Inhibitory effects of fermented soybean tempeh on the anti-adhesive properties of Actinomyces viscosus and plaque growth in vitro Allimalar Sathiaseelan, Chong Seng Shit, Tsun-Thai Chai Pharmacognosy Magazine 2019 15(66):371-376 Background: Tempeh, the Southeast Asian traditional food, has garnered great attention for its antibacterial property against Gram-positive and Gram-negative pathogens, and antidiarrheal effect. We have previously reported the potentiality of tempeh hexane fraction (HXF) in ceasing Actinomyces viscosus biofilm formation in vitro. Objective: Here, we investigated the efficacy of tempeh HXF on other cariogenic virulence traits of A. viscosus such as adhesive properties, acid production, and plaque growth. Materials and Methods: Anti adhesion of HXF was assessed based on its effects on the number of cells adhering to the surface of tooth in sucrose-dependent (SD) and sucrose-independent (SI) medium. The potential of HXF to inhibit the capability of A. viscosus to generate acids was investigated by pH drop assay. The HXF at different concentrations were used to determine the LC50based on brine shrimp lethality assay. Finally, the prospect of HXF as an inhibitor of plaque formation was investigated using artificial saliva-coated denture as an in vitro batch model. Results: HXF significantly decreased colony-forming unit of SD (1.07 log reduction) and SI (0.56 log reduction)-mediated adsorption of bacterial cells onto the tooth surface over 4- and 12-h incubation, respectively. Acid production was reduced after treated with HXF in a dose-dependent manner. Finally, a substantial reduction in plaque coverage area >55% was found on the HXF treated-denture. Conclusion: The anti-biofilm effect of HXF was associated with the suppression of A. viscosus adhesion to tooth surfaces and reduction in acid production. Furthermore, in vitro anti-plaque potential of HXF was demonstrated.

Selection of suitable reference genes for reverse transcription-quantitative polymerase chain reaction normalization in Artemisia annua L. plants at different stages of growth and development Javed Ahmad, Nazima Nasrullah, Naved Quadri, Umara Nissar, Shashi Kumar, Malik Zainul Abdin Pharmacognosy Magazine 2019 15(66):377-385 Background: Artemisinin, an antimalarial compound suggested by the WHO to treat drug-resistant malaria, was obtained from Artemisia annua L. plants. However, due to the low level of artemisinin in the plant causes limitation to its commercialization, so to increase the concentration of artemisinin, two transgenic lines were developed by us, overexpressing the key genes of artemisinin biosynthetic pathway, namely, 3 S-hydroxy-3-methyl glutaryl-CoA reductase (HMGR), amorpha-4,11-diene synthase (ADS) (Trans. 1) and HMGR, ADS, and CYP71AV1 (cytochrome P450 monooxygenase) (Trans. 2). Objectives: Our main aim for this study was to select the suitable reference gene for the normalization of reverse transcription-quantitative polymerase chain reaction ( Reverse transcription RT-qPCR) data in different tissues at various developmental stages in A. annua L. plants. Materials and Methods: Six candidate reference genes, namely; β-actin (ACT), elongation factor 1-alpha (EF1α), TAP-42 interacting protein (TAP42), SAND family protein, β-tubulin, and protein phosphatase 2A (PP2A) for their expression stability in the root, stem, leaf, and flower of A. annua L. plants at vegetative, preflowering, and flowering stages were analyzed using geNorm, NormFinder and BestKeeper, the Excel-based research tools. Results: The genes ACT/PP2A, PP2A/TAP42, and EF1&#945/PP2A were appropriate as reference genes in the leaf tissues at vegetative, preflowering, and flowering stages, respectively. In addition, EF1&#945/PP2A genes at vegetative and flowering stage, while EF1&#945/TAP42 gene at preflowering stage was found suitable reference genes for normalization of expression data in the stem. In the root samples, ACT/EF1&#945, EF1&#945/PP2A, and ACT/TAP42 sets were found to be reliable reference genes at vegetative, preflowering, and flowering stages, respectively, whereas, PP2A/TAP42 gene set was found suitable for flower tissues at flowering stage. Conclusion: These results will be helpful in the normalization of expression data in RT-qPCR to find the reliable outcome.

