%, and also decreased plasma and PMF triglyceride levels by 29% and 28%, respectively. Also, VA at 100 and 200 mg/kg caused a dose-dependent increase in plasma HDL-cholesterol levels by 41% and 59%, respectively. However, there were no significant differences (p > 0.05) in the PMF HDL-cholesterol and phospholipid levels of the treated rats when compared to hypercholesterolemic rats. There were significant decreases (p < 0.05) in the LPO levels of extract-treated rats. Precisely, VA at 100 and 200 mg/kg decreased the levels of plasma and PMF LPO by 38%, 42% and 35%, 45%, respectively. In addition, VA augmented the cholesterol-induced decrease in PMF glutathione levels of the rats. Taken together, these results suggest the lipid-lowering effects of VA and, probably serve as a new potential natural product for the treatment of hyperlipidemia.     

In vitro activity of eight antibiotics againstSalmonella andShigella species

The minimum inhibitory concentrations (MICs) of ampicillin, chloramphenicol, trimetoprimsulfamethoxazole (TMP-SMX), ofloxacin, ciprofloxacin, pefloxacin, enoxacin and fleroxacin forSalmonella spp. (n=72) andShigella spp. (n=52) were established.S. typhi isolates were all susceptible to all of the antibiotics tested. In non-typhi salmonellae andShigella spp., resistance to ampicillin, chloramphenicol and TMP-SMX with various rates were encountered, but all isolates were susceptible to fluoroquinolones.     

Hematological and gill histopathological parameters of three tropical fish species from a polluted lagoon on the west coast of Mauritius

Gill histopathology and hematological primary indices, including blood delta-aminolevulinic acid dehydratase (deltaALA-D) activity and nucleocytoplasmic ratio of erythrocytes, were assessed in three tropical marine fish species, Scarus ghobban, Epinephelus merra, and Siganus sutor, from the presumably contaminated lagoon of Bain des Dames, Mauritius. Concurrently, the nonpolluted region of Blue Bay/Pt d'Esny was used as a reference site for comparison of fish physiological responses and seawater quality. Bain des Dames showed high seawater mercury content (6.4+/-0.5 microg/L), traces of iron (70+/-40 microg/L), and fluctuating biochemical oxygen demand values (0.488+/-0.171 mg/L day(-1)). Gill histopathological analysis revealed lesions such as epithelial hyperplasia and inflammation. Similarly, a generalized increase in blood deltaALA-D activity (131.27-355.76 nmol PBG/ml RBC.h) was recorded. Fish from Bain des Dames showed species-specific hematological responses including normocytic macrocytic blood cells (S. ghobban), macrocytic anemia (S. sutor), and active erythropoiesis (E. merra). Though the species displayed impaired health, attributed to changes in water quality, the blood responses seemingly indicate adaptation to hypoxic conditions arising from both gill degradation and perhaps oxygen-level fluctuations.     

Antioxidant and antidiabetic activities of extracts from Cirsium japonicum roots

This study investigated the antioxidant activity of methanol (MeOH) and water extracts from roots of Cirsium japonicum in vitro. MeOH extract showed a stronger free radical scavenging activity than water extract. However, both of extracts showed a concentration dependent hydroxyl radical scavenging activity, reducing power and metal chelating ability. MeOH extract had greater phenolic and flavonoid contents than water extract. The antidiabetic activity of these two extracts was evaluated by the α-glucosidase inhibition assay. The water extract showed a considerable α-glucosidase inhibitory activity. To our knowledge, this may be the first time to report the antioxidant and antidiabetic activities in Cirsium japonicum roots.     

两株非典型羊种布鲁氏菌的鉴定报告

本文报告了应用常规试验方法难以定种的两株非典型布氏菌株,而应用六群布氏菌噬菌体的代表株裂解试验和氧化代谢试验鉴定为变异深刻的羊种布氏菌。     

Effects of N-acetylcysteine on alcohol abstinence and alcohol-induced adverse effects in rats

