scholarly journals Anti-Hepatocellular Carcinoma (HepG2) Activities of Monoterpene Hydroxy Lactones Isolated from the Marine Microalga Tisochrysis Lutea

Marine Drugs ◽  
2020 ◽  
Vol 18 (11) ◽  
pp. 567
Author(s):  
Katkam N. Gangadhar ◽  
Maria João Rodrigues ◽  
Hugo Pereira ◽  
Helena Gaspar ◽  
F. Xavier Malcata ◽  
...  
Keyword(s):  
Hepg2 Cells ◽  
Preparative Thin Layer Chromatography ◽  
Omega 3 ◽  
Nmr Analysis ◽  
Tisochrysis Lutea ◽  
Marine Microalga ◽  
Bioassay Guided Fractionation ◽  
First Time ◽  
Layer Chromatography

Tisochrysis lutea is a marine haptophyte rich in omega-3 polyunsaturated fatty acids (e.g., docosahexaenoic acid (DHA)) and carotenoids (e.g., fucoxanthin). Because of the nutraceutical applications of these compounds, this microalga is being used in aquaculture to feed oyster and shrimp larvae. In our earlier report, T. lutea organic crude extracts exhibited in vitro cytotoxic activity against human hepatocarcinoma (HepG2) cells. However, so far, the compound(s) accountable for the observed bioactivity have not been identified. Therefore, the aim of this study was to isolate and identify the chemical component(s) responsible for the bioactivity observed. Bioassay-guided fractionation through a combination of silica-gel column chromatography, followed by preparative thin layer chromatography (PTLC), led to the isolation of two diastereomers of a monoterpenoid lactone, namely, loliolide (1) and epi-loliolide (2), isolated for the first time in this species. The structural elucidation of both compounds was carried out by GC-MS and 1D (1H and 13C APT) and 2D (COSY, HMBC, HSQC-ed, and NOESY) NMR analysis. Both compounds significantly reduced the viability of HepG2 cells and were considerably less toxic towards a non-tumoral murine stromal (S17) cell line, although epi-loliolide was found to be more active than loliolide.

scholarly journals In Vitro Anti-Trypanosoma Cruzi Activity of Halophytes From Southern Portugal Reloaded: A Special Focus on Sea Fennel (Crithmum Maritimum L.)

2020 ◽  
Author(s):  
Catarina Pereira ◽  
Carolina Moraes ◽  
Caio Franco ◽  
Raphaël Grougnet ◽  
Euzébio Barbosa ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Preparative Thin Layer Chromatography ◽  
Selectivity Index ◽  
Special Focus ◽  
Medicinal Species ◽  
Southern Portugal ◽  
Parasitic Activity ◽  
First Time ◽  
Layer Chromatography

Abstract Marine halophytes are an outstanding reservoir of natural products and several species have anti-infectious traditional uses. However, little is known about their potential against neglected tropical ailments, such as Chagas disease. This work evaluated for the first time the in vitro anti-Trypanosoma cruzi activity of extracts from the aromatic and medicinal species Helichrysum italicum subsp. picardii (Boiss. & Reut.) Franco (Asteraceae, everlasting) and Crithmum maritimum L. (Apiaceae, sea fennel). For that purpose, decoctions, tinctures and essential oils from everlasting’s flowers and sea fennel’s stems, leaves and flowers were tested against intracellular amastigotes of two T. cruzi strains. Sea fennel’s flowers decoction displayed significant anti-trypanosomal activity and no toxicity towards the host cell (EC50 = 17.7 µg/mL, selectivity index > 5.65). This extract was partitioned using liquid-liquid extraction affording 5 fractions that were re-tested in the same model of anti-parasitic activity. Fraction 1 was the most active and selective (EC50 = 0.47 μg/mL, selectivity index = 59.6) and was submitted to preparative thin layer chromatography. The major compound present, likely responsible for the observed anti-trypanosomal activity, was identified as falcarindiol. Target fishing studies showed falcarindiol as a ligand of T. cruzi spermidine synthase, pointing to a potential enzyme-inhibiting anti-trypanosomal mechanism of action.

scholarly journals Structural Elucidation of Irish Ale Bioactive Polar Lipids with Antithrombotic Properties

Biomolecules ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 1075 ◽  
Author(s):  
Alexandros Tsoupras ◽  
Ronan Lordan ◽  
Eoin O'Keefe ◽  
Katie Shiels ◽  
Sushanta Kumar Saha ◽  
...  
Keyword(s):  
Inflammatory Mediator ◽  
Fatty Acid Content ◽  
Platelet Activating Factor ◽  
Preparative Thin Layer Chromatography ◽  
Polar Lipids ◽  
Liquid Chromatography Mass Spectrometry ◽  
Omega 3 ◽  
Human Platelet Aggregation ◽  
Layer Chromatography ◽  
By Products

