scholarly journals Bioprocessing of Squid Pens Waste into Chitosanase by Paenibacillus sp. TKU047 and Its Application in Low-Molecular Weight Chitosan Oligosaccharides Production

Polymers ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 1163 ◽  
Author(s):  
Chien Thang Doan ◽  
Thi Ngoc Tran ◽  
Van Bon Nguyen ◽  
Trung Dung Tran ◽  
Anh Dzung Nguyen ◽  
...  
Keyword(s):  
Biological Activities ◽  
Radical Scavenging Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Radical Scavenging ◽  
Maximum Activity ◽  
Degree Of Polymerization ◽  
Chitosan Oligosaccharide ◽  
Scavenging Activity ◽  
Paenibacillus Sp

Chitosan oligosaccharide (COS) has become of great interest in recent years because of its worthy biological activities. This study aims to produce COS using the enzymatic method, and investigates Paenibacillus sp. TKU047, a chitinolytic-producing strain, in terms of its chitosanase productivity on several chitinous material-containing mediums from fishery process wastes. The highest amount of chitosanase was produced on the medium using 2% (w/v) squid pens powder (0.60 U/mL) as the single carbon and nitrogen (C/N) source. The molecular mass of TKU047 chitosanase, which could be the smallest one among chitinases/chitosanases from the Paenibacillus genus, was approximately 23 kDa according to the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method. TKU047 chitosanase possessed the highest activity at 60 °C, pH 7, and toward chitosan solution with a higher degree of deacetylation (DDA) value. Additionally, the hydrolysis products of 98% DDA chitosan catalyzed by TKU047 chitosanase showed the degree of polymerization (DP) ranging from 2 to 9, suggesting that it was an endo-type activity chitosanase. The free radical scavenging activity of the obtained chitosan oligosaccharide (COS) was determined. The result showed that COS produced with Paenibacillus sp. TKU047 chitosanase expressed a higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity than that from the commercial COSs with maximum activity and IC50 values of 81.20% and 1.02 mg/mL; 18.63% and 15.37 mg/mL; and 15.96% and 15.16 mg/mL, respectively. As such, Paenibacillus sp. TKU047 may have potential use in converting squid pens waste to produce chitosanase as an enzyme for bio-activity COS preparation.

scholarly journals Production of a Thermostable Chitosanase from Shrimp Heads via Paenibacillus mucilaginosus TKU032 Conversion and its Application in the Preparation of Bioactive Chitosan Oligosaccharides

Marine Drugs ◽  
2019 ◽  
Vol 17 (4) ◽  
pp. 217 ◽  
Author(s):  
Chien Thang Doan ◽  
Thi Ngoc Tran ◽  
Van Bon Nguyen ◽  
Anh Dzung Nguyen ◽  
San-Lang Wang
Keyword(s):  
Antioxidant Activities ◽  
Radical Scavenging Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Radical Scavenging ◽  
Chitosan Oligosaccharide ◽  
Chitosan Oligosaccharides ◽  
Potential Applications ◽  
Shrimp Heads ◽  
Fraction I

Chitosanase has attracted great attention due to its potential applications in medicine, agriculture, and nutraceuticals. In this study, P. mucilaginosus TKU032, a bacterial strain isolated from Taiwanese soil, exhibited the highest chitosanase activity (0.53 U/mL) on medium containing shrimp heads as the sole carbon and nitrogen (C/N) source. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, a chitosanase isolated from P. mucilaginosus TKU032 cultured on shrimp head medium was determined at approximately 59 kDa. The characterized chitosanase showed interesting properties with optimal temperature and thermal stability up to 70 °C. Three chitosan oligosaccharide (COS) fractions were isolated from hydrolyzed colloidal chitosan that was catalyzed by TKU032 chitosanase. Of these, fraction I showed the highest α-glucosidase inhibitor (aGI) activity (65.86% at 20 mg/mL); its inhibitory mechanism followed the mixed noncompetitive inhibition model. Fractions II and III exhibited strong 2,2-diphenyl1-picrylhydrazyl (DPPH) radical scavenging activity (79.00% at 12 mg/mL and 73.29% at 16 mg/mL, respectively). In summary, the COS fractions obtained by hydrolyzing colloidal chitosan with TKU032 chitosanase may have potential use in medical or nutraceutical fields due to their aGI and antioxidant activities.

