TLC Bioautography Research Articles

α-Glucosidase Enzyme Inhibitory Activity of Extracts from the Fermentation of Endophytic Bacteria from Periwinkle (Catharanthus roseus (L.) G. Don) Leaves

Catharanthus roseus (L.) G. Don contain bioactive compounds that act as inhibitors against the α-glucosidase enzyme. This study aims to test the α-glucosidase enzyme inhibitor activity of fermented extract of endophytic bacteria from tapak dara leaves and identify the bacterial species. Endophytic bacteria were isolated using a direct planting method, resulting in eight isolates. The bacteria were fermented for 24-72 hours in nutrient broth media. Ethyl acetate was used to extract the fermented bacteria. Screening for α-glucosidase enzyme inhibitory activity was conducted using the TLC bioautography method. Four bacterial isolates (TD 1, TD 3, TD 4, TD 7) produced clear zones, qualitatively indicating that the isolates have inhibitory activity against the α-glucosidase enzyme. The IC50 results from the eight isolates showed that three isolates provided strong IC50 values: isolate TD 1 at 19.93 µg/ml, isolate TD 2 at 92.34 µg/ml, and isolate TD 4 at 13.94 µg/ml, compared to acarbose at 48.85 µg/ml). Molecular identification showed that isolate TD 1 was Pseudomonas oryzihabitans and isolate TD 4 was Staphylococcus warneri. This study concluded that endophytic bacteria from tapak dara leaves have the potential as a source of α-glucosidase enzyme inhibitor compounds for the development of antidiabetic drugs.

Exploring the Antibacterial and Antibiofilm Efficacy of Psammogeton biternatum Edgew and Identification of a Novel Quinoline Alkaloid using X-ray Crystallography.

The prevalence of resistance to harmful human pathogens is steadily rising, emphasizing the urgent need to identify novel antimicrobial compounds. For this purpose, plants stand out as a significant source of bioactives worthy of exploration. Among these, alkaloids, a vast and structurally diverse category of plant secondary metabolites, have emerged as a foundation for crucial antibacterial medications such as metronidazole and the quinolones. In the current work, the crude methanol leaf extract of Psammogeton biternatum Edgew collected from District Bannu, Pakistan, was subjected to TLC (indirect) bioautography and X-ray crystallography for the isolation of potential antibacterial agents. From the crude extract, a novel quinoline alkaloid called quinoline dione ((3R,3aS,5aR)-3,5a,9-trimethyl-3a,4,5,5a-tetrahydro-2H-isoxazolo[2,3-a] quinoline-2,8(3H)-dione (C14H17NO3)) was isolated. The crystal information (M = 247.296 g/mol) is as follows: orthorhombic, P212121, a = 7.7339(14) Å, b = 10.7254(19) Å, c = 15.730(2) Å, V = 1304.8(4) Å3, Z = 4, T = 296 K, μ(Mo Kα) = 0.088 mm-1, ρ calc = 1.259 g/cm3, 13928 reflections measured (5.86° ≤ 2Θ ≤ 51.98°), 2478 unique (R int = 0.1613, R σ = 0.1335). The final R 1 was 0.1098 (I ≥ 2u(I)), and wR 2 was 0.2183. The antibacterial activity for both crude extract of leaves and quinoline dione was determined by a well diffusion method. The quinoline dione alkaloid demonstrated excellent inhibition zones against methicillin-resistant Staphylococcus aureus (18 mm), Bacillus subtills (17 mm), Escherichia coli (20 mm), and Pseudomonas aeruginosa (23 mm) compared to the crude extract. The antibiofilm potential was recorded against Pseudomonas aeruginosa by the 96-well microtiter plate method. A dose-dependent biofilm inhibition response was recorded, which increased with the increase in concentration. Moreover, quinoline dione showed a greater antibiofilm effect as compared to the crude extract, which may be linked to the presence of a particular active functional group positioned on the compound isolated in its pure form. Through in silico studies, i.e., molecular docking, quinoline dione shows strong binding energies with the LasR transcriptional regulator (6MVN) at -9.3 and LasR transcriptional activator (3IX4) at -9.2 kcal/mol, as well as moderate affinities with other targets such as AHL synthase LasI (PDB ID 1RO5) and OprM channel (PDB ID 3D5K), indicating its potential as a quorum sensing inhibitor. Thus, the antibacterial and antibiofilm potential of quinoline dione was confirmed.