The structures of bioactive polar lipids (PLs) of Irish ale with potent antithrombotic and cardioprotective properties were elucidated. Ale PL was fractionated by preparative thin layer chromatography (TLC) into subclasses, and their antithrombotic effect was assessed against human platelet aggregation induced by the pro-inflammatory mediator, platelet-activating factor (PAF). The fatty acid content and the overall structures of ale PL were elucidated by liquid chromatography mass spectrometry (LC-MS). Phosphatidylcholines (PC) and molecules of the sphingomyelin (SM) family exhibited the strongest anti-PAF effects, followed by phosphatidylethanolamines (PE). PC contained higher amounts of omega-3 polyunsaturated fatty acids (n-3 PUFA) and thus the lowest n-6/n-3 ratio. Bioactive diacyl and alkyl-acyl PC and PE molecules bearing n-3 PUFA at their sn-2 position, especially docosahexaenoic acid (DHA) and α-linolenic acid (ALA) but mostly oleic acid (OA), were identified in both PC and PE subclasses. Eicosapentaenoic acid (EPA) was present only in bioactive PC molecules and not in PE, explaining the lower anti-PAF effects of PE. Bioactive sphingolipid and glycolipid molecules with reported anti-inflammatory and anti-tumour properties, such as specific ceramides and glucosylcerebrosides with sphingosine, phytosphingosine and dihydrosphingosine bases but also specific monogalactodiglycerides and SM species bearing ALA at their sn-2 position, were identified in the SM subclass, providing a rational for its strong bioactivities against the PAF pathway. Further studies are required on the health benefits of bioactive PL from beer and brewery by-products.

Antifungal and Antibacterial Activities of Pentanema divaricatum and Its Active Constituent

2008 ◽  
Vol 63 (9-10) ◽  
pp. 649-652 ◽  
Author(s):  
Nastaran Momen-Roknabadi ◽  
Ahamd R. Gohari ◽  
Hamid R. Monsef-Esfehani ◽  
Farideh Attar ◽  
Reza Hajiaghaee ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Aspergillus Niger ◽  
Preparative Thin Layer Chromatography ◽  
Antibacterial Activities ◽  
Diffusion Method ◽  
Active Constituent ◽  
Disk Diffusion Method ◽  
Bioassay Guided Fractionation ◽  
Layer Chromatography ◽  
And Staphylococcus Aureus

The antimicrobial activity of ethanol and chloroform extracts of Pentanema divaricatum Cass. was studied using the conventional disk diffusion method. The extracts’ highest antimicrobial activity was observed against Aspergillus niger. Bioassay-guided fractionation of the crude extract by preparative thin layer chromatography (PTLC) showed one antimicrobial fraction which was especially effective against Aspergillus niger. By conventional spectroscopy the active fraction was identified as 4α,5α-epoxy-10α,14H-1-epi-inuviscolide. This compound represented the most potent antimicrobial candidate, with MIC values of < 25 μg/disk against A. niger strains and 200 μg/disk against Bacillus cereus and Staphylococcus aureus.

scholarly journals An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpart

2021 ◽  
Vol 33 (1) ◽  
Author(s):  
Doaa E. Ahmed ◽  
Fatma B. Rashidi ◽  
Heba K. Abdelhakim ◽  
Amr S. Mohamed ◽  
Hossam M. M. Arafa
Keyword(s):  
Gene Expression ◽  
Hepatocellular Carcinoma ◽  
Membrane Potential ◽  
Hepg2 Cells ◽  
Mitochondrial Membrane ◽  
Caspase 3 ◽  
Caspase 9 ◽  
First Time ◽  
Bcl2 Gene