scholarly journals Synthesis, Antiproliferative Activity and Radical Scavenging Ability of 5-O-Acyl Derivatives of Quercetin

Molecules ◽  
2021 ◽  
Vol 26 (6) ◽  
pp. 1608
Author(s):  
Stephen Lo ◽  
Euphemia Leung ◽  
Bruno Fedrizzi ◽  
David Barker
Keyword(s):  
Antiproliferative Activity ◽  
Biological Activities ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Breast Cancer Cell Lines ◽  
Scavenging Activity ◽  
Plant Materials ◽  
Wide Range ◽  
Derivatives Of ◽  
Acyl Derivatives

Quercetin is a flavonoid that is found in many plant materials, including commonly eaten fruits and vegetables. The compound is well known for its wide range of biological activities. In this study, 5-O-acyl derivatives of quercetin were synthesised and assessed for their antiproliferative activity against the HCT116 colon cancer and MDA-MB-231 breast cancer cell lines; and their radical scavenging activity against the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical species. Four derivatives were found to have improved the antiproliferative activity compared to quercetin whilst retaining radical scavenging activity.

Synthesis and Biological Activities of Hydroxytyrosol Ester Derivatives

2020 ◽  
Vol 42 (1) ◽  
pp. 109-109
Author(s):  
Hao Zang Hao Zang ◽  
Qian Xu Qian Xu ◽  
Luyun Zhang Luyun Zhang ◽  
Guangqing Xia Guangqing Xia ◽  
Jiaming Sun and Junyi Zhu Jiaming Sun and Junyi Zhu
Keyword(s):  
Ascorbic Acid ◽  
Antioxidant Activities ◽  
Biological Activities ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Acetylcholinesterase Inhibition ◽  
Hydroxyl Group ◽  
Scavenging Activity ◽  
Antioxidant Power ◽  
Better Than

A series of hydroxytyrosol (HT) derivatives were synthesized by modification of alcohol hydroxyl group of HT, twenty-five target compounds were obtained and characterized by NMR and HRMS. The antioxidant activities of those compounds were evaluated in three different assays. Except 3e and 3y, all other compounds demonstrated significant 2,2and#39;-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) free radical cation scavenging activity ranging from IC50 3.4 to 24.4 μM, which were more potent than L-ascorbic acid (IC50=24.8 μM). Compounds 3b-3d, 3f-3k, 3m-3x were better than Trolox (18.3 M). Moreover, the ferric reducing antioxidant power (FRAP) of all compounds were discovered to be more potent than L-ascorbic acid (40.7 mmol/g), except 3e, all other compounds (141.5-202.1 mmol/g) were better than Trolox (94.7 mmol/g). Compounds 3a-3d, 3f-3j, 3l-3m, 3o, 3q, 3t, 3v-3y exhibited more potent hydroxyl radical scavenging activity (IC50=245.1-475.1 M) than L-ascorbic acid (554.4 M) and Trolox (500.4 M). Compounds 3q, 3t and 3y exhibited more potent -Glucosidase inhibition activity (39.1-52.4 M) than Acarbose (60.9 M). Compounds 3a, 3d, 3f-3m, 3s-3t, 3v-3y showed some acetylcholinesterase inhibition activities, compounds 3a, 3d, 3f-3j, 3l-3m, 3o-3p, 3s-3t, 3w showed some butyrylcholinesterase inhibition activities.

scholarly journals GC-MS Assisted Phytoactive Chemical Compounds Identification and Profiling with Mineral Constituents from Biologically Active Extracts of Aerva javanica (Burm. f) Juss. ex Schult.