Alcoholism is rampant in modern society and some antioxidant compound could perhaps be useful to reduce the damage done by alcohol consumption and abstinence. The present study was undertaken to investigate the association of N-acetylcysteine (NAC) intake, alcoholism, and alcohol abstinence on lipid profile, in vivo low-density lipoprotein (LDL) oxidation, oxidative stress, and antioxidant status in serum and liver of rats. Initially, male Wistar 30 rats were divided into two groups: (C, N = 6) given standard chow and water; (E, N = 24) receiving standard chow and aqueous ethanol solution in semi-voluntary research. After 30 days of ethanol exposure, (E) group was divided into four subgroups (N = 6/group): (E-E) continued drinking 30% ethanol solution; (E-NAC) drinking ethanol solution containing 2 g/L NAC; (AB) changed ethanol solution to water; (AB-NAC) changed ethanol to aqueous solution 2 g/L NAC. After 15 days of the E-group division, E-E rats had higher serum alanine transaminase, lower body weight, and surface area, despite higher energy intake than C. E-E rats had also lower feed efficiency, dyslipidemia with enhanced triacylglycerol, very low-density lipoprotein (VLDL), lipid hydroperoxide (LH) and in vivo oxidized-LDL (ox-LDL). AB, E-NAC, and AB-NAC rats ameliorated serum oxidative stress markers and normalized serum lipids. E-E rats had higher hepatic LH and lower reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio than C, indicating hepatic oxidative stress. AB and E-NAC rats normalized hepatic LH, GSSG, and the GSH/GSSG ratio, compared to E-E. AB-NAC rats had the lowest serum ox-LDL, hepatic LH levels, and the highest GSH reductase activity in hepatic tissue. In conclusion, the present study brought new insights into alcohol consumption, because ethanol exposure enhanced serum in vivo ox-LDL, as well as serum and hepatic oxidative stress. N-acetylcysteine offers promising therapeutic value to inhibit ethanol-induced adverse effects. Ethanol withdrawal had beneficial effects on serum lipids, but was more effective when coupled with NAC supplementation. Ethanol abstinence and NAC intake interact synergistically, improving serum lipids and hepatic antioxidant defenses.     

Leishmania species identification using FTA card sampling directly from patients' cutaneous lesions in the state of Lara, Venezuela

A molecular epidemiological study was performed using FTA card materials directly sampled from lesions of patients with cutaneous leishmaniasis (CL) in the state of Lara, Venezuela, where causative agents have been identified as Leishmania (Viannia) braziliensis and L. (Leishmania) venezuelensis in previous studies. Of the 17 patients diagnosed with CL, Leishmania spp. were successfully identified in 16 patients based on analysis of the cytochrome b gene and rRNA internal transcribed spacer sequences. Consistent with previous findings, seven of the patients were infected with L. (V.) braziliensis. However, parasites from the other nine patients were genetically identified as L. (L.) mexicana, which differed from results of previous enzymatic and antigenic analyses. These results strongly suggest that L. (L.) venezuelensis is a variant of L. (L.) mexicana and that the classification of L. (L.) venezuelensis should be reconsidered.     

In situ bioassay with Eisenia andrei to assess soil toxicity in an abandoned uranium mine

The goal of this study was to develop an in situ bioassay with Eisenia andrei, deploying it in several locations of an abandoned mining area. Our objectives were two-fold: (i) we intended to validate the in situ soil bioassay procedures, while (ii) providing ecologically relevant data to complement the ongoing risk evaluation based on laboratorial assays. To promote cost- and time-effectiveness, the in situ exposure was short (48 h) and the endpoints analysed included oxidative stress biomarkers and metal content in soil and organisms. The bioassay was carried out under different experimental conditions, simulating local (natural soil) vs. control conditions (LUFA soil), and irrigation with artificial rainwater vs. irrigation with diluted acidic effluent. Variation in the data was mostly due to soil type, rather than irrigation water, and substantial spatial heterogeneity was observed. Oxidative stress biomarkers did not fully work as sensitive parameters to environmental contamination. Earthworm metal burdens suggested a potential concern in terms of bioaccumulation of some metallic elements.     