Modulation of sodium arsenite-induced Toxicity in mice by ethanolic seed extract of Trigonella foenum graecum Surjyo Jyoti Biswas, Goutam Ghosh, Ved Prakash Dubey Pharmacognosy Magazine 2019 15(66):386-395 Background: Trigonella foenum graecum (TG) Linn. (Methi) is widely used as a spice and known for its pharmacological properties. Objective: The current study was conducted to examine the efficacy of TG Linn., family: Fabaceae, against sodium arsenite-induced toxicity in mice. Materials and Methods: Sixty mice (Mus musculus) weighing about 25 g were randomized into six groups; each of ten mice: Group I served as untreated control; Group II received only sodium arsenite (100 ppm) in drinking water for 2 months. The Group III mice fed chronically with sodium arsenite for 2 months as in Group II and then fed a vehicle of 1:20 alcohol to distilled water (1:20) for 15 and 30 days, respectively; Group IV to VI mice were treated as in Group II and then fed with 50, 150, and 250 mg/kg of TG seed extract, once daily for 15 and 30 days. Results: The IC50of the seed extract was 66.78 μg/mL, and it reduced the activities of toxicity marker enzymes such as gamma glutamyl transferase, lactate dehydrogenase, lipid peroxidation, aspartate transaminase, alanine aminotransaminase, acid phosphatase, alkaline phosphatase, and pro-inflammatory cytokines such as tumor necrosis factor-alpha and interleukin-6 (P < 0.05 to P < 0.001) and elevated the activities of catalase, superoxide dismutase, and G6PD (P < 0.05 to P < 0.001), a similar trend was also noted with hematological variables. Further, the normal architecture of the kidney was retained in the TG-fed series than arsenic (As)-treated series. Urinary excretion of As was high in treated groups compared to controls (P < 0.05 to P < 0.001), and 150 mg/kg dose offered better protection than the other two doses. Conclusion: TG seed extract has revealed potent antioxidant properties and consequently can be used as a protective agent in As-induced toxicity.

Protective Effect of Madhuca longifolia Leaves in 7, 12-Dimethylbenz(a)anthracene Induced Mammary Carcinoma in Sprague Dawley Rat model Maheswari Chinnadhurai, Faisal Al-Otaibi, Kenneth Nelson, Geetha Kandasamy, Meena Shanmugam, R Venkatnarayanan Pharmacognosy Magazine 2019 15(66):396-401 Objectives: To evaluate the pretreatment with Madhuca longifolia leaves on 7, 12-Dimethylbenz(a)anthracene (DMBA)-induced mammary carcinoma in Sprague Dawley rat. Materials and Methods: Thirty female Sprague Dawley rats were divided into five groups, and each group is having six rats. Group I rats received vehicle (1 mL of emulsion of sunflower oil and physiological saline) subcutaneously and 1 mL of 2% dimethyl sulfoxide per orally. Groups II, III, IV, and V were induced mammary carcinogenesis by giving single dose of subcutaneous injection of 25 mg of DMBA. Group III, IV, and V rats were administered with MEML 100, 200 mg/kg and Vincristine 0.5 mg/kg dissolved in 1 ml of 2% dimethylsulfoxide given 1 week before the administration of the carcinogen, respectively, and continued for 16 weeks. At the end of experiment, the animals were sacrificed and biochemical estimations were done in all groups. Mammary tissues in all groups were dissected out and used for histopathological studies. Results: Oral administration of 200 mg/kg of MEML to DMBA-treated rats effectively prevented the tumor incidence, total number of tumors, and tumor volume and brought back the biochemical markers to normal, which was comparable with standard group. In lower dose 100 mg/kg, the effect was very less compared to normal and standard groups. Our data showed that MEML 200 mg/kg significantly restored the breast tissue biochemically and histologically which was comparable with standard. Conclusion: Our results concluded that the leaves of Madhuca longifolia may be used in the treatment of mammary carcinoma.