2018 ◽  
Vol 46 (2) ◽  
pp. 517-524
Author(s):  
Kandhan KARTHISHWARAN ◽  
Subban KAMALRAJ ◽  
Chelliah JAYABASKARAN ◽  
Shyam S. KURUP ◽  
Sabitha SAKKIR ◽  
...  
Keyword(s):  
Biological Activities ◽  
Radical Scavenging Activity ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Biologically Active ◽  
Scavenging Activity ◽  
Medicinal Uses ◽  
Chemical Structures ◽  
Aerial Parts ◽  
Mineral Constituents

Aerva javanica (Burm. f) Juss. ex Schult. (Family: Amaranthaceae) family is one of the traditional medicinal plant growing in the United Arab Emirates. Apart from studies related to some medicinal properties, phytochemical, GC MS compound characterization and biological activities still to be investigated. An experiment was conducted to determine the possible bioactive components with their chemical structures and elucidation of phytochemicals from the aerial parts of the plant. The macro and micro-mineral constituents and antioxidant activities were also evaluated. Aerial parts of A. javanica were extracted sequentially with hexane, chloroform, ethyl acetate, acetone, methanol by cold percolation method. Free radical scavenging and antioxidant properties of methanolic extract were evaluated by using in vitro antioxidant assays such as hydroxyl radical scavenging activity, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, superoxide radical scavenging activity and ABTS radical scavenging activity. Primary phytochemical and micro-macro element was tested using standard protocol. The chemical characterization was done with the help of Gas Chromatography-Mass Spectrometry (GC–MS), and the mass spectra of the total compounds in the extract were matched with the National Institute of Standard and Technology (NIST) library. Mineral constituents were identified and estimated by ICP-OES. Ninety-nine metabolites were obtained by GC-MS anslysis; indole was found to be major components followed by 2-Chlorallyl diethyldithiocarbamate (CDEC), Carbaril, Bis(2-ethylhexyl) phthalate, Quinoline, 4H-Cyclopenta[def]phenanthrene,2-[Bis(2-chloroethylamino)]-tetrahydro-2H-1,3,2-oxazaphosphorine-2-oxide, Phenobarbital, 1H-Indole, 2-methyl-, 2,3,7,8-Tetrachlorodibenzo-p-dioxin Disulfide, diphenyl. The presence of various bioactive compounds in the extract validates the traditional medicinal uses of this plant.

scholarly journals Phytochemical, in-vitro biological and chemo-preventive profiling of Arisaema jacquemontii Blume tuber extracts

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Saira Tabassum ◽  
Muhammad Zia ◽  
Esperanza J. Carcahe de Blanco ◽  
Riffat Batool ◽  
Roohi Aslam ◽  
...  
Keyword(s):  
Cell Lines ◽  
Biological Activities ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Reducing Power ◽  
Scavenging Activity ◽  
Zone Of Inhibition ◽  
Meoh Extract ◽  
Etoac Extract

Abstract Background Arisaema jacquemontii is traditionally used in treatment of different diseases. In this study, phytochemical, in vitro biological and chemo-preventive screening of A. jacquemontii was carried out to explore its pharmacological potential. Methods The dried tuber of A. jacquemontii was extracted in 11 organic solvent mixture of different polarity. The extracts were screened for phytochemical assays (phenolics and flavonoids), antioxidants potential (free radical scavenging activity, total antioxidant activity, reducing power), biological activities (antibacterial, antifungal, cytotoxic, antileishmanial, protein kinase inhibition), and chemopreventive activities using different cell lines through standard protocols. Results Significant amount phenolic contents were determined in EtOH and MeOH extracts (210.3 ± 3.05 and 193.2 ± 3.15 μg GAE/mg, respectively). Maximum flavonoid content was determined in MeOH extract (22.4 ± 4.04 μg QE/mg). Noteworthy, DPPH scavenging activity was also recorded for MeOH extract (87.66%) followed by MeOH+EtOAc extract (85.11%). Considerable antioxidant capacity (7.8 ± 0.12 μg AAE/mg) and reducing power (3.1 ± 0.15 μg AAE/mg) was observed in extract of MeOH. The LC50 against brine shrimp and leishmanial parasite was found 9.01 and 12.87 μg/mL for n-Hex and CHCl3 extracts, respectively. The highest zone of inhibition against Streptomyces hyphae formation (12.5 ± 1.77 mm) by n-Hex extract. Growth zone of inhibition 13.8 ± 1.08 mm was recorded for EtOAc and MeOH extracts, respectively against Micrococcus luteus while 10.0 ± 0.11 mm for MeOH extract against Aspergillus flavus. In-vitro cytotoxic assay showed that n-Hex extract had higher cytotoxicity against DU-145 prostate cancer and HL-60 cancer cell lines. NF-kB and MTP potential showed 34.01 and 44.87 μg/mL for n-Hex and CHCl3 extracts, respectively in chemo-preventive potential. Conclusion The study concludes that Arisaema jacquemontii bears significant phytochemical activity and pharmacological activities, this plant can be further explored for isolation of active component against a number of aliments.

