Activity Of Ethyl Acetate Fraction Research Articles

Bioassay-guided isolation and identification of antimutagenic compounds from Morina coulteriana and evaluation of its therapeutic potential.

BackgroundIn the expanding field of cancer research, antimutagenic studies hold a promising ground. In recent times these studies have shown a shift towards the diverse flora of nature and its potential to treat cancer. Genus Morina has been extensively used in Chinese, Tibetan, and Indian traditional medicine to cure numerous diseases. The plant Morina coulteriana has been traditionally used to cure eye diseases. Nevertheless, it has been explored for the first time for its antimutagenic/antigenotoxic potential and phytochemical profile. PurposeThe present study aimed to determine the antimutagenic/antigenotoxic potential of Morina coulteriana and the bioassay-guided identification of its bioactive constituents. MethodsThe methanol crude extract and its subsequent fractions were tested for antimutagenic/antigenotoxic activity in vitro (Ames assay) and in vivo (Chromosomal aberration assay in Balb/c mice). The active (ethyl acetate) fraction was further investigated to isolate and identify its bioactive compounds utilizing chromatographic purification and detailed spectroscopic techniques respectively. ResultsThe results revealed that the plant and its fractions were neither mutagenic nor toxic. The ethyl acetate fraction showed significant antimutagenic/antigenotoxic activity (66.13%) in vitro and (88.15%) in vivo. Further, four identified major compounds which were also tested in vitro. Our results revealed that the plant is a potent antimutagen. It worked in a dose-dependent manner and its administration decreased the mutagenic load on the cells both in vitro and in vivo experiments. The 2 isolated compounds viz.; 4hydroxyacetophenone, a phenol and picein, a phenolic glycoside were reported for the first time from the genus Morina. ConclusionThe findings of the present investigation clearly show that the plant M. coulteriana Royle has a strong antimutagenic potential. Its administration decreased the mutagenic load on the cells both in vitro and in vivo experiments Thus the present study provides a platform for the exploration of new plants with potential antimutagenic properties.

Bioactivity Profiling and Phytochemical Analysis of Carissa carandas Extracts: Antioxidant, Anti-Inflammatory, and Anti-Urinary Tract Infection Properties.

Carissa carandas L. (Apocynaceae) is widely distributed in tropical and subtropical regions of Asia including Pakistan, India, Afghanistan, and Sri Lanka. C. carandas is considered as an integral component of traditional medicinal systems to combat several health ailments. The present study aimed to assess this plant's phytochemical contents and biological potential by performing sequential extraction, adopting a bioassay-guided approach. C. carandas powder was extracted with n-hexane to remove fatty substances and then residues were sequentially extracted with dichloromethane, methanol, and 50% methanol. All the sequential crude extracts were evaluated for phytochemical contents (total phenolics, flavonoids, and anthocyanins), in vitro antioxidant activity (FRAP, DPPH), in vitro anti-inflammatory activity (serum and egg albumin denaturation), in vivo anti-inflammatory activity (carrageenan- and formaldehyde-induced paw edema), and in vitro antimicrobial activity. Active crude extract was then partitioned using the liquid-liquid separation method followed by further separation of the active fraction by RP-HPLC. The active fraction was then subjected to LC-ESI-MS/MS analysis for tentative identification of bioactive metabolites responsible for its bioactive properties, followed by HPLC quantification. The analysis revealed methanol extract to have more phytochemical contents, radical scavenging properties, reduced inflammation in both models (in vitro and in vivo), and antimicrobial properties against urinary tract infection-causing agents as compared to dichloromethane and 50% methanol extracts. The ethyl acetate fraction obtained after liquid-liquid partitioning (LLP) of the active methanol extract exhibited more activity as compared to C. carandas methanol extract. RP-HPLC sub-fractionation yielded seven sub-fractions, but a slight decrease in biological potential was recorded. Therefore, LLP fraction B was subjected to further analysis. LC-ESI-MS/MS analysis led to the tentative identification of phenolic acids (chlorogenic acid, quinic acid), flavonoids (quercetin), and anthocyanins (peonidin-3-arabinoside, delphinidin-3-galactoside, delphinidin-3-rutinoside) in the active LLP ethyl acetate fraction. Chlorogenic acid, ellagic acid, and quinic acid were quantified as 17.6 µg/mg, 5.90 µg/mg, and 3.30 µg/mg, respectively, on a dry weight basis by HPLC. C. carandas may be considered a promising therapeutic plant, and the results of the current study provide more evidence to support the assertions made in ancient medical traditions. These findings highlight its promising applications in health, medicine, cosmetics, preservatives, and as a natural coloring agent.

