Petroleum Ether and Chloroform Soluble Fractions of Whole Plant Extract of Acanthus ilicifolis Linn. Possesses Potential Analgesic and Antioxidant Activities.

Background: Medicinal plants are the major sources of traditional treatment of disease and new drug discovery due to major side effects of synthetic drug. Objectives: The aim of study was to evaluate analgesic and antioxidant activities of petroleum ether and chloroform soluble fractions of whole plant extract of Acanthus ilicifoius. Materials and Methods: The plant extract, standard diclofenac, and distilled water as control was administered post orally in Swiss albino mice and observe the analgesic activity by acetic acid (0.6%) induced writhing method. The plant extract was also subjected to perform reducing power assay, DPPH free radical scavenging activity, and FRAP assay to evaluate antioxidant activity. Results: The pet ether, and chloroform soluble fraction of plant extract revealed significant analgesic activity on mice model. Notably, the pet ether and chloroform fraction showed (40.14 ± 2.32) % and (40.12 ± 0.9) % analgesic inhibition, whereas standard diclofenac revealed (52.79 ± 2.62) % analgesic inhibition. In antioxidant activity assay, the plant extract showed mild to moderate antioxidant activities compare to standard ascorbic acid. Conclusion: From the results, it could be concluded that, the pet ether and chloroform fractions of whole plant extract of A. ilicifolius possesses potential analgesic and antioxidant properties

Molecular docking supported investigation of antioxidant, analgesic and diuretic effects of Costus speciosus rhizome

ABSTRACT. The aim of the current study was to analyze the polyphenols and determines the antioxidant, analgesic and diuretic properties of the methanolic extract of C.speciosus rhizome. DPPH and ferric reducing antioxidant power (FRAP) assays were used to determine the antioxidant activity. Acetic acid-induced writhing and formalin-induced licking experiments were used to assess the analgesic effect. The total phenolic, flavonoid and flavonol contents were found 51.73± 0.25 mg GAE/g dry weight, 3.41± 0.07mg QE/g dry weights and 44.19± 2.24 mg QE/g dry weight, respectively. The plant extract exhibited weak antioxidant activity in the DPPH and FRAP assays, with an IC50 value of 1699±62 μg/mL and an EC50 value of 125±2 μg/mL, respectively. The extract significantly reduced the number of writhes at both doses (200 and 400 mg/kg body weight) as compared to the control. The extract (400 mg/kg) also significantly reduced the percent inhibition of licking by 31.96 and 62.69% compared to the control in the early and late phase, respectively. Compared to the standard drug furosemide, the plant extract also showed a weak diuretic effect. The docking study supported the analgesic activity of rhizome extract. The potent analgesic activity of the plant extract justifies the traditional and medicinal aspects.   KEY WORDS: Costus speciosus, Analgesic activity, Diuretic effect, Molecular docking Bull. Chem. Soc. Ethiop. 2022, 36(3), 627-640.                                                                 DOI: https://dx.doi.org/10.4314/bcse.v36i3.12 &nbsp

Polyphenols and extracts from Zingiber roseum (Roxb.) Roscoe leaf mitigate pain, inflammation and pyrexia by inhibiting cyclooxygenase-2: an in vivo and in silico studies

Zingiber roseum (Roxb.) Roscoe, a perennial herb from the Zingiberaceae family, has a long history of traditional use in the treatment of several ailments including pain, inflammation, fever, cough, arthritis, skin diseases, and liver infections. This study sought to confirm the efficacy of Zingiber roseum (Roxb.) Roscoe leaves methanol extract (ZrlME) as reported in traditional usage by evaluating its analgesic, anti-inflammatory, and antipyretic capabilities. In addition, in silico molecular docking of the metabolites identified in ZrlME was studied to verify the experimental outcomes. ZrlME demonstrated strong dose-dependent analgesic efficacy against all analgesic tests. ZrlME (400 mg/kg) showed higher anti-inflammatory activity than the standard in the carrageenan-induced paw edema test model. A significant reduction of rectal temperature (3.97°F↓) was also recorded at the same dose of ZrLME after 24 h of treatment. Seven polyphenolic metabolites were identified and quantified by HPLC-DAD analysis, including 3, 4- dihydroxy benzoic acid, (-) epicatechin, rutin hydrate, p-coumaric acid, trans-ferulic acid, rosmarinic acid, and myricetin. Strong binding affinities (ranges from −5.8 to −8.5 Kcal/mol) between the aforesaid polyphenols and cyclooxygenase-2 were discovered. Moreover, molecular dynamics simulations (MDS) demonstrated that these polyphenols exhibit significant COX-2 inhibitory activity due to their high stability in the COX-2 active site. In computational prediction, the polyphenols were also found to be nontoxic, and a variety of biological activities, such as antioxidant, analgesic, anti-inflammatory, antipyretic, and hepatoprotective, were observed. The results of this study revealed that ZrlME possesses notable analgesic, anti-inflammatory, and antipyretic properties

