Free Radicals and Antioxidants

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Online ISSN: 2231-2536
Introduction High level of polyunsaturated fatty acids of lipid bilayer membrane coupled with increased oxygen consumption exposed brain cells to oxidative stress. Oxidative stress has been implicated as the etiological factor of many neurodegenerative diseases. Chemopreventive agents both synthetic and natural have the potential of inducing endogenous antioxidants with the capability of attenuating oxidant induced damage. This wok examined the ability of tert-butylated hydroxyl quinone (tBHQ), ethoxyquin, coumarin and selenium to induce NAD(P)H quinine oxoreductase 1 (NQO1) in neuroblastoma cell line. Method SHSY-5SY neuroblastoma cells were culture in medium containing 100 μM of the individual chemopreventive agents or in combination with 50 nM sodium selenite. The cells were harvested after 24 h exposure with the chemopreventive agents. Protein levels were determined by immunoblotting, while enzyme assay was done using dichloro-phenol-indo-phenol as a substrate in the presence of NADH and FAD to determine reductase activity, while dicumarol was used as an inhibitor of NQO1. Results Coumarin, ethoxyquin and tBHQ-induced NQO1 marginally (1.2–1.5-fold). Treatment of cells with selenium as sodium selenite together with tBHQ-induced NQO1 by about 3-fold compared with control. The enzyme activity was significantly increased by all chemopreventive agents (p < 0.05). Conclusion Inducers of endogenous antioxidants that can pass blood–brain barrier provide hope of delaying and attenuating neurodegenerative diseases associated with either increased free radical production or genetic predisposition.
Kinetics studies on the effect of pyruvate on ROS production in phagocytosing and TLR4-stimulated granulocytes obtained from Type 2 diabetes patients. Each point represents the average of 15 experiments Æ standard deviation; RLU 1⁄4 Relative Light Units; panels A and B show the results obtained from experiments with opsonized particles, and panels C and D show the results obtained from experiments with LPS (TLR4)-stimulated granulocytes. 
Effect of pyruvate on reactive oxygen species (ROS) production by LPS/TLR4 and phagocytosing granulocytes obtained from type 2 diabetes patients.
Aim & background Pyruvate is considered as an anti-inflammatory and anti-oxidant produced from glucose metabolism. We examined the effects of sodium pyruvate on reactive oxygen species (ROS) production induced by opsonized particles in phagocytosing or toll-like receptor 4 (TLR4)-stimulated granulocytes obtained from type 2 diabetic (T2DM) patients compared with those from healthy individuals. Methods Luminol-dependent chemiluminescence was used to quantify the ROS generated. The phagocytosis by granulocytes obtained from T2DM patients or healthy individuals was evaluated in the absence of stimulation and in the presence of opsonized particles (zymosan complex recovered using the complement fragment C3b, ZC3b) or LPS as a TLR4 activator. Pyruvate (10−4 M) markedly inhibited ROS generation in unstimulated and ZC3b-stimulated granulocytes from T2DM patients and healthy individuals. Results Our results showed 370.0% and 199.0% activation of ROS generation during phagocytosis in healthy subjects and T2DM patients, respectively. In the presence of pyruvate, these percentages were reduced to 81.0% and 80.0%, respectively. Thus, pyruvate exhibited similar suppressive activity during granulocyte phagocytosis in healthy individuals and T2DM patients (p > 0.05). In contrast, pyruvate did not inhibit or down-regulate ROS generation in granulocytes stimulated with LPS. LPS-induced ROS production in granulocytes from healthy subjects (309%) and T2DM patients (62.5%). In the presence of pyruvate, the ROS generation in LPS-stimulated granulocytes was greater (64.0%) in the cells obtained from T2DM patients compared with cells obtained from healthy individuals (38.0%) (p < 0.05; chi-square test). Conclusion The dual effect of pyruvate might be associated with a metabolic signaling pathway that depends on the oxidizing profile of the target cell. However, the effects of pyruvate must be further studied before using this compound as an anti-inflammatory or anti-oxidant therapeutic resource.
Objective The present study was designed to evaluate the in vitro antioxidant and DNA nicking potential of various extracts and fractions of Pterospermum acerifolium (L.) Willd (Family: Stericuliaceae). Research design and methods Antioxidant properties of the extracts and fractions was assayed by DPPH scavenging activity, non-site-specific and site-specific OH radical scavenging activity mediated 2-deoxy-d-ribose degradation, total antioxidant activity and lipid peroxidation assay by using rat liver homogenate. DNA nicking assay was studied by PT257R/T plasmid. Estimation of total phenolic content and total flavonoid content was done. Further HPTLC fingerprint of active fractions was performed. Results Ethyl acetate fractions of leaves, flowers and bark exhibited potent antioxidant property and DNA protective effect compare to all the other extracts and fractions. Total phenolic and flavonoid content determination was also showed ethyl acetate fractions were rich in phenolic and flavonoid contents. HPTLC fingerprint revealed the total number of peaks present in the active ethyl acetate fractions of leaves, flowers and bark. Conclusion The present study indicated that, ethyl acetate fractions of leaves, flowers and bark showed effective antioxidant and DNA protection activity and it could be the initiation for various other pharmacological studies on those fractions.
Background and objectives Acerola (Malpighia emarginata DC.) is a tropical fruit known for its nutritional and functional properties due to its great contents vitamin C, carotenoids and anthocyanins. The antioxidant potential from plants extracts associated to hepatoprotective activity has been widely studied. However, the effect of antioxidants in natural fruit juice has not been fully searched. The aim of this work was to investigate the antioxidant properties of acerola juice and its hepatoprotective potential against acute ethanol-induced stress. Materials and methods Ripe acerola were collected, processed into juice and initially analyzed to antioxidant properties in vitro. Afterwards, in vivo hepatoprotective activity was evaluated in mice. The animals received the juice by gavage as pretreatment for 15 consecutive days and then, were submitted to ethanol-induced stress in single dose (5 g/kg). The activities of serum enzymes as well as lipid peroxidation degree were evaluated. The activities of serum marker enzymes for liver damage as well as lipid peroxidation degree were evaluated. Results Acerola juice presents great vitamin C (1799.5 mg/100 g FW) and total phenolic (188.4 mg GAE/100 g FW), anthocyanins (9.2 mg/100 g FW), flavonols (7.8 mg/100 g FW) contents and high activity of superoxide dismutase (1053.6 UA/g DM) with a total antioxidant activity of 137.5 μmol Trolox/g FW. The juice treatment inhibited lipid peroxidation and reduced the activities of aminotransferases (p ≤0.05), in mice liver. Conclusion These results indicate that acerola juice is able to prevent the hepatic damage induced by ethanol, probably as a result of an enhancement of the antioxidant status in the animals.
Evaluation of scavenging activities of ethyl acetate fraction of E. jambolana or standard antioxidant against hydroxyl radical, hydrogen peroxide radical, nitric oxide radicals production and lipid peroxidation inhibitory potentiality along with ABTS and DPPH radicals scavenging activities through in vitro study model. 
Introduction:Eugenia jambolana of Myrtaceae family is widely distributed and used as diabetic therapeutic traditional medicine in rural India. Antidiabetic potentiality of ethyl acetate fraction of hydromethanolic (40:60) extract of seed of E. jambolana was investigated following in-vivo models in experimental diabetic rat and antioxidative efficacy following in-vitro models. Materials and Methods: Alteration in carbohydrate metabolism during hyperglycaemia was assessed by increased fasting blood glucose, glycated hemoglobin levels along with diminished body weight, level of serum insulin and glycogen contents in liver, skeletal and cardiac tissues of diabetic rat. In-vitro antioxidative potentiality was measured by inhibitory activity of lipid peroxidation, scavenging activity against hydrogen peroxide, nitric oxide, hydroxyl, ABTS and DPPH radicals along with preliminary phytochemical analysis. Bioactive phytoingredients were isolated from ethyl acetate fraction through column chromatography, HPTLC fingerprinting and RP-HPLC analysis. Results: Oral administration of 20 mg ethyl acetate fraction or 0.6 mg glibenclamide in 0.5 mL water/100 g body weight/rat for twice a day at fasting state to diabetic rats for 28 days significantly (p<0.05) resettled carbohydrate metabolomics towards the control levels. This fraction established its primary antioxidant attribute by scavenging hydroxyl, nitric oxide, hydrogen peroxide, ABTS and DPPH radicals along with the inhibition of lipid peroxidation with IC50 values 26.49,28.14,18.45,13.65,10.46, 28.88 μg/mL respectively. Phytochemical screening confirmed isolated compounds were chemically gallic acid in nature. Two separate spots of ethyl acetate fraction were recorded after scanning of HPTLC fingerprinting. RP-HPLC study also shows two completely resolved peaks. Its biosafety profile was established following guidelines. Conclusion: On the basis of experimental studies ethyl acetate fraction of E. jambolana proved its antihyperglycemic and antioxidant nature.
The in vitro antioxidant activity of compounds 6aek in nitric oxide (NO) method.
