Chemical composition, antioxidant, anti-inflammatory and antitumor activities of <em>Eucalyptus globulus</em> Labill.

734-742Eucalyptus globulus L. is used in folk medicine throughout the world and its essential oils are widely used in modern pharmaceutical, food, and cosmetic industries. In this study, E. globulus leaves were extracted using three solvents (methanol, chloroform and hexane). The polyphenolics were quantified with HPLC and the volatiles analyzed by GC/MS with antioxidant, anti-inflammatory and antitumor activities. Results have shown a hierarchy of antioxidant and anti-inflammatory activity for three extracts as hexane > chloroform > methanol extracts with DPPH, FRAP, and oxygen radical absorbance capacity, respectively. A similar order of results was observed for antitumor activity by potato disc and colorimetric assays. The GC/MS analysis led to the identification of 1, 8-cineole (eucalyptol) as a major constituent of methanol (48.2%), chloroform (35.5%), and hexane (5.8%) extracts. Different phenolic acids (gallic acid, ellagic acid, syringic acid, and vanillic acid) and flavonoids (quercetin, rutin, and catechin) were highly abundant in methanol extract. The methanol extract of E. globulus exhibited the maximum antioxidant, anti-inflammatory, and antitumor activities. These results demonstrate E. globulus leaf extracts may be used as a potential source of bioactive compounds with remarkable antioxidant, anti-inflammatory, and antitumor activities

Purification and characterization of buffalo liver L-arginase and its kinetic properties with dihydropyrimidine and metal ions

414-419<span style="font-size:11.0pt;font-family: " times="" new="" roman";mso-fareast-font-family:"malgun="" gothic";mso-bidi-font-family:="" mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:="" hi"="" lang="EN-GB">Arginase (L-arginine amidinohydrolase, EC.3.5.3.1) from animal tissues such as, liver and kidney has been partially characterized by many researchers. In this study, we purified arginase to homogeneity from buffalo liver with about ~2857 purification fold<span style="font-size:11.0pt;font-family: " times="" new="" roman";mso-fareast-font-family:advtimes;mso-bidi-font-family:mangal;="" mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:hi"="" lang="EN-GB"> and a 20% recovery by chromatographic and spectroscopic analysis were obtained. The molecular mass determined by gel filtration and SDS-PAGE was found to be 118 kDa and 47 kDa, respectively. The optimal pH and temperature of the arginase was 9.5 and 40<span style="font-size:11.0pt;font-family:Symbol; mso-ascii-font-family:" times="" new="" roman";mso-fareast-font-family:"malgun="" gothic";="" mso-hansi-font-family:"times="" roman";mso-bidi-font-family:mangal;mso-ansi-language:="" en-gb;mso-fareast-language:en-us;mso-bidi-language:hi;mso-char-type:symbol;="" mso-symbol-font-family:symbol"="" lang="EN-GB">°<span style="font-size:11.0pt;font-family: " times="" new="" roman";mso-fareast-font-family:"malgun="" gothic";mso-bidi-font-family:="" mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:="" hi"="" lang="EN-GB">C, respectively. Kinetic parameters (Km and Vmax) showed activation of arginase in the reaction medium with decrease in Km (7.14, 5.26, 4.0 and control 3.22 mM) and Vmax (0.05, 0.035, 0.027 and control 0.021 mg/mL/min), while co-factor activity of arginase was optimized using metal ions like Mn2+ and Mg2+ at 2 mM, which revealed an increase in Vmax values (0.011, 0.013, 0.015 and control 0.010 mg/mL/min) and a decrease in Km values (2.22, 2.12, 1.88 and control 1.66 mM). The kinetic data suggested that the arginase activity is enhanced in the presence of dihydropyrimidine derivative and metal ions, indicating essential mode of activation.</span

<span style="font-size:11.0pt;font-family: "Times New Roman","serif";mso-fareast-font-family:"Malgun Gothic";mso-bidi-font-family: Mangal;background:white;mso-ansi-language:EN-GB;mso-fareast-language:EN-US; mso-bidi-language:HI" lang="EN-GB">Determination of <span style="font-size:11.0pt;font-family:"Times New Roman","serif";mso-fareast-font-family: Calibri;mso-bidi-font-family:Mangal;mso-ansi-language:EN-GB;mso-fareast-language: EN-US;mso-bidi-language:HI" lang="EN-GB">polyphenols<span style="font-size:11.0pt;font-family:"Times New Roman","serif";mso-fareast-font-family: "Malgun Gothic";mso-bidi-font-family:Mangal;background:white;mso-ansi-language: EN-GB;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN-GB"> and<span style="mso-bidi-font-weight:bold"> <span style="font-size:11.0pt;font-family:"Times New Roman","serif"; mso-fareast-font-family:Calibri;mso-bidi-font-family:Mangal;mso-ansi-language: EN-GB;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN-GB">antioxidant activity of <i style="mso-bidi-font-style:normal">Vitis labrusca</i> cv. <span style="font-size:11.0pt;font-family:"Times New Roman","serif"; mso-fareast-font-family:"Malgun Gothic";mso-bidi-font-family:Mangal;mso-ansi-language: EN-GB;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN-GB">baile berries</span></span></span></span></span></span>