scholarly journals Tetrapleura tetraptera of Ghanaian Origin: Phytochemistry, Antioxidant and Antimicrobial Activity of Extracts of Plant Parts

2020 ◽  
pp. 78-96
Author(s):  
Christopher Larbie ◽  
Felix Charles Mills- Robertson ◽  
Emmanuel Bright Quaicoe ◽  
Rita Opoku ◽  
Naomi Chounbayor Kabiri ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Biological Activities ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Stem Bark ◽  
Scavenging Activity ◽  
Methanolic Extracts ◽  
Healthcare Needs ◽  
Tetrapleura Tetraptera ◽  
Phytochemical Constituents

Introduction: The role of medicinal plants in meeting the healthcare needs of the populace, particularly in developing countries cannot be overemphasized. They provide holistic treatment and wellbeing due to a plethora of phytochemicals found in them. Among such is Tetrapleura tetraptera (Schum. & Thonn.) Taub., a multipurpose tree plant with varied biological activities. The current study was aimed at assessing the basic phytochemical constituents, free radical scavenging activity as well as the antimicrobial effect on various solvent extracts of leaves, fruit and stem bark of T. tetraptera of Ghanaian origin. Materials and Methods: Aqueous, 50% hydroethanolic and methanolic extracts were made from the leaves, fruits, and stem bark of T. tetraptera. They were assessed for the qualitative and quantitative phytochemical constituents, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, antimicrobial activity as well as the spectroscopic properties (Ultraviolet – visible and Fourier-transform infrared spectroscopy). Compounds in the methanolic extracts were evaluated using the gas chromatography–mass spectrometry. Results: Tannins, reducing sugars, coumarins and flavonoids were present in all extracts while hydroethanolic and methanolic extracts were rich in phenols, tannins and flavonoids, especially for the stem bark. Leaf extracts had better antimicrobial activity against Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Candida albicans and Proteus mirabilis. Conclusion: Generally, leaves and stem bark extracts of T. tetraptera are rich in biologically active phytochemicals, supporting their use in ethnomedicine and could be exploited further for other biological activities.

scholarly journals BIOLOGICAL ACTIVITIES OF SOME SELECTED NEPALESE MEDICINAL PLANTS AND ISOLATION OF CHEMICAL CONSTITUENTS FROM CALLICARPA MACROPHYLLA

2020 ◽  
pp. 91-98
Author(s):  
KHAGA RAJ SHARMA ◽  
KHEMINDRA RANA
Keyword(s):  
Secondary Metabolites ◽  
Column Chromatography ◽  
Plant Extract ◽  
Biological Activities ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Significant Inhibition ◽  
Scavenging Activity ◽  
Ic50 Value ◽  
Bauhinia Purpurea