Phytochemical Analysis and In Vivo Antimalarial Activities of Ethyl Acetate Fraction of Spilanthes filicaulis on Mice Subjected to Plasmodium berghei.

Background and Objectives: Malaria airs a life-threatening risk in Tropical African countries, stemming from infection by Plasmodium species. This region is richly endowed by nature with a wealth of diverse and largely unexplored plants that hold the potential for managing this protozoan parasite. The currently accessible over-the-counter drugs for disease management often present affordability challenges for the average person, exacerbated by the parasite's increasing resistance to them. This study investigated the phytoconstituents present in the ethyl acetate fraction of Spilanthes filicaulis (EFSF) and explored the antimalarial effects of EFSF on mice infected with Plasmodium berghei. Methods: Standard methods and gas chromatography-mass spectrometry (GC-MS) were used to identify phytoconstituents. Chloroquine phosphate-sensitive P. berghei (NK-65) was intraperitoneally inoculated into Swiss mice. The invivo antimalarial activity of EFSF was assessed at dose levels of 250, 500, and 750 mg/kg, using 4-day suppressive and curative antimalarial models. Parameters evaluated in the inoculated mice included rectal temperature (RT), body weight (BW), packed cell volume (PCV), level of parasitemia, and mean survival time (MST). Results: Steroids, alkaloids, flavonoids, tannins, saponins, terpenoids, and cardiac glycosides were the identified phytochemicals present in EFSF, and GC-MS alongside reveals the presence of 20 bioactive compounds predominantly fatty acids and alcohol esters. Significant prevention of reductions in RT, BW, and PCV was observed in the EFSF-treated groups dose dependently relative to the untreated group. In addition, EFSF-treated groups significantly (p < 0.05) suppressed parasitemia and exhibited chemosuppression of 79.46% and 77.38% in 4-day suppressive, whereas suppression of 59.74% and 58.66% in curative treatment, respectively, at 500 and 750 mg/kg thus consequently extending the MST of infected treated mice compared with the untreated group. Interpretation and Conclusion: Put together, the EFSF exhibited enhanced antimalarial efficacy against mice infected with P. berghei thus affirming that plants still maintain lead way as a potential source of novel antimalarial remedies.

In vitro Antibacterial and in vivo Antinociceptive Activities of Ethyl Acetate Fraction of Dipterocarpus turbinatus Leaves

The study was aimed to assess the antibacterial and analgesic properties of the ethyl acetate extract from the leaves of Dipterocarpus turbinatus. The antibacterial effectiveness of the extract obtained from the leaves of D. turbinatus was evaluated using the disc diffusion method against both positive and negative Gram strains. Furthermore, the analgesic qualities were assessed by doing the formalin-incited paw licking test in mice model. The ethyl acetate fraction demonstrated significant antibacterial activity against all tested microorganisms (Lactobacillus casei, Corynebacterium sp., Bacillus cereus, Staphylococcus aureus, Escherichia coli, Bacillus azotoformans, and Salmonella typhi) at concentrations of 0.1 mg/disc, 0.3 mg/disc, and 0.5 mg/disc using the disc diffusion method. The extract's zone of inhibition measured between 12 and 25 mm was less than the Kanamycin's diameter of 32–36 mm. The analgesic activity of the substance was evaluated in Swiss Albino mice using the tail immersion and formalin test procedures, revealing a modest level of analgesic activity. In the formalin test, the extract had a strong dose-dependent impact at dosages of 200 mg/kg and 400 mg/kg, as shown by a larger inhibition percentage compared to the standard. This result was statistically significant (***p&lt;0.001) when compared to the control group. During the tail immersion test, the extract showed a significant impact at dose 400 mg/kg (**p&lt;0.01). The plant extract has a modest level of antibacterial and analgesic properties. Bangladesh Pharmaceutical Journal 27(2): 207-214, 2024 (July)

Phytochemical analysis, in-vitro and in-silico study of antiproliferative activity of ethyl acetate fraction of Launaea cornuta (Hochst. ex Oliv. & Hiern) C. Jeffrey against human cervical cancer cell line.