Investigation of in vitro biological activity of young Hordeum vulgare leaf in correlation with its bioactive compounds.

Young shoots of cereals are widely regarded as superfoods with health benefits attributed to their potential antioxidant activity and antioxidant-related effects (e.g. anticancer). The current study aimed to examine the chemical characteristics of Hordeum vulgare methanolic and aqueous extracts and assess their antioxidant activity using the DDPH and ORAC. Furthermore, the inhibitory effect of xanthine oxidase was screened. TLC bioautography was employed to determine the polarity of the compounds present in the extracts that exhibited the most potent free radical scavenging activity. Total flavonoid content of the methanolic and aqueous extracts was 0.14mgQE/g and 0.012mgQE/g, respectively. The antioxidant activity of the methanolic extract was found to be more potent, with a value of 0.97 ± 0.13mmolTE/g than the aqueous extract which had no activity. This study presents novel findings on the xanthine inhibitory activity of H. vulgare. The methanolic extract demonstrated moderate inhibition of xanthine oxidase with a value of 23.24%. The results of our study were compared with the phytochemical and pharmacological analysis of Triticum aestivum, and further comparison was made with the data reported in the literature. Inconsistencies were observed in the chemical and pharmacological properties of H. vulgare, which could be a result of using herbal material harvested in different vegetative phases and various methods used for extraction. The findings of our study indicate that the timing of the harvest and extraction method may play crucial role in attaining the optimal phytochemical composition of H. vulgare, hence enhancing its pharmacological activity.

The potency of yellow pigment extract from the marine bacterium Pseudomonas oryzihabitans SAB E-3 as an antioxidant agent

Abstract. Putri DA, Astuti RI, Wahyudi AT. 2024. The potency of yellow pigment extract from the marine bacterium Pseudomonas oryzihabitans SAB E-3 as an antioxidant agent. Biodiversitas 25: 2565-2575. Metabolic processes in cells can produce byproducts in the form of Reactive Oxygen Species (ROS). Excessive amounts of ROS are one of the causes of oxidative stress. Antioxidants are known to act as free radical-scavenging compounds. Pigment-producing bacteria associated with sponges have been widely reported to have antioxidant compounds. In this study, the marine bacterium Pseudomonas oryzihabitans SAB E-3 produced intracellular yellow pigments with potential antioxidant activity. Therefore, this study aimed to assess the in vitro antioxidant and mitochondrial activity at the cellular level and identify chemical compounds. The intracellular pigments produced by P. oryzihabitans SAB E-3 were extracted using methanol, acetone, chloroform, and ethyl acetate. The methanol pigment extract contained the highest number of phenolic and flavonoid compounds, which may have contributed to its antioxidant activity. Furthermore, the active fraction obtained from the bioautographic TLC and harvested using preparative TLC had stronger antioxidant activity against DPPH radicals (75.71 µg/mL) than the crude pigment extract. However, the ABTS radical-reducing activity was lower (226.59 µg/mL) than the crude pigment extract. The maximum wavelength of the pigment extract was 453 nm, indicating the presence of carotenoids. In addition, the lowest concentration of the crude pigment extract (50 µg/mL) and selected active fractions (18.75 µg/mL) increased yeast Schizosaccharomyces pombe ARC039 tolerance to oxidative stress and induced mitochondrial activity. Active fraction one extract analyzed using LC-MS/MS contained piceatannol, resveratrol, isorhapontigenin, isoliquiritigenin, liquiritin, and 2-Omethylisohemigossylic acid lactone, which may contribute to its antioxidant activity. Therefore, the yellow pigment extract and the active fraction produced by P. oryzihabitans SAB E-3 are potential candidates for natural antioxidant sources.