Aim & background To study the synthesis of a series of (4-benzoyl-phenoxy)-acetic acid derivatives (6a–k) and to test their antioxidant activity. Methods The newly synthesized compounds were characterized by IR, 1H NMR and LC–MS analyses. All these compounds were screened for their in vitro antioxidant activity by employing 1,1-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO) and hydrogen peroxide (H2O2) radical scavenging assays. Results Compounds 6h with chloro substituent in benzoyl ring and 6f with no substituent in benzoyl ring showed good radical scavenging activity in all the three methods compared to the standard drug ascorbic acid. Whereas compound 6k with methoxy substituent in benzoyl ring showed good antioxidant activity only in hydrogen peroxide method and remaining compounds showed moderate to mild radical scavenging activity. Conclusion Compounds 6f, 6h and 6i showed excellent activity, almost equivalent to that of standard and the remaining compounds showed moderate to mild scavenging activity.
Objective Limonia acidissima L. an underutilized edible fruit was evaluated for its antioxidant activity, free radical scavenging ability, proximate and amino acid analysis using established in vitro assay models. Methods 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, trolox equivalent antioxidant capacity (TEAC) assay, hydroxyl radical scavenging activity (HRSA), ferric reducing antioxidant power (FRAP) assay, nitric oxide radical (NO) scavenging activity, total antioxidant activity (TAA) were carried out. The total phenolic (TP) and flavonoid contents (TF) of the extracts were determined and expressed as gallic acid and quercetin equivalents. Results The highest percentage of phenol and flavonoid contents were observed in methanol and the lowest content was found in chloroform extract. Also, methanol extract recorded higher activity in DPPH, HRSA, FRAP and TAA whereas, ethyl acetate extract of the fruit was found to be active for ABTS+ radical scavenging activity. Further, water extract of the fruit exhibited potentially high nitric oxide radical scavenging activity than other solvent extracts. Moreover, the phenolic and flavonoid contents of the fruit extract significantly correlated with antioxidant capacity. Amino acid analysis revealed that among, all essential amino acids, the concentrations of isoleucine, phenylalanine and tryptophan were found to be present in higher amounts. Conclusion Positive correlation was observed between polyphenolic contents and the antioxidant capacities. It is evident from the study that the fruit possess potent antioxidant activity with enormous health benefits and thus may be used in food and pharmaceutical applications.
Introduction: The genus Callistemon is known in folk medicine for its anticough, antibronchitis, and insecticidal effects and its volatile oils have been used as antimicrobial and antifungal agents. Methods: The essential oils obtained by hydrodistillation of the leaves of Callistemon comboynensis (Cc) was investigated by GC/MS. Antioxidant activity of Cc was investigated using 1,1-diphenyl-2-picrylhydrazyl (DPPH). The antimicrobial activity of the essential oil of Cc was evaluated against both gram positive (Bacillus subtilis and Staphylococcus aureus), gram negative (Proteus vulgaris, Pseudomonas aeruginosa) and a pathogenic fungus Candida albicans.Results: It was found that Cc afford 0.22% volatile oil. The major components of the volatile oil of Cc are 1, 8-cineol (53.03%), eugenol (12.1%), methyl eugenol (9.2%) and α-pinene (8.3%). The oil had pronounced antibacterial and antifungal activities on all the tested microbes. Nevertheless, Cc leaf oil extract exhibited high antioxidant activity (91.1 ± 0.3 %) at a concentration of 1000 μ 1, comparable to 100 μ 1 gallic acid (95.7 ± 2).
Introduction In recent years, natural antioxidants have seen an unprecedented importance and demand in bio-pharmaceuticals, nutraceuticals besides their use as food additives. Antioxidants act as potential prophylactic and therapeutic agents against various diseases caused by free-radicals. Plants offer tremendous source of antioxidants and are therefore being evaluated for their potentials. Eulophia nuda is an important medicinal plant used by local healers in India; however its antioxidant properties have not yet been investigated. Methods Aqueous (AqE), methanol (ME), aqueous–methanol (AqME) and acetone (AE) extracts of shade dried tubers were obtained and were concentrated in vacuo. Total phenols, flavonoids, ascorbic acid and carotenoids were estimated from all extracts using standard methods. Antioxidant activities of extracts were determined by total antioxidant activity, FRAP, ABTS, DPPH, and OH radical scavenging assays besides lipid peroxidation inhibition. Extracts were evaluated for protection of Fenton's reagent induced DNA damage. Results The results confirmed the plant as a rich source of phenols, flavonoids, vitamin C and carotenoids. Among four extracts, AqME showed highest antioxidant activities as evidenced by maximum scavenging of ABTS (98%), DPPH (87%), and OH radicals (99%) at 1 mg ml−1 concentration and showed maximal inhibition of lipid peroxidation. All extracts protected the DNA from hydroxyl-radical-induced damage. Again, AqME was proved to be best in providing protection to DNA against damage caused by free-radicals. Conclusion The results provides scientific basis for its traditional usage as natural antioxidant and phyto-therapeutic agent. The plant possesses high amount of phenolic compounds and showed a broad-spectrum antioxidant properties including DNA protection.
Natural polyphenols, gallic acid, tannic acid, quercetin and salicylic acid, were investigated for their antimicrobial and antioxidant activities against Streptococcus mutans. Ascorbic acid, well known for its antimicrobial and antioxidant activities, was used as a criterion for the polyphenols. The antimicrobial effect was assessed using the plate dilution assay and the minimum inhibitory concentration (MIC) of each polyphenol was then determined from the antimicrobial activity results. Salicylic acid was the weakest antimicrobial with the highest MIC (3.8 mg/mL), and tannic acid was the strongest antimicrobial with the lowest MIC of 0.4 mg/mL. Antioxidant capacities were evaluated using the DMPD and ABTS decolorizing assays. These polyphenols show high antimicrobial activity and inoxidizability. Antioxidant activity for quercetin according to the DMPD method was inconclusive because it had color interference with the DMPD radicals. Although some conflicting results were observed between the DMPD and ABTS methods, the polyphenols with high antioxidant capacities still showed high antimicrobial activities, which suggest that the antioxidant capacity attributes to the antimicrobial effects.
Composition of the diets
Changes in body weight of control and experimental groups of rats
Effect of J. montana ethanolic extract on weight gain, food intake, and feed efficiency
J. montana extract in the form of ethanolic formulation is rich in polyphenols, mono- and sesquiterpenes, Essential oils, flavonoids, tri- tetra- and pentamethoxy quercetin derivatives. The present study was designed to investigate the anti-obesity, antiatherogenic, anti-diabetic and antioxidant activities of J. montana using obese diabetic rats' model. Rats received either regular diet, high-fat diet or high-fat diet with additional J. montana (150 and 300 mg/kg bw) for 8 weeks. In the preventive experiment, J. montana co-administered with a high fat diet significantly inhibited body weight gain, blood glucose, triglyceride, total cholesterol, LDL-C, vLDL-C, HDL-C, free fatty acid and atherogenic index levels in a dose dependent manner. J. montana-treated rats at doses of 150 and 300 mg/kg improved the insulin resistance index when compared to the high fat diet (HFD) control. Finally, the present study is designed to evaluate the effect of ethanolic extract of J. montana on figh fat diet induce obesity in rats. J. montana treatment (150 and 300 mg/kg bw) for 8 consecutive weeks prior to obese rats administration significantly prevented the decrease in the levels of hepatic oxidative stress biomarkers reduced Glutathione (GSH), Glutathione peroxidase (GPx), Glutathione reductase (GR), Superoxide dismutase (SOD) and Catalase (CAT). The J. montana extract, also exhibited its capacity to prevent the elevated thiobarbituric acid reactive substances (TBARS) in the liver tissue. In conclusion, the anti-obesity actions of J. montana are considered attributable to increased expression of energy expenditure-related fatty liver degradation, and decreased fatty acid synthesis and fat intake in the liver. Taken together, J. montana has potential as a preventive agent for type 2 diabetes mellitus (and possibly obesity) and deserves clinical trial in the near future.
The present study was carried out to evaluate the anti-inflammatory and antioxidant activities of Moringa peregrina seeds ethanolic and aqueous extracts. The anti-inflammatory effect Moringa peregrina extracts was examined in rats, using fresh egg albumin-induced oedema, and diclofenac (100 mg/kg) was used as reference drug for comparison. The methods selected to design a convenient system to assess the antioxidant activity of Moringa peregrina extracts including, reducing power, chelating activity on Fe2 +, free radical-scavenging, total antioxidant, superoxide radical scavenging, hydrogen peroxide scavenging and hydroxyl radical scavenging activities in an attempt to understand its mechanism of action, which may pave the way for possible therapeutic applications. In addition, the results were compared with natural and synthetic antioxidants, such as α- tocopherol, ascorbic acid, butylated hydroxytoluene(BHT), butylated hydroxyanisole (BHA) and trolox. The results indicated that Moringa peregrina ethanolic and aqueous extracts (100-300 mg/kg p. o.) also dose-dependently and significantly inhibited (p < 0.05-0.001) fresh egg albumin-induced acute inflammation. Ethanolic and aqueous extracts of Moringa peregrina exhibited a strong reducing power, Fe2 + chelating effect, free radical scavenging activity, hydrogen peroxide scavenging ability, and hydroxyl radical scavenging activity Antioxidant activity of the ethanolic and aqueous extracts increased with increased concentrations in the range of 20-140 μg/ ml the extracts also exhibited a strong superoxide radical scavenging activity. This study showed that Moringa peregrina extracts exhibited antioxidant activity in all tests and the extracts could be considered as a source of natural antioxidants. Taken together, Moringa peregrina has potential as an anti-inflammatory and antioxidant agent against inflammation and free radicals and deserves clinical trial in the near future.
