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   2018| January-March  | Volume 10 | Issue 1  
    Online since February 19, 2018

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Antioxidant, Anti-inflammatory, Analgesic Properties, and Phytochemical Characterization of Stem Bark Extract and Fractions of Anthocleista nobilis
Kenneth Gerald Ngwoke, Amaka Godsaveus Akwagbulam, Ernest Oghenesuvwe Erhirhie, Daniel Lotanna Ajaghaku, Festus Basden Chiedu Okoye, Charles Okechukwu Esimone
January-March 2018, 10(1):81-87
DOI:10.4103/pr.pr_73_17  PMID:29568192
Background: Anthocleista nobilis (Loganiaceae) is used by Mbano people of Imo State, Nigeria, for the treatment of various ailments Objective: The aim of this study is to evaluate the antioxidant, anti-inflammatory, and analgesic properties of the methanol extract, fractions, and subfractions of A. nobilis. Materials and Methods: The powdered stem bark was extracted with methanol and sequentially fractionated into n-hexane, ethyl acetate, and butanol fractions. The constituents of the fractions were analyzed using high-pressure liquid chromatography (HPLC), and the components were identified by dereplication. Antioxidant potential of the extracts and fractions was investigated using 2,2-diphenyl-1-picrylhydrazyl free-radical scavenging method. Anti-inflammatory and analgesic activities of the extract and fractions were also investigated using xylene-induced inflammation and acetic acid-induced writhing models, respectively. Results: A total of five compounds isovitexin (Rt = 18.77 min), isovitexin-2''-O-xyl (Rt = 19.68 min), p-Hydroxybenzoic acid (Rt = 11.88 min), Sarasinoside L (Rt = 19.64 min), isovitexin (Rt = 18.77), and apigenin monoglycoside (Rt = 19.64 min) were identified by HPLC analysis and dereplication. The ethyl acetate fraction and subfraction elicited the best anti-inflammatory activity. The ethyl acetate subfraction also inhibited acetic acid-induced pain by 79% and 85.0% at the doses of 100 mg/kg and 200 mg/kg, respectively, which was better than 71.1% and 81.3% observed for diclofenac at similar doses. Conclusion: A. nobilis could be a potential source of anti-inflammatory and analgesic lead compounds.
  3,850 42 -
Alleviation of Heat Stress by a Polyherbal Formulation, Phytocee™: Impact on Zootechnical Parameters, Cloacal Temperature, and Stress Markers
Ramasamy Selvam, Subramaniyam Suresh, Marimuthu Saravanakumar, CV Chandrasekaran, D'Souza Prashanth
January-March 2018, 10(1):1-8
DOI:10.4103/pr.pr_138_17  PMID:29568180
Background: The range of thermoneutral zone of chickens is narrow, and they become easily susceptible to environmental stress, a common and major concern for poultry causing a production loss. Objective: The present study was designed to comparatively evaluate anti-stress activity of Phytocee™ and Vitamin C in chickens reared under heat stress. Materials and Methods: A total of 600-day-old chicks of Cobb 400 were randomly assigned to 4 groups with 6 replicates comprising 25 birds each (n = 150). G1 served as a normal control (NC) and supplemented with Vitamin C at 100 g/ton of feed. G2 served as a heat stress control (HSC), subjected to heat stress (34°C–36°C) without Vitamin C supplementation. G3 and G4 served as positive control and treatment group (TC), subjected to heat stress and supplemented with Vitamin C and Phytocee™ at 100 g/ton of feed, respectively. The impact on zootechnical parameters and cloacal temperature was assessed at regular intervals, and blood was collected at the end of the experiment for evaluation of stress parameters, namely heterophil lymphocyte ratio (H:L ratio) and serum corticosterone. Results: Exposure of chickens to heat stress caused a significant decrease in body weight, worsening of feed conversion ratio, higher mortality, and poor production efficiency. Moreover, serum corticosterone level, H:L ratio, and cloacal temperature were significantly increased in HSC as compared to NC. However, supplementation of Phytocee™ in feed significantly ameliorated the negative impact of heat stress in broiler birds. Conclusion: The supplementation of Phytocee™ demonstrated an anti-stress effect in chickens through restoration of serum corticosterone level, H:L ratio, and thermoregulatory mechanism.
