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Journal of Pharmaceutical Negative Results
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   Table of Contents - Current issue
Coverpage
January-December 2017
Volume 8 | Issue 1
Page Nos. 1-57

Online since Friday, April 21, 2017

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EDITORIAL  

New Editorial Board (2017-18) – Widening the scope of the “Journal of Pharmaceutical Negative Research (JPNR)” into biomedical research p. 1
Subramani Parasuraman
DOI:10.4103/jpnr.JPNR_9_17  
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ORIGINAL ARTICLES Top

Ultraviolet spectrophotometric method: Not possible for the simultaneous estimation of active constituents in Phyllanthus amarus p. 2
Rakhi Khabiya
DOI:10.4103/0976-9234.204909  
Objective: Our objective was to develop and validate UV spectrophotometric method for simultaneous estimation of phyllanthin, hypophyllanthin, and niranthin in Phyllanthus amarus. P. amarus is highly valued in the treatment of liver ailments and has been shown to posses antihepatitis B virus surface antigen activity. Several analytical methods including high-performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC) are reported for respective phytoconstituents. However, there is no simultaneous estimation of phyllanthin, hypophyllanthin, and niranthin, active constituent of P. amarus. Hence, attempts have been made to develop UV spectrophotometric method because of the simplicity and economical advantage of this technique over other analytical techniques as HPTLC, HPLC, etc. Materials and Methods: Standard solution of phyllanthin, hypophyllanthin, and niranthin were prepared in methanol and scanned under a Shimadzu UV spectrophotomer, under full UV (ultraviolet) range (200-400 nm) to get the spectra. Result: After some experiments, it was found that it is impossible to quantify phyllanthin, hypophyllanthin, and niranthin simultaneously by UV spectrophotometric method. This was evident from the UV spectra as they were overlapping with each other. Conclusion: Same class of drugs may have almost the same functional groups, and gradient reverse-phase liquid chromatography and HPTLC will be more useful to separate such complicated mixtures. Therefore, we propose to develop either HPLC or HPTLC method for simultaneous estimation of these compounds.
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In vitro cytotoxicity evaluation of stem bark and leaves of Anogeissus pendula: Lack of potent activity p. 7
Deeksha Singh, US Baghel, Rakesh Yadav
DOI:10.4103/0976-9234.204915  
Objective: Anogeissus pendula Edgew (Combretaceae) which is also known as “button tree” is one of the prominent species of Anogeissus genus which has several ethnomedicinal uses. Phytochemical investigation revealed the presence of alkaloids and phenolic compounds, such as flavonoids, lignins, tannins. So, pharmacological evaluation of this plant may lead to discovery of new activity. Therefore, the objective of the present study is to evaluate the in vitro cytotoxicity of stem bark and leaves hydroalcoholic extract of A. pendula. Materials and Methods: In vitro cytotoxicity was evaluated by MTT (3-(4,5-dimethyl thiazol-2-yl)-5-diphenyl tetrazolium bromide) assay against A431 (human epithelial carcinoma) and HepG-2 (human liver carcinoma) cell lines. The assay is based on the principle that the dead cells do not reduce the tetrazolium into formazan. Various concentrations (62.5-1000 μg/mL) of hydroalcoholic extracts were used and the concentration of test drug needed to inhibit cell growth by 50% (IC50) values is generated from the dose–response curves for each cell line. Microscopic examination was also carried out for morphological analysis of cells treated with extracts of A. pendula. Results: The hydroalcoholic extracts of stem bark and leaves revealed dose dependent, but insignificant activity against the cell lines i.e.A431 and HepG-2 as the IC50 was found to be greater than 1000 μg/mL for both the extracts against both the cell lines. There were no morphological changes in cells which supports the insignificant cytotoxicity. Conclusion: The results suggest that the hydroalcoholic extracts of stem bark and leaves of A. pendula have insignificant cytotoxicity.