Introduction: Cervical cancer is one of the leading causes of death among women globally due to the limitation of current treatment methods and their associated adverse side effects. Launaea cornuta is used as traditional medicine for the treatment of a variety of diseases including cancer. However, there is no scientific validation on the antiproliferative activity of L. cornuta against cervical cancer. Objective: This study aimed to evaluate the selective antiproliferative, cytotoxic and antimigratory effects of L. cornuta and to explore its therapeutical mechanisms in human cervical cancer cell lines (HeLa-229) through a network analysis approach. Materials and methods: The cytotoxic effect of L. cornuta ethyl acetate fraction on the proliferation of cervical cancer cells was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) bioassay and the antimigratory effect was assessed by wound healing assays. Compounds were analysed using the qualitative colour method and gas chromatography-mass spectroscopy (GC-MS). Subsequently, bioinformatic analyses, including the protein-protein interaction (PPI) network analysis, Gene Ontology (GO), and Kyoto Encyclopaedia of Genes and Genomes (KEGG) analysis, were performed to screen for potential anticervical cancer therapeutic target genes of L. cornuta. Molecular docking (MD) was performed to predict and understand the molecular interactions of the ligands against cervical cancer. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to validate the network analysis results. Results: L. cornuta ethyl acetate fraction exhibited a remarkable cytotoxic effect on HeLa-229 proliferation (IC50 of 20.56 ± 2.83μg/mL) with a selectivity index (SI) of 2.36 with minimal cytotoxicity on non-cancerous cells (Vero-CCL 81 (IC50 of 48.83 ± 23.02). The preliminary screening revealed the presence of glycosides, phenols, saponins, terpenoids, quinones, and tannins. Thirteen compounds were also identified by GC-MS analysis. 124 potential target genes associated with the effect of L. cornuta ethyl acetate fraction on human cervical cancer were obtained, including AKT1, MDM2, CDK2, MCL1 and MTOR were identified among the top hub genes and PI3K/Akt1, Ras/MAPK, FoxO and EGFR signalling pathways were identified as the significantly enriched pathways. Molecular docking results showed that stigmasteryl methyl ether had a good binding affinity against CDK2, ATK1, BCL2, MDM2, and Casp9, with binding energy ranging from -7.0 to -12.6kcal/mol. Tremulone showed a good binding affinity against TP53 and P21 with -7.0 and -8.0kcal/mol, respectively. This suggests a stable molecular interaction of the ethyl acetate fraction of L. cornuta compounds with the selected target genes for cervical cancer. Furthermore, RT-qPCR analysis revealed that CDK2, MDM2 and BCL2 were downregulated, and Casp9 and P21 were upregulated in HeLa-229 cells treated with L. cornuta compared to the negative control (DMSO 0.2%). Conclusion: The findings indicate that L. cornuta ethyl acetate fraction phytochemicals modulates various molecular targets and pathways to exhibit selective antiproliferative and cytotoxic effects against HeLa-229 cells. This study lays a foundation for further research to develop innovative clinical anticervical cancer agents.

In Vitro Cytotoxic Activity and In Vivo Antiproliferative Activity of Ethyl Acetate Fraction of Bunium bulbocastanum Seed Against Prostatic Neoplasia

AbstractSeveral in vitro investigations of the therapeutic characteristics of B. bulbocastanum have shown that it has cytotoxic, antifungal, and antibacterial effects. It also exhibits antioxidant and anticancer effects. When the ethyl acetate fraction of B. bulbocastanum was examined for its phytochemical composition, it was found to be rich in phenolic compounds and had significant cytotoxic effects on PC-3 cell lines. The prostate length-to-weight ratio was significantly higher in vivo in the ethyl acetate fraction-treated group. Compared to the disease control group, histopathological examination of the ethyl acetate-treated group revealed a reduction in inflammation and malignant lesions, confirming its antiproliferative efficacy. According to serum biochemistry, acid phosphatase and PSA levels in the ethyl acetate fraction treatment group were significantly lower than those in the disease control group. When compared to the disease control group, malondialdehyde levels in the ethyl acetate fraction treatment group were likewise reduced dramatically. However, in the group treated with the ethyl acetate fraction, glutathione levels increased considerably. The ethyl acetate fraction of B. bulbocastanum may have cytotoxic and antiproliferative potential, both in vitro and in vivo.

Further Polycyclic Quinones of Micromonospora sp.

