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   2015| April-June  | Volume 7 | Issue 2  
    Online since February 16, 2015

 
 
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ORIGINAL ARTICLES
Anti-inflammatory and antiarthritic activity of UNIM-301 (a polyherbal unani formulation) in Wistar rats
Surender Singh, Rohit Kumar, Hitesh Jain, YK Gupta
April-June 2015, 7(2):188-192
DOI:10.4103/0974-8490.150515  PMID:25829793
Background: UNIM-301 is a polyherbal formulation used in the Unani system of medicine for the treatment of joint pain and rheumatoid arthritis (RA). Objective: The objective was to evaluate the anti-inflammatory and antiarthritic activity of UNIM-301 in carrageenan-induced paw edema and complete Freund's Adjuvant (CFA) induced arthritis. Materials and Methods: The anti-inflammatory and antiarthritic activity of UNIM-301 was evaluated using carrageenan-induced paw edema and CFA induced animal arthritis models, respectively, in doses of 250, 500, and 1000 mg/kg body weight. Anti-inflammatory activity of UNIM-301 was evaluated using carrageenan-induced paw edema model using a digital plethysmometer. Anti-arthritic activity was evaluated using CFA induced arthritis, and joint sizes were measured at regular intervals using a micrometer screw gauge. Serum was collected and subjected to estimation of pro-inflammatory cytokine. Indomethacin 3 mg/kg body weight) was used as a standard drug in both the models. The acute and chronic toxicity study was carried out to evaluate the safety of the test drug. Results: UNIM-301 treatment produced a dose-dependent reduction in paw edema and paw thickness in carrageenan-induced paw edema and CFA-induced arthritis, respectively, as compared to control. UNIM 301 also reduced the expression of pro-inflammatory mediator in a dose-dependent manner as compared to control. Conclusion: The result of the present study suggests that anti-inflammatory and anti-arthritic activity of UNIM-301, which might be accredited to inhibitory activity on pro-inflammatory cytokines to its various individual constituents.
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Cytotoxic activity of ten algae from the Persian Gulf and Oman Sea on human breast cancer cell lines; MDA-MB-231, MCF-7, and T-47D
Nasrollah Erfani, Zahra Nazemosadat, Mahmoodreza Moein
April-June 2015, 7(2):133-137
DOI:10.4103/0974-8490.150539  PMID:25829786
Background: Seaweeds have proven to be a promising natural source of bioactive metabolites for drug development. Objective: This study aimed to monitor the ethanol extract of ten algae from the Persian Gulf and Oman Sea, for their in vitro cytotoxic activity on three human breast cancer cell lines. Materials and Methods: Three human breast cancer cell lines including MDA-MB-231(ER ), MCF-7(ER + ), and T-47D (ER + ) were treated by different concentrations of total ethanol (90%) algae extracts and the cytotoxic effects were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Doxorubicin (Ebewe, Austria) was used as a positive control. After 72 h of incubation, the cytotoxic effect of the algae was calculated and presented as 50%-inhibitory concentration (IC 50 ). Results: The results indicated Gracilaria foliifera and Cladophoropsis sp. to be the most active algae in terms of cytotoxic effects on the investigated cancer cell lines. The IC 50 values against MDA-MB-231, MCF-7, and T-47D cells were, respectively, 74.89 ± 21.71, 207.81 ± 12.07, and 203.25 ± 30.98 mg/ml for G. foliifera and 66.48 ± 4.96, 150.86 ± 51.56 and >400 mg/ml for Cladophoropsis sp. The rest of the algal extracts were observed not to have significant cytotoxic effects in the concentration range from 6.25 mg/ml to 400 mg/ml. Conclusion: Our data conclusively suggest that G. foliifera and Cladophoropsis sp. may be good candidates for further fractionation to obtain novel anticancer substances. Moreover, stronger cytotoxic effects on estrogen negative breast cancer cell line (MDA-MB-231(ER )) in comparison to estrogen positive cells (MCF-7 and T-47D) suggest that the extract of G. foliifera and Cladophoropsis sp. may have an estrogen receptor/progesterone receptor-independent mechanism for their cellular growth inhibition.