671-675<span style="font-size:11.0pt;font-family: " times="" new="" roman","serif";mso-fareast-font-family:calibri;mso-bidi-font-family:="" mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:="" hi"="" lang="EN-GB">Grape juice and grape skin extracts are important commercial source of polyphenolic compounds which exert different functional <span style="font-size:11.0pt; font-family:" times="" new="" roman","serif";mso-fareast-font-family:calibri;="" mso-bidi-font-family:mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;="" mso-bidi-language:hi"="" lang="EN-GB">properties such as color potential, antimicrobial, antioxidant activity, and health benefits. In this paper we describe a sensitive and specific assay for determination of bioactive polyphenolic compounds in Campbell Early (<i style="mso-bidi-font-style: normal">Vitis labrusca cv. baile). Five polyphenolic components were separated on an Agilent Zorbax Extend C18 Column (250 mm × 4.6 mm × 5 μm) and detected by a diode array detector. The mobile phase was composed of (a) aqueous phosphoric acid (0.2%, v/v); and (b) acetonitrile using a gradient elution. Analytes were performed at 25°C with a flow rate of 0.8 ml/min and UV detection at 280<span style="mso-bidi-font-family: Mangal;mso-bidi-font-weight:bold;font-style:normal">, 360, and 520 nm. All calibration curves showed good linear regression (r2 ≥ 0.9999) within tested ranges. Overall intra- and inter-day variations were less than 1.90%, and the average recoveries were 95.5-105% for analytes. The antioxidant activity determined by DPPH radical assay, ranged from 86-105 for extracts, and 165-252 for studied standards <span style="font-size:11.0pt;font-family: " times="" new="" roman","serif";mso-fareast-font-family:"malgun="" gothic";mso-bidi-font-family:="" mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:="" hi;mso-bidi-font-weight:bold"="" lang="EN-GB">(µM trolox/100 g dry wt.).<span style="font-size:11.0pt; font-family:" times="" new="" roman","serif";mso-fareast-font-family:"malgun="" gothic";="" mso-bidi-font-family:mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;="" mso-bidi-language:hi"="" lang="EN-GB"> <span style="font-size:11.0pt; font-family:" times="" new="" roman","serif";mso-fareast-font-family:"malgun="" gothic";="" mso-bidi-font-family:mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;="" mso-bidi-language:hi"="" lang="EN-GB">The proposed method would be sensitive enough and reliable for quality control in functional food and modernization of Campbell Early (<i style="mso-bidi-font-style: normal">Vitis labrusca cv. baile) as potent antioxidant agent.</span

Characterization of organic acids, sugars, antioxidant and anti-inflammatory activities of <em>Citrus reticulate </em> Blanco cv. Nagpur

604-610The citrus fruits, commonly found in many regions of the world with a considerable number of species, have numerous beneficial effects, particularly for human health. The citrus organic acids and sugars play pivotal role in flavour and consumer acceptance of citrus fruit beverages. Here, some nutritive parameters related to the fruit Citrus reticulate Blanco cv. Nagpur (locally called santara) quality including sugars, organic acids, soluble solids, total acidity, and the content of bioactive compounds, such as phenolics and flavonoids were determined. The antioxidant and anti-inflammatory activities have been checked using in vitro assays. Four organic acids (citric, ascorbic, malic, and oxalic acids) and three sugars including glucose, sucrose, and fructose were determined. Citric acid was found as major organic acid whereas sucrose as major sugar in extracted orange juice and wine from this citrus fruit wine and juice. The HPLC quantification and analysis showed total 15 phenolic and flavonoid compounds including hydroxybenzoic acids, hydroxycinnamic acids, and flavanones in studied citrus juice and wine. The major phenolics and flavonoids found were ferulic acid, hesperidin, narirutin, and naringin in santara juice and wine. The santara juice showed significant antioxidant activity towards DPPH, FRAP, and H2O2 assays (IC50±SEM [μg/mL]: 85.6±1.6, 60.2±1.6, and 35.5±1.1, respectively) and also exhibited greater inhibition towards lipoxidase (92.2%), β-glucuronidase (65.3%), hyaluronidase (48.1%), and diene-conjugates (40.6%) for anti-inflammation compared to orange wine. The high fruit quality of the studied citrus species indicates the importance of this fruit in human nutrition with prominent biological effects

Antioxidant, Anti-inflammatory and Xanthine oxidase inhibitory Activity of Tephrosia purpurea L