Acute toxicity of widely used pharmaceuticals in aquatic species: Gambusia holbrooki, Artemia parthenogenetica and Tetraselmis chuii

Pharmaceuticals are continuously dispersed into the environment as a result of human and veterinary use, posing relevant environmental concerns. This study evaluated the acute toxicity of three therapeutic agents (diazepam, clofibrate, and clofibric acid) and a detergent (sodium dodecyl sulfate; SDS) in three aquatic species, namely the euryhaline fish Gambusia holbrooki, the hypersaline crustacean Artemia parthenogenetica, and the marine algae Tetraselmis chuii. The ranking of 50% lethal concentrations (LC50) for the two animal species and 50% inhibitory concentration (IC50) for the algal species was, in decreasing order, clofibric acid > SDS > diazepam > clofibrate for G. holbrooki, clofibric acid > clofibrate > SDS > diazepam for A. parthenogenetica, and clofibric acid > clofibrate > SDS > diazepam for T. chuii. These differences show that the intrinsic nature of test organisms must be considered when evaluating the toxicity of these agents to aquatic ecosystems.     

Regiospecific profiles of fatty acids in triacylglycerols and phospholipids from Adzuki beans (Vigna angularis)

Regiospecific distributions of fatty acids (FA) of triacylglycerols (TAG) and phospholipids (PL) isolated from five cultivars of adzuki beans (Vigna angularis) were investigated. The lipids comprised mainly PL (72.2-73.4 wt-%) and TAG (20.6-21.9 wt-%), whilst other components were detected in minor proportions (0.1-3.4 wt-%). The principal profiles of the FA distribution in the TAG and PL were evident in the beans among the five cultivars: unsaturated FA were predominantly distributed in the sn-2 position, whilst saturated FA primarily occupied the sn-1 or the sn-3 position in the these lipids. The results would be useful information to both producers and consumers for manufacturing traditional adzuki confectionaries such as wagashi in Japan.     

十种清热解毒中药提取物抗泛耐药鲍曼不动杆菌感染的研究

目的 研究金果榄、矮脚苦蒿和姜黄等十种清热解毒中药抗泛耐药鲍曼不动杆菌(XDR-AB)感染的活性,筛选出具有体内抗感染活性的中药提取物,为发现新型抗XDR-AB感染的抗菌药物提供研究基础。方法 分别采用水、50%乙醇、95%乙醇提取十种清热解毒中药并制成不同浓度的药物浸膏,与课题组前期优化的XDR-AB感染秀丽隐杆线虫模型共孵育,通过线虫的存活率判断中药提取物的体内抗XDR-AB活性。结果 随着姜黄和土荆皮提取物浓度的增大,XDR-AB感染的线虫存活率不断提高。姜黄的水提取物、50%乙醇提取物和95%乙醇提取物在浓度1 000μg/mL时,可使XDR-AB感染的秀丽隐杆线虫存活率分别提高至54.2%(与阴性对照组比较,P<0.001)、18.8%、13.3%;土荆皮的水提取物、50%乙醇提取物和95%乙醇提取物在浓度1 000μg/mL时,可使XDR-AB感染的秀丽隐杆线虫存活率分别提高至47.4%(与阴性对照组比较,P<0.001)、23.8%、15.8%。结论 姜黄和土荆皮水提取物具有较好的抗XDR-AB感染活性,可对其主要化学成分进行体外抗菌药效验证,以期发现新型抗X...     

Low level occupational exposure to styrene: its effects on DNA damage and DNA repair

The present study aimed to evaluate the effects of styrene exposure at levels below the recommended standards of the Threshold Limit Value (TLV-TWA(8)) of 20 ppm (ACGIH, 2004) in reinforced-fiberglass plastics workers. Study subjects comprised 50 exposed workers and 40 control subjects. The exposed workers were stratified by styrene exposure levels, i.e. group I (<10 ppm, <42.20 mg/m(3)), group II (10-20 ppm, 42.20-84.40 mg/m(3)), and group III (>20 ppm, >84.40 mg/m(3)). The mean styrene exposure levels of exposed workers were significantly higher than those of the control workers. Biomarkers of exposure to styrene, including blood styrene and the urinary metabolites, mandelic acid (MA) and phenylglyoxylic acid (PGA), were significantly increased with increasing levels of styrene exposure, but were not detected in the control group. DNA damage, such as DNA strand breaks, 8-hydroxydeoxyguanosine (8-OHdG), and DNA repair capacity, were used as biomarkers of early biological effects. DNA strand breaks and 8-OHdG/10(5)dG levels in peripheral leukocytes of exposed groups were significantly higher compared to the control group (P<0.05). In addition, DNA repair capacity, determined by the cytogenetic challenge assay, was lower in all exposed groups when compared to the control group (P<0.05). The expression of CYP2E1, which is involved in styrene metabolism, in all styrene exposed groups, was higher than that of the control group at a statistically significant level (P<0.05). Levels of expression of the DNA repair genes hOGG1 and XRCC1 were significantly higher in all exposed groups than in the control group (P<0.05). In addition to styrene contamination in ambient air, a trace amount of benzene was also found but, the correlation between benzene exposure and DNA damage or DNA repair capacity was not statistically significant. The results obtained from this study indicate an increase in genotoxic effects and thus health risk from occupational styrene exposure, even at levels below the recommended TLV-TWA(8) of 20 ppm.     