Catecholamine homeostasis in <em>Tetrahymena</em> species and high throughput toxicity testing of selected chemicals and ultrafine particles.

96-well plate formate was used for the determination of toxicity to the CAs, 5-FU, PFCs and TiO2 nanoparticles in Tetrahymena species. Every compound of CAs depleted the total number of cells and consequently exhibited moderate toxicity. Dopamine strongly affected the natural noradrenaline synthesis. Moreover, this exogenous exposure of dopamine and L-DOPA at 0.12 ppm caused the formation of a novel metabolite by Tetrahymena. The molecular weight of this new metabolite was found to be 150 m/z. In addition, dopamine D1 type receptor was explored for the first time in T. thermophila by the use of fluorescent ligands. 5-FU exerted moderate toxicity to Scenedesmus spec. and Tetrahymena spec., and can be transferred from algae to protozoan via genetic material. PFCs caused slight depletion of cell numbers and produced moderate cell membrane damage. Tetrahymena cell endocytose ultrafine particles which later exocytose after aggregation to larger particles in the food vacuole

<em>Tetrahymena</em> spp. (Protista, Ciliophora) as test species in rapid multi-level ecotoxicity tests.

This review summarizes the application of Tetrahymena spp. in ecotoxicology, in order to promote a more integrated, multi-level ecotoxicological assessment approach regarding the effects of chemical stressors on several biological levels (from molecule to ecosystem). Such a multi-level testing approach in one species facilitates the establishment of missing causal relationships between biochemical responses and ecological effects. The review illustrates that Tetrahymena spp. represent excellent ecotoxicological test species due to their important role in the microbial foodweb, wide distribution and abundance, sequenced genome in T. thermophila, large background knowledge and scientific publications in cellular biology, ecology and ecotoxicology. Several bioassays have already been developed on different biological organisation levels, such as enzyme assays (biochemical level), behavioral tests (individual level), population growth tests (population level) and microcosms (community level). Moreover, specific mode-of-action based assays are available (e. g. genotoxicity), or are in development (e.g. endocrine disruption and neurotoxicity). Tetrahymena spp. combine traits of (1) a single cell, thus might replace or complement specific cell-line testing approaches, with traits of (2) a whole organism and population, thus allowing to study complete metabolic pathways and its consequences on population growth and genetic adaptation. Assays involving Tetrahymena spp. might easily be adapted for a rapid multi-level in situ or ex situ toxicity biosensor test system for ecologically relevant risk assessment

5-Fluorouracil accumulation in green microalgae and its biogenetic transfer into ciliate protozoan.

The study has demonstrated that anticancer drug 5-fluorouracil causes acute toxicity and interferes with the growth of green microalgae, Scenedesmus vacuolatus. It accumulates in microalgae biomass with bioaccumulation factor of 1.84 &times; 10(4) and further integrates into the DNA and RNA of microalgae. In addition, the labelled microalgae genome is transferred into protozoan Tetrahymena pyriformis on feeding and is retained in the food vacuoles of predator organisms. This biotransfer of labelled 5-fluorouracil via genomic material was evaluated using radioactivity in Tetrahymena cell pellets though radioactivity did not detect anticancer drug in the genome of the predator organism

Identification of dopamine receptor in tetrahymena thermophila by fluorescent ligands.