Comparison of the 2D gel electrophoresis of serum proteins of normal subjects (Control) and chronic pancreatitis patients (Test) in 12% SDS-PAGE (second dimension) in which 350 µg of protein was loaded into IEF. The figure is the representative of 5 individual experiments conducted by using pooled serum samples. 
a. Cropped 2D gel images of selected proteins in normal subjects (Control) and chronic pancreatitis patients (Test) in which Spot 1 and Spot 2 are selected for the study. 
c. Mascot score histogram of proteins in spot 2. Ions score is –10*Log (p), where p is the probability that the observed match is a random event. Individual ions score >42 indicate identity or extensive homology (p < 0.05). Protein scores are derived from ions scores as a non-probabilistic basis for ranking protein hits. 
b. Probability based mowse score of proteins in spot 1. Ions score is –10*Log (p), where p is the probability that the observed match is a random event. Individual ions score >43 indicate identity or extensive homology (p < 0.05). Protein scores are derived from ions scores as a non-probabilistic basis for ranking protein hits. 
Background and Aim: Serum proteome analysis is a novel tool to detect the protein markers for any pathological disease. Diagnosis of chronic pancreatitis is mainly based on the level of serum marker enzymes amylase and lipase. The aim of the present study is to evaluate other new serum protein markers for chronic pancreatitis in human subjects by proteome analysis. Subjects and Methods: Serum proteome analysis was carried out in groups of pooled serum samples from chronic pancreatitis patients (n-25) registered in the Department of Surgical Gastroenterology and Proctology, Stanley Medical College and Hospital in the age group of 30 – 50. Normal serum samples (n-20) were pooled in groups from age and sex matched normal healthy volunteers. Serum proteome analysis was carried out by conducting 2D-electrophoresis, identifying differentially expressed proteins, isolating and excising the bands of interest, digesting the proteins by trypsin, separating of digested peptides by 2D Liquid Chromatography Mass spectroscopy (2D LC-MS) and identifying peptides by Mass Finger Printing Technique. The proteins were then confirmed by protein search engine programme MASCOT. Quantitative analysis of such protein(s) was carried out individually in all the samples for confirmation. Results: Among the 45 differentially expressed proteins, we have considered Haptoglobin-2 (Hp-2) as one of the marker, since Hp was quantitatively elevated significantly in all the samples considered for the study. Hp-2 isoform was also confirmed by native PAGE by using specific Benzidine-H2O2 stain. Being a natural acute phase reactant, Hp-2 level might have been elevated as a compensatory mechanism to counteract the tissue damage and to overcome the oxidative stress as an antioxidant because pancreatic tissue damage and inflammation is associated with the harmful effects of free radicals. Conclusion: Hp-2 isoform, a positive acute phase reactant with antioxidant property is identified as a marker for chronic pancreatitis by serum proteome analysis.
Oxidative stress resulting from biomolecular oxidative damage due to the imbalance between reactive species production and antioxidant response has become an universal constraint of life-history evolution in animals and a modulator of phenotypic development and trade-offs. Redox balance is an important selective pressure faced by most organisms, and a myriad of mechanisms have evolved to regulate and adjust this balance. This diversity of mechanisms means that organisms have a great deal of flexibility in how they deal with reactive species challenges across time, conditions, and tissue types, as well as that different organisms may evolve different strategies for dealing with similar challenges. In the following paragraphs, we review the adaption of biological membranes as structural antioxidant defense against reactive species evolved by animals. In particular, it is our goal to describe the physiological mechanisms underlying the structural adaption of cellular membranes to oxidative stress, to explain the meaning of this adaptive mechanism, and to review the state of the art about the link between membrane composition and longevity of animal species.
Levels of lipid peroxidation of participants administered with quinine. 
The erythrocyte membrane is susceptible to lipid peroxidation due to its high content of polyunsaturated lipids. The present study ascertained levels of lipid peroxidation of non-parasitized erythrocytes of non-malarious male participants administered with quinine. Determination of erythrocyte lipid peroxidation was by a monitor of levels of thiobarbituric acid reactive substances, measured spectrophotometrically by taking the sample absorbance at 432 nm against a reagent blank at 24 °C. Fifteen (15) non-malarious male (59–79 kg) participants of confirmed HbAA genotype between the age brackets of 21–34 yr enrolled for this study. Three (3) participants per group were administered with quinine according to the corresponding doses: 1.5, 3.0, 4.5 and 6.0 mg/kg (control; n = 3). At regular time interval of 3 h for 12 h, blood samples were withdrawn for determination of erythrocyte membrane lipid peroxidation. Erythrocyte membrane of participants administered with quinine exhibited relatively high levels of lipid peroxidation in a dose dependent manner. Generally, within 0–6 h, the four adminstered doses of quinine caused a time dependent elevation of lipid peroxidation. Further increase in time (t > 6 h) showed decreasing levels of lipid peroxidation in participants administered with quinine. Generally, the levels of erythrocyte lipid peroxidation in the presence of quinine showed 2 phase profile. The first stage showed increasing levels of lipid peroxidation at t < 6 h after drug administration, followed by the second phase which showed decreasing levels of lipid peroxidation. The results showed that administration of quinine to non-malarious individuals elicited flux in erythrocyte lipid peroxidation.
Weight of rabbits before and after V. amygdalina aqueous leaf extract administration. Values are expressed as mean ± SD, n = 5. Values were significant at P ≤ 0.05, indicates point of extract administration.  
The mean level of glucose, malondialdehyde (MDA) and glutathione (GSH) in normal or alloxan-induced diabetic rabbits after administration of the aqueous leaf extract of V. amgydalina. 
Introduction: Diabetes is a disease that constitutes multiple sources of free radicals, thus oxidative stress is expected to have a double impact. Since oxidative stress is mediated by hyperglycemia-induced generation of free radical, it is supposed that compounds with hypoglycemic and antioxidative properties would be useful antidiabetic agents. Our study investigated the use of V. amygdalina as a potential hypoglycemic and antioxidative agent by testing its effect on the antioxidant biomarkers and lipid peroxidation.Methods: Dried leaves of V. amgydalina were extracted for 48 hrs and freeze dried. Acute toxicity was investigated in 25 rats. Thirty six rabbits were divided into 6 groups: groups I - III were normal; diabetes was induced in groups (IV – VI). Groups I and IV were normal and diabetic controls respectively. The animals were treated with aqueous leaf extract of V. amygdalina. Blood samples were collected and used for the study.Results: The reduction in body weight in the diabetic groups was regained following administration of the extract of V. amgydalina. The extract is considered safe and had little or no effect on blood glucose, MDA and GSH levels of the normal rabbits. Extract significantly reduced glucose and MDA concentrations but increased GSH levels in the diabetic rabbits. Similarly, the extract had no effect on the activities of SOD, CAT and GPx in normal rabbits, however in diabetic rabbits, the enzymes activities increased dose-dependently.Conclusion: This finding provides basis for the use of V. amgydalina as potential antidiabetic antioxidant agent and may be useful for its hypoglycemic property.
Introduction: Free radical induced oxidative stress is involved in the pathogenesis of various diseases and disorders. Antioxidants play an important role against this oxidative stress to protect our body. The present study was carried out to evaluate the in vitro antioxidant properties of methanol extract of C.maxima aerial parts (MECM). Methods: MECM was assayed on different in vitro free radical models like, DPPH, nitric oxide, superoxide, hydrogen peroxide and lipid peroxide radical models. Reductive ability of the extract was also tested by the complex formation with potassium ferricyanide. Further total phenolic and flavonoid contents of the crude extract were also measured. Butylated hydroxy toluene was taken as standard. Result: The extract showed good dose dependent free radical scavenging activity in all the models. Reductive ability was also found to increase with increase in extract concentration. Determination of total phenolic and total flavonoids content showed that 1 gm of dry extract contains 66.70±3.60 mg equivalent of pyrocatechol and 26.50±1.40 mg equivalent of quercetin. Conclusion: All the results of the in vitro antioxidant assays revealed potent antioxidant and free radical scavenging activity of the aerial part of C.maxima, equivalent to that of standard BHT and this antioxidant property may be attributed to its high phenolic and flavonoid contents.