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Psidium guajava Linn. leaf extract affects hepatic glucose transporter-2 to attenuate early onset of insulin resistance consequent to high fructose intake: An experimental study
R Mathur, Shagun Dutta, T Velpandian, SR Mathur
April-June 2015, 7(2):166-175
DOI:10.4103/0974-8490.151459  PMID:25829790
Background: Insulin resistance (IR) is amalgam of pathologies like altered glucos metabolism, dyslipidemia, impaired glucose tolerance, non-alcoholic fatty liver disease, and associated with type-II diabetes and cardiometabolic diseases. One of the reasons leading to its increased and early incidence is understood to be a high intake of processed fructose containing foods and beverages by individuals, especially, during critical developmental years. Objective: To investigate the preventive potential of aqueous extract of Psidium guajava leaves (PG) against metabolic pathologies, vis-à-vis, IR, dyslipidemia, hyperleptinemia and hypertension, due to excess fructose intake initiated during developmental years. Materials and Methods: Post-weaning (4 weeks old) male rats were provided fructose (15%) as drinking solution, ad libitum, for 8 weeks and assessed for food and water/fructose intake, body weight, fasting blood sugar, mean arterial pressure, lipid biochemistry, endocrinal (insulin, leptin), histopathological (fatty liver) and immunohistochemical (hepatic glucose transporter [GLUT2]) parameters. Parallel treatment groups were administered PG in doses of 250 and 500 mg/kg/d, po × 8 weeks and assessed for same parameters. Using extensive liquid chromatography-mass spectrometry protocols, PG was analyzed for the presence of phytoconstituents like Myrecetin, Luteolin, Kaempferol and Guavanoic acid and validated to contain Quercetin up to 9.9%w/w. Results: High fructose intake raised circulating levels of insulin and leptin and hepatic GLUT2 expression to promote IR, dyslipidemia, and hypertension that were favorably re-set with PG. Although PG is known for its beneficial role in diabetes mellitus, for the first time we report its potential in the management of lifelong pathologies arising from high fructose intake initiated during developmental years.
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A study of Nigella sativa induced growth inhibition of MCF and HepG2 cell lines: An anti-neoplastic study along with its mechanism of action
Y Padmanabha Reddy, KB Chandrasekhar, Mohammed Jaffar Sadiq
April-June 2015, 7(2):193-197
DOI:10.4103/0974-8490.150541  PMID:25829794
Objective: To evaluate the anticancer potential of seeds of Nigella sativa using MCF and HepG2 cell lines along with its mechanism of action. Materials and Methods: (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and acridine orange/ethidium bromide nuclear staining technique were selected to evaluate anticancer potential and mechanism of action of test extract. Results: Aqueous extract of N.sativa at a test dose of 180 mg and 300 mg was identified to be the best as anticancer agent against MCF and HepG2 cell lines among different solvent test extract where doxorubicin and cisplatin were employed as standard references. Discussion: Further study including separation and characterization of active principles in the aqueous extract shall prove beneficial.