The crude acetone extract of a marine Micromonospora sp. strain associated with Eudistoma vannnamei was fractioned with hexane and ethyl acetate. The crude extract and both soluble fractions were assayed against several bacteria strains. The new polycyclic quinones 12-hydroxy-9-propyltetracene-6,1-dione (1), 5,12-dihydroxy-4-methoxy-9-propyltetracene-5,12-dione (2), and 4,6-dihydroxy-3-methoxycarbonyl- methyl-6a-(oxobutyl)-5,12-anthraquinone (3), along with the known 4,6-dihydroxy-3-methoxycarbonyl-methyl-6a-(oxo-3-methyl-butyl)-5,12-anthraquinone (4) and 4,6-dihydroxy-3-methoxycarbonyl-methyl-6a-(oxopentyl)-5,12-anthraquinone (5) were isolated from the hexane-soluble fraction, while from the active ethyl acetate fraction were isolated the known 4,6,11-trihydroxy-9-propyltetracene-5,12-dione (6), 4-methoxy-9-propyltetracene-6,11-dione (7), 7,8,9,10-tetrahydro-9-hydroxy-4-methoxy-9-propyltetracene-6,11-dione (8), and 10β-carbomethoxy-7,8,9,10-tetrahydro-4,6,7α,9α,11-pentahydroxy-9-propyltetracene-5,12-dione (9). The structures of the new compounds were established by interpretation of HRMS and NMR techniques. A study of molecular docking was performed with the compounds from the active ethyl acetate fraction to correlate tentatively with the antimicrobial activity. Molecular docking, RMSD, RMSF, and MM-GBSA evaluations were performed to investigate the inhibitory activity of 6-8 against the protein PDB-codex 1MWT, being considered a promising target for studying drug development responsible for inhibiting replication of Staphylococcus aureus. Penicillin G was used as the standard inhibitory. Anthracyclinones 6-8 were the best hydrolase inhibitor with affinity energy -8.1 to -7.9 kcal/mol compared to penicillin G, which presented -6.9 kcal/mol. Both 8 and 7 present potent inhibitory effects against hydrolase through molecular dynamics simulation and exhibit favorable drug-like properties, promising new hydrolase blockers to fight bacterial infections from Staphylococcus aureus.

Aaptamine-rich Fraction from the Indonesian Marine Sponge, Aaptos suberitoides, Exhibits a Cytotoxic Effect on DLD-1 Colorectal Cancer Cells.

This study aimed to investigate the cytotoxicity effect of the ethyl acetate extract of Aaptos suberitoides on colorectal cancer cells (DLD-1) and murine fibroblast cells (NIH-3T3). A. suberitoides was collected from Putus Island, Bunaken National Park, North Sulawesi, Indonesia, and was processed with maceration and ethyl acetate extraction. The sponge extract was characterized based on Thin Layer Chromatography (TLC) and then identified by using LCMS/MS analysis. DLD-1 and NIH-3T3 cells were treated with the ethyl acetate extract and then followed by 3- [4, 5-dimethylthiazol-2-yl] -2.5 diphenyl tetrazolium bromide (MTT) assay to assess their cytotoxicity effect. LCMS/MS analysis showed that the most abundant compounds in this extract were identified as aaptamine (1). Furthermore, this study revealed that the active ethyl acetate fraction of A. suberitoides has cytotoxic effects in colorectal cancer DLD-1 cells with an IC50 value of 9.597 µg/mL, higher than NIH-3T3 cells with an IC50 value of 12.23 µg/mL Thus, the active ethyl acetate fraction of A. suberitoides is considered more toxic to cancer cells than normal cells. This study provides the first evidence to support the role of the ethyl acetate extract of A. suberitoides sponge extracts to be developed as a colorectal anticancer agent.

Anticancer Activity of Ethyl Acetate Fraction and Ethanol Leaf Extract of Olax subscorpioidea against DMBA-Induced Female Rats

Anticancer Activity of Ethyl Acetate Fraction and Ethanol Leaf Extract of Olax subscorpioidea against DMBA-Induced Female Rats

Evaluation of the anxiolytic activity of ethyl acetate fraction of Calotropis gigantea leaves in rats

Evaluation of the anxiolytic activity of ethyl acetate fraction of Calotropis gigantea leaves in rats

Free radical scavenging polyphenols isolated from Phyllanthus niruri L. ameliorates hyperglycemia via SIRT1 induction and GLUT4 translocation in in vitro and in vivo models