ANTIOXIDANT AND ANTIBACTERIAL ACTIVITIES OF ETHYL ACETATE EXTRACT OF ACTINOMYCETES ISOLATED FROM TERMITE NESTS

Actinomycetes are an important source for the discovery of bioactive secondary metabolite compounds. Over 10,000 bioactive metabolite compounds have been isolated from terrestrial actinomycetes with various biological activities, such as antibiotic, antiviral, anti-inflammatory, antitumor, anticancer, and antioxidant. One source of origin for actinomycetes is soil, including termite nests. Utilization of natural sources such as actinomycetes from termite nests as antioxidants and antibacterials can be an alternative source of production of new secondary metabolite compounds. This research aims to evaluate the antioxidant and antibacterial activities of actinomycete extracts from termite nests. Antioxidant activity using the DPPH free radical scavenging method, and antibacterial activity against S. aureus. Antioxidant and antibacterial activity were performed by using the TLC dot-blot, TLC bioautography, and microdilution methods to determine the inhibitory concentration of 50% (IC50), antioxidant activity index (AAI), and minimum inhibitory concentration (MIC). From the 33 extracts tested, 16 extracts showed antioxidant activity with IC50 values range of 76.64-126.22 µg/mL or AAI value > 0.05 (moderate), and 8 extracts had moderate antibacterial activity against S. aureus (MIC values range of 256-512 µg/mL). Future research for scaling-up of actinomycete culture, isolating of active compounds, determining of the chemical structure of active compounds, and further testing as antioxidants and antibacterials still need to be carried out.

Antibacterial activity of Shallots (Allium xwakegi Araki.) cultivars in Palu Valley against Salmonella Typhi ATCC 27870 through in vitro and in silico evaluation

Background and Objectives: Shallots, recognized for their minimal toxicity, cost-effectiveness, and widespread avail- ability, are increasingly considered a viable source of biological activity. This study evaluates the antibacterial efficacy of a specific shallot cultivar from Palu Valley, Indonesia, against Salmonella typhi, the pathogen responsible for typhoid fever. Materials and Methods: Utilizing thin-layer chromatography (TLC-bioautography) and gas chromatography-mass spec- troscopy (GC-MS), the study identifies active compounds in shallot ethanol extract and employs molecular docking to assess interactions between receptors and ligands. Results: Findings indicate significant antibacterial activity, with a notable inhibition zone diameter of 31.5 mm at spot Rf 0.28 in TLC bioautography and an optimum concentration of 2% yielding an average clear zone diameter of 28.27 mm in the agar diffusion test. GC-MS analysis reveals 41 compounds, predominantly dodecanoic acid and 1,2,3-propanetriyl ester. Ad- ditionally, molecular docking reveals the lowest binding affinity (-7.3 kcal/mol) for Ergost-8-En-3-Ol, 14-Methyl-, (3.Beta,5. Alpha.) against DNA gyrase. Conclusion: This study confirms Palu Valley shallot extract's potent antibacterial effect against Salmonella typhi, highlight- ing its therapeutic potential.

Rhizosphere actinobacteria isolated from Pometia pinnata and its antimicrobial activity

Abstract. Janatiningrum I, Zahra A, Fitriyanti, Anggia V. 2024. Rhizosphere actinobacteria isolated from Pometia pinnata and its antimicrobial activity. Biodiversitas 25: 1007-1014. Actinobacteria are currently the largest antibiotic-producing bacteria due to their extreme environments such as soil, roots and rhizosphere. The diversity of rhizosphere actinobacteria is affected by plant exudates. Pometia pinnata is a plant that has long been used for traditional medicine in Indonesia. This study was aimed at isolating and evaluating isolate and evaluate the actinobacteria rhizosphere of P. pinnata, to screen their antibacterial and antifungal properties. Isolation of actinobacteria was done by using serial dilution method and grown on media. About 34 actinobacteria were successfully isolated from the rhizosphere of P. pinnata and 50% of them showed index antimicrobial activity for Staphylococcus aureus ATCC 6538 (0.52), Streptococcus pneumoniae ATCC 1705 (2.87), and Escherichia coli ATCC 25922 (1.28). Several microbial isolated microbes (20.8%) were found capable in inhibiting fungal growth. The highest antibacterial activity against all bacteria was shown by RM12 isolate. The aqueous fraction of RM12 isolate demonstrated a significant inhibitory activity against S. aureus ATCC 6538 and E. coli ATCC 25922 at concentrations of 250 ppm, 500 ppm and 1000 ppm. TLC bioautography showed that aqueous fraction RM 12 do not belong to the polyphenol or phenol compound group. The RM12 was characterized by using 16S ribosomal RNA sequence found to belong to actinobacteria genera to Streptomyces bungoensis.