Abstract Background Glufosfamide (β-d-glucosylisophosphoramide mustard, GLU) is an alkylating cytotoxic agent in which ifosforamide mustard (IPM) is glycosidically linked to the β-d-glucose molecule. GLU exerted its cytotoxic effect as a targeted chemotherapy. Although, its cytotoxic efficacy in a number of cell lines, there were no experimental or clinical data available on the oncolytic effect of oxazaphosphorine drugs in hepatocellular carcinoma. Therefore, the main objective of the current study is to assess the cytotoxic potential of GLU for the first time in the hepatocellular carcinoma HepG2 cell line model. Methods Cytotoxicity was assayed by the MTT method, and half-maximal inhibitory concentration (IC50) was calculated. Flow cytometric analysis of apoptosis frequencies was measured by using Annexin V/PI double stain, an immunocytochemical assay of caspase-9, visualization of caspase-3, and Bcl2 gene expression were undertaken as apoptotic markers. Mitochondrial membrane potential was measured using the potentiometric dye; JC-1, as a clue for early apoptosis as well as ATP production, was measured by the luciferase-chemiluminescence assay. Results Glufosfamide induced cytotoxicity in HepG2 cells in a concentration- and time-dependent manner. The IC50 values for glufosfamide were significantly lower compared to ifosfamide. The frequency of apoptosis was much higher for glufosfamide than that of ifosfamide. The contents of caspase-9 and caspase-3 were elevated following exposure to GLU more than IFO. The anti-apoptotic Bcl2 gene expression, the mitochondrial membrane potential, and the cellular ATP levels were significantly decreased than in case of ifosfamide. Conclusions The current study reported for the first time cytotoxicity activity of glufosfamide in HepG2 cells in vitro. The obtained results confirmed the higher oncolytic activity of glufosfamide than its aglycone ifosfamide. The generated data warrants further elucidations by in vivo study.

Inhibition of the ATPase Domain of Human Topoisomerase IIa on HepG2 Cells by 1, 2-benzenedicarboxylic Acid, Mono (2-ethylhexyl) Ester: Molecular Docking and Dynamics Simulations

2019 ◽  
Vol 19 (6) ◽  
pp. 495-503 ◽  
Author(s):  
Jemimah Naine Selvakumar ◽  
Subathra Devi Chandrasekaran ◽  
George Priya C. Doss ◽  
Thirumal D. Kumar
Keyword(s):  
Molecular Docking ◽  
Hepg2 Cells ◽  
Flow Cytometric Analysis ◽  
Dynamics Simulation ◽  
Benzenedicarboxylic Acid ◽  
Bioassay Guided Fractionation ◽  
Marine Streptomyces ◽  
Dynamics Simulations ◽  
Molecular Docking And Dynamics

Background: The major attention has been received by the natural products in the prevention of diseases due to their pharmacological role. Objective: The major focus of the study was to search for highly potential anti-cancer compounds from marine Streptomyces sp. VITJS4 (NCIM No. 5574). Methods: Cytotoxic assay was examined by MTT assay on HepG2 cells. Bioassay-guided fractionation of the ethyl acetate extract from the fermented broth led to the isolation of the compound. The lead compound structure was elucidated by combined NMR and MS analysis, and the absolute configuration was assigned by extensive spectroscopic analysis. Results: On the basis of spectroscopic data, the compound was identified as 1, 2 benzenedicarboxylic acid, mono 2-ethylhexyl (BMEH). The compound exhibited in vitro anticancer potential against liver (HepG2) cancer cells. Based on the flow cytometric analysis, it was evident that the BMEH was also effective in arresting the cell cycle at G1 phase. Further, the Western blotting analysis confirmed the down-regulation of Bcl-2 family proteins, and activation of caspase-9 and 3. The molecular docking and dynamics simulation were performed to reveal the activity of the compound over a time period of 10ns. From the molecular dynamics studies, it was found that the stability and compactness were attained by the protein by means of the compound interaction. Conclusion: This study highlights our collaborative efforts to ascertain lead molecules from marine actinomycete. This is the first and foremost report to prove the mechanistic studies of the purified compound 1, 2-benzene dicarboxylic acid, mono(2-ethylhexyl) ester isolated from marine Streptomyces sp.VITJS4 against HepG2 cells.

Antioxidant and Immunomodulatory Constituents of Henna Leaves

2004 ◽  
Vol 59 (7-8) ◽  
pp. 468-476 ◽  
Author(s):  
Botros R. Mikhaeil ◽  
Farid A. Badria ◽  
Galal T. Maatooq ◽  
Mohamed M. A. Amer
Keyword(s):  
Methanolic Extract ◽  
Radical Scavenging ◽  
Lymphocyte Transformation ◽  
Coumaric Acid ◽  
Lawsonia Inermis ◽  
Physical Chemical ◽  
Bioassay Guided Fractionation ◽  
Free Radical Scavenging Assay ◽  
First Time

AbstractThe immunomodulatory bioassay-guided fractionation of the methanolic extract of henna (Lawsonia inermis L.; syn. Lawsonia alba L.) leaves resulted in the isolation of seven compounds; three have been isolated for the first time from the genus, namely p-coumaric acid, 2-methoxy-3-methyl-1,4-naphthoquinone and apiin, along with the previously isolated compounds: lawsone, apigenin, luteolin, and cosmosiin. Structural elucidation of the isolated compounds was based upon their physical, chemical as well as spectroscopic characters. Their immuomodulatory profile was studied using an in vitro immunoassay, the lymphocyte transformation assay. The ABTS [2,2′-azino-bis (3-ethyl benzthiazoline-6-sulfonic acid)], free radical scavenging assay depicted that all isolated compounds exhibited antioxidant activity comparable to that of ascorbic acid.

scholarly journals In Vitro Anti-Trypanosoma cruzi Activity of Halophytes from Southern Portugal Reloaded: A Special Focus on Sea Fennel (Crithmum maritimum L.)