  2,540 43 -
Quantitative Phytochemical Constituents and Antioxidant Activities of the Mistletoe, Phragmanthera capitata (Sprengel) Balle Extracted with Different Solvents
Franklin Uangbaoje Ohikhena, Olubunmi Abosede Wintola, Anthony Jide Afolayan
January-March 2018, 10(1):16-23
DOI:10.4103/pr.pr_65_17  PMID:29568182
Objective: The investigation aimed to evaluate the in vitro quantitative phytochemical compositions and antioxidant capacity of the dried leaves of Phragmanthera capitata extracted with different solvents. Methodology: Different phytochemical assays were used to evaluate the phenols, flavonoids, and proanthocyanidin contents of the acetone, methanol, ethanol, and aqueous extracts of the sample. Antioxidant activity was measured using 2,2diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), Ferric reducing antioxidant power (FRAP), nitric oxide, and total antioxidant capacity (TAC) assays. Results: Methanol had the highest extraction yield (21.50%) followed by aqueous (10.87%). Acetone and ethanol yielded 6.87% and 4.3%, respectively. However, acetone and ethanol extracts had more phenolic content (218.6 ± 3.38 and 214.8 ± 3.56 mg gallic acid equivalent (GAE)/g, respectively) that were significantly higher (P < 0.05) than the methanol and aqueous extracts (203.8 ± 1.95 and 175.5 ± 0.32 mg GAE/mg, respectively). Acetone extract also had more flavonoid and proanthocyanidin contents (679.8 ± 6.26 mg quercetin equivalent/g and 645.7 ± 6.35 mg catechin equivalent/g, respectively) that were significantly higher than the other solvent extracts (P < 0.05). All the solvent fractions showed great antioxidant activities with the acetone fraction having the highest capacity based on ABTS, DPPH, and TAC assays (half maximal inhibitory concentration [IC50] = <5 μg/mL, 24.5 μg/mL, and 85 μg/mL respectively). The methanol extract however had FRAP and nitric oxide antioxidant activities (IC50 = 302 μg/mL and <25 μg/mL respectively). Conclusion: The findings of this study provide evidence that P. capitata has potential as a natural source of antioxidant with acetone showing to be the best extractant of the polyphenolics with corresponding higher antioxidant activities.
  2,520 54 -
Cytotoxic Compounds from Wrightia pubescens (R.Br.)
Mariquit M De Los Reyes, Glenn G Oyong, Vincent Antonio S. Ng, Chien-Chang Shen, Consolacion Y Ragasa
January-March 2018, 10(1):9-15
DOI:10.4103/pr.pr_45_17  PMID:29568181
Background: Mixtures of ursolic acid (1) and oleanolic acid (2) (1:1 and 1:2), oleanolic acid (2), squalene (3), chlorophyll a (4), wrightiadione (5), and α-amyrin acetate (6) were isolated from the dichloromethane (CH2Cl2) extracts of the leaves and twigs of Wrightia pubescens (R.Br.). Objectives: To test for the cytotoxicity potentials of 1–6. Materials and Methods: The antiproliferative activities of 16 against three human cancer cell lines, breast (MCF-7) and colon (HT-29 and HCT-116), and a normal cell line, human dermal fibroblast neonatal (HDFn), were evaluated using the PrestoBlue® cell viability assay. Results: Compounds 4, 1 and 2 (1:2), 2, 1 and 2 (1:1), and 5 exhibited the most cytotoxic effects against HT-29 with half maximal inhibitory concentration (IC50) values of 0.68, 0.74, 0.89, 1.70, and 4.07 μg/mL, respectively. Comparing 2 with its 1:1 mixture with 1 (IC50 = 1.70 and 7.18 μg/mL for HT-29 and HCT-116, respectively) and 1:2 mixture with 1 (0.74 and 3.46 μg/mL for HT-29 and HCT-116, respectively), 2 also showed strong cytotoxic potential against HT-29 and HCT-116 (0.89 and 2.33 μg/mL, respectively). Unlike the mixtures which exhibited low effects on MCF-7 (IC50 = 20.75 and 30.06 μg/mL for 1:1 and 1:2, respectively), 2 showed moderate activity against MCF-7 (10.99 μg/mL). Compound 6 showed the highest cytotoxicity against HCT-116 (IC50 = 4.07 μg/mL). Conclusion: Mixtures of 1 and 2 (1:1 and 1:2), 2, 3, 4, 5, and 6 from the CH2Cl2extracts of the leaves and twigs of W. pubescens (R.Br.) exhibited varying cytotoxic activities. All the compounds except 6 exhibited the strongest cytotoxic effects against HT-29. On the other hand, 6 was most cytotoxic against HCT-116. Overall, the toxicities of 16 were highest against HT-29, followed by HCT-116 and MCF-7. All the compounds showed varying activities against HDFn (IC50 <30 μg/mL). Abbreviation Used: IC50: Half maximal inhibitory concentration.