Exploration of antioxidant capacity of extracts of Perna viridis, a marine bivalve Venkateskumar Krishnamoorthy, Leow Yu Chuen, Vengadan Sivayogi, Sathasivam Kathiresan, Mohd Baidi Bahari, Gunasunderi Raju, Subramani Parasuraman Pharmacognosy Magazine 2019 15(66):402-409 Background: The biopotential of Malaysian green mussels (Perna viridis) has not been fully explored. The aim of the study is to screen the antioxidant capacity of extracts of Malaysian green mussels. Materials and Methods: Mussels were extracted by using solvents such as water, methanol, and ethanol, and methods such as microwave-assisted extraction (MAE) and animal tissue homogenization (ATH) were employed. The percentage yield, total protein content (TPC), total phenolic content, trace element analysis, and biochemical screening of extracts were carried out. The antioxidant capacity of the extracts was assessed by 2,2,-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay, 2,2'-azino-bis[3-ethylbenzothiazoline-6-sulfonic acid] (ABTS) assay, hydrogen peroxide scavenging assay, and ferric-reducing antioxidant power (FRAP) methods. Results: The yield of extracts prepared by MAE methods was higher than the extracts prepared by ATH methods. The total phenolic contents and TPC were higher in the extracts prepared by ATH method than those prepared by MAE methods. Biochemical screening results revealed the presence of alkaloids, phenolic compounds, and saponins. The percentage radical scavenging and inhibitory concentration50(IC50) values of all extracts were found to be lesser than the standard. Results showed that extracts by ATH methods displayed high antioxidant activity than extracts by MAE methods. The methanolic extracts of P. viridis showed better antioxidant capacity than other extracts in DPPH and ABTS assay methods, whereas the ethanolic extracts displayed high antioxidant activity than other extracts in hydrogen peroxide and FRAP assay methods. The antioxidant activity of the extracts could be attributed to the hydrogen-donating ability of bioactive peptides, phenolic compounds, alkaloids, reducing sugars, and trace elements present in such extracts. Conclusion: The results of the present study lay the platform to isolate newer antioxidants from Malaysian green mussels. It is concluded that extensive mechanistic studies are imminent to ascertain the molecular mechanism involved in the antioxidant activity of such extracts.

Comparative in vitro and in silico characterization of anticancer compounds piceatannol, biochanin-A and resveratrol on breast cancer cells Pardhasaradhi Mathi, Neelima Musunuru, Udayapriya Adurthi, Mahendran Botlagunta Pharmacognosy Magazine 2019 15(66):410-418 Background: Biochanin-A and Piceatannol are phytochemical constituents extracted from Sophora interrupta. Although both the compounds were isolated from a single plant, these compounds were not compared against anticancer activity. Objective: A systematic comparative analysis of biochanin-A, piceatannol, and resveratrol was performed to investigate cancer cell viability, motility, metabolic changes in Michigan Cancer Foundation-7 breast cancer cells, and structure compound interaction with the vascular endothelial growth factor (VEGF) receptors were studied. Materials and Methods: Cancer cell viability was studied using 3 (4, 5 dimethyl thiazol 2yl) 2, 5 diphenyltetrazo- lium bromide and acridine orange (AO)/ethidium bromide (EtBr) assay. The wound-healing assay was performed by measuring cell migration from the scratch area. Metabolic changes of the compounds in culture conditions were recorded using Fourier-transform infrared (FT-IR) spectroscopy. Molecular docking and dynamic simulations were performed using Schrödinger software. Results: Our results showed that the half-maximal growth inhibitory concentration for biochanin-A is 150 μM/ml and piceatannol and resveratrol showed 150 μM/ml, which is evident from the uptake of AO and EtBr dyes by live/dead cells. Moreover, drug-treated cells were unable to fill the cleared area from the scratch area, which suggests that all compounds effectively inhibit cell motility. FT-IR fingerprint showed a marked difference in the percentage of transition and dynamic structural changes between untreated and treated samples. Strong hydrogen-bond interaction with VEGF receptor-1 (VEGFR1) and VEGFR2 proteins and their interactions were stable throughout the simulation period. Moreover, these compounds inhibited sprouting of a new blood vessel from the chicken aorta and microvessels formation in the in ovo chorioallantoic membrane assay. Conclusion: Taken together, we conclude that anticancer and anti-angiogenic activity, structure-function relationship of biochanin-A is like well-known anticancer compound resveratrol and its metabolic product piceatannol in breast cancer cells.