Background: Inflammation is typically characterized by increased permeability of endothelial tissue resulting in oedema. Much of pyrimidine derivatives have been highly useful for treating acute and self-limited inflammatory conditions. This study was performed to explore the acute toxicity, anti-inflammatory activity and biochemical effect of natural heterocycle pyrimidine glucoside vicine obtained from Vicia faba L. and its aglucone (divicine) and their effect on some oxidative stress biomarkers in Albino rats. Also, the anti-inflammatory effect of the tested compounds were compared with diclofenac sodium (100 mg/kg bw). On the other hand, this article was extended to assess the impact of a very high dose of diclofenac sodium (100 mg/kg bw) on liver enzymes, which was used by many authors to explain the negative effects of diclofenac when used at this dose as a standard anti-inflammatory drug. Methods: The median lethal dose (LD50) and histopathological effects of vicine and divicine was determined in order to assess their safety. Also, the inflammatory effect of the tested compounds was examined at doses (50–200 mg/kg bw) in rats, using fresh egg albumin-induced oedema and compared with standard anti-inflammatory drugs, diclofenac sodium (100 mg/kg bw). Finally, vicine and divicine treatment (50—200 mg/kg bw) was evaluated for 10 consecutive days prior in Albino rats in the serum levels of hepatic enzymes, glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT), gamma glutamyl transferase (γ-GT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), super oxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (GPx) and thiobarbituric acid reactive substances (TBARs). Results:Vicine and divicine were found to have an LD50 of 2100 and 1950 mg/ kg bw orally, respectively and a significant (p < 0.001) dose-dependent anti-inflammatory activity with the highest percentage inhibition (59.56%) and (45.65%) for vicine and divicine, respectively, at 150 mg/kg against the fresh egg albumin-induced oedema in rats after 180 min. Also, the standard anti-inflammatory diclofenac sodium (100 mg/kg) showed 100% inhibition against the fresh egg albumin-induced oedema in rats after 180 minutes. In addition, administration of vicine and divicine orally to the rats at doses of 50—200 mg/kg bw for 10 days showed non-significant changes in liver enzymes and serum TBARs as compared with the control group. But administration of diclofenac sodium orally at a concentration of 100 mg/kg bw daily for 10 consecutive days to rats showed significant increase in liver enzymes and serum TBARs and significant decrease in blood GSH, SOD and GPx. Histopathology of the rat liver morphology of all the treated groups showed no disorder except for the photomicrograph of the cross section of liver from rats treated with diclofenac sodium 100 mg/kg for 10 consecutive days, shows pronounced dilatation of the hepatic portal vein. Conclusion: The observation allows conclusion that vicine and divicine at concentration of 50—150 mg/kg bw exhibited good anti-inflammatory effect and have no hepatotoxic activity.
DPPH radical scavenging activity of different Ajuva iva extracts. CE: methanol extract; EAE: ethyl acetate extract; CHE: chloroform extract, Rut: rutin, quer: quercetine and GA: gallic acid. Data are presented as IC 50 values mean ± SD (n = 3).  
Total polyphenols and flavonoid contents of Ajuga iva root extracts Value are expressed as mean ± SEM, n = 5 
Percentage inhibition of the linoleic acid oxidation by the A. iva extracts (CE: methanol extract; CHE: chloroform extract; EAE: ethyl acetate extract), BHT and blanks (H 2 O and Methanol) after 24h. Results are means of three different experiments.  
Comparison the EC 1 values of Ajuga iva extracts (CE: methanol extract; CHE: chloroform extract; EAE: ethyl acetate extract), with standards; quercetin, gallic acid, and ascorbic acid. Data are the mean values ± SD, n=3. EC 1 : concentration of antioxidant having a ferric reducing ability equivalent to that of 1 mmol/L FeSO 4 ,7H 2 O.  
Introduction: Antioxidant effects of Ajuga iva L extracts (AIE), a traditionally used plant in Algerian folk medicine, were investigated. This plant is widely used to treat diabetes, gastric ulcer, dysuria, painful joints of the limbs and other free radicals related disorders. Methods: In order to determine the antioxidant activities of AIE, the shoot extracts were prepared using solvents of varying polarity. Xanthine oxidoreductase (XOR) was purified from bovine milk. Anti-radical activities were determined by enzymatic and non enzymatic methods: The enzymatic methods were realised by either production of uric acid or reduction of cytochrome c. The non enzymatic methods were conducted using in vitro techniques: NBT test, ß-carotene linoleate acid, 1,1- diphenyl-2 picrylhydrazyl (DPPH) radical-scavenging, ferric reducing /antioxidant power (FRAP) and Ferrous ion chelating activity Results: All extracts showed inhibitory properties on xanthine oxidase, the IC50(μM / quercetin equivalen) ranges from 3.878 ± 0.717 to 5.835 ± 0.468), with an additional superoxide scavenging capacity. These extracts showed a potent DPPH radical scavenging activity and a powerful scavenging activity of superoxide anion more than that of gallic acid using NBT test. They possess a similar inhibition ratio of the linoleic acid oxidation to that of the BHT, and gave a reduction power better than rutin in Ferric Reducing Ability of Plasma assay (FRAP). However, they were weaker than EDTA in Fe2+ chelating activity. Conclusion:Ajuga iva L. appears to be a valuable plant and could be used to treat conditions where inhibition and free-radicals scavenging of XO is warranted.
Endotoxin-ROS-Liver damage axis.  
Overview of ethanol absorption, formation and circulation of free radicals and lipid peroxidation products (LPP).  
Almost 95% of alcohol is metabolized by the liver but a small fraction is also metabolized in other organs like brain, heart, kidney, pancreas, skeletal muscle etc. Alcohol is metabolized by oxidative and non-oxidative pathways. Alcohol metabolism results in the generation of reactive molecule acetaldehyde and reactive oxygen species (ROS), which in turn leads to oxidative stress. The reactive metabolic products of alcohol and ROS cause lipid peroxidation, the lipid peroxidation products (LPP) like malondialdehyde (MDA) enter the circulation. In most of the studies on alcoholic liver disease the serum levels of ROS/LPP were estimated, but these products are also contributed by other tissues. Even though the maximum amount of ROS/LPP is produced by the liver, after reaching other organs through circulation, ROS/LPP further multiply and again reach the circulation. At this point by measuring the ROS/LPP in blood and claiming it's of liver, are we estimating it correctly?
In-vitro concentration dependent percentage inhibition of Superoxide radical by different extracts of Thespesia populnea seeds.  
Total phenolic and alkaloid content (mg/gm) of Thespesia populnea seeds extracts
Introduction: Anti-oxidants are vital substances which possess the ability to protect the body from damage caused by free radical induced oxidative stress. There is an increasing interest in natural anti-oxidants, e.g., polyphenols, present in medicinal and dietary plants, which might help prevent oxidative damage. Methods: In this present study we investigated preliminary phytochemical, total phenolic, alkaloid content and In-vitro antioxidant activity of hexane, ethyl acetate, ethanol (70%) and methanol extracts of Thespesia populnea Seeds. Results:Thespesia populnea Seeds revealed the presence of steroids, flavonoids and alkaloids, glycosides, tannins, quinones and carbohydrates. The extracts do not contain the amino acids, oils and the hexane fraction do not contain triterpenes and tannins. The ethyl acetate extract have more phenolic content than other extracts and the methanolic extract has more alkaloidal than other extracts. The selected plant extracts were produced concentration dependent percentage inhibition of superoxide radical and produced maximum activity at a concentration of 160 μg and there after the percentage inhibition were raised gradually to its maximum level with higher concentrations. Conclusion: Among the four types of T.populnea seeds extracts, the methanolic extract showed better activity than aqueous extracts at 160 μg concentrations.
Introduction: In the present study, hexane, Chloroform, ethyl acetate, butanol and methanol extracts of fruits of Evolvulus alsinoides Linn (Convolvulaceae) were examined total phenolics (TP) and in vitro antioxidative capacity. Methods: For the determination of total phenolics (TP) and in vitro antioxidative capacity, established assay methods such as 1, 1 - diphenyl – 2- picryl hydroxyl (DPPH) radical assay, reducing power, ferric ion chelating assay, superoxide anion, and nitric oxide scavenging activity assays were used with reference to synthetic antioxidant butyl hydroxyl toluene (BHT). One way analysis of variance (ANOVA) and Duncan's Multiple Range Test (DMRT) were carried out. Results: The TP ranged from 1686 ± 1.527 mg GAE /100 of Dry weight (DW) to 1255 ± 0.020 mg GAE/100 of Dry weight (DW). The data obtained from the study showed high levels of antioxidant activity of the fruit extracts. Conclusion: From the findings, it was observed that there was a well correlation between antioxidant activity and total phenolic/flavonoid contents. These results may be useful to further analyze wild edible fruits that contain most antioxidant activity in order to identify the active constituents.
Effect of the stem bark of Alstonia scholaris methanol extract on Carrageenan-induced rat paw edema
Effect of stem bark of Alstonia scholaris methanol extract on Dextran-induced rat paw edema
Background: The plant Alstonia scholaris R.Br. (Labiatae) is widely cultivated throughout India. The bark of this tree is medicinally used as an astringent, tonic, anthelmintic, antiperiodic and febrifuge. Methods: The effect of Methanolic extract of stem bark of Alstonia scholaris R.Br. was investigated to evaluate the antioxidant activity by TLC method and anti-inflammatory activity by Carrageenan induced rat paw odema, dextran induced rat paw odema and cotton pellet induced granuloma. Result: Anti-inflammatory activity of the tested extract and its fractions was comparable with that of the standard drug Indomethacin (10 mg/kg). Conclusion: The present investigation for the first time, confirms the potential anti-inflammatory activity in the methanol extract of Alstonia scholaris stem bark. Antioxidant property of the extract was confirmed quantitatively by β-carotene–linoleate oxidation method by thin layer chromatography.