T. purpurea root extract was evaluated for antioxidant, anti-inflammatory, and xanthine oxidase (XO) inhibitory activities. Antioxidant activity was measured using ABTS and FRAP methods, anti-inflammatory activity was measured by Diene-conjugate and β-glucuronidase assay. In vitro XO inhibitory activity was measured by using cow milk xanthine oxidase enzyme. The average antioxidant activity of T. purpurea root extracts (1-2 µg/mL) in the reacting system revealed significant activity viz; 42.2 (ABTS) and 36.5 (FRAP) percent. The anti-inflammatory activities reveled, 45.40 and 70.50 percent inhibition. The result for XO inhibitory activity by plant extracts reveled, 95.5 % inhibition to that, off control (allopurinol) 92 % inhibition. The kinetic parameters of XO inhibition, revealed noncompetitive mode of inhibition, where, Km and Vmax of T. purpurea root extracts (25 to 100 µg/mL)) were,  0.25 mM/mL and 0.040, 0.036, 0.032 and 0.030 (µg/min) while for positive control Km and Vmax is 0.30 mM/mL and 0.045 (µg/min) respectively. Results suggest that, T. purpurea root can be exploited against diseases associated, with free radical formation and xanthine oxidase activity; further by isolation and structural elucidation of active phytochemicals from T. purpurea root

Determination of Anthocyanin Content and Antioxidant Capacity of Different Grape Varieties

The aim of this study was to determine anthocyanin content and antioxidant activity of 20 different grape varieties, using HPLC and DPPH, FRAP and ABTS assays, respectively. The identified anthocyanins were malvidin-3-glucoside, delphinidin-3-glucoside, petunidin-3-glucoside, cyanidin-3 glucoside, and peonidin-3-glucoside on the basis of their retention times. Comparatively the total anthocyanin content varied from 181.2 mg/100 g FW (‘Vidal Black’) to 716.4 mg/ 100 g FW (‘Catawba’). High anthocyanin content was found in ‘Catawba’ (716.4 mg/100 g), ‘Ruby Seedless’ (634.5 mg/100 g) and ‘Campbell Early’ (611.1 mg/100 g) extracts. The antioxidant activities of grape extracts varied from 32.8% (‘Campbell Early’) to 87.6% (‘Hongiseul’) by DPPH, 79.1% (‘Campbell Early’) to 197% (‘Hongiseul’) in case of FRAP and 11.1% (‘Chasselas Rouge’) to 74.5% (‘Flouxa’) by ABTS antioxidant assay. The results suggested that the anthocyanin content in studied grape varieties showed statistically significant correlation with free radical scavenging activity. Thus depending on results we say that these grape varieties may serve as a potential source of nutraceuticals and functional food development

Screening of ferulic acid related compounds as inhibitors of xanthine oxidase and cyclooxygenase-2 with anti-inflammatory activity

The ferulic and gallic acid related compounds from natural origin were studied against xanthine oxidase and cyclooxygenase-2 along with their anti-inflammatory activity. The compounds gallic acid, ferulic acid, caffeic acid and p-coumaric acid revealed promising anti-inflammatory activity (30–40% TNF-α and 60–75% IL-6 inhibitory activity at 10 μM). Bioavailability of compounds were checked by in vitro cytotoxicity using CCK-8 cell lines and confirmed to be nontoxic, but found toxic at higher concentration (50 μM). Gallic, ferulic, caffeic acid was demonstrated potential dual inhibition toward xanthine oxidase and cyclooxygenase-2 as calculated by IC50: 68, 70.2, and 65 μg/ml (xanthine oxidase) and 68.5, 65.2, and 62.5 μg/ml (cyclooxygenase-2), respectively. The structure activity relationship and in silico drug relevant properties (HBD, HBA, PSA, c Log P, ionization potential, molecular weight, EHOMO and ELUMO) further confirmed that the compounds were potential candidates for future drug discovery study, which was expected for further rational drug design against xanthine oxidase and cyclooxygenase. Keywords: Ferulic acid, Gallic acid, Xanthine oxidase, Cyclooxygenase, SAR, Gou

Quercetin-3-Glucoside Extracted from Apple Pomace Induces Cell Cycle Arrest and Apoptosis by Increasing Intracellular ROS Levels

Cervical cancer is a life-threatening disease and the fourth most common cancer among women worldwide. Apple pomace is a multifunctional phenolic compound possessing effective biological activity against cervical cancer cells. This study aimed to investigate the anticancer effects of quercetin-3-glucoside (Q3G) extracted from apple pomace in HeLa cell lines and analyze its molecular mechanisms. High-performance liquid chromatography revealed that Q3G, coumaric acid, phloridzin, quercetin, and phloretin are the major polyphenolic compounds constituting apple pomace. Among them, Q3G possessed the greatest antioxidant and anti-inflammatory effects in vitro and exhibited significant cytotoxic effects in HeLa cells in a dose-and time-dependent manner. Flow cytometric analysis indicated that Q3G induced cell cycle arrest at the S phase in a time-dependent manner by altering cyclin-dependent kinase 2. Moreover, it induced apoptosis via chromosomal DNA degradation and increased reactive oxygen species generation. Furthermore, Q3G treatment altered the apoptosis-associated protein expression in the cells by activating caspase-9/-3, downregulating anti-apoptosis protein B-cell lymphoma (Bcl)-2 expressions and up regulating the pro-apoptotic Bcl-2-associated X protein. BH3-interacting domain death agonist cleavage occurred prior to the degradation of an anti-apoptotic Mu-2-related death-inducing gene involved in cell death signaling. Consequently, apple pomace Q3G holds promise as an anti-inflammatory and anticancer agent for treating cervical cancer