Statistical approach towards optimization of extraction process of karanjin from Pongamia pinnata seeds Sartaj Akhtar Ansari, Lubna Abidin, Tinku Gupta, Mohd Mujeeb, Aftab Ahmad, Varish Ahmad, Showkat Rasool Mir Pharmacognosy Magazine 2019 15(66):419-425 Background: Pongamia pinnata is a valuable herb with loads of pharmacological activities owing to its phytochemical profile. Karanjin is one phytocompound found in the seeds of P. pinnata. Optimization of karanjin extraction process becomes a high priority task because of its high significance. Objective: Use of Box–Behnken design for optimization of extraction of karanjin from P. pinnata seeds. Materials and Methods: Design expert software was used for optimization purpose. Extraction temperature, extraction time and solvent-to-drug ratio were taken as input variables which affected the karanjin content. Quantification of karanjin in different extracts was done through high-performance liquid chromatography using methanol and water (80:20% v/v) as mobile phase. Results: Ultrasound-assisted extraction stood out to be the most efficient mode for extraction of karanjin using methanol as solvent. Extraction temperature of 57.85°C, extraction time of 25.45 min, and solvent-to-drug ratio of 86.4709% v/w were established as optimum conditions for extraction of karanjin from P. pinnata seeds. Under such extraction conditions, 8.33%w/w karanjin was extracted. Conclusion: From our study, it was concluded that non-thermal methods are a better choice for extraction of karanjin and methanol is the most efficient solvent for the same. All the three input variables significantly affected karanjin content which was confirmed by model fitting and analysis of regression coefficients. Our research shows the relevance of a statistical approach in phytocompound research area which makes the extraction process cheap and less laborious.

Metabolic Profile Elucidation of Ventilago calyculata Aqueous Extract Attenuating Sequelae of Aspirin Retarded Wound Healing Shweta Kumar, Rajesh Singh Pawar, Deepti Jain Pharmacognosy Magazine 2019 15(66):426-432 Background: Impaired wound healing due to aspirin is a common cause of delay in healing. Ventilago calyculata Tul. (Rhamnaceae) is used extensively in the Indian traditional medicines for skin problems. Objective: The objective of this work was to test the potential of formulations prepared from the extracts of V. calyculata against aspirin-retarded wound healing in rats and investigate the probable mechanism of action. Materials and Methods: The effect of topical administration of fractionates of V. calyculata against aspirin-delayed wound healing was assessed in Wistar albino rats. The chemo-profiling (liquid chromatography-mass spectroscopy [LC-MS]) of the extract with the most potent wound healing activity was carried out. Further, the mechanism of the most potent Ventilago calyculata aqueous extract (VCA) was determined by viability and plasma membrane integrity assays in H2O2-challenged wild type Saccharomyces cerevisiae BY4743) and knock-out strain (Δtrx2) strains. Results: The results of our investigations showed that in excision wound model; all formulations had statistically significant (P < 0.01) wound healing activity compared to the negative control. However, aqueous extract treatment (HF3) exhibited maximum activity, and the chemo-profiling of VCA by LCMS suggested that the potent activity may be due to individual and/or synergistic effect of the identified pharmacologically active phytoconstituents. The study on yeast indicates that VCA was able to act intracellularly also as it was able to overcome the growth inhibitory effect of the H2O2significantly (P < 0.01). Conclusion: We propose that treatment with HF3(VCA) is a therapeutically beneficial method of decelerating wound retardation caused by aspirin intake in patients on long-term aspirin therapy.