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Assessment of in vitro cytotoxicity and in vivo antitumor activity of Sphaeranthus amaranthoides burm.f
S Gayatri, C Uma Maheswara Reddy, K Chitra, V Parthasarathy
April-June 2015, 7(2):198-202
DOI:10.4103/0974-8490.150544  PMID:25829795
Background: A number of plants have been used in Indian system of medicine such as ayurveda, unani and siddha, but most of these plants were not explored properly. Sphaeranthus amaranthoides (SA) Burm.f., is one such plant used as an energizer in siddha. Objective: To evaluate the anticancer effect of chloroform extract of the whole plant of SA Burm.f. against Ehrlich ascites carcinoma bearing Swiss albino mice. Materials and Methods: The anticancer effect of chloroform extract was investigated in Swiss albino mice bearing Ehrlich ascites carcinoma at two different dose levels. Acute toxicity studies were also performed to determine the safety of the extract. Mice injected with Ehrlich ascites carcinoma cells were treated with the extract of doses 200 and 400 mg/kg body weight and standard 5-fluorouracil 20 mg/kg body weight for 15 consecutive days. Animals were sacrificed on day 15 for determination of anticancer activity by evaluating tumor volume, nonviable and viable tumor cell count and hematological parameters. Results: Mice treated with the extract showed a significant decrease in tumor volume and viable cell count and an increase in nonviable cell count and mean survival time. The hematological parameters were also found to be restored to a normal level. Conclusion: The results indicate that the chloroform extract was producing anticancer activity comparable with that of the standard 5-fluorouracil.
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Evaluation of Antidiabetic and Antihyperlipidemic Effects of Hydroalcoholic Extract of Leaves of Ocimum tenuiflorum (Lamiaceae) and Prediction of Biological Activity of its Phytoconstituents
Subramani Parasuraman, Subramani Balamurugan, Parayil Varghese Christapher, Rajendran Ramesh Petchi, Wong Yeng Yeng, Jeyabalan Sujithra, Chockalingam Vijaya
April-June 2015, 7(2):156-165
DOI:10.4103/0974-8490.151457  PMID:25829789
Objective: The aim was to evaluate the anti-diabetic and anti-hyperlipidemic effects of hydroalcoholic extract of leaves of Ocimum tenuiflorum (Lamiaceae) and prediction of biological activities of its phytoconstituents using in vivo anti-diabetic model and in silico analysis respectively. Materials and Methods: The leaves of O. tenuiflorum were extracted with 60% ethanol, and the extract was used for further pharmacological screening. The acute toxicity of the extract was evaluated as per the guidelines set by the Organization for Economic Co-operation and Development, revised draft guidelines 423. The oral anti-diabetic activity of the hydroalcoholic extract of O. tenuiflorum (125, 250 and 500 mg/kg) was studied against streptozotocin (STZ) (50 mg/kg; i.p.) + nicotinamide (120 mg/kg; i.p.) induced diabetes mellitus. The animals were treated with the investigational plant extract and standard drug (glibenclamide) for 21 consecutive days and the effect of hydroalcoholic extract of O. tenuiflorum on blood glucose levels was measured at regular intervals. At the end of the study, blood samples were collected from all the animals for biochemical estimation, then the animals were sacrificed and the liver and kidney were collected for organ weight analysis. Prediction for pharmacological and toxicological properties of phytoconstituents of O. tenuiflorum was carried out using online web tools such as online pass prediction and lazar toxicity prediction. Results: The hydroalcoholic extract of O. tenuiflorum showed significant anti-diabetic and anti-hyperlipidemic activity at 250 and 500 mg/kg, and this effect was comparable with that of glibenclamide. Predicted biological activities of phytoconstituents of O. tenuiflorum showed presence of various pharmacological actions, which includes anti-diabetic and anti-hyperlipidemic activities. Prediction of toxicological properties of phytoconstituents of O. tenuiflorum did not show any major toxic effects. Conclusion: The hydroalcoholic extract of O. tenuiflorum showed significant anti-diabetic and anti-hyperlipidemic activity against STZ + nicotinamide induced diabetes mellitus in rats. Further studies are required to confirm the anti-diabetic and anti-hyperlipidemic activities of individual phytoconstituents of O. tenuiflorum.