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Poor antimicrobial activity on seven cuban plants p. 11
Orlando A. Abreu, Ileana Sánchez, Guillermo Barreto, Ana C. Campal
DOI:10.4103/0976-9234.204910  
Background: Plant Kingdom still constitutes a source for antimicrobials, screening bioassays of plants extracts is one of the first steps in the search of new antimicrobial compounds. Cuba has a rich flora with a high degree of endemism that practically has not been investigated. Method: Twelve acetone extracts (1mg/ml) of seven Cuban plants (four endemic), were tested by agar disc diffusion method against selected strains of bacteria and fungi: Escherichia coli (three strains), Staphylococcus aureus and Candida albicans. Results: In general null or poor antimicrobial effect was observed, except in Caesalpinia bahamensis subs bahamensis heartwood, that showed activity of interest against bacteria even at 0,5mg/ml; while modest antibacterial effects was found in endemics: Hypericum styphelioides (leave, stem), H. limosum (leave, stem), Vaccinium leonis (leave) and V. ramonii (leave, stem); Agdestis clematidea show lowest effect. S. aureus was the most sensitive bacteria (six species and 11 extracts) and only a negligible activity was found against C. albicans in four extract of four species. Discussion: Morphology of Gram positive bacteria, in contrast to Gram negative that have a more resistant outer membrane, seem to be the cause of S. aureus sensitiveness. Starting from these results, further current antimicrobial research approach related virulence factors mechanisms interference can be carried out in these plants. Conclusions: In this experimental conditions, except in one specie, null or poor antimicrobial effect was found in tested plants, studies on these plants have to be continues, since in other conditions maybe could be found some bacteriostatic or bactericide activity.
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Total phenolic content and anti-oxidant potential of Ficus deltoidea using green and non-green solvents p. 15
Mun Hui Sin, Awang Soh Mamat, Muhammad Shahzad Aslam, Muhammad Syarhabil Ahmad
DOI:10.4103/0976-9234.204913  
The aim of the present study is to determine the total phenolic content (TPC) and anti-oxidant activity of green and non-green solvent extracts. Solvents have an important role in chemical or pharmaceutical industry. The usage of non-green solvent may cause detrimental effects on human safety and health. Therefore, green solvents are encouraged to be utilized because they are environmental friendly and easily acquired during the agricultural crops processing. For this study, the green solvent used in the extraction of Ficus deltoidea's leaves was water whereas non-green solvents used were ethanol and methanol. Folin-Ciocalteu method was used for measuring the presence and amount of phenolic content in each of the sample. The results showed that aqueous extract had the highest TPC (368.42 ± 6.37 mg GAE/g), followed by methanolic (295.03 ± 16.65 mg GAE/g) and ethanolic (263.45 ± 5.28 mg GAE/g) extracts. Furthermore, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay was used in determining the anti-oxidant activity of the extracts. The lowest IC50 value based on the graph of percentage inhibition against concentration of sample was shown by ethanolic extract (16.5 µg/mL), followed by methanolic extract (22.0 µg/mL), and aqueous extract (23.5 µg/mL). For the three samples, the correlations between TPC and IC50 were negative and weak. Hence, the anti-oxidant activity of an extract cannot be predicted based on its TPC. The present study has shown that F. deltoidea considerably has the potential as a source of natural anti-oxidants.
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Low inhibition of alpha-glucosidase and xanthine oxidase activities of ethanol extract of Momordica charantia fruit p. 20
A Khatib, V Perumal, QU Ahmed, BF Uzir, S Murugesu
DOI:10.4103/0976-9234.204906  
Objective: To evaluate the α-glucosidase and xanthine oxidase inhibitory activities of different ethanolic and aqueous extracts of Momordica charantia fruit. Materials and Methods: The M. charantia fruits were extracted using different concentrations of ethanol in water (0, 20, 40, 60, 80 and 100 %, v/v). The samples from different ethanolic and aqueous extracts of M. charantia fruit (0, 20, 40, 60 and 100 % ethanol in water [v/v] extracts) were tested for α-glucosidase and xanthine oxidase inhibitory activities. Quercetin and allopurinol were used as positive controls for α-glucosidase and xanthine oxidase inhibitory assay, respectively. The samples were also tested for phytochemicals screening such as alkaloid, phenol, steroid, tannin, coumarins and flavonoid. The significant difference was determined by one-way ANOVA test with a Tukey comparison at confidence interval of 95%. Results: Phytochemical screening showed the presence of coumarin, alkaloid, steroid and phenol in the extracts. The six different M. charantia extracts showed low inhibition on α-glucosidase and xanthine oxidase activities. Conclusions: The antidiabetic activity of the ethanolic and aqueous extracts of M. charantia fruit was not through the inhibition of α-glucosidase and xanthine oxidase.