Evaluation of antioxidant and antacid activities of crude methanol and peptide extracts of Momordica charantia, Luffa cylindrica, and Jatropha curcas

Background: There are many uses for therapeutic-based compounds in the food and pharmaceutical industries, and natural products are one of the best sources of these compounds. Momordica charantia (Family Cucurbitaceae), Luffa cylindrica (Family Cucurbitaceae), and Jatropha curcas (Family Euphorbiacece) have been documented for various medicinal applications. Objective: This study aimed to assess the anti-ulcer and antioxidant properties of M. charantia, L. cylindrica and J. curcas extracts and their crude peptide. Methods: The plant samples were collected, identified, and authenticated at Forest Herbarium, Ibadan, Nigeria. The samples were allowed to air dry, were pulverized, and then were extracted using methanol. The peptidic principles were extracted using Dichloromethane: Chloroform: Water mixtures in the ratio 1:1:0.5 and concentrated in vacuo. The peptidic extracts were screened for peptide and peptide-related compounds by TLC bioautography using Ninhydrin, Pyridine, and modified G-250 reagents. The extracts were assessed for antioxidant (2,2-diphenyl-1-picrylhydrazyl, DPPH), free radical scavenging activity, total phenolic content (TPC), total flavonoid content (TFC), and antiulcer activity. The antiulcer activity was evaluated using the artificial gastric acid neutralizing activity. Results: It was observed that all M. charantia, L. cylindrica, and J. curcas and their crude peptide extracts contain phenols and flavonoids. The TPC values of the crude methanol extracts were greater than those of the corresponding peptide extracts for all three species. The TFC of M. charantia and L. cylindrica crude methanol extracts were lower than their crude peptide extracts, while the TFC of J. curcas crude methanol extract was higher than the TFC value for its crude peptide extract. The DPPH radical scavenging activity of the crude extracts and crude peptide extract of M. charantia, L. cylindrica, and J. curcas was low when compared to the standard (Gallic acid). The M. charantia crude peptide extract had the lowest IC50 value of 282.4 μg/mL, indicating that it had better scavenging efficacy compared to the other extracts that were examined, while the lowest IC50 value was obtained from J. curcas methanol extracts (411.7 μg/mL). The result from this study shows that M. charantia, L. cylindrica, and J. curcas and their crude peptide possess artificial gastric acid neutralizing activity, which implies that they can be used as antacids to reduce the acidity of the stomach. Hence, the plants were evaluated for their efficacy in managing peptic ulcers by their ability to reduce the acidity of the stomach by raising the pH of the stomach. This is the first report on the antacid potential of Momordica charantia, Luffa cylindrica, and Jatropha curcas in literature. Conclusions: The results from this study support the ethnomedicinal claims that M. charantia, L. cylindrica, and J. curcas can be used in the management of peptic ulcers, and these plants could serve as sources of lead compounds to produce natural antiulcer drugs. There were traces of the presence of linear peptides and cyclic peptides (cysteine-rich cyclotides). Keywords: Antiulcer, Antioxidant, Peptides, Momordica charantia, Luffa cylindrica, Jatropha curcas

Silver-doped graphite oxide composites used as antimicrobial agents against Staphylococcus aureus, Escherichia coli and Tatumella terrea evaluated by direct TLC bioautography

Silver-doped graphite oxide composites presented mixed Ag and Ag2O crystalline phases accompanied by C/O and ID/IG average ratios of 2.13 and 1.16, associated with a good oxidation degree and high structural disorder or defects in the carbon material, respectively. The low-silver-doped GrO composite showed homogenous silver particle dispersion and a low particle size distribution (96 nm). However, high-silver-doped GrO composites generated materials with high relative crystallinity, silver particle agglomeration, and Ag metal phase promotion. At 3 and 5 mg ml−1 per chromatographic plate, the silver-doped graphite oxide composites were tested by direct TLC bioautography against Staphylococcus aureus, Escherichia coli, and Tatumella terrea microorganisms, where the minimum inhibitory concentration was 3 mg ml−1 per chromatographic plate. At 3.0 mg ml−1 per chromatographic plate, high-silver-doped GrO composites exhibited a 39%, 3.2-fold, and 83% higher retention factor (R f ) compared with the composites with low-silver-doped GrO composites against S. aureus, E. coli, and T. terra microorganisms, respectively. However, both composites showed similar inhibition capacities at 5.0 mg ml−1 per chromatographic plate against the three microorganisms. This behavior may be associated with both composites reaching the threshold limit. In general, the silver acetate amount used in the silver-doped GrO composites influenced the dispersion, crystalline phase promotion, particle size distribution, and the silver particle release capacity, which modified the electrostatic adsorption type between the composites and the bacterial cell walls.