Ameliorative potential of a traditionally used plant Fraxinus micrantha against oxidative stress and paracetamol-induced hepatotoxicity Hasandeep Singh, Sarabjit Kaur, Saroj Arora, Balbir Singh Pharmacognosy Magazine 2019 15(66):433-439 Background: Liver disorders are one of the serious health issues. The treatment of liver disorders and their associated complications must be done with care due to the adverse effects of present-day medications. Therefore, there is an urgent need to develop effective and non-toxic herbal drugs for hepatoprotection. Objective: The objective of the present study is to investigate the hepatoprotective potential of traditionally used plant Fraxinus micrantha against paracetamol (PCM)-induced hepatotoxicity in rats. Methods: Hepatotoxicity was induced by a standardized single oral dose of paracetamol (PCM) at 3 g/kg body weight. Standard drug (silymarin 50 mg/kg) and test drugs (chloroform and methanol extracts) were administered orally to rats for 7 days. All the animals were sacrificed on the 8th day, and blood was withdrawn by retro-orbital vein puncture, collected in fresh centrifugation tubes, and centrifuged at 10,000 rpm for 10 min to separate serum for various estimations. Livers were isolated immediately for various biochemical and histopathological studies. Results: Treatment with chloroform extract (200, 400, and 800 mg/kg) and methanol extract (200 and 400 mg/kg) ameliorated the elevated levels of hepatic markers, but a significant reduction in these levels was observed by treatment with methanol extract at 800 mg/kg. In addition, treatment with chloroform and methanol extracts resulted in the amelioration of oxidative stress along with the histopathological changes. High-performance liquid chromatographic analysis of methanol extract of F. micrantha revealed the presence of various polyphenols such as rutin, naringenin, kaempferol, physicion, quercetin, and gallic acid. Conclusion: The results of the present study revealed that the observed hepatoprotective effect of methanol extract of F. micrantha may be due to the presence of various polyphenols and also provides pharmacological evidence for the use of F. micrantha bark in folk medicine for the treatment of liver diseases.

Antithrombocytopenic potential of bioactivity guided fractions of traditionally used Psidium guajava Linn. leaves in busulfan induced-thrombocytopenic rats Adil Ahmad, Mohammad Ibrahim, Karishma Chester, Washim Khan, Sayeed Ahmad, SH Ansari Pharmacognosy Magazine 2019 15(66):440-448 Background: Psidium guajava L. (family: Myrtaceae) is a traditionally used medicinal plant and possesses various therapeutic properties. This study was aimed to explore the antithrombocytopenic potential of bioactive fraction of P. guajava L. Materials and Methodology: Hydroalcoholic extract of dried leaves was prepared through soxhlation, and further, it was fractionated. Hydroalcoholic extract and its fractions were tested for antithrombocytopenic potential in busulfan-treated rats. Bioactive fractions were metabolically characterized through ultra-performance liquid chromatography–mass spectrometry (UPLC-MS) and. Results: A total of 30.3% w/w hydroalcoholic extract was obtained from which 5.7%, 4.2%, 8.6%, and 11.8% w/w of n-hexane, dichloromethane, n-butanol, and aqueous fractions were collected, respectively. n-butanol and aqueous fractions were identified as the potent bioactive fractions as antithrombocytopenic agents. Using toluene:ethyl acetate:formic acid (6:3:1 v/v/v) as a solvent system in TLC, 12 and 9 metabolites were separated in n-butanol and aqueous fractions, respectively, whereas 11 and 12 metabolites were tentatively identified through UPLC-MS in aqueous and n-butanol fractions, respectively. Five metabolites (showing m/z values of 303.05, 302.5, 313.2, 573.51, and 341.32 eluted at 2.80, 3.81, 7.94, 8.55 and 8.69 min, respectively) were found in both aqueous and n-butanol fractions. Conclusion: Chromatographically characterized n-butanol and aqueous fractions of hydroalcoholic extract of P. guajava increased platelet count, and it can be further explored for the development of new phytopharmaceutical.