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Effectiveness of photodynamic therapy and probiotics as an adjunct to scaling and root debridement in the treatment of chronic periodontitis p. 25
Lo S Ann, Gun C Yin, Siti Fatimah, Ng S En, Daniel D Dicksit, CG Kalyan, Khairiyah A Muttalib, Srinivas S Ramachandra
DOI:10.4103/0976-9234.204908  
Objective: This study aimed to evaluate the effectiveness of a single application of photodynamic therapy (PDT) and probiotics as an adjunct to scaling and root debridement (SRD) to treat chronic periodontitis. The objectives of the study were to compare the probing pocket depths and clinical attachment loss (CAL) in experimental sites (E-sites) treated with PDT and probiotics with SRD against SRD alone (controls) at the baseline and 3 months post intervention. Materials and Methods: In 22 chronic periodontitis patients aged 25–45 years, 136 sites with pocket depths of 4–6 mm were identified. In each patient, one E-site was randomly selected while one served as control. E-sites were treated with SRD + PDT + probiotics whereas control sites received only SRD. Clinical parameters of pocket depths and CAL were measured at the baseline and at 3 months. Results: In the E-sites, mean pocket depth was 3.62 ± 1.40 mm at the baseline and 3.12 ± 1.48 mm after 3 months. In the control sites, the mean pocket depth was 3.43 ± 1.46 mm at the baseline and 2.99 ± 1.45 after 3 months. In the E-sites, the mean CAL before treatment was 5.31 ± 1.39 mm and 5.10 ± 1.46 mm after treatment. In the control sites, mean CAL before treatment was 5.15 ± 1.41 mm and 4.87 ± 1.55 mm after treatment. There was no significant improvement in pockets depths and CAL between the E-sites and control sites. Conclusion: A single application of PDT + probiotics with SRD did not show any additional clinical benefits when compared with SRD alone. Further studies with multiple applications of PDT and probiotics are needed.
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Lack of in vitro anticancer and antimicrobial activities from Karanda (Carissa carandas) fruit extracts p. 31
Yuttana Sudjaroen
DOI:10.4103/jpnr.JPNR_5_17  
Background: Carissa carandas L. (Apocynaceae), commonly known as Karanda, is a widely used medicinal plant. In Thailand, Karanda fruits are favorite fruits especially in central region due to attractive shape and color with health promoting activities. Aims: To evaluate its antimicrobial and anticancer activities for claim promoting activities. Cytotoxicity of extracts was also evaluated with normal cells to claim for health safety. Materials and Methods: Anticancer activities of Karanda fruits extracted with dichloromethane (KD) and methanol (KM) were performed by resazurin microplate assay and tested with five cell lines, including KB-oral cavity cancer, MCF7-breast cancer, NCI-H187-small lung cancer, HepG2-hepatocarcinoma, and Caco2-colon adenocarcinoma cell lines. Cytotoxicity of KD and KM were performed as above and used HDFn-neonatal dermal fibroblast as normal cells. Antimicrobial activities of KD and KM against herpes simplex virus type I (HSV-1) and Mycobacterium tuberculosis were tested according by green fluorescent protein-based assay; Candida albicans were tested according to resazurin microplate assay, and Plasmodium falciparum K1 strain was tested according to microculture radioisotope techniques. Results: Both extracts were not possessed anticancer activity to any cancer cell lines at maximum concentration = 100 µg/mL. In the same way to anticancer activity assays, both extracts were not inhibited HSV-1, P. falciparum K1 strain, and M. tuberculosis (maximum concentration = 50 µg/mL). Also, both extracts were nontoxic to normal cells. Conclusion: KD and KM extracts of ripped Karanda fruits had no significant anticancer and antimicrobial activities with noncytotoxicity.