The sphincteric anal pressure was measured through a manometric equipment with a balloon catheter (Proctossystem Viotti, Sao Paulo, Brazil). Values were expressed in cm of H 2 O. Group CO: control animals treated with saline (NaCl 0.9%); Group CO + SOD: animals treated with SOD; Group DM: diabetic animals untreated with SOD; Group DM + SOD: diabetic animals treated with SOD. a P < 0.05 versus Control; b P < 0.01 versus DM. 
Effects of SOD Treatment on Body weight, Glucose level, Lipid Peroxidation (TBARS) and SOD activity in the intestine tissue of DM rats.
Comet assay in the intestine from diabetic and non-diabetic rats treated or untreated with superoxide dismutase (SOD).
Introduction: The gastrointestinal system is frequently affected in patients with diabetes mellitus (DM). Some of these alterations are due to oxidative stress and the production of free radicals. The present study was designed to evaluate whether treatment with superoxide dismutase (SOD) exerts protection on established bowel alterations in experimental diabetes mellitus induced by streptozotocin. We measured the lipid peroxidation, the superoxide dismutase activity and the DNA damage. Materials and Methods: We used the anorectal pressure to evaluate the nitrosative stress and used an inflammatory score to measure the macroscopic and microscopic bowel alterations. Results: The oxidative stress and the DNA damage was elevated in DM group and reduced with the SOD administration. The use of SOD also ameliorates the inflammatory bowel alterations and the anorectal pressure. Conclusion: SOD administration showed beneficial effects in all parameters of large bowel alterations in DM rats.
Alzheimer's disease (AD) is a neurodegenerative process associated with oxidative stress. In the past, it was claimed that all neuronal lesions involved in the onset and progression of AD were related to oxidative stress.Today, we know that intracellular amyloid beta (Ab) could play a central role in the pathophysiology of the disease. Ab binds to heme groups in mitochondrial membranes causing electron transport chain impairment and loss of respiratory function. The experimental evidence of such oxidative stress leads to the basis for treatment of AD with antioxidants. Many clinical trials have been developed to clarify whether antioxidants are beneficial in AD treatment. However, the results obtained in no way confirm that antioxidants are an effective AD therapy. More research is necessary to clarify this point.
An in vitro study was carried out to investigate levels of oxidative stress indicators of sickle erythrocytes incubated in aqueous extracts of Anacardium occidentale, Psidium guajava and Terminalia catappa for 12 h. At regular time intervals of 3 h, portions of the incubation mixtures were withdrawn and spectrophotometric method was used to assay for levels of erythrocyte malondialdehyde (MDA) and methaemoglobin (Met.Hb%). The control analysis showed that within the experimental time, erythrocyte MDA increased from 2.45±0.35 to 3.13±0.59 mmol/ml (p > 0.05; p value = 0.801176). Erythrocyte MDA concentrations in the presence of the three extracts were higher than the control samples at t = 3 h (p > 0.05; p value = 0.963253). Compared with the control samples at the given time (t) intervals, extract of T. catappa exhibited the highest capacity to cause reduction of erythrocyte MDA ([T. catappa] = 800 mg%; [MDA] = 2.89±0.33 mmol/ml; t = 12h). Erythrocyte Met.Hb% increased from 2.42±0.55 to 2.51±0.49% (p > 0.05; p value = 0.995171) in the control samples within 12 h. Incubation of sickle erythrocytes with extract of [P guajava] = 800 mg% for 9 h caused reduction of Met.Hb% from 2.49±0.49% to 2.29±0.45% (p > 0.05; p value = 0.983519). Extracts of A. occidentale, P. guajava and T. catappa exhibited variable capacities to hinder lipid peroxidation, but did not cause corresponding reduction in erythrocyte Met. Hb%, as shown by negative correlation between the two oxidative stress indicators in the presence of T. catappa and higher concentrations of A. occidentale and P. guajava.
Background Andrographis serpyllifolia, used as a medicinal plant in traditional practices in India, where in cancer patients are treated using the leaves of A. serpyllifolia. The crude extracts of A. serpyllifolia not only contain a wide variety of phenolic compounds but also show an excellent antioxidant activity. Therefore, this plant might be a good candidate for further development for its antioxidant remedies. However, the biological activities of the phenolic extracts of A. serpyllifolia on cancer have not been studied to date.Objective To investigate the antioxidant and antiproliferative properties of the phenolics of A. serpyllifolia.Materials and methodsFree and (ASFP) and bound (ASBP) phenolics of A. serpyllifolia were isolated and determined antioxidant and antiproliferative abilities that are required for anticancer properties.ResultsIndividual phenolic constituents present in each of these fractions and their precise contribution to both antioxidant and antiproliferative activities were determined to justify the traditionally observed result of anticancer properties. Treatment of HeLa cells with ASFP and ASBP showed antiproliferative activity with increased malondialdehyde (MDA) as well as decreased levels of reduced glutathione (GSH). The present experimental data suggest that components within the ASFP may have inherent properties that suppress cancer cell proliferation. The phenolic fractions were also screened for their potential antioxidant activities using DPPH, reducing power, DNA protection, inhibition of lipid peroxidation and protein carbonyls model systems. ASFP exhibited highest antioxidant activity in all the model systems employed to study antioxidant activity. The positive correlation between polyphenolic content of A. serpyllifolia to its antioxidant activity was seen.Conclusion Potent antiproliferative and DNA protective activity of ASFP and ASBP may contribute significantly against cancer pathogenesis.
Background:Diospyros peregrina (Ebenaceae) is a medium-sized evergreen tree having ethnomedicinal significance as an aphrodisiac, astringent, bactericide and for the treatment of diarrhoea, cholera, dysentery, fever, malaria and diabetes. Objectives: To evaluate the anti-oxidant and anti-cancer activity of different extracts and fractions of matured unripe fruits of Diospyros peregrina on various in vitro models. Materials and Methods: Anti-oxidant and free radical scavenging activity of various extracts and fractions were determined by DPPH, ABTS and Alkaline DMSO methods. In vitro cytotoxicity of different extracts and fractions of matured unripe fruits of Diospyros peregrina was evaluated on MCF-7 and Hep-G2 cell lines by MTT and SRB methods. Results: Among the extracts and fractions studied for antioxidant activity n-butanol fraction (NBF) and aqueous extract (AE) showed potential scavenging effect against DPPH, ABTS and superoxide free radicals. In vitro cytotoxicity assays of extracts and fractions by MTT and SRB methods revealed that dichloromethane fraction (DCMF) exhibited maximum cytotoxicity on both MCF-7 and Hep G2 cell lines. Conclusion: The higher anti-oxidant potential of n-butanol fraction (NBF) and aqueous extract (AE) may be due to the presence of phenolic compounds, flavonoids or flavonoid glycosides. Dichloromethane fraction (DCMF) exhibited maximum cytotoxicity on both MCF-7 and Hep G2 cell lines. The cytotoxic activity of DCM fraction may be attributed due the presence of triterpenoids.
Cellular damage or oxidative injury arising from free radicals or reactive oxygen species (ROS) now appears the fundamental mechanism underlying a number of human neurodegenerative disorders, diabetes, inflammation, viral infections, autoimmune pathologies and digestive system disorders. Free radicals are generated through normal metabolism of drugs, environmental chemicals and other xenobiotics as well as endogenous chemicals, especially stress hormones (adrenalin and noradrenalin). Accumulated evidence suggests that ROS can be scavenged through chemoprevention utilizing natural antioxidant compounds present in foods and medicinal plants. India is blessed with enormous biodiversity resources, but plagued with several diseases, including those with ROS as the etiological factor. The present study was designed to evaluate the antioxidants activities of unripe fruits of Ficus glomerata Roxb and three complementary test systems, namely superoxide anion scavenging activity, reducing power, hydroxyl radical scavenging activity were used for the antioxidant analysis. In this study Ficus glomerata exhibited strong antioxidant activity.