  1 4,378 48
Serum total antioxidant capacity in oral carcinoma patients
KS Rathan Shetty, Arunava Kali, KS Rachan Shetty
April-June 2015, 7(2):184-187
DOI:10.4103/0974-8490.150513  PMID:25829792
Context: Oxidative stress is a major determinant in carcinogenesis and serum total antioxidant capacity (TAC) indirectly reflects the level of oxidative stress. Although oral cancer is the third most frequent cancer in Indian population, there are no standard noninvasive marker for early detection and monitoring therapeutic response in oral cancer patients. Aims: This study was carried out to investigate implications of serum TAC in oral cancer by evaluating pre- and post-operative levels in oral cancer patients. Settings and Designs: It was a prospective single blinded study. Subjects and Methods: Serum TAC was measured using ferric-reducing antioxidant power assay. Pre- and post-operative values were estimated and compared in 30 oral cancer patients who underwent surgery. Statistical Analysis Used: Paired t-test was used to compare pre- and post-operative values. Results: Compared to the normal value, both pre- and post-operative serum TAC were significantly low in oral cancer patients. However, there was no statistically significant difference between pre- and postoperative levels. Conclusions: In this study, low mean serum TAC was detected in oral cancer patients. However, its diagnostic and prognostic significance in oral cancer needs further investigation.
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SHORT COMMUNICATION
In vivo anti-inflammatory and antiarthritic activities of aqueous extracts from Thymelaea hirsuta
Zora Azza, Mounia Oudghiri
April-June 2015, 7(2):213-216
DOI:10.4103/0974-8490.150510  PMID:25829798
Background: The aerial parts of Thymelaea hirsuta (TH) are used as a decoction in the treatment of different pathologies in folk medicine in Morocco. Objective: The aqueous extracts were evaluated for its anti-inflammatory activity and in inhibition of adjuvant induction arthritis in male Wistar rats. Materials and Methods: The anti-inflammatory activity was carried out using carrageenan-induced rat paw edema model, and the antiarthritic activity was carried out using complete Freund's adjuvant-induced arthritis model. Results: The plant extract (500 mg/kg body weight) exhibited significant activity in acute inflammation produced 60% of inhibition after 4 h as compared with that of the standard anti-inflammatory drug, the diclofenac (100 mg/kg) which showed 40% of inhibition. In arthritis model, the extract produced 85% inhibition after 18 days when compared with the diclofenac (10 mg/kg; 72%). Conclusion: These results indicate that the aqueous extract of TH had an anti-inflammatory activity and inhibited the induction of adjuvant arthritis in male Wistar rats.
  1 3,869 34
ORIGINAL ARTICLES
Nucleotide sequence of Phaseolus vulgaris L. alcohol dehydrogenase encoding cDNA and three-dimensional structure prediction of the deduced protein
Kassim Amelia, Chin Yin Khor, Farida Habib Shah, Subhash J Bhore
April-June 2015, 7(2):203-208
DOI:10.4103/0974-8490.150532  PMID:25829796
Background: Common beans (Phaseolus vulgaris L.) are widely consumed as a source of proteins and natural products. However, its yield needs to be increased. In line with the agenda of Phaseomics (an international consortium), work of expressed sequence tags (ESTs) generation from bean pods was initiated. Altogether, 5972 ESTs have been isolated. Alcohol dehydrogenase (AD) encoding gene cDNA was a noticeable transcript among the generated ESTs. This AD is an important enzyme; therefore, to understand more about it this study was undertaken. Objective: The objective of this study was to elucidate P. vulgaris L. AD (PvAD) gene cDNA sequence and to predict the three-dimensional (3D) structure of deduced protein. Materials and Methods: positive and negative strands of the PvAD cDNA clone were sequenced using M13 forward and M13 reverse primers to elucidate the nucleotide sequence. Deduced PvAD cDNA and protein sequence was analyzed for their basic features using online bioinformatics tools. Sequence comparison was carried out using bl2seq program, and tree-view program was used to construct a phylogenetic tree. The secondary structures and 3D structure of PvAD protein were predicted by using the PHYRE automatic fold recognition server. Results: The sequencing results analysis showed that PvAD cDNA is 1294 bp in length. It's open reading frame encodes for a protein that contains 371 amino acids. Deduced protein sequence analysis showed the presence of putative substrate binding, catalytic Zn binding, and NAD binding sites. Results indicate that the predicted 3D structure of PvAD protein is analogous to the experimentally determined crystal structure of s-nitrosoglutathione reductase from an Arabidopsis species. Conclusions: The 1294 bp long PvAD cDNA encodes for 371 amino acid long protein that contains conserved domains required for biological functions of AD. The predicted deduced PvAD protein's 3D structure reflects the analogy with the crystal structure of Arabidopsis thaliana s-nitrosoglutathione reductase. Further study is required to validate the predicted structure.