Objective: The main objectives of this study was to analyze the phytochemicals, determine the total flavonoid content, brine shrimp toxicity, antibacterial activity, evaluate the antioxidant, antimicrobial, anti-diabetic activities of nine medicinal plants Callicarpamacrophylla, Bauhinia purpurea, Plumeriarubra, Girardiniadiversifolia, Acacia nilotica, Woodfordiafruticosa (Bark) Woodfordiafruticosa (flower), Terminaliaalata, and Premnabarbata. Methods: The cold percolation method was adopted for the extraction of secondary metabolites in methanol. The preliminary phytochemical analysis was performed by colour differentiation methods. The radical scavenging activity was evaluated by DPPH (2,2-diphenyl-1-picrylhydrazyl) method. The antidiabetic activity was performed by α-amylase enzyme inhibition activity. The chemical constituent was isolated by column chromatography from the biologically active plant fraction. Results: The phytochemical investigation has shown plants are the rich source of secondary metabolites as quinones, saponins, terpenoids and glycosides. Among the nine tested plants, Terminaliaatalia showed the highest radical scavenging activity 96.41±0.47 with IC50 value 6.353 µg/ml, followed by Girardiniadiversifolia 97.26±0.67 with IC50 value 11.52 µg/ml whereas ascorbic acid has 39.85 µg/ml as standard. Bauhinia purpurea showed significant inhibition to the α-amylase enzyme having inhibitory concentration IC50 17.05±13.00 SD in a dose-dependent manner. Woodfordiafruticosa demonstrated significant toxicity to A. salina with LC50 value of 457.08 µg/ml. Callicarpamacrophylla bark showed a potential inhibitory activity against the growth of Straphylococcusaureus as compared to standard chloramphenicol. Active plant extract of Callicarpamacrophylla was subjected for column chromatography. Conclusion: Out of nine plant samples Terminaliaatalia showed the highest radical scavenging activity. The plant extract of Bauhinia purpurea showed significant inhibition to the α-amylase enzyme inhibition. Woodfordiafruticosa demonstrated significant toxicity to A. salina, whereas Callicarpamacrophylla showed the potent antibacterial activity. The active plant extract was subjected for column chromatography and different fractions were collected in solvent polarity basis. Conclusion: The phytochemical investigations showed that plant extracts are the rich sources of secondary metabolites such as alkaloids, flavonoids, saponins, glycosides, polyphenols, coumarins and reducing sugars which showed they are supposed to be responsible for different biological activities. IC50 values showed the varied degree of antioxidant property of which Plumeriarubra and Acacia nilotica exhibit good antioxidant property with IC50 value close to the standard ascorbic acid.

scholarly journals Bagged Aronia Melanocarpa Tea: Phenolic Profileand Antioxidant Activity / Aronia Melanocarpa Filter Čaj: Fenolni Profil I Antioksidativna Aktivnost

2014 ◽  
Vol 31 (4) ◽  
pp. 245-252 ◽  
Author(s):  
Jovana Veljković ◽  
Jelena Brcanović ◽  
Aleksandra Pavlović ◽  
Snežana Mitić ◽  
Biljana Kaličanin ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Caffeic Acid ◽  
Antioxidant Activities ◽  
Biological Activities ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Scavenging Activity ◽  
Antioxidant Power ◽  
Aronia Melanocarpa ◽  
Tea Infusions

Summary While there is a large number of scientific papers reporting chemical composition and biological activities of Aronia melanocarpa, there is a lack information regarding the commercially available bagged tea. In order to supply new information on the antioxidant activity of the Aronia melanocarpa tea infusions, the aim of this study was to evaluate individual phenolic compounds which could be responsible for antioxidant activities of these beverages. Selected anthocyanins (cyanidin-3-O-galactoside, cyanidin-3-O-glucoside, cyanidin- 3-O-arabinoside, and cyanidin-3-O-xyloside), gallic acid, caffeic acid, rutin, morin, and protocatechuic acid were simultaneously detected from commercially available tea infusions using a High Performance Liquid Chromatographic (HPLC) method. The antioxidant activity was measured using five in vitro spectrophotometric methods: 1,1-diphenyl- 2-picrylhydrazyl radical scavenging activity (DPPH), 2,2'-azino-bis (3-ethylbenzthiazoline- 6-sulphonic acid) radical cation scavenging activity (ABTS), ferric reducing-antioxidant power (FRAP) and reduction power (RP)Fe(III) to Fe(II). Obtained results showed that anthocyanins, predominantly of cyanidin-3-O-galactoside, are the major class of polyphenolic compounds in tea infusions. Among phenolic acids the most abundant is caffeic acid. A significant correlation between DPPH and ABTS and FRAP and RP suggested that antioxidant components in these beverages were capable scavenging free radicals and reducing oxidants. Generally, these beverages had relatively high antioxidant capacities and could be important dietary sources of antioxidant phenolics for the prevention of diseases caused by oxidative stress.