Introduction: Ingredients of Indian cuisine are well known for their antioxidant properties which can help prevent many diseases. The fruits of Dillenia indica and Garcinia penducalata are commonly used by the Assamese speaking population of Assam, India, but their antioxidant properties have not been investigated. Methods: Methanol, petroleum ether and water extracts of the shade dried fruits of Dillenia indica and Garcinia penducalata were obtained and the IC50 values of their DPPH, hydroxyl, oxygen and nitric oxide scavenging activities were estimated along with their reductive ability, vitamin C and total phenolic content. Vitamin C was used as a standard reference for the antioxidant scavenging activities. Result: The IC50 values for DPPH, hydroxyl, oxygen and nitric oxide inhibition of vitamin C were 43.7 ug/ml, 50.44 ug/ml, 61.04 ug/ml and 41.82 ug/ml, respectively while the IC50 value for its reductive ability was 47.17 ug/ml. The IC50 values for the DPPH, hydroxyl, oxygen, nitric oxide and reductive ability of the methanolic extract of Dillenia indica were 31.25 ug/ml, 51.82 ug/ml, 51.44 ug/ml, 39.73 ug/ml and 40.18 ug/ml, respectively. These IC50 values were superior in comparison to its petroleum ether and water extracts. The methanolic extract of Garcinia penducalata had the highest amount of phenolic content and its IC50 values for DPPH, oxygen and nitric oxide scavenging activities were 50.23 ug/ml, 66.06 ug/ml and 63.02 ug/ml, respectively. Conclusion: The higher amount of phenolic content in the methanolic extract of Dillenia indica may contribute to its better in vitro anti oxidant property.
Background: The present study was aimed to evaluate in vitro anti-oxidant activity of the Ruellia tuberosa (Acanthaceae) roots. Materials and Methods: Anti-oxidant activity was evaluated by using DPPH free radical scavenging activity and reducing power by FeCl3. The methanolic extract (ME), water extract (WE), ethyl acetate extract (EAE) and ethyl acetate fraction of methanol extract (EAFME) of root were tested. The Ascorbic acid was used as positive control. Total phenolic and total flavonoid content were also determined by Folin-Ciocalteu reagent and complementary colorimetric methods (aluminum chloride method and 2, 4-dinitrophenylhydrazine method respectively. Results: The EAFME of root showed the highest concentration of phenolic, flavonoid content, free radical scavenging activity and reducing power. The various extract showed a significant anti-oxidant activity when (P < 0.05) compared with standard. Conclusion: It is concluded that the R tuberosa root possess anti-oxidant activity. Further studies are suggested to isolate the active principle responsible for the activity.
Effect of Manjisthadi churna and Dhatrinisha churna on SOD level of Liver tissues of different groups of rats (Units/mg protein).  
Effect of Manjisthadi churna and Dhatrinisha churna on Catalase level of Liver tissues of different groups of rats (Units/mg protein).  
Effect of Manjisthadi churna and Dhatrinisha churna on Reduced Glutathione level of Liver tissues of different groups of rats (nmoles/mg protein).  
Effect of Manjisthadi churna and Dhatrinisha churna on Glutathione Peroxidase level of Liver tissues of different groups of rats (Units/mg protein).  
Effect of Manjisthadi churna and Dhatrinisha churna on Lipid Peroxidation level of Liver tissues of different groups of rats (Units/mg protein).  
Background: Dhatrinisha churna and Manjisthadi churna have been traditionally used in the Ayurvedic system of medicine and by traditional medical practicioners of India to treat hyperlipidemia. Objective: To evaluate the antioxidant activity of Dhatrinisha churna and Manjisthadi churna in experimental hyperlipidemic rats. Materials and Methods: Hyperlipidemia was induced by administrating exogenous cholesterol. Assessment of anti-oxidant status for liver was carried out by assay of various physiological enzymatic (like SOD, catalase, gluatathione peroxidase) and non enzymatic (GSH, lipid peroxidative indices) antioxidant. Result: Significant increase in the level of enzymes like SOD, catalase, glutathione peroxidase, GSH and decrease in lipid peroxidase were observed in drug treated animals. Conclusion: In conclusion, these observations show that Manjisthadi churna and Dhatrinisha churna exert their protective effect by improving antioxidant status and decreasing the lipid peroxidation, thus establishing them as an effective antioxidant.
Diagram of the extraction process.  
Total phenolic contents in Viola tricolor flowers and leaves/roots fractions 
IC 50 (μg/mL) values of the DPPH and TBARS assay in Viola tricolor flowers and leaves/roots fractions 
Rutin concentration in Viola tricolor flowers fractions 
Inhibition Percentage (IP %) of V. tricolor fractions . (<) = n-butanol, (·) = dichloromethane and (+) = ethyl acetate fractions from flowers. (ù) = Ethyl Acetate, () = n-butanol and (X) = dichloromethane fractions from the leaves/roots. (o) = L-Ascorbic acid.  
Background: Natural products with anti-oxidant properties have gained importance in recent years due to their ability to neutralize free radicals or their actions. Viola tricolor belong to Violaceae family and is known as heartsease. The aims of the study was to evaluate the phenolic contents and anti-oxidant capacity in the different flowers and leaves/ roots fractions of Viola tricolor, as well as was identified and quantify rutin in the flowers fractions. Materials and Methods: Total phenolic content was determined using the Folin-Ciocalteu assay, the anti-oxidant capacity by the 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) scavenging method and thiobarbituric acid reactive species (TBARS) assay. The presence of rutin was investigated by High Performance Liquid Chromatography (HPLC). Results: In the flowers fractions the phenolic content varied from 12.84 ± 0.072 to 0.23 ± 0.042 mg/g; and 7.49 ± 0.002 to 0.63 ± 0.002 mg/g for leaves/roots. IC50 DPPH values ranged from 13.40 to 14.18 mg/ml in flowers, and 32.84 to 284.87 μg/ml for the leaves/roots. The fractions showed inhibition against TBARS, following order: ethyl acetate fractions > butanolic fractions > dichloromethane fractions. HPLC results indicate a very high content of rutin (177.46 mg/g) in this species that could explain the good anti-oxidant activities and the high phenolic contents in these fractions. Conclusions: These findings suggest that V. tricolor contains anti-oxidant principles and rutin contribute, in party, for the action.
Introduction: Michelia champaca (Magnoliaceae) is a medicinal plant of paramount importance which is traditionally used against a number of diseases including inflammatory conditions. In the present study, crude hexane, ethyl acetate extracts and compounds 1-3 isolated from the extracts of M. champaca flower were investigated for possible antioxidant and antibacterial activity. Method: The DPPH radical scavenging assay was carried out by the protocol given by Nickavar et al 2006.[1] The extracts and the compounds were also investigated for antibacterial activity through disc diffusion method against four different bacterial strains viz. Bacillus subtilis, Staphylococcus aureus, Salmonella typhi and Shigella dysenteriae using ampicillin as control. Results: In the present study the ethyl acetate extract, hexane extract exhibited strong antioxidant activity and the IC50 values in DPPH radical scavenging assay were found to be 160 μg/ml and 250 μg/ml, respectively while the IC50 values of isolated compounds 1-3 were 200 μg/ml, 220 μg/ml and 150 μg/ ml, respectively. In this bioassay the extracts showed significant results compared to the individual isolated compounds. Ethyl acetate extract was more effective against all the bacterial strains followed by the hexane extract, compound-3, compound-1 and compound-2. Results of the present study suggest that M. champaca flower extracts and the isolated compounds possess strong antioxidant and antibacterial activity.
Antibacterial activity of oven-dried culinary herbs in comparison with commercial herbs 
Introduction: Although the antioxidant and antibacterial properties of Labiatae herbs are well known, the effects of different drying methods are yet to be determined. In this study, the antioxidant and antibacterial properties of fresh and oven-dried herbs of oregano, marjoram, rosemary, sage, basil, thyme, peppermint, and spearmint were investigated, in comparison with commercial brands of dried herbs. Methods: Antioxidant properties of total phenolic content, total flavonoid content, caffeoylquinic acid content, free radical scavenging activity, and ferric reducing power were assessed using the Folin-Ciocalteu, aluminium chloride, molybdate, DPPH radical scavenging, and potassium ferricyanide assays, respectively. Antibacterial properties were assessed using the disc-diffusion assay based on minimum inhibitory dose (MID). Bacteria tested were Gram-negative Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhi, and Gram-positive Bacillus cereus, Micrococcus luteus, and Staphylococcus aureus. The three drying treatments were oven drying at 50 °C (OD50), oven drying at 80 °C (OD80), and oven drying at 50 °C with microwave pre-treatment (MOD50). Results: Fresh and commercial rosemary, and oven-dried oregano had the strongest antioxidant properties. Generally, MOD50 herbs had the strongest antioxidant properties followed by OD50 and OD80 herbs. Oven-dried rosemary had lower phenolic content and antioxidant activity than commercial rosemary, while oven-dried oregano, spearmint, thyme, peppermint, and basil had higher values. All herbs showed no antibacterial activity against Gram-negative E. coli, P. aeruginosa, and S. typhi. Rosemary, sage, peppermint, and spearmint inhibited the growth of Gram-positive B. cereus, M. luteus, and S. aureus. Compared to green and black teas of Camellia sinensis, rosemary and sage have stronger antibacterial properties. Conclusion: Labiatae herbs can have enhanced antioxidant and antibacterial effects when used in combination. Further research is needed to study the synergistic behaviour of these herbs.