The antioxidant activity of endophytic fungal extract of nutmeg plant (Myristica fragrans Houtt.) from Cimande West Java, Indonesia

The bioproduction of bioactive metabolites from nutmeg endophytic fungi from Cimande, Wesh Java Indonesia was successful. In this research, bioactive antioxidant metabolites were extracted from nutmeg endophytic fungi. Endophytic fungi from nutmeg plant parts were grown on CMMA media. Each fungus was isolated by growing it on PDA media. A total of 21 endophytic fungi isolates were grown on PDB media before being extracted with ethyl acetate as a solvent. The morphology of 21 endophytic fungal isolates was identified based on morphological characteristics. There are 11 Phomopsis sp., 1 Fusarium oxysporum, 5 Colletotrichum sp., 1 Schizophyllum sp., 1 Dematiaceae sp., 1 Coelomycetes sp., and 1 Xylaria sp. The bioautographic TLC method and DPPH-free radical scavenging activity were used to determine antioxidant activity. The antioxidant activity analysis showed that 14 of the 21 endophytic fungus extracts were active. One nutmeg pericarp endophytic fungus showed strong antioxidant activity, with an IC50 of 30.0 g mL-1 and AAI of 1.04. Furthermore, the IC50 values for two fungal extracts from the leaf and pistil nutmeg were less than 100 g.mL-1. The three fungi with IC50 less than 100 g.mL-1 were identified as Phomopsis sp.

Exploring biotechnological approaches on in vitro callus induction and evaluation of biomedicinal properties of Cleome gynandra L.

In the present study, indirect organogenesis method was adopted for the production of callus from stem and leaf explants of Cleome gynandra using MS media. Production of callus using stem and leaf explants revealed that the highest dry biomass was achieved with the addition of IBA and 2,4-D in MS medium resulting in values of 180.33 and 190 mg, respectively. Antibacterial study inferred that methanol extract of C. gynandra callus exerted significantly pronounced inhibitory zones (8–16 mm) with minimal bacteriostatic (50–125 µg ml−1) and bactericidal (75–150 µg ml−1). Biofilm inhibitory activity revealed that callus extract surpassed leaf extract and showed > 80% biofilm inhibition, as confirmed by microscopic analysis. The callus extract also exhibited a relatively higher total phenolic content, while the leaf extract showed stronger antioxidant activity. Haemolysis analysis indicated that it was comparatively lower in callus extract and thus substantiating its biocompatibility. Phytochemical profile showed a meagre difference between the extracts. Cytotoxicity of the callus extracts against Artemia nauplii registered LC50 value of 53.71 µg ml−1. Partial purification and TLC bioautography evidenced Methanol: Acetone: Petroleum ether (8:1:1) as the most effective solvent system against the bacterial pathogens. GC-MS analysis substantiated benzofuran, sesquiterpenoids, fatty acids, diterpenes, hydrocarbons and alkanes as the major chemical constituents.

Antibacterial Activity of Ethanol Extract of Purple Kencana Leaves (Ruellia tuberosa L.) Using TLC-Bioautography and Agar Diffusion Methods

Tunjuk Langit Plant (Helminthostachys zeylanica (L.) Hook) contains a lot of secondary metabolite compounds, one of which is found in the root which contain the flavonoids ugonin and quercetin. The purpose of this study was to determine the antibacterial activity of compound 4 isolates IFATL-05 endophytic fungi from Tunjuk Langit (Helminthostachys zeylanica (L.) Hook) root against bacterial gastrointestinal tract infection. The results of purification of endophytic fungi with IFATL-05 code on macroscopic examination showed blackish-white fungus color, round with scalloped colony shapes, wavy edges and drop-like elevation angles. The process of separating the compounds using KLTP with n-hexane:ethyl acetate (1:1) as an eluent resulted in four bands of active compounds in UV lamps of 254 and 366 nm (TL-1, TL-2, TL-3 and TL-4). Test results for compound 4 isolates of IFATL-05 endophytic fungi from Tunjuk Langit root (Helminthostachys zeylanica (L.) Hook) using the Bioautographic TLC method using n-hexane:ethyl acetate (6:1) as eluent on Escherichia coli, Salmonella thypi, Shigella dysenteriae and Vibrio cholerae with Rf value = 0.90.