Antioxidant activity and hepatoprotective effects of whey protein and Spirulina in rats

Objectives

The aims of the present study were to evaluate the antioxidant, radical scavenging, and metal-chelating activity of whey protein concentrate (WPC) and Spirulina alone or in combination in vitro and to evaluate their hepatoprotective effects against CCl₄ in vivo.

Methods

Five concentrations (20, 40, 60, 80, and 100 mg/100 mL) of WPC, Spirulina, and their combination were tested in vitro. In the in vivo study, eight groups of male rats comprised the control group and the groups treated with WPC, Spirulina alone, or in combination with or without CCl₄ were used.

Results

The in vitro study showed that WPC and Spirulina showed antioxidant, radical scavenging, and metal-chelating activities in dose-dependent manner. The in vivo study showed that both agents succeeded in preventing liver damage induced by CCl₄. This prevention was more pronounced in rats receiving the combination of WPC and Spirulina.

Conclusion

Whey protein concentrate and Spirulina have free radical scavenging properties and antioxidant activity.     

Phytotoxic effects of copper on nitrogen metabolism and plant growth in Brassica pekinensis Rupr

Copper (Cu) from various anthropogenic and natural sources is one of the major heavy metal contaminants in the environment. To study Cu-induced nitrogen (N) metabolism damage in the popular vegetable Chinese cabbage (Brassica pekinensis Rupr.), aquatic culture experiments with this plant were performed. Two Cu levels (0.3 micromol/L (control) and 10.3 micromol/L) and two N levels (0.05- and 1-fold Hoagland's solution) were used for the aquatic culture experiments. The results demonstrated adverse effects of Cu on N metabolism and plant growth. Cu exposure elevated Cu concentration and decreased nitrate reductase (NR) activity in the roots and shoots and reduced total chlorophyll content. It also shortened root length and produced fewer leaves and lower plant biomass. However, Cu exposure increased total free amino acid content in the leaves. The results also demonstrated effects of N deficiency on N metabolism and plant growth. N deficiency increased the root/shoot ratio of biomass and the total free amino acid content and decreased the nitrate contents and NR activity in roots and leaves. In addition, there were interactive effects between Cu exposure and N level on plant biomass, root/shoot ratio, and chlorophyll and nitrate content in leaves. The results suggested that Cu toxicity to the plant was at least partly due to an influence on N metabolism.     

Extensive examination of different tick species for infestation with Coxiella burnetii in Slovakia

Between 1987–1989 almost 7000 adult Ixodes ricinus, Dermacentor reticulatus, Dermacentor marginatus, Haemaphysalis concinna, Haemaphysalis punctata and Haemaphysalis inermis ticks collected in all 38 districts of Slovakia were screened for the presence of Coxiella burnetii. The proportion of ticks containing C. burnetii as indicated by the haemocyte test was less than 3%. Attempts to recover C. burnetii by inoculation of yolk sacs of embryonated hen eggs from pools of 1–6 specimens of haemocyte test positive ticks resulted in the isolation of 10 rickettsial strains. Six strains were recovered from I. ricinus, the remaining ones from single pools of D. reticulatus, D. marginatus. H. concinna and H. inermis ticks.In addition to the previous recovery of C. burnetii from H. punctata ticks, the agent was thus isolated from all important ticks living in Slovakia. The agent was found in tick habitats regardless of the latitude and altitude in the entire country. These results are not consistent with the negligible number of Q fever cases occurring in past years in Slovakia.Presented at the 4th International Symposium on Rickettsiae and Rickettsial Diseases, Pietany, C.S.F.R., 1–6 October, 1990.