scholarly journals In Depth Natural Product Discovery from the Basidiomycetes Stereum Species

2020 ◽  
Vol 8 (7) ◽  
pp. 1049 ◽  
Author(s):  
Mengqing Tian ◽  
Peiji Zhao ◽  
Guohong Li ◽  
Keqin Zhang
Keyword(s):  
Secondary Metabolites ◽  
Natural Product ◽  
Pancreatic Lipase ◽  
Biological Activities ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Malarial Parasite ◽  
Scavenging Activity ◽  
Structural Types ◽  
Unidentified Species

Natural metabolites from microorganisms play significant roles in the discovery of drugs, both for disease treatments in humans, and applications in agriculture. The Basidiomycetes Stereum genus has been a source of such bioactive compounds. Here we report on the structures and activities of secondary metabolites from Stereum. Their structural types include sesquiterpenoids, polyketides, vibralactones, triterpenoids, sterols, carboxylic acids and saccharides. Most of them showed biological activities including cytotoxic, antibacterial, antifungal, antiviral, radical scavenging activity, autophagy inducing activity, inhibiting pancreatic lipase against malarial parasite, nematocidal and so on. The syntheses of some metabolites have been studied. In this review, 238 secondary metabolites from 10 known species and various unidentified species of Stereum were summarized over the last seven decades.

scholarly journals Evaluation of Antiradical Activity Grape Seeds and Olive (Olea europaea) Pits Extracts

2013 ◽  
Vol 6 (1) ◽  
pp. 185-190
Author(s):  
F. Tajalli
Keyword(s):  
Antiradical Activity ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Maximum Activity ◽  
Fruit Juices ◽  
Scavenging Activity ◽  
Grape Seeds ◽  
Antiradical Properties ◽  
Environmental Treatment ◽  
Dpph Method

The antiradical activity of grape seeds and olive pits extracts were investigated. The aim of this study is evaluating the radical scavenging activity of methanol extracts of olive pits and grape seeds and to recover a functional and antiradical compound from olive oil and fruit juices factories waste and also for environmental treatment. The antiradical properties of ripe and unripe olive pits (RIOP and URIOP) and grape seeds (IGS) that are respectively used in Iranian oil industries and fruit juices producers are examined. All seeds and pits extracts showed DPPH radical scavenging activity ranging from 24.51 to 97.06. For this purpose a methanolic extract was prepared from each of the RIOP, URIOP and IGS and their radical scavenging ability is determined with DPPH method. For this trial the effect of 3 different dilutions (100, 200 and 300 ?g/L) of RIOP, URIOP, IGS extracts was used, separately. It was appeared that in above 100 ppm concentrations the antiradical properties reaches to its maximum activity. Also, IGS extract shows better effects in 100 ppm concentrations in comparison with RIOP, URIOP and ascorbic acid. The study shows that grape seeds can be used as a rich source of functional and antiradical compound and anticancer drugs production. Keywords: Iranian olive pits; DPPH; Antiradical activity; Radical scavenging; Grape seeds.  © 2014 JSR Publications. ISSN: 2070-0237 (Print); 2070-0245 (Online). All rights reserved.  doi: http://dx.doi.org/10.3329/jsr.v6i1.16654 J. Sci. Res. 6 (1), 185-190 (2014)

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