Molecular and Pharmacokinetic Aspects of the Acetylcholinesterase-Inhibitory Potential of the Oleanane-Type Triterpenes and Their Glycosides.

The acetylcholinesterase-inhibitory potential of the oleanane-type triterpenes and their glycosides from thebark of Terminalia arjuna (Combreatceae), i.e.,arjunic acid, arjunolic acid, arjungenin, arjunglucoside I, sericic acid and arjunetin, is presented. The studies are based on in silico pharmacokinetic and biomimetic studies, acetylcholinesterase (AChE)-inhibitory activity tests and molecular-docking research. Based on the calculated pharmacokinetic parameters, arjunetin and arjunglucoside I are indicated as able to cross the blood-brain barrier. The compounds of interest exhibit a marked acetylcholinesterase inhibitory potential, which was tested in the TLC bioautography test. The longest time to reach brain equilibrium is observed for both the arjunic and arjunolic acids and the shortest one for arjunetin. All of the compounds exhibit a high and relatively similar magnitude of binding energies, varying from ca. -15 to -13 kcal/mol. The superposition of the most favorable positions of all ligands interacting with AChE is analyzed. The correlation between the experimentally determined IC50 values and the steric parameters of the molecules is investigated. The inhibition of the enzyme by the analyzed compounds shows their potential to be used as cognition-enhancing agents. For the most potent compound (arjunglucoside I; ARG), the kinetics of AChE inhibition were tested. The Michaelis-Menten constant (Km) for the hydrolysis of the acetylthiocholine iodide substrate was calculated to be 0.011 mM.

Optimization of aqueous extraction of henna leaves (Lawsonia inermis L.) and evaluation of biological activity by HPLC-based profiling and molecular docking techniques

Due to the antimicrobial capabilities of henna leaf extract (Lawsonia inermis L.), this study intends to identify environmentally friendly ideal conditions for extracting bioactive compounds from henna leaf powder. For this purpose, the maceration method with a 100% aqueous solvent was selected for further analysis. Optimization of the extraction method was carried out by central composite design (CCD) protocol as a subset of Response Surface Methodology (RSM). The temperature of 40–90 °C, extraction time of 2–24 h, and the solvent-plant weight ratio ranging from 10 to 100 mL/mg were all considered independent variables. Also, the amount of lawsone, as an active component, and the total phenolic content were tracked as responses. Finally, a yield of 30.59% was achieved under the best extraction condition at 40 °C, 2 h, and a solvent-plant weight ratio of 1:100. The biological activity of an aqueous extract was evaluated using an HPLC-based activity profiling and TLC bioautography method. Peaks collected as fractions between 3 and 30 min were tested for antibacterial activity against Staphylococcus aureus ATCC 25923, and micro fractions from 5, 6, 7.5, 9, 10.5, 12, 13.5, 15, 19, and 30 min exhibited significant inhibitory effects. All spotted fractions on the TLC plate inhibited the growth of Escherichia coli ATCC 25922, except for micro fractions from 3, 18, and 21 min. Moreover, molecular docking analysis of quinones in the extract identified 3a, 4a-Dihydroxy-a-tetralone (C-2) as the most potent compound in interacting with bacteria sterol 14-demethylase protein, with docking scores of − 6.715 and − 7.196 kcal/mol, respectively.

Significance of Astragaloside IV from the Roots of Astragalus mongholicus as an Acetylcholinesterase Inhibitor-From the Computational and Biomimetic Analyses to the In Vitro and In Vivo Studies of Safety.

The main aim of the study was to assess the acetylcholinesterase-inhibitory potential of triterpenoid saponins (astragalosides) found in the roots of Astragalus mongholicus. For this purpose, the TLC bioautography method was applied and then the IC50 values were calculated for astragalosides II, III and IV (5.9 μM; 4.2 μM, and 4.0 μM, respectively). Moreover, molecular dynamics simulations were carried outto assess the affinity of the tested compounds for POPC and POPG-containing lipid bilayers, which in this case are the models of the blood-brain barrier (BBB). All determined free energy profiles confirmed that astragalosides exhibit great affinity for the lipid bilayer. A good correlation was obtained when comparing the logarithm of n-octanol/water partition coefficient (logPow) lipophilicity descriptor values with the smallest values of free energy of the determined 1D profiles. The affinity for the lipid bilayers changes in the same order as the corresponding logPow values, i.e.,: I > II > III~IV. All compounds exhibit a high and also relatively similar magnitude of binding energies, varying from ca. -55 to -51 kJ/mol. Apositive correlation between the experimentally-determined IC50 values and the theoretically-predicted binding energies expressed by the correlation coefficient value equal 0.956 was observed.