  1,905 36 -
Thermogenic Blend Alone or in Combination with Whey Protein Supplement Stimulates Fat Metabolism and Improves Body Composition in Mice
Paula de Lima Vieira-Brock, Brent M Vaughan, David L Vollmer
January-March 2018, 10(1):37-43
DOI:10.4103/pr.pr_53_17  PMID:29568185
Background: Certain food ingredients promote thermogenesis and fat loss. Similarly, whey protein improves body composition. Due to this potential synergistic effect, a blend of thermogenic food ingredients containing African mango, citrus fruit extract, Coleus forskohlii, dihydrocapsiate, and red pepper was tested alone and in combination with a whey protein supplement for its effects on body composition in sedentary mice during high-fat diet. Objective: The objective of this study was to evaluate the interaction of thermogenic foods on improving body composition during consumption of an unhealthy diet. Materials and Methods: C57BL/6J young adult male mice (n = 12) were placed on a 60% high-fat diet for 4 weeks and subsequently randomly assigned to receive daily dosing by oral gavage of vehicle, the novel blend alone or with whey protein supplement for another 4 weeks. Body composition, thermal imaging of brown adipose tissue (BAT), mitochondrial BAT uncoupling protein 1 (UCP1), and plasma levels of leptin were assessed. Results: Novel blend alone and in combination with protein supplement attenuated body weight gain, fat, and increased surface BAT temperature in comparison to vehicle control and to baseline (P < 0.5). The combination of novel blend and whey protein supplement also significantly increased UCP1 protein expression in BAT mitochondria in comparison to vehicle control and novel blend alone (P < 0.5). Conclusions: These data indicate that this novel blend stimulates thermogenesis and attenuates the gain in body weight and fat in response to high-fat diet in mice and these effects were improved when administered in combination with whey protein supplement.
  1,885 39 -
Identification of Antidiabetic Compounds from Polyphenolic-rich Fractions of Bulbine abyssinica A. Rich Leaves
Samuel Wale Odeyemi, Anthony Jide Afolayan
January-March 2018, 10(1):72-80
DOI:10.4103/pr.pr_55_17  PMID:29568191
Background: Bulbine abyssinica has been reported to possess a variety of pharmacological activities traditionally. Previous work suggested its antidiabetic properties, but information on the antidiabetic compounds is still lacking. Objective: The present research exertion was aimed to isolate and identify biologically active polyphenols from B. abyssinica leaves and to evaluate their efficacy on carbohydrate digesting enzymes. Materials and Methods: Fractionation of the polyphenolic contents from the methanolic extract of B. abyssinica leaves was executed by the silica gel column chromatography to yield different fractions. The antioxidant activities of these fractions were carried out against 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,2-diphenyl-1-picrylhydrazyl radicals, and ferric ion-reducing antioxidant power (FRAP). In vitro antidiabetic experimentation was performed by evaluating the α-amylase and α-glucosidase inhibitory capacity. The isolated polyphenols were then identified using liquid chromatography and mass spectroscopy (LC/MS). Results: Out of the eight polyphenolic fractions (BAL 1–8), BAL-4 has the highest inhibitory activity against ABTS radicals whereas BAL-6 showed potent ferric ion-reducing capacity. BAL-5 was the most effective fraction with antidiabetic activity with IC50of 140.0 and 68.58 ± 3.2 μg/ml for α-amylase and α-glucosidase inhibitory activities, respectively. All the fractions competitively inhibited α-amylase, BAL-5 and BAL-6 also inhibited α-glucosidase competitively, while BAL-4 and BAL-1 exhibited noncompetitive and near competitive inhibitions against α-glucosidase, respectively. The LC/MS analysis revealed the presence of carvone in all the fractions. Conclusions: The present study demonstrates the antioxidant and antidiabetic activities of the isolated polyphenols from B. abyssinica.
  1,846 42 -
Isolation of 5,7-Dihydroxy, 6,8-Dimethyl Flavanone from Syzygium aqueum with Its Antioxidant and Xanthine Oxidase Inhibitor Activities
Muhamad Insanu, Zelika Mega Ramadhania, Evelyne Nadia Halim, Rika Hartati, Komar Ruslan Wirasutisna
January-March 2018, 10(1):60-63
DOI:10.4103/pr.pr_59_17  PMID:29568189
Background: Syzygium aqueum Burm.f. Alston (water apple) belonging to Myrtaceae family was originated from tropical areas. It was traditionally used as a medicinal plant. Objective: The objective of the study was to isolate the active compound from the methanolic extract of S. aqueum leaves. Methods: Extraction was done using continuous extraction with methanol as a solvent. The extract was then fractionated using liquid–liquid extraction, vacuum liquid chromatography, and radial chromatography. Recrystallization was done for purification. The structure of the compound was determined by Ultraviolet-Visible and (1D and 2D) nuclear magnetic resonance (NMR) spectrometer. Results: The isolate showed maximum wavelengths at 347 (band I) and 296 (band II) nm. After addition of NaOH and CH3COONa, the maximum wavelengths of band II moved to 340 and 339 nm, respectively. There was no change in wavelengths after addition CH3COONa/H3BO3and AlCl3. The1H-NMR spectrum showed 16 protons, whereas13C-NMR spectrum showed 15 carbons. Based on those data, the isolate was determined as 5,7-dihydroxy-6,8-dimethyl flavanone (demethoxymatteucinol). At a concentration of 100 and 50 μg/mL, it could inhibit 25.13% of xanthine oxidase (XO) activity and scavenge 11.87% of diphenyl-picrylhydrazyl, respectively. Conclusion: Demethoxymatteucinol was isolated for the first time from S. aqueum and it had mild antioxidant and XO inhibitory activities.