Type 2 diabetes milletus (T2DM) is a complex multifaceted disorder characterized by insulin resistance in skeletal muscle. Phyllanthus niruri L. is well reported sub-tropical therapeutically beneficial ayurvedic medicinal plant from Euphorbiaceae family used in various body ailments such as metabolic disorder including diabetes. The present study emphasizes on the therapeutic potential of Phyllanthus niruri L. and its phytochemical(s) against insulin resistance conditions and impaired antioxidant activity thereby aiding as an anti-hyperglycemic agent in targeting T2DM. Three compounds were isolated from the most active ethyl acetate fraction namely compound 1 as 1-O-galloyl-6-O-luteoyl-β-D-glucoside, compound 2 as brevifolincarboxylic acid and compound 3 as ricinoleic acid. Compounds 1 and 2, the two polyphenols enhanced the uptake of glucose and inhibited ROS levels in palmitate induced C2C12 myotubes. PNEAF showed the potent enhancement of glucose uptake in palmitate-induced insulin resistance condition in C2C12 myotubes and significant ROS inhibition was observed in skeletal muscle cell line. PNEAF treated IR C2C12 myotubes and STZ induced Wistar rats elevated SIRT1, PGC1-α signaling cascade through phosphorylation of AMPK and GLUT4 translocation resulting in insulin sensitization. Our study revealed an insight into the efficacy of marker compounds isolated from P. niruri and its enriched ethyl acetate fraction as ROS scavenging agent and helps in attenuating insulin resistance condition in C2C12 myotubes as well as in STZ induced Wistar rat by restoring glucose metabolism. Overall, this study can provide prospects for the marker-assisted development of P. niruri as a phytopharmaceutical drug for the insulin resistance related diabetic complications.

Antibacterial and Anti Diabetic Activities of Ethyl Acetate Fractions of Lactobacillus Isolated from Cow and Goat Milk

ABSTRACT: Antibiotics are one of our most important weapons in fighting with bacterial infections and diabetics. Drugs were derived from natural sources that play an important role in the treatment and prevention of human diseases. Traditional fermented foods containing lactic acid bacteria such as milk, yogurt, curd, etc., have historically been consumed by human. Lactobacilli are well known friendly bacteria for their probiotic activities against pathogens. In this investigation cow and goat milk samples were selected and collected for the study. The gram stain was used for morphological identification of Lactobacillus sp. in the two samples. The two isolated Lactobacillus sp. from goat and cow milk samples were subjected to the extraction of secondary metabolites using the solvent ethyl acetate. The two extracts of Lactobacillus spp. those were isolated from cow and goat milk samples, showed antibacterial activity against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Pseudomonas aeruginosa. The zone of inhibition was assessed by mm. The good results of zones were observed at 1000 and 2000 µg in cow milk and 2000 µg in goat milk extraction. These metabolites were also showed good results in antidiabetic activity using the alpha amylase inhibition activity. The IC50 value has showed moderate activity in the two crude extractions when compared to acarbose standard drug. Therefore, in this investigation the Lactobacillus sp. producing secondary metabolites was good source for the activity of antibacterial and antidiabetic activity.

Phytochemical profile, acute oral toxicity, antioxidant, and antispasmodic effects of ethyl acetate and aqueous residual fractions of Diospyros mespiliformis Hochst. ex A. DC (Ebenaceae) leaves on isolated duodenum of rat

Introduction: Diospyros mespiliformis Hochst. ex A. Rich (Ebenaceae) is a nutritional, artisanal and medicinal plant. It is used in alternative medicine in Burkina Faso for the treatment of conjunctivitis, menorrhagia, dysentery, and especially diarrhea. Aims: Our study aimed to evaluate the chemical profile, the antioxidant and anti-inflammatory activities, the safety of use, and the spasmolytic effects of the fractions obtained from the aqueous decoction of the leaves of Diospyros mespiliformis. Methods: Phytochemical screening by HPTLC and the determination of compounds of interest were carried out. The antioxidant activity was evaluated according to the ABTS, DPPH, FRAP, and LPO methods. The inhibitory activity of Phospholipase A2 and 15-lipoxygenase was evaluated. Acute oral toxicity was carried out on female mice (NMRI). The ex vivo spasmolytic effect of the fractions was tested on isolated rat duodenum using ACh and BaCl2 as contracting agents. Results: At the end of these tests, the fractions contain flavonoids, tannins, sterols, triterpenes, and saponosides. The content of total phenolics was respectively for the ethyl acetate fraction (EAF) and the residual aqueous fraction (RAF) 84.15±1.73 mg EAT/g and 89.67±2.35 mg EAT/g. That of flavonoids was respectively 45.91 ± 0.98 mg EQ/g and 10.46±0.28 mg EQ/g for the two fractions. The 50% inhibitory concentrations (IC50) of EAF for the ABTS, DPPH, and FRAP tests were 2.26±0.16 μg/mL, 22.34±7.23 μg/mL and 1136.25±0.90 mol EAA/g respectively, and an inhibition % of 43.80 ± 6.31% for LPO. For RAF, the IC50 were in the same range as EAF of 16.82±0.23 μg/mL, 18.58±2.91 μg/mL, 1138.4±1.27 mol EAA/g and an inhibition of 45.14±10.35% for LPO. The fractions had an inhibitory effect on phospholipase A2 compared to Betamethasone. Finally, the most active EAF caused a spasmolytic effect with Emax of 87.4±15.71% and 90.40±7.84%, respectively, during contractions induced by BaCl2 and ACh. Conclusion: Finally, this work provided scientific data and could justify the use of D. mespiliformis leaves in the treatment of diarrhea.&#x0D; Keywords: Diospyros mespiliformis, Leaves, Antioxidants, Anti-inflammatory, Safety of use, Spasmolytic