Two types dopamine receptor present in the cell membrane of vertebrates. But in this study D1 receptor was identified in the invertebrate ciliates protozoan, Tetrahymena thermop0hila by use of fluorescent ligands. D1 specific agonist SKF-38393 binds specifically to Tetrahymena. The specific binding of SKF-38393 was encountered by equimolar addition of D1 antagonist thus showed no binding of ligands. In addition, it was also proved that the D1 specific agonist did not cross bind with the D2 type receptor due to the equimolar addition of D2 selective antagonist spiperone. Interestingly this study also showed that the dopamine receptor present in the endoplasmic reticulum and endosomes of Tetrahymena as well as cell membrane which was revealed by laser scanning microscope. Therefore, this evidence supports the existence of a D1 receptor in the ciliate protozoan

Method for toxicity test of titanium dioxide nanoparticles in ciliate protozoan <em>Tetrahymena</em>.

Titanium dioxide (TiO2) nanoparticles have a high surface-to-mass ratio and rapidly aggregate in water causing great difficulties for toxicity test exposed to aquatic organisms or other cell lines. This study uses a cell viability kit for routine toxicity test of TiO2 as well as other nanoparticles which accumulate in the aquatic environment. Tetrahymena immediately endocytoses TiO2 nanoparticles and stores them in food vacuoles until the particles undergo exocytosis as larger aggregates. However, during the process of endocytosis and exocytosis, TiO2 particles interfere with cell growth and consequently induce acute toxicity. It exerted high cell growth inhibition at 20h incubation and induces significant cytotoxic effects. Surprisingly, the effect of TiO2 decreases at 40h incubation, due to the recovery of cell growth and reduction of the cytotoxicity of the particles

Growth inhibition and biodegradation of catecholamines in the ciliated protozoan Tetrahymena pyriformis.

A 96-well plate culture methodology for the unicellular eukaryote Tetrahymena pyriformis, strain GL was used for the determination of toxicity and metabolism of catecholamines. Catecholamines exhibited moderate acute toxicity to Tetrahymena cells where dopamine and L-DOPA showed higher toxic potential at EC10 (0.39 ppm and 0.63 ppm, respectively) and EC20 (1.1 ppm and 1.0 ppm respectively) after 48 h exposure. All tested catecholamines were highly degradable in the PPY-medium due to the oxidizing environment during incubation. Also the catecholamines were naturally synthesized and released by Tetrahymena cells into the culture medium and increasingly accumulated with time where noradrenalin exhibited the highest degree of accumulation. However, the exogenous exposure of catecholamines to the cells caused the depletion of natural noradrenalin synthesis even with the addition of very low physiological concentration (0.12 ppm). Dopamine caused the higher effect on inhibiting noradrenalin synthesis. Treatment with a higher concentration (8.0 ppm) of dopamine in 96-well plates caused strong excitation of the cells and ascertained a new metabolite in vivo while the other representative catecholamines were not responsible for the production of this metabolite. This dopamine metabolite is relatively non-polar as compared to noradrenalin, adrenaline and dopamine and eluting later through the reverse phase C-18 column

PHYTOCHEMICAL SCREENING, PLANT GROWTH INHIBITION, AND ANTIMICROBIAL ACTIVITY STUDIES OF XYLOCARPUS GRANATUM

Phytochemical analysis of the methanolic extract of Xylocarpus granatum Koen indicated the presence of carbohydrates, saponins, tannins and flavonoid types of compounds. Alkaloids and glycosides were found to be absent from the extract. The primary methanolic extract exhibited a potent growth inhibitory effect. Inhibition of both the rootlet and shoot showed a dosedependent response. The residual methanolic extract also has a growth inhibitory effect. Bothmethanolic extracts have a greater inhibitory effect on rootlet growth than shoot growth. The residual methanolic extract has a lesser inhibitory effect than the primary methanolic extract.Removal of the non-polar compounds (by n-hexane and chloroform) from the primary methanolic extract reduced the inhibitory activity on both the rootlet and shoot growth, which suggests that the non-polar fractions may contain growth inhibitory principles. The primary extract demonstrated antibacterial activity against the gram positive bacteria Staphylococcus aureus and Bacillus subtilis and the gram negative bacteria Proteus vulgaris. The primary methanolic extract was found to be inactive against Escherichia coli andPseudomonas aeruginosa. The primary methanolic extract was more active against grampositive bacteria than gram-negative bacteria. The residual methanolic extract was also found to be inactive against all the tested microorganisms