  1,796 53 -
Anticonvulsant and Antioxidant Effects of Musa sapientum Stem Extract on Acute and Chronic Experimental Models of Epilepsy
Aditya J Reddy, Ashok Kumar Dubey, Shailendra S Handu, Prashant Sharma, Pramod K Mediratta, Qazi Mushtaq Ahmed, Seema Jain
January-March 2018, 10(1):49-54
DOI:10.4103/pr.pr_31_17  PMID:29568187
Background: Musa sapientum (banana) plant extract has been shown to possess antioxidant activity in previous studies. Neuronal injury resulting from oxidative stress is an important factor involved in pathogenesis of epilepsy. Objective: The present study aimed to evaluate the anticonvulsant activity of M. sapientum stem extract (MSSE) in acute and chronic experimental models in mice and its effects on various markers of oxidative stress in the brain of pentylenetetrazole (PTZ)-kindled animals. Material and Methods: Maximal electroshock seizures (MES) and PTZ-induced convulsion models were used for acute studies. For the chronic study, the effect of MSSE on the development of kindling was studied. For the evaluation of the effects of MSSE on oxidative stress in brain, malondialdehyde (MDA) and reduced glutathione (GSH) levels were estimated in the brains of the kindled animals. Results: MSSE significantly increased the latency to onset of myoclonic jerks and the duration of clonic convulsions following PTZ administration. The MSSE pretreated group showed significantly reduced mean seizure score on PTZ-induced kindling. There was a significant increase in the brain MDA levels and decrease in GSH levels in response to PTZ-induced kindling. On MSSE pretreatment, there was a significant decrease in the MDA levels in the brains, though the increase in the GSH levels was not significant. Conclusion: The results from this study suggest the presence of significant anticonvulsant activity in MSSE, in both acute and chronic PTZ-induced seizure models, which could be due to its antioxidant activity, as is reflected by the change in oxidative stress markers in brain.
  1,646 47 -
Method Development and Validation for the Determination of Caffeine: An Alkaloid from Coffea arabica by High-performance Liquid Chromatography Method
P Naveen, HB Lingaraju, M Deepak, B Medhini, K Shyam Prasad
January-March 2018, 10(1):88-91
DOI:10.4103/pr.pr_79_17  PMID:29568193
Objective: The present study was investigated to develop and validate a reversed phase high performance liquid chromatography method for the determination of caffeine from bean material of Coffee arabica. Materials and Methods: The separation was achieved on a reversed-phase C18 column using a mobile phase composed of water: methanol (50:50) at a flow rate of 1.0 mlmin-1. The detection was carried out on a UV detector at 272 nm. The developed method was validated according to the requirements for International Conference on Harmonisation (ICH) guidelines, which includes specificity, linearity, precision, accuracy, limit of detection and limit of quantitation. Results: The developed method validates good linearity with excellent correlation coefficient (R2 > 0.999). In repeatability and intermediate precision, the percentage relative standard deviation (% RSD) of peak area was less than 1% shows high precision of the method. The recovery rate for caffeine was within 98.78% - 101.28% indicates high accuracy of the method. The low limit of detection and limit of quantitation of caffeine enable the detection and quantitation of caffeine from C. arabica at low concentrations. Conclusion: The developed HPLC method is a simple, rapid, precisely, accurately and widely accepted and it is recommended for efficient assays in routine work.