Uric acid–reducing efficacy of Plectranthus amboinicus (Lour.) Spreng in mice with potassium oxonate-induced hyperuricemia

Plectranthus amboinicus (Lour.) Spreng is a type of Lamiaceae perennial herb that is widely used in folk treatments for respiratory and skin diseases. Notably, research on the hypouricemic effect of P. amboinicus is limited. This study investigated the uric acid–reducing efficacy of P. amboinicus in mice with potassium oxonate-induced hyperuricemia by performing an in vitro xanthine-oxidase-inhibition-activity-guided test of P. amboinicus extract (PAE) and its fractions. The ethyl acetate fraction (EAF) of PAE exhibited optimal efficacy in terms of the phenolic content, DPPH scavenging activity, and xanthine oxidase inhibition. We further demonstrated the uric acid–reducing activity of EAF in a mouse model of potassium bromate–induced hyperuricemia. The results can serve as a useful preclinical reference for researching the gout prevention effects of functional foods.&#x0D; Keywords: Plectranthus amboinicus (Lour.) Spreng, hyperuricemia, uric acid–reducing activity

Isolation of Biomolecules from the Leaves of Lecaniodiscus cupanoides (Sapindaceae), a Plant used in Traditional Medicine in Benin

The aim of the present study was to identify the structure of the bioactive molecules in the active ethyl acetate fraction of the hydroethanolic extract of Lecaniodiscus cupanoides (Sapindaceae), a Beninese plant used in the treatment of microbial infections. We prepared the hydroethanolic extract from powdered dried leaves. We fractionated the hydroethanol extract using the liquid-liquid extraction method with solvents of increasing polarity. The active ethyl acetate fraction obtained after bioguided fractionation of the hydroethanol extract on bacterial strains was purified by a series of atmospheric pressure column chromatographic methods coupled with thin layer chromatography. At the end of this purification process, three compounds, including a flavonoid and two fatty acids, were isolated and identified by interpretation of 1H NMR, 13C NMR and mass spectrometry spectra. These were: 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-2,3-dihydrochromen-4-one;(E)-3-(3,4-dihydroxyphenyl)-2-(3-(3,4-dihydroxyphenyl) acryloyloxy) propanoic acid and (E)octadec -9- enoic acid (oleic acid).&#x0D; The added value of this work lies in the fact that these three molecules have never before been identified in this plant, and therefore represent a potential avenue for the development of a therapeutic arsenal to combat microbial infections.

Bioactivity-Guided Isolation and Antihypertensive Activity of Citrullus colocynthis Polyphenols in Rats with Genetic Model of Hypertension.