Phytochemical Screening of Nyctanthes arbor-tristis Plant Extracts and Their Antioxidant and Antibacterial Activity Analysis.

Nyctanthes arbor-tristis, alias "Vishnu Parijat," is a medicinal plant used to treat various inflammation-associated ailments and to combat innumerable infections in the traditional system of medicine. In the present study, we collected the samples of N. arbor-tristis from the lower Himalayan region of Uttarakhand, India, and carried out their molecular identification through DNA barcoding. To examine the antioxidant and antibacterial activities, we prepared the ethanolic and aqueous extracts (from flowers and leaves) and performed their phytochemical analysis by using different qualitative and quantitative approaches. The phytoextracts showed marked antioxidant potential, as revealed by a comprehensive set of assays. The ethanolic leaf extract showed marked antioxidant potential towards DPPH, ABTS, and NO scavenging (IC50 = 30.75 ± 0.006, 30.83 ± 0.002, and 51.23 ± 0.009 μg/mL, respectively). We used TLC-bioautography assay to characterize different antioxidant constituents (based on their Rf values) in the chromatograms ran under different mobile phases. For one of the prominent antioxidant spots in TLC bioautography, GC-MS analysis identified cis-9-hexadecenal and n-hexadecanoic acid as the major constituents. Furthermore, in antibacterial study, the ethanolic leaf extract showed marked activity against Aeromonas salmonicida (113.40 mg/mL of extract was equivalent to 100 μg/mL of kanamycin). In contrast, the ethanolic flower extract showed considerable antibacterial activity against Pseudomonas aeruginosa (125.85 mg/mL of extract ≡100 μg/mL of kanamycin). This study presents the phylogenetic account and unravels the antioxidant-related properties and antibacterial potential of N. arbor-tristis.

Antibacterial Activity, TLC-Bioautography Analysis, and Determination of Bioactive Components in Ethyl Acetate Extract of Robusta Coffee Leaf (Coffea canephora L.) From Aceh, Indonesia

Infectious diseases affect many people in the world, including Indonesia. One of the causes of infection is bacteria, including scherichia coli and Staphylococcus aureus. Robusta coffee (Coffea canephora L.) is one of the plants that is widely encountered on Indonesian plantations. The leaves are usually discarded after being trimmed to maximize the growth of coffee beans. The results of several studies show that robusta coffee leaves demonstrate antibacterial and antioxidant activity due to the presence of flavonoids, alkaloids, tannins, saponins, polyphenols, and chlorogenic acids. The aim of this research to investigate antibacterial activities, total flavonoid content and inspect the phytochemicals of the bioactive compounds as antibacterial with TLC Bioautography in the ethyl acetate extract of robusta coffee leaves. The identification of secondary metabolites was conducted by using the TLC-Bioautography method against Escherichia coli and Staphylococcus aureus. The determination of total flavonoid levels was carried out by using Spectrophotometry UV-Vis. Phytochemical testing showed that the extract contained alkaloids, steroids, phenolics, tannins, and flavonoids. The ethyl acetate extract inhibits the growth of Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922 (P=0.00) with 100% extract as the most potent. The results of TLC-Bioautography showed the presence of polyphenolic compounds (Rf 0.36) which was confirmed by a clear zone in the disc diffusion test. The total flavonoid content in robusta coffee leaves was 57.255±0.0348 mg QE/g extract. Ethyl acetate extract from coffee leaves (Coffea canephora L). inhibits the growth of S. aureus and E. coli due to the presence of polyphenolic compounds

Evaluation of hydromethanolic extract and aglycone fraction of Actiniopteris semiflabellata for antioxidant activity

Actiniopteris semiflabellata Pic-Ser grows in Yemen and is used by the local population for wounds and burns. The goal of the current study was to assess the Actiniopteris semiflabellata's chemical composition and antioxidant activity. Phenols, flavonoids, triterpenes and sterols were identified in the hydromethanolic extract and in the aglycone fraction, while carbohydrates were only found in the hydromethanolic extract. In the hydromethanolic extract and the aglycone fraction, the total phenolic content was evaluated as 17.73 mg/g and 7.22 mg/g respectively, while the total flavonoid content was 11.21 mg/g and 4.13 respectively. Studied extract and fraction exhibited antioxidant activity by TLC bioautography and DPPH radical scavenging activity assay. The IC50 values for the hydromethanolic extract and the aglycone fraction were 79.64 and 148.60 µg/ml respectively. These results confirm the traditional use and are useful for further study of this plant, in addition, it may be a good source of natural antioxidants.