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Investigation of mast cell stabilization and antiulcer activity of protein extract of Perna viridis p. 37
K Venkateskumar, S Parasuraman, PV Christopher, Syed Ali, Leow Yu Chuen, Winnie Tang, Siaw Yee Sii, Tan Chun Yin
DOI:10.4103/jpnr.JPNR_4_17  
Objective: To study the mast cell stabilization and antiucler activity of protein extract of Perna viridis using rodent models. Materials and Methods: The total protein from P. viridis was extracted, purified, and screened for mast cell stabilization and antiulcer activity. Intestinal mesentery of male rats was used to study the peritoneal mast cell stabilization of protein extract of P. viridis. The rat intestinal mesentery was exposed to 10, 30, 100, 300, and 600 μg/mL of protein extract of P. viridis and the peritoneal mast cell stabilization was compared with that of standard (ketotifen) at a dose of 20 μg/mL. Antiulcer activity of protein extract of P. viridis was studied at the dose levels of 100, 200, and 400 mg/kg and the effect was compared with the omeprazole (20 mg/kg) using ethanol-induced ulcer model. At the end of the study, ulcer index and percentage of ulcer inhibition was calculated. Results: The total protein content in extract was found to be 32 μg/mL. The protein extracts of P. viridis showed significant mast cell stabilization only at high dose (600 μg/mL) and did not show any significant antiulcer activity for doses of 100 and 200 mg/kg in administered animals, but the significant antiulcer activity was observed at a dose of 400 mg/kg. Conclusion: The present study findings suggests that the protein extract of P. viridis did not exhibited significant mast cell stabilization and antiulcer activity at optimal doses.
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The evaluation of quercetin and luteolin efficacy on cutaneous leishmaniasis in mice infected with Leishmania major p. 43
Nasrin Hamidizadeh, Sara Ranjbar, Qasem Asgari, Gholamreza Hatam
DOI:10.4103/jpnr.JPNR_1_17  
Introduction: Almost 1.5-2 million people are infected with leishmaniasis and 60,000 die due to the disease every year. The treatment of cutaneous leishmaniasis with the existing medications is not effective and accompanied with various side effects. Expensiveness and not being available in different forms are other limitations associated with these drugs. In addition to, parasite resistance restricts the use of these medications; therefore, identification of new treatment methods is highly essential. Humans have always used medicinal plants for the treatment of diseases. This study aimed to assess the impact of quercetin and luteolin on leishmaniasis wounds created on mice's tails. Materials and Methods: A total of 56 infected Balb/C mice were divided into eight groups, each containing seven animals. The first three groups received 3.5 mg/kg luteolin through oral, intradermal, and intraperitoneal routes. The other three groups received 14 mg/kg quercetin through oral, intradermal, and intraperitoneal routes. The treatments were administered twice a week for 4 weeks in comparison to meglumine antimonite as control groups. Results: The results showed that there was no statistically significant difference between the recovery of mice in the luteolin and quercetin groups and those in the meglumine antimonite group. However, the ratio of recovered Balb/C mice in the luteolin and quercetin group is 1.75-2 times more than meglumine group, respectively. Conclusion: Quercetin and luteolin seems to be candidate medications with fewer side effects for recovery from cutaneous leishmaniasis. They can also be used as complementary medications together with other standard drugs, such as meglumine, antimonite, and cryotherapy.
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LETTERS TO THE EDITOR Top

The 'HOPE' for primary prevention of cardiovascular disease: An Asian perspective p. 49
Sunil B Pai, Ashwin Kamath
DOI:10.4103/0976-9234.204917  
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Antibacterial efficacy after incorporation of CPP-ACP (GC Tooth MousseTM) into glass Ionomer cement against Streptococcus Mutans: A preliminary in vitro study p. 51
Niraj Gokhale, Parneeka Sood, Shivayogi Hugar, Parin Shah, Divyata Kohli, Chandrashekhar Badkar
DOI:10.4103/0976-9234.204907  
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Poor antimicrobial activity of methanol and ethanol extract of leaves of Solanum trilobatum Linn. p. 53
Subramani Parasuraman, Leow Yu Chuen, Siaw Sze Hoong, David Lim De Wei, Lim Nian Zou, Sunderesan Loshini
DOI:10.4103/jpnr.JPNR_7_17  
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Underreporting of mild adverse events during plateletpheresis donations p. 56
Sonam Kansay
DOI:10.4103/0976-9234.204911  
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