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Analysis of common bean (Phaseolus vulgaris L., genotype BAT93) calmodulin cDNA using computational tools
Kassim Amelia, Jasvin Singh, Farida Habib Shah, Subhash J Bhore
April-June 2015, 7(2):209-212
DOI:10.4103/0974-8490.150536  PMID:25829797
Background: Common bean (Phaseolus vulgaris L.) is an important part of the human diet and serves as a source of natural products. Identification and understanding of genes in P. vulgaris is important for its improvement. Characterization of expressed sequence tags (ESTs) is one of the approaches in understanding the expressed genes. For the understanding of genes expression in P. vulgaris pod-tissue, research work of ESTs generation was initiated by constructing cDNA libraries using 5-day and 20-day old bean-pod-tissues. Altogether, 5972 cDNA clones were isolated to have ESTs. While processing ESTs, we found a transcript for calmodulin (CaM) gene. It is an important gene that encodes for a calcium-binding protein and known to express in all eukaryotic cells. Hence, this study was undertaken to analyse and annotate it. Objective: The objective of this study was to analyze and annotate P. vulgaris CaM (PvCaM) gene cDNA and its deduced protein (amino acids) sequence. Materials and Methods: Both strands of PvCaM cDNA clone were sequenced using M13 forward and reverse primer to elucidate the nucleotide sequence. The cDNA sequence and deduced protein sequence were analyzed and annotated using bioinformatics tools available online. The secondary structures and three-dimensional (3D) structure of PvCaM protein were predicted using the Phyre automatic fold recognition server. Results: Results showed that PvCaM cDNA is 818 bp in length. The cDNA analysis results showed that it contains an open reading frame that encodes for 149 amino acid residues. The deduced protein sequence analysis results showed the presence of conserved domains required for CaM function. The predicted secondary structures and 3D structure are analogous to the Solanum tuberosum CaM. Conclusions: This study analyzed and annotated PvCaM cDNA and protein. However, in order to obtain a complete understanding of PvCaM protein, further study on its expression, structure and regulation is essential.
  - 3,814 30
Development of phytovesicles containing triterpenoids from Samadera indica
Vidya Viswanad, Subin Mary Zachariah, Aiswarya Sathi, NA Aleykutty
April-June 2015, 7(2):176-183
DOI:10.4103/0974-8490.151461  PMID:25829791
Introduction: Samadera indica belonging to Simaroubaceae family is being used traditionally for many diseases including arthritis, edema, itching, skin diseases, constipation, and general debility. Objective: The effectiveness of any drug delivery system depends upon its ability to deliver the active components at therapeutic level. In this study, a novel phyto vesicular formulation for the enhanced topical delivery of methanol extract of S. indica in order to treat skin infections was developed. Materials and Methods: The methanol extract fraction of leaves of S. indica which showed more antifungal activity was purified to separate an antifungal compound. Phytovesicles were formulated using the more antifungal fraction in order to treat topical and deep seated fungal infections. Pytovesicles were prepared using 1:2 molar ratio of antifungal triterpenoid from S. indica (AFTSI)-phosphatidylcholine by film hydration method. Results and Discussion: Chloroform 100% fraction of methanol extract of S. indica showed more activity against the fungus Candida albicans. Further purification gave a fraction with minimum inhibitory concentration value of 15.6 mg/ml against C. albicans and showed positive test for triterpenoids. The fraction was named as AFTSI. A compound (20 mg) was isolated from this fraction at an R F value. The phytovesicle gel formulated using AFTSI showed enhanced skin permeability and antifungal activity. Conclusion: The study demonstrated that the phytovesicular gel developed using methanol extract of S. indica would be beneficial for treating deep seated fungal infections.