Levels of human erythrocyte glutathione S-transferase (GST) activity were ascertained in the presence of separate increasing concentrations (0.2, 0.4, 0.6, and 0.8 mg% w/v) of five antimalarial drugs {Pyrimethamine/Sulphadoxine (PS), Halofantrine-HCl (H-HCl), Quinine, Artemether/umefantrine (AL) and Chloroquine Phosphate (CP)} in vitro. Determination of erythrocyte GST activity was carried out by spectrotrophotometric method with the standard substrate (1-chloro-2, 4-dinitrobenzene) and co-substrate (reduced gluthathione) at maximum absorbance (λmax) = 340 nm. Human erythrocyte GST activity ranged between 3.27 ± 0.13 and 3.40 ± 0.05 iu/gHb. The addition of increasing concentrations of four antimalarial drugs to assay mixture engendered decreased levels of GST activity in a concentration dependent manner, which was in the order: CP > AL > Quinine > PS. Specifically, 0.8 mg% CP exhibited the highest capacity to cause decreased GST activity from values of 3.41 ± 0.06 to 2.18 ± 0.09 iu/gHb, representing 33.7% relative inhibition of GST activity. In contrast, concentrations of H-HCl between 0.2 and 0.6 mg% caused elevation of GST activity above the values of the control samples. However, the increased levels of GST activity was not significantly different (p > 0.05) compared with the control samples. The five antimalarial drugs exhibited variable capacities to alter human erythrocyte GST activity, which suggest the capability of these drugs to perturb the redox status of human erythrocytes.
Introduction:Antioxidants are micronutrients that have gained importance in recent years due to their ability to neutralize free radicals or their actions. Sesbania sesban (L) Merr is an ancient plant which is traditionally used as an antioxidant folklore plant. The present research deals with the phytochemical screening and in vitro evaluation of antioxidant activity of the leaves of Sesbania sesban (L) Merr.Methods:The ethanolic extract of the plant Sesbania sesban (L) Merr was subjected for the phytochemical screening. The preliminary screening reports the presence of Saponin, Tanin, Phenolic compound, Flavonoid in ethanolic extracts. DPPH scavenging activity or the Hydrogen donating capacity was quantified in presence of stable DPPH radical on the basis of Blois method. NO scavenging activity was performed in the presence of nitric oxide was generated from sodium nitroprusside and measured by the Greiss reaction according to the method of Marcocci. Ascorbic acid was used as standard for the both.Results:The scavenging was found to dose dependent. Thus extract has been established the as an antioxidant. The reducing capacity serves as significant indicator of antioxidant activity. The reducing power increased with the increasing concentration of sample.Conclusion:The folklore use of Sesbania sesban (L)Merr has been proved in present research work. Further studies along with isolation and molecular mechanism on extract of Sesbania sesban (L) Merr may lead to significant out come.
Measurement of DPPH free radical scavenging activity of solvent extracts of Mokkathotapapada leaves of Piper betel L. Cv. Kapoori, a local cultivar 
In vitro antioxidant activity of solvent extracts of Mokkathotapapada leaves of Piper betel L. Cv. Kapoori, a local cultivar against Nitric oxide method
Introduction: Ingredients of Indian cuisine are well known for their antioxidant properties which can help prevent many diseases. The Betel (Piper betel L.) is the leaf of a vine belonging to the Piperaceae family; it is figured out both as a mild stimulant and for its medicinal properties. The betel plant originated from South and South East Asia. The leaves of Piper betel L. have been traditionally known for its various therapeutic uses. Piper betel L. Cv. Kapoori is a local cultivar obtained from Chintalapudi, Guntur district of Andhra Pradesh, India, but their antioxidant properties have not been investigated. Methods: Organic solvent extracts of Mokkathotapapada leaves of Piper betel L. Cv. Kapoori was determined by DPPH free radical scavenging activity, reducing power by FeCl3, nitric oxide free radical scavenging activity, super oxide scavenging activity.Also used specific standard for each test. Results: Solvent extracts of Mokkathotapapada leaves of Piper betel L. Cv. Kapoori were compared with the light of respective standard drugs and the extracts were found to be equipotent with the standards in all the tested methods. Conclusion: Presence of phenols and phenolics (Chavicol, Chavibetol, Chavibetol acetate and Eugenol) in the Mokkathotapapada leaves of Piper betel L. may be credulous to be responsible for its antioxidant activity.
Renal disorders have always remained a major area of concern for physicians since a long time. It is the 9th leading cause of death in United States. Incidence of kidney diseases leading to kidney failure is increasing day by day. A large number of chemicals in common use are potential renal toxins. The use of herbal drugs for the prevention and treatment of various diseases is constantly developing throughout the world. Many studies have been carried out that strongly support the contribution of antioxidants to the prevention of cardiovascular diseases, cancer, osteoporosis, neurodegenerative diseases, and diabetes mellitus, and suggest a role in the prevention of peptic ulcer. Polyphenols display a number of pharmacological properties in the renal area, acting as diuretic, anti inflammatory, antispasmodic, and antioxidant agents. The antioxidant properties of phenolic compounds have been widely studied, but it has become clear that their mechanisms of action go beyond the modulation of oxidative stress. Various polyphenolic compounds have been reported for their nephroprotective activity with a good level of renal protection. Therefore, considering the important role of polyphenolic compounds in the prevention or reduction of renal disorders induced by various nephrotoxic chemical agents, in this review, we have summarized the literature on some potent nephroprotective plants, such as, Achyrocline satureioides, Zingiber officinalis, Teminalia chebula etc having antioxidant properties.
This paper evaluates critically the science base that underpins the argument that oxidative damage is a significant causative factor in the development of human diseases and that antioxidants are capable of preventing or ameliorating these disease processes. Lignans, as polyphenol compounds, have been reported to serve as good and healthy nutrients. By reason of possessing many biological activities, it takes a major part in curing degenerative diseases. Secoisolariciresinol diglucoside (SDG), the chief lignan found in flaxseed, is believed to reduce serum cholesterol levels, delay the onset of type II diabetes and decrease formation of breast, prostate and colon cancers. These health benefits of SDG may be partially attributed to its antioxidant properties. Retrospectively of the above information, in the present study, SDG was synthesized for the first time and screened for its antioxidant properties. The synthetic route of SDG used the coupling of two molecules of 3,4-dimethoxy toluene with dibromobutanediol as the key step to yield the SDG skeleton. The synthesized SDG was found to have potential antioxidant activity. It scavenges the DPPH radical at EC50 of 76.67 μg/ml which is comparable to that of antioxidant references namely Ascorbic acid, Std-SDG and α-tocopherol (60.64, 66.57, 83.24 μg/ml) respectively. Further, its dose dependent reducing power was observed. The synthesized SDG showed 1.25 times higher activity compared to α-tocopherol and lesser activity (0.70, 0.75 times) compared to Ascorbic acid and Std-SDG respectively. Additionally, it was found to protect DNA from hydroxyl radicals generated by an oxidant agent as Fenton's reagent at a concentration of 0.5 mg/ml.
IntroductionPhytochemicals are extensively found at different levels in many medicinal plants. To investigate the phenolic compound content and in vitro antioxidant activity of phenolic extract from Prasium majus L (Lamiaceae).Methods The present investigation comprises, estimation of total polyphenol, flavonoid, tannin, in vitro antioxidant assays such as total antioxidant capacity, DPPH, ABTS, β-carotene and ferric reducing power.ResultsP. majus exhibited 64.25 mg GAE g−1 extract of polyphenol phenol content, and better scavenging activity of DPPH (IC50 = 7.95 μg mL−1), ABTS+ (IC50 = 373.78 μg mL−1), and β-carotene (IC50 = 122.56 μg mL−1).Conclusion Our results clearly demonstrated that phenolic extract P. majus has antioxidant capacity. Therefore is a valuable source of natural antioxidants.
Introduction: The current study was carried out to investigate the phytochemical constituents and in vitro antioxidant potential of the aerial parts of Justicia beddomei (Clarke) Bennett belonging to the family Acanthaceae. The aerial part of J. bebbomei is traditionally used as a hepatoprotective, antitussive and antispasmodic. Methods: The dried powdered aerial parts of J. beddomei were extracted using petroleum ether, chloroform, ethyl acetate and methanol using a soxhlet extractor and preliminary phytochemical screening was performed using standard protocols. All the extracts were evaluated for their potential antioxidant activities using tests such as DPPH, hydroxyl radical, superoxide anion radical scavenging abilities, β-carotene-linoleic acid model, reducing power ability and total phenolic and flavonoid contents. Results: Preliminary screening revealed the presence of bioactive components especially phenolics and flavonoids in all the extracts. The phenolic and flavonoid content was found to be highest in methanolic extract and lowest in petroleum ether extract. All extracts showed strong antioxidant activity when compared with the standard. Conclusion: The results of the present study indicate that the aerial part extracts of J. beddomei is a good source of antioxidant constituents.