  1,634 38 -
Algae from Portuguese Coast Presented High Cytotoxicity and Antiproliferative Effects on an In vitro Model of Human Colorectal Cancer
Celso Alves, Susete Pinteus, Ana Rodrigues, André Horta, Rui Pedrosa
January-March 2018, 10(1):24-30
DOI:10.4103/pr.pr_151_16  PMID:29568183
Background: The marine environment has shown to be an interesting source of new antitumor agents, representing an important tool in cancer research. Objective: The aim of this study was to evaluate the antitumor activities of 12 algae from Peniche coast (Portugal) on an in vitro model of human colorectal cancer (Caco-2 cells). Materials and Methods: The antitumor potential was accessed by evaluating Caco-2 cell's viability and proliferation through the 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide and calcein-AM methods. Results: The dichloromethane extracts of Asparagopsis armata and Sphaerococcus coronopifolius induced the highest decrease on cell's viability (1 mg/mL; 24 h), 98.96% ± 0.39% and 98.08% ± 0.89%, respectively, followed by the methanolic extracts of S. coronopifolius (96.47% ± 1.26%) and A. armata (92.68% ± 1.17%). Regarding cell proliferation, the highest decrease of Caco-2 cell's proliferation (1 mg/mL; 24 h) was induced by the dichloromethane extract of A. armata (100% ± 0.48%), S. coronopifolius (99.04 ± 0.51%), and Plocamium cartilagineum (95.05% ± 1.19%). The highest potency was shown by the dichloromethane extract of S. coronopifolius in both, cytotoxicity and antiproliferative tests, with an IC50of 21.3 and 36.5 μg/mL, respectively. Conclusion: The extracts of A. armata and S. coronopifolius are promising sources of new bioactive molecules with application in cancer therapeutics.
  1,559 42 -
Antiproliferative and Apoptotic Activities of the Medicinal Plant Ziziphus obtusifolia
Edna D Molina-Romo, Adriana Garibay-Escobar, Olivia Valenzuela-Antelo, Eduardo Ruiz-Bustos, Javier Hernández Martínez, Carlos Vélazquez, Luisa A Rascón-Valenzuela, Ramón Enrique Robles-Zepeda
January-March 2018, 10(1):55-59
DOI:10.4103/pr.pr_38_17  PMID:29568188
Background: Ziziphus obtusifolia is a spiny shrub found in Northwest Mexico desert, with traditional medicinal use to treat several diseases including cancer. Objective: The aims of the present study were to evaluate the antiproliferative and apoptotic activities of the aerial parts of this plant. Materials and Methods: The methanol extract and its fractions were prepared using several solvents. The antiproliferative activity was evaluated by the (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium) (MTT) assay on HeLa, A549, RAW 264.7, M12.C3.F6, and L-929 cell lines, and the apoptotic activity using Annexin V and (5,50,6,60-tetra-chloro-1,10,3,30-tetra-ethylbenzimidazol-carbocyanine iodide) staining. The most active fraction was further separated by column chromatography. Results: The most active fraction was hexane with an IC50of 90.4 μg/mL against RAW 264.7, 94 μg/mL against M12.C3.F6, 165.5 μg/mL against HeLa and 187.7 μg/mL against A549 cell lines. In apoptotic activity assays the methanol extract and its n-hexane fraction were found to induce mitochondrial depolarization in HeLa cells (83 and 87% respectively), and both induced the externalization of the phosphatidylserine increasing the percentage of cells in early apoptosis from 1.4% in untreated control cells, to 1.9% and 3.5% for methanol extract and n-hexane fraction-treated cells, respectively, statistically different for the total percentage of apoptotic cells (P < 0.05). Conclusions: These results show that Z. obtusifolia has antiproliferative and apoptotic activities in vitro and confirms its use in traditional medicine. The methanol extract and its fractions using several solvents were evaluated in the antiproliferative activity by the MTT assay on HeLa, A549, RAW 264.7, M12.C3.F6, and L-929 cell lines, and the apoptotic activity using Annexin V and (5,50,6,60-tetra-chloro-1,10,3,30-tetra-ethylbenzimidazol-carbocyanine iodide) staining. The most active fraction against cell lines was hexane. In apoptotic activity assays, the methanol extract and its n-hexane fraction were found to induce mitochondrial depolarization. This results we showed that Ziziphus obtusifolia has antiproliferative and apoptotic activities in vitro.