Isolation and screening of Thermoactinomycetaceae family members as an extremophilic poor investigated and promising natural source of antimicrobial substances.

Recent evidences have shown that methicillin-resistant Staphylococcus aureus (MRSA) can cause severe infections and is resistant to almost all commercially available antibiotics. Therefore, screening unknown sources of biological compounds such as the Thermoactinomycetaceae family as extremophilic bacteria may be helpful to find new antimicrobial agents. Various samples were collected from different ecosystems, including desert, volcano, compost, and forest. They were cultured on Soil extract agar and Water agar. The antimicrobial activity of the isolates was evaluated using agar overlay and well diffusion methods. Members of the Thermoactinomycetaceae family were selected for further study: Their ability to grow at different temperatures, NaCl concentrations, and pH values, enzyme production ability, antimicrobial secondary screening, fractionation of their supernatants and so on. According to molecular identification of active isolates against MRSA, three strains, including Laceyella sacchari UTMC 2705, Thermoactinomyces sp. UTMC 2721, and Laceyella sp. UTMC 2731, belonged to Thermoactinomycetaceae were identified. The minimum inhibitory concentrations of their extracts were tested against some pathogenic bacteria, showing their antimicrobial activity with a broad spectrum. The results of TLC bioautography of the extracts showed that the most active fractions were semi-polar. Also, the results of HPLC analysis showed the existence of several UV-active compounds in their extracts. The present study highlighted the importance and potential of Thermoactinomycetaceae members as a less-known source of antibiotics against pathogenic bacteria.

In silico screening, TLC bioautography and in vivo studies of Zanthoxylum armatum DC. (Indian Prickly Ash) extract as a potential neuroprotective agent

Zanthoxylum armatum DC., a popular spice, and folklore medicine, is widely distributed throughout the Northeastern region of India and practiced by indigenous tribes as a remedy for digestive and nervous system disorders. The study aimed to assess the neuro-protective potentiality of Zanthoxylum armatum extract through different learning and memory paradigms against scopolamine-induced cognitive impairment. For the isolation of the compounds, coarsely powdered shade-dried fruits were extracted with ethanol by the cold maceration method. In silico studies were performed for the isolated compounds. TLC bioautographic enzyme assay was developed to screen acetylcholinesterase and butyrylcholinesterase inhibition by plant extract using donepezil, a known acetylcholinesterase inhibitor. Memory retention activities of ethanolic extract (100, 200, and 400 mg/kg; p.o.) were determined in-vivo , along with an estimation of whole-brain cholinesterase and antioxidant activity. Chromatographic analysis from chloroform fraction resulted in the isolation of three bio-active phytoconstituents, and structural elucidation identified them as β-sitosterol (ZA-01), Stigmasterol (ZA-02), and Lupeol (ZA-03). In silico molecular docking studies revealed stigmasterol has the minimum binding energy with acetylcholinesterase among all the isolated bioactive. The stability of the protein-ligand complex was found satisfactory after performing molecular dynamics simulation studies. Pre-treatment with extract significantly improved transfer latency in maze tests decreased rat brain acetylcholinesterase activity, and showed antioxidant potentiality against scopolamine-induced memory impairment. The combination of neuroprotective, anticholinesterase, and antioxidant effects shown by Z. armatum extract may be what makes it a nootropic drug. These are good signs for the search for natural nootropics from Z. armatum extract. Even so, more research with extracts and isolated compounds will be needed to make the traditional uses of the plant more scientifically sound. • Zanthoxylum armatum is evaluated for its potential as neuroprotective agent. • β-sitosterol, Stigmasterol and Lupeol were isolated from the plant extract. • In silico molecular docking and molecular dynamics simulations were performed. • TLC bioautography, in vivo animal tests showed good neuroprotective action of Zanthoxylum armatum extract.