Background and Objectives: Citrullus colocynthis belongs to the Cucurbitaceae family and is a wild medicinal plant used in folk literature to treat various diseases. The purpose of the current study was to explore the antihypertensive and antioxidant potentials of Citrullus colocynthis (CC) polyphenol-rich fractions using a spontaneous hypertensive rat (SHR) model. Materials and Methods: The concentrated aqueous ethanol extract of CC fruit was successively fractioned using solvents of increasing polarity, i.e., hexane, chloroform, ethyl acetate and n-butanol. The obtained extracts were analyzed for total phenolic content (TPC), total flavonoid content (TFC) and total flavonol content (TOF). Moreover, the CC extracts were further evaluated for radical scavenging capacity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays and antioxidant activity using inhibition of linoleic acid peroxidation and determination of reducing potential protocols. The phytochemical components were characterized by HPLC-MWD-ESI-MS in positive ionization mode. Results: The results showed that ethyl acetate fraction (EAF) exhibited a higher content of phenolic compounds in term of TPC (289 mg/g), TFC (7.6 mg/g) and TOF (35.7 mg/g). EAF showed higher antioxidant and DPPH and ABTS scavenging activities with SC50 values of 6.2 and 79.5 µg/mL, respectively. LCMS analysis revealed that twenty polyphenol compounds were identified in the EAF, including phenolic acids and flavonoids, mainly myricetin and quercetin derivatives. The in vivo antihypertensive activity of EAF of CC on SHR revealed that it significantly decreased the mean arterial pressure (MAP), systolic blood pressure (SBP), diastolic blood pressures (DBP) and pulse pressure (PP) as compared to normal and hypertensive control groups. Moreover, EAF of CC significantly reduced the oxidative stress in the animals in a dose-dependent manner by normalizing the levels of superoxide dismutase (SOD), malondialdehyde (MDA), reduced glutathione (GSH), nitric oxide (NOx) and total antioxidant capacity (TAC). Furthermore, the treatment groups, especially the 500 mg of EAF per kg body weight (EA-500) group, significantly (p ≤ 0.05) improved the electrocardiogram (ECG) pattern and pulse wave velocity (PWV). Conclusion: It was concluded that the EAF of CC is a rich source of polyphenols and showed the best antioxidant activity and antihypertensive potential in SHR.

Antiproliferative activity of ethyl acetate fraction of Euphorbia ingens against prostate cancer cell line: An in silico and in vitro analysis

Current prostate cancer (PCa) treatments often lead to severe side effects, prompting the exploration of safer alternatives. Euphorbia ingens is a medicinal plant used for cancer treatment in African communities; however, there is no scientific validation of its anticancer activity. This study therefore evaluated the antiproliferative activity of E. ingens on the DU-145 human PCa cell line. The 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) bio-assay was used to determine the antiproliferative activity of E. ingens ethyl acetate-, and water-fractions. Only the ethyl acetate fraction was considered to be active. It inhibited DU-145 cell growth selectively without toxicity to non-cancerous Vero E6 cells. The IC50 was 9.71 ± 0.4 µg/ml and had a selectivity index of 8.26, indicating promising efficacy and cellular safety. Qualitative phytochemical screening identified the presence of phenols, terpenoids, flavonoids, tannins, sterols, and saponins in the ethyl acetate fraction. An additional 18 chemical compounds with potential synergistic roles were identified through gas chromatography-mass spectrometry analysis. Network pharmacology was then employed to predict molecular targets and mechanisms of action for drug-like chemical compounds, implicating key targets such as ESR1, IL6, MMP9, CDK2, MAP2K1, AR, PRKCD, CDK1, CDC25B, and JAK2 with regulation of PI3K/Akt, MAPK, and p53 signaling pathways suggested to be the potential mechanism of action. Further, gene expression analysis of selected targets through reverse transcription-quantitative polymerase chain reaction revealed downregulation of AR and BCL2 levels, along with upregulation of p53 and caspase-3 in fraction-treated DU-145 cells compared to the 0.2 % DMSO-negative control. In conclusion, the findings of this study validate the traditional use of E. ingens in cancer management. The identified drug-like compounds, their targets, and associated signaling pathways could serve as a foundation for developing novel strategies for prostate cancer management. However, we recommend additional in vitro and in vivo studies to further substantiate these findings.

Limosilactobacillus reuterias sustainable biological control agent against toxigenic Fusarium verticillioides.

Corn contamination with Fusarium verticillioides (Sacc.) Nirenberg is a worldwide problem that affects yield and grain quality resulting in severe economic losses and implications for food safety. Control of F. verticillioides is a challenge, but lactic acid bacteria (LAB) has high potential as a biological control agent. In this study, the antifungal effect of Limosilactobacillus reuteri (formerly Lactobacillus reuteri) LR-92 against F. verticillioides 97L was investigated. Cell-free supernatant (CFS) from L. reuteri showed concentration-dependent fungicidal and fungistatic activity against F. verticillioides 97L. The antifungal compounds from CFS showed heat stability and pH dependence, and antifungal activity was not affected by treatment with proteolytic enzymes. High-performance liquid chromatography analysis indicated that L. reuteri LR-92 produces lactic and acetic acids. After liquid-liquid extraction, electrospray ionization mass spectrometry analysis of the active ethyl acetate fraction containing antifungal compounds revealed the production of 3-phenyllactic acid, cyclo-(L-Pro-L-Leu), cyclo-(L-Pro-L-Phe), and cyclo-(L-Phe-trans-4-OH-L-Pro). L. reuteri LR-92 has potential as a biocontrol agent for F. verticillioides and contributes to food safety.