  1,533 46 -
Evaluation of Antithrombotic Activities of Solanum xanthocarpum and Tinospora cordifolia
Onila Lugun, Sobharani Bhoi, Priyanka Kujur, D Vinay Kumar, William R Surin
January-March 2018, 10(1):98-103
DOI:10.4103/pr.pr_80_17  PMID:29568195
Background: Solanum xanthocarpum and Tinospora cordifolia have been reported to exhibit anti-inflammatory, antiarthritic, antioxidant, antiallergic, and hepatoprotective activities. The origins of many of the currently available antithrombotic treatments are from natural products and natural sources. Objective: To investigate the antithrombotic activities of methanolic leaf extracts of S. xanthocarpum(SXME) and T. cordifolia(TCME). Materials and Methods: Antithrombotic activities were assessed by thrombin inhibition assay, thrombin generation assay, platelet adhesion assay on collagen-coated surface, and platelet PAC1-FITC binding by flow cytometry. Results: SXME significantly inhibited thrombin activity at 5–20 mg/ml concentrations, whereas TCME inhibited thrombin activity at 500 μg/ml–5 mg/ml concentrations. Further, SXME inhibited thrombin generation at 2–20 mg/ml concentrations, whereas TCME exhibited significant inhibition at 200 μg/ml, suggesting that TCME has higher efficacy as compared to SXME. Moreover, SXME did not inhibit platelet adhesion on collagen-coated surface, whereas TCME inhibited platelet adhesion on collagen-coated surface at 5 mg/ml. Indomethacin showed significant inhibition in platelet adhesion at 300 μM. Further, SXME inhibited thrombin-induced platelet activation (PAC1-FITC binding) significantly at 1 mg/ml by about 80%, whereas TCME inhibited thrombin-induced platelet activation (PAC1-FITC binding) by about 40% at 1 mg/ml. Conclusion: These results strongly suggested that SXME and TCME possess antithrombotic activities. However, further studies are essential to find out the active constituent responsible for antithrombotic effect.
  1,494 51 -
Analytical Method Development and Validation for the Simultaneous Estimation of Abacavir and Lamivudine by Reversed-phase High-performance Liquid Chromatography in Bulk and Tablet Dosage Forms
Sufiyan Ahmad Raees Ahmad, Lalit Patil, Mohammed Rageeb Mohammed Usman, Mohammad Imran, Rashid Akhtar
January-March 2018, 10(1):92-97
DOI:10.4103/pr.pr_96_17  PMID:29568194
Objective: A simple rapid, accurate, precise, and reproducible validated reverse phase high performance liquid chromatography (HPLC) method was developed for the determination of Abacavir (ABAC) and Lamivudine (LAMI) in bulk and tablet dosage forms. Methods: The quantification was carried out using Symmetry Premsil C18 (250 mm × 4.6 mm, 5 μm) column run in isocratic way using mobile phase comprising methanol: water (0.05% orthophosphoric acid with pH 3) 83:17 v/v and a detection wavelength of 245 nm and injection volume of 20 μl, with a flow rate of 1 ml/min. Results: In the developed method, the retention times of ABAC and LAMI were found to be 3.5 min and 7.4 min, respectively. The method was validated in terms of linearity, precision, accuracy, limits of detection, limits of quantitation, and robustness in accordance with the International Conference on Harmonization guidelines. Conclusion: The assay of the proposed method was found to be 99% – 101%. The recovery studies were also carried out and mean % recovery was found to be 99% – 101%. The % relative standard deviation from reproducibility was found to be <2%. The proposed method was statistically evaluated and can be applied for routine quality control analysis of ABAC and LAMI in bulk and in tablet dosage form.
  1,501 38 -
In vitro Evaluation of Antioxidant Potential of Isolated Compounds and Various Extracts of Peel of Punica granatum L.
Janani Jacob, P Lakshmanapermalsamy, Ramanaiah Illuri, Damaji Bhosle, Gopala Krishna Sangli, Deepak Mundkinajeddu
January-March 2018, 10(1):44-48
DOI:10.4103/pr.pr_36_17  PMID:29568186
Background: Punica granatum L. (Lythraceae) peel has been proven to exhibit widespread pharmacological application against multitude of diseases due to the presence of bioactive principles. Objective: The objective is to isolate the bioactive compounds from the pericarp of P. granatum and to evaluate the antioxidant activity of various extracts. Materials and Methods: Dried peel of P. granatum was extracted with aqueous acetone and chromatographed on Diaion HP-20. Enriched fractions were rechromatographed on Sephadex LH-20 and purified on preparative high-performance liquid chromatography to identify individual compounds. The dried peel was extracted with different solvents to evaluate the antioxidant activity of the extracts. Results: On the chemical investigation, three compounds were isolated and characterized as punicalagin, 2,3-(S)-hexahydroxydiphenoyl-D-glucose, and punicalin, using various spectroscopic techniques. Conclusion: Results indicate that the isolated compounds have possessed antioxidant activity, and aqueous, methanol, and aqueous acetone extract showed significant scavenging of 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) radicals.