List of endophytes from different parts of Crotalaria pallida on PDA media
Phytochemical analysis of ethanol extract of different plant parts
EC 50 values of endyphytic ethanol extract
Introduction: The species Crotalaria pallida, which belongs to the Fabaceae family (Sub-family Faboideae), the members of which are herbs, shrubs and trees found in both temperate and tropical areas. All parts of the plants were incubated to know and isolate the endophytes. Antioxidants play an important role in protecting cellular damage by reactive oxygen species. Phenolic compounds from plants or endophytes have been reported to possess strong antioxidant properties. Results: Four different endophytic fungi isolated from Crotalaria pallida were tested for various phytochemicals. Aspergillus niger and Fusarium oxysporum yielded the tannin, flavonoids, tepenoids, phenol and saponins from ethanol extract. All four different endophytic extracts were used to evaluated in vitro antioxidant activity by ABTS, DPPH and FRAP method. Antioxidant compounds like total phenol and flavonoid were also determined. The ethanol extracts of A. niger and F. oxysporum showed potent antioxidant activity against ABTS, FRAP and DPPH radicals with EC50. The total amount of phenol and flavonoid quantified were of 19.20, 19.23 gallic acid equivalent per gram of two endophytic fungi, A. niger and F. oxysporum and 7.25 and 6.41 μg/mg of quercetin equivalent respectively. Conclusions: The antioxidant potential may be directly linked to the phenolic compounds present in the endophytes, A. niger and F. oxysporum of Crotalaria pallida. The outcome of the present investigation clearly indicates that A. niger and F. oxysporum showed potential phytochemicals and they can used as antioxidants.
Background The study focused to long-term effect of aspartame on membrane bound enzymes, oxidative stress markers and histopathology in brain regions of Wistar albino rats. Hence it is essential to observe whether the chronic aspartame administration (75 mg/kg b. wt) could release methanol and induce oxidative stress in the rat brain. Many reports are available on the use of aspartame as it releases methanol during metabolism. Methods To mimic the human methanol metabolism the methotrexate treated rats were included to study the aspartame effects and the gamma glutamyl transpeptidase, NO, H2O2 and the membrane bound enzymes were observed in brain discrete regions. Results There was a significant increase in all the parameters except with a significant decrease in membrane bound ATPases and creatine kinase. Luxol fast blue (LFB) staining were performed on brain cerebellum region which showed histopathological changes in aspartame treated MTX animals which showed a marked decrease in the density of white matter, intensity of staining and no. of stained neuron cells when compared to control and MTX control animals. Conclusion Moreover, the increases in some of these enzymes were due to methanol per se and its metabolite may be responsible for the generation of oxidative stress and histopathology in brain regions.
Relationship between Total Phenol Content and Total antioxidant capacity of different garlic extracts. GAE ¼ Gallic Acid Equivalent, TE ¼ Trolox Equivalent.
IntroductionGarlic is well known for its health protective abilities. Many studies have also proven garlic as an oxidative stress fighter with unique antioxidant potential. These studies have extracted raw garlic in conventional manner i.e. using organic solvents. Such antioxidant capacities cannot be well implicated for health purposes.Methods This study deals with measurement of antioxidant capacity of raw as well as cooked garlic extracted by chemical as well as physiological method (in vitro gastrointestinal digestion). The Total antioxidant capacity was measured by methods like DPPH Radical Scavenging Ability, ABTS Radical Scavenging Ability, Ferric Reducing Antioxidant Power and Reducing Power Assay. Total Phenol was also evaluated.ResultsResults show a wide difference between the antioxidant capacity of conventional and physiological extracts. The in vitro digested extracts of raw garlic show highest antioxidant capacity of all raw and cooked garlic extracts. Loss of phenolic compounds and antioxidant potential on cooking can also be clearly observed in both chemical and physiological extracts.Conclusion It can be thus concluded that the physiological method of antioxidant extraction is more applicable and reliable than the conventional chemical extraction methods that do not resemble the biological behavior of antioxidants.
Free radicals induce numerous diseases by lipid peroxidation, protein peroxidation, and DNA damage. It has been reported that numerous plant extracts have antioxidant activities to scavenge free radicals. In the present study, we examines the in vitro radical scavenging and antioxidant capacity of Crude extract of Carthamus tinctorius by using different in vitro analytical methodologies such as total antioxidant activity determination by ferric thiocyanate, hydrogen peroxide scavenging, 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH) scavenging, 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activity and superoxide anion radical scavenging by riboflavin–methionine-illuminate system. Also, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and α-tocopherol were used as the reference antioxidant radical scavenger compounds.Extract inhibited 94.50% lipid peroxidation of linoleic acid emulsion at 20 μg/mL concentration. On the other hand, the above mentioned standard antioxidants indicated an inhibition of 93.75%, 96.66% and 83.33% on peroxidation of linoleic acid emulsion at 60 μg/mL concentration, respectively. In addition, hydrogen peroxide scavenging, DPPH scavenging, ABTS+ radical scavenging and superoxide anion radical scavenging. Also, those various antioxidant activities were compared to BHA, BHT and α-tocopherol as references antioxidant compounds. The present study shows that Extract is the effective natural antioxidant component.
A phytochemical analysis of the acetone extract of the aerial parts of the plants Salvia splendens and Salvia lanigra yielded a long chain alkyl p-coumaric acid ester; eicosanyl-cis-p-coumarate (1), which has not previously been isolated from Salvia genus, together with two triterpenoids; oleanolic acid (2) and echinocystic acid (3) and two flavonoids; 7-methoxyapigenin (4) and luteolin-7-O-glucoside (5). The structures of these compounds were assigned by spectroscopic analysis and comparison with literature data of known compounds. The antioxidant potential of the p-coumaric acid ester (1) was evaluated, in vitro, by using DPPH for free radical scavenging activity. A moderate free radical scavenging ability was observed.
Areca nut (Areca catechu L.) or Pinang is one of the most widely used psychoactive substance with several hundred million users worldwide, predominantly in Southern Asia. This study evaluates the antioxidant activity and the total phenolic compound of methanolic and aqueous extract of seeds (ripe and unripe seeds), root and adventitious root. The antioxidant activity was determined using the 1, 1 -diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay and total phenolic using the Folin-Ciocalteu method. The results from this study showed that the antioxidant activities of the water and methanolic extracts of the seeds as determined by the 1, 1 -Diphenyl-2-picrylhydrazyl (DPPH) presented higher percentage inhibition than the root and adventitious root. The methanol extracts gave higher antioxidant activities than the aqueous extracts and comparable to that BHT and Vitamin C. The results also showed that unripe seeds methanolic extract possessed higher content of phenolic (186.2 ± 0.04 mg GAE/g) and total flavonoid (18.13 ± 0.007 mg/g) than other parts of A. catechu. We also evaluated the in vitro effect of various A.catechu methanolic and aqueous extract on the activity of Phase II metabolizing enzyme, glutathione-S-transferase (GST) in rat liver. Unripe seeds methanolic showed the effective GST specific activity inhibition with an IC50 of 115.05 μg/mL with maximum inhibition > 70%. These results suggest that areca nut extracts have the potential to prevent oxidative damage in normal cells due to their antioxidant characteristics.
Background Newcastle disease (ND) is highly contagious disease caused by Newcastle disease virus (NDV) causes mass mortalities in poultry and triggers poultry economy. Methods NDV induced oxidative stress representing marker enzymes and lipid peroxidation levels were evaluated in tissues of control and virus infected animals by using standard protocols and methods. Results Lipid peroxidation levels were significantly increased in cerebrum and cerebellum of NDV infected chickens on 10 (dpi) as compared to controls. The activities of superoxide dismutase, catalase and glutathione metabolism representing enzymes were significantly decreased in cerebrum and cerebellum. Extensive histopathological malformations were observed in cerebrum and cerebellum. Conclusion The results of the present study, thus demonstrated that region specific alterations in the antioxidant defense mechanism due to NDV infection suggesting its critical role in cellular injury prominently in cerebrum and cerebellum and moderately in optic lobes of brain. The findings could help to advance the therapeutic armamentarium for control of Newcastle disease in poultry.
Study population 
Positive correlations among parameters in the subjects 
Introduction: Malaria and typhoid are two diseases of public health importance that are associated with fever in the tropics. Objective: To assess antioxidant responses during malaria, typhoid and typhoid + malaria infections, the activities of glutathione transferase (GST), catalase (CAT) and the concentrations of glutathione (GST). Materials and Methods: Spectrophometric techniques were used to assay for the biomarkers of oxidative stress in plasma and erythrocytes of patients in Abeokuta, Nigeria (n = 115). Result: The presence of either or both parasitic infections resulted in significant alterations in the antioxidant response of the subjects (p < 0.05). Depletion of erythrocyte GSH, increase in plasma GSH and increased expression of CAT in both plasma and erythrocyte, characterized the antioxidant response of the subjects. While the highest erythrocyte CAT activity was observed in typhoid-infected males, the highest plasma CAT activity was observed in females infected with typhoid (p < 0.05). GST activity in malaria infection was not significantly different from control (p > 0.05), whereas the activity of the enzyme reduced in typhoid infection and increased when typhoid and malaria infections were present (p < 0.05). Conclusion: Our findings indicate that sex differences might play a significant role in the antioxidant response of subjects in typhoid infection.
Top-cited authors
Satyajit Sarker
  • Liverpool John Moores University
Ipek Süntar
  • Gazi University
Dr Lutfun Nahar
  • Liverpool John Moores University & Palacký University
Eric Wei Chiang Chan
  • UCSI University
Dharmendra Khatri
  • Institute of Chemical Technology, Mumbai