GC-MS chemical profiling, antioxidant, anti-diabetic, and anti-inflammatory activities of ethyl acetate fraction of Spilanthes filicaulis (Schumach. and Thonn.) C.D. Adams leaves: experimental and computational studies.

Introduction: This study aimed to investigate the chemical profile of GC-MS, antioxidant, anti-diabetic, and anti-inflammatory activities of the ethyl acetate fraction of Spilanthes filicaulis leaves (EFSFL) via experimental and computational studies. Methods: After inducing oxidative damage with FeSO4, we treated the tissues with different concentrations of EFSFL. An in-vitro analysis of EFSFL was carried out to determine its potential for antioxidant, anti-diabetic, and anti-inflammatory activities. We also measured the levels of CAT, SOD, GSH, and MDA. Results and discussion: EFSFL exhibited anti-inflammatory properties through membrane stabilizing properties (IC50 = 572.79μg/ml), proteinase inhibition (IC50 = 319.90μg/ml), and inhibition of protein denaturation (IC50 = 409.88μg/ml). Furthermore, EFSFL inhibited α-amylase (IC50 = 169.77μg/ml), α-glucosidase (IC50 = 293.12μg/ml) and DPP-IV (IC50 = 380.94μg/ml) activities, respectively. Our results indicated that induction of tissue damage reduced the levels of GSH, SOD, and CAT activities, and increased MDA levels. However, EFSFL treatment restores these levels to near normal. GC-MS profiling shows that EFSFL contains 13 compounds, with piperine being the most abundant. In silico interaction of the phytoconstituents using molecular and ensembled-based docking revealed strong binding tendencies of two hit compounds to DPP IV (alpha-caryophyllene and piperine with a binding affinity of -7.8 and -7.8Kcal/mol), α-glucosidase (alpha-caryophyllene and piperine with a binding affinity of -9.6 and -8.9Kcal/mol), and to α-amylase (piperine and Benzocycloheptano[2,3,4-I,j]isoquinoline, 4,5,6,6a-tetrahydro-1,9-dihydroxy-2,10-dimethoxy-5-methyl with a binding affinity of -7.8 and -7.9Kcal/mol), respectively. These compounds also presented druggable properties with favorable ADMET. Conclusively, the antioxidant, antidiabetic, and anti-inflammatory activities of EFSFL could be due to the presence of secondary metabolites.

Evaluation of Rhodiola rosea Root Fraction as a Functional Cosmetic Ingredient

To develop functional cosmetic material using &lt;i&gt;Rhodiola rosea&lt;/i&gt;, the biological activities such as anti-oxidant, anti-inflammatory, astringent, anti-tyrosinase, and anti-elastase activity were investigated with methanol, &lt;i&gt;n&lt;/i&gt;-hexane, chloroform, ethyl acetate, n-butanol, and water fractions of &lt;i&gt;R. rosea&lt;/i&gt; Root. DPPH and ABTS radical scavenging activities of ethyl acetate fraction were increased in a dosedependent manner; the activities were higher than other fractions. The total phenolic content of ethyl acetate fraction also showed a relatively high value of 500.485±4.848 mgGAE/g. The ethyl acetate fraction also significantly decreased LPS-induced NO production and IL-6 secretion compared with untreated control cells at above 5-100 μg/mL concentrations. The astringent activity of ethyl acetate fraction was 94.31% at a concentration of 500 μg/mL, and the activity of ethyl acetate fraction was similar to that of tannic acid. The mushroom tyrosinase inhibition abilities of hexane, ethyl acetate, and butanol were 10.44%, 17.53%, and 14.55%, respectively, at 100 μg/mL. Furthermore, an elastase inhibition rate showed a relatively high effect at 100 μg/mL, 45.46% of inhibition rate in the ethyl acetate and 45.07% of inhibition rate in the butanol fraction. These results indicate that R. rosea Root would be used as a natural material in the functional cosmetic and beauty-food industries.