  1,497 35 -
Comparative Molluscicidal and Schistosomicidal Potentiality of Two Solanum Species and Its Isolated Glycoalkaloids
Muhammad A Alsherbiny, Shymaa A El Badawy, Hesham Elbedewy, Shahira M Ezzat, Fatma S Elsakhawy, Mostafa A Abdel-Kawy
January-March 2018, 10(1):113-117
DOI:10.4103/pr.pr_71_17  PMID:29568198
Schistosomiasis is the most noteworthy parasitic disease after malaria. Furthermore, the significant activity of the genus Solanum against Schistosoma worms and its intermediate host snails reinforced the study of Solanum seaforthianum Andr. (SS) and Solanum macrocarpon L. (SM) for their molluscicidal and schistosomicidal potentiality. In this study, different extracts, fractions and isolated compounds of both Solanum species are evaluated for the molluscicidal and schistosomicidal potentialities. The niclosamide was used as positive molluscicide control against Biomphalaria alexandrina snails. Different extracts, fractions, or isolated compounds were used at a concentration of 100 μg/ml and dead snails were counted in each case. On the other hand, washed and sterilized Schistosoma mansoni adult worms were used in three replicates, and three worm pairs were placed in each well with 2 ml test solution of 100 μg/ml concentration. Positive (praziquantel [PZQ] 0.2 ug/ml) and negative controls were concurrently used and examined daily for 3 days for viability. The mortality rate was calculated and then both LC50and LC90were determined in triplicates. Highest potency was indicated to total glycoalkaloid (TGA) fraction of SM followed by TGA of SS. On the other hand, TGA fractions of both species showed higher potency than other extracts and isolated compounds. Meanwhile, solasodine-free aglycone showed declined activity compared to its glycosides. Promising molluscicidal and schistosomicidal activities were displayed which are attributed to the glycoalkaloid content. Therefore, this study can efficiently contribute toward validation of the traditional use of SS and SM in schistosomiasis control.
  1,486 29 -
Anti-inflammatory Effect of an Extract of Agave americana on Experimental Animals
Arup Kumar Misra, Sushil Kumar Varma, Ranjeet Kumar
January-March 2018, 10(1):104-108
DOI:10.4103/pr.pr_64_17  PMID:29568196
Background: Medicinal plants have the potential treasure to treat medical conditions for a long time based on error and trial method. Agave americana has been known as century plant or American aloe. The leaves contain genins, saponins, steroidal flavonoids, isoflavones, and coumarins. It has long been used for anti-inflammatory effects. Objectives: This study was designed to evaluate the anti-inflammatory activity of the extract of A. americana. Materials and Methods: A. americana leaves extract is collected and the extract is done by percolation method. Inflammation is induced in experimental animals' carrageenan-induced paw edema model and cotton pellet-induced granuloma model. The extract of A. americana and standard, aspirin and indomethacin are given orally for the anti-inflammatory effects. The anti-inflammatory effects were measured at regular intervals of time to see the percentage inhibition of paw edema and reduction of weight of granuloma with percentage protection to compare the effect of the extract with the control and standard. Results: The extract of A. americana showed improved in percentage inhibition of paw edema in graded doses of hydroalcoholic extract of A. americana (HEAA) as compared with control in carrageenan-induced paw edema model. The percentage inhibition of 400 mg/kg of HEAA at the 3rd h is almost comparable with the standard, aspirin. On the other hand, graded doses (200 and 400 mg/kg) showed a highly significant reduction (P < 0.001) in the weights of granuloma in comparison with the control in cotton pellet-induced granuloma model. The percentage inhibition of weight of granuloma by 400 mg/kg of HEAA is quite comparable to the standard, Indomethacin. Conclusion: Anti-inflammatory activity is seen with the graded dose of HEAA. The plant contains flavonoids and genins which have been shown to have anti-edematous effects in the acute phase of inflammation. Thus, the plant contributes to its anti-inflammatory activity. Thus, higher doses of A. americana leaves need to be used to have the better understanding of the mechanism of anti-inflammation.
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Antioxidant Activities of Basella alba Aqueous Leave Extract In Blood, Pancreas, and Gonadal Tissues of Diabetic Male Wistar Rats
Dennis Seyi Arokoyo, Ibukun Peter Oyeyipo, Stefan Simon Du Plessis, Yapo Guillaume Aboua
January-March 2018, 10(1):31-36
DOI:10.4103/pr.pr_84_17  PMID:29568184
Background: Oxidative stress is frequently identified as a key element in the pathophysiology of many complications of diabetes mellitus, including reproductive complications. The antioxidant potential of medicinal plants have been suggested for therapeutic focus of diseases in recent reports. Objective: To investigate the effect of Basella alba (Ba) aqueous leave extract on diabetes-induced oxidative stress. Materials and Methods: Forty male Wistar rats (8–10 weeks) were randomly divided into four groups (n = 10) and treated as follows; Control (C + Ns) and Diabetic (D + Ns) animals received oral normal saline 0.5 ml/100 g body weight daily, while Healthy Treatment (H + Ba) and Diabetic Treatment (D + Ba) rats were given Ba extract at an oral dose of 200 mg/kg body weight daily. Treatment was by gavage and lasted 4 weeks in all groups. Diabetes was induced in D + Ns and D + Ba rats by single intraperitoneal injection of streptozotocin (55 mg/kg) and fasting blood sugar (FBS) recorded weekly in all rats afterwards. Animals were euthanized at the end of the experiment and blood samples, pancreas, testes, and epididymis were preserved for analysis of oxidative stress biomarkers. Results: Oral administration of aqueous leave extract of Ba significantly (P < 0.0001) lowered FBS in D + Ba rats. There was significantly higher blood superoxide dismutase activity and serum ferric reducing antioxidant power, but lower serum concentration of conjugated dienes and thiobarbituric acid reactive substances in D + Ba compared to D + Ns rats (P < 0.05). Conclusion: Ba exerts antioxidant effects in the gonads by enhancing antioxidant parameters in circulating blood, but not necessarily in the gonadal tissues.