  - 3,428 31
Chemical constituents and bioactivities of Glinus oppositifolius
Consolacion Y Ragasa, Esperanza C Cabrera, Oscar B Torres, Adiel Inah Buluran, Dinah L Espineli, Dennis D Raga, Chien-Chang Shen
April-June 2015, 7(2):138-147
DOI:10.4103/0974-8490.150520  PMID:25829787
Objectives: To isolate the secondary metabolites from the dichloromethane (DCM) extracts of Glinus oppositifolius; to test for the cytotoxicity of a new triterpene, oppositifolone (1); and to test for the hypoglycemic, analgesic, and antimicrobial potentials of 1, DCM and aqueous leaf extracts of G. oppositifolius. Methods: The compounds were isolated by silica gel chromatography and identified by nuclear magnetic resonance spectroscopy. The cytotoxicity potential of 1 was tested using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Triterpene 1, DCM, and aqueous leaf extracts were tested for hypoglycemic potential using the oral glucose tolerance test; analgesic potential using the tail-flick assay, and antimicrobial potential using the disc diffusion method. Results: The DCM extracts of G. oppositifolius afforded 1, squalene, spinasterol, oleanolic acid, phytol, and lutein from the leaves; squalene and spergulagenin A from the stems; and spinasterol from the roots. Triterpene 1 was cytotoxic against human colon carcinoma 116 with an IC 50 value of 28.7 but did not exhibit cytotoxicity against A549. The aqueous leaf extract at 200 mg/kg body weight (BW) exhibited hypoglycemic activity with a pronounced % blood glucose reduction of 70.76% ±17.4% within 0.5 h after introduction. The DCM leaf extract showed a lower % blood glucose reduction of 18.52% ±13.5% at 200 mg/kg BW within 1.5 h after introduction, while 1 did not exhibit hypoglycemic activity. The samples did not exhibit analgesic property and were inactive against multiple drug resistant bacterial pathogens. Conclusion: The compounds responsible for the hypoglycemic activity of G. oppositifolius which are fast acting (0.5 h) are found in the aqueous leaf extract.
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Comparing and authenticating on anatomical aspects of Abrus cantoniensis and Abrus mollis by microscopy
Zhifeng Zhang, Luyang Lu, Yuan Liu, Linsen Qing
April-June 2015, 7(2):148-155
DOI:10.4103/0974-8490.150526  PMID:25829788
Background: Abrus cantoniensis is popularly used as traditional Chinese medicine and a cool tea in South of China. However, due to diminishing source of A. cantoniensis, it is usually interchanged or adulterated with other species of Abrus genus because of the limited knowledge in identification and differentiation. Especially, Abrus mollis is widely mixed on herbal markets and pharmaceutical preparation. Objective: To ensure safety and efficacy, a detailed comparison was undertaken to carry out an anatomical and micro-morphological study of two species of A. cantoniensis and A. mollis. Materials and Methods: Microscopic characteristics of roots, leaves and stems, including transverse sections and the crude drug powder, were observed using a light microscope according to the usual microscopic techniques. Results: The basic diagnostic features of A. cantoniensis include that stem is extremely thin; xylem vessels of root are radially arranged in 10 or more bundles; pith is hollow in stem, and the palisade tissue is made up of two layers of palisade cells. Furthermore, scanning electron microscopy was used to compare nonglandular hairs and the stomata of the leaflet surface. A table of the key authentication parameters based on the analyzed microscopic characteristics was drawn up. Conclusion: The study demonstrated that the microscopy and related techniques provided a systematic method that is convenient, feasible, and can be unambiguously applied to the authentication of the species of Abrus.
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