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Assessment of In vivo Antiviral Potential of Datura metel Linn. Extracts against Rabies Virus
Soumen Roy, Lalit Samant, Rajeshkumar Ganjhu, Sandeepan Mukherjee, Abhay Chowdhary
January-March 2018, 10(1):109-112
DOI:10.4103/pr.pr_121_17  PMID:29568197
Objective: The soxhlet, cold, and ayurvedic extracts of Datura metel Linn. were evaluated for in vivo antirabies activity. Materials and Methods: Soxhlet and cold extraction method were used to extract Datura (fruit and seed) extracts, and ayurvedic extraction of Datura was prepared. In vivo toxicity assay was performed as per the OECD 420. LD50dose was calculated by Reed and Muench method. The in vivo antirabies activity was screened in Swiss albino mice with the virus challenge dose of 10 LD50 (intracerebrally) in both preexposure (PE) and postexposure treatment with oral administration of Datura extracts in Swiss albino mice and observed for 21 days. The virus load in the mice brain was evaluated by TCID50 titration method. Results: Datura (ayurvedic preparation) was found to be nontoxic up to 2000 mg/kg in Swiss albino mice, i.e., 60 mg/30 g of mice, when administered (0.5 ml) orally and observed till 21 days. Up to 20% survival rate on the test group (PE of Datura extracts) up to 14 days postinfection as compared to the virus control group (10 LD50) was observed. No survival rate was observed in the postexposure group of Datura extract; however, the survival time was increased by 4 days as compared to the virus control group. Viral load of the infected mice brain sample was estimated in vero cell line, and 3 log reduction in the virus titer was observed in text group as compared to the virus control, suggesting that Datura extract has an in vivo antirabies activity. Conclusion: To the best of our knowledge, this is the first study of in vivo antiviral activity of an ayurvedic preparation of D. metel Linn. against rabies virus. Datura extracts have a potential in vivo antirabies activity.
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The Protective effect of different extracts of three Artemisia species against H2O2-Induced Oxidative Stress and Apoptosis in PC12 Neuronal Cells
Leila Hosseinzadeh, Alireza Malekshahi, Farahnaz Ahmadi, Seyed Ahmad Emami, Marziyeh Hajialyani, Mahdi Mojarrab
January-March 2018, 10(1):64-71
DOI:10.4103/pr.pr_98_17  PMID:29568190
Background: Oxidative stress causes cell damage and is involved in many neurological diseases. The antioxidant properties of plant materials for the maintenance of health and protecting against different diseases stimulated scientist to investigate different herbs. Different Artemisia species have exhibited antioxidant activity. This study aims to investigate whether different Artemisia species could protect the PC12 cells against oxidative stress mediated by H2O2. Methods: For this purpose, different extracts of three Artemisia species (Artemisia aucheri, Artemisia turanica, and Artemisia turcomanica) were prepared using petroleum ether, dichloromethane, ethyl acetate, ethanol, and Water: Ethanol mixture (1:1 volume ratio). The protective effect of the prepared extracts against H2O2-induced cytotoxicity and reactive oxygen species production were compared. The effect of treatment of PC12 cells with different extracts on total glutathione (GSH) level, caspase-3 activity, and mitochondrial membrane potential were also compared. Results: The A. aucheri extracts could not rescue the PC12 cells from oxidative stress consequences. The A. turanica and A. turcomanica extracts were found potent in suppressing the toxicity and apoptosis of PC12 cells mediated by H2O2and significantly antagonized the H2O2-induced GSH depletion. The hydroethanolic and ethyl acetate extracts of A. turanica and the petroleum ether and hydroethanolic extracts of A. turcomanica more efficiently suppressed cytotoxicity and loss of GSH caused by H2O2. Conclusion: This study shows the protective effects of Artemisia extracts on PC12 cell line and suggested that these species could be also considered as promising neuroprotective agents in treatment of different neurodegenerative diseases.
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