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2235
Author(s):  
Catarina G. Pereira ◽  
Carolina Borsoi Moraes ◽  
Caio H. Franco ◽  
Clarissa Feltrin ◽  
Raphaël Grougnet ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Preparative Thin Layer Chromatography ◽  
Selectivity Index ◽  
Special Focus ◽  
Medicinal Species ◽  
Southern Portugal ◽  
Parasitic Activity ◽  
The One ◽  
First Time

Marine halophytes are an outstanding reservoir of natural products and several species have anti-infectious traditional uses. However, reports about their potential use against neglected tropical ailments, such as Chagas disease, are scarce. This work evaluated for the first time the in vitro anti-Trypanosoma cruzi activity of extracts from the aromatic and medicinal species Helichrysum italicum subsp. picardii (Boiss. & Reut.) Franco (Asteraceae, everlasting) and Crithmum maritimum L. (Apiaceae, sea fennel). For that purpose, decoctions, tinctures, and essential oils from everlasting’s flowers and sea fennel’s stems, leaves, and flowers were tested against intracellular amastigotes of two T. cruzi strains. The extract from the sea fennel flower decoction displayed significant anti-trypanosomal activity and no toxicity towards the host cell (EC50 = 17.7 µg/mL, selectivity index > 5.65). Subsequent fractionation of this extract afforded 5 fractions that were re-tested in the same model of anti-parasitic activity. Fraction 1 was the most active and selective (EC50 = 0.47 μg/mL, selectivity index = 59.6) and was submitted to preparative thin-layer chromatography. One major compound was identified, falcarindiol, which was likely the one responsible for the observed anti-trypanosomal activity. This was confirmed using a commercially sourced molecule. Target-fishing studies showed falcarindiol as a ligand of T. cruzi spermidine synthase, pointing to a potential enzyme-inhibiting anti-trypanosomal mechanism of action. Overall, this work shows that sea fennel can provide effective anti-parasitic molecule(s) with potential pharmacological applications in the treatment of CD.

scholarly journals Microbial biotransformation of beclomethasone dipropionate by Aspergillus niger

2014 ◽  
Vol 50 (4) ◽  
pp. 903-909
Author(s):  
Saeed Ahmad ◽  
Farhan Hameed Khaliq ◽  
Asadullah Madni ◽  
Muhammad Nabeel Shahid ◽  
Irfan Pervaiz
Keyword(s):  
Aspergillus Niger ◽  
Beclomethasone Dipropionate ◽  
Microbial Transformation ◽  
Preparative Thin Layer Chromatography ◽  
Spectroscopic Techniques ◽  
Potential Drug ◽  
Drug Molecules ◽  
First Time ◽  
Layer Chromatography ◽  
Old Drugs

In the present research, the steroidal anti-asthmatic drug beclomethasone dipropionate was subjected to microbial biotransformation by Aspergillus niger. Beclomethasone dipropionate was transformed into various metabolites first time from microbial transformation. New drug metabolites produced can act as new potential drug molecules and can replace the old drugs in terms of safety, efficacy, and least resistance. They were purified by preparative thin layer chromatography technique, and their structures were elucidated using modern spectroscopic techniques, such as 13C NMR, 1H NMR, HMQC, HMQC, COSY, and NOESY, and mass spectrometry, such as EI-MS. Four metabolites were purified: (i) beclomethasone 17-monopropionate, (ii) beclomethasone 21-monopropionate, (iii) beclomethasone, and (iv) 9beta,11beta-epoxy-17,21-dihydroxy-16beta-methylpregna-1,4-diene-3,20-dione 21-propionate.

scholarly journals Bacterial Lipid II Analogs: Novel In Vitro Substrates for Mammalian Oligosaccharyl Diphosphodolichol Diphosphatase (DLODP) Activities

Molecules ◽  
2019 ◽  
Vol 24 (11) ◽  
pp. 2135 ◽  
Author(s):  
Ahmad Massarweh ◽  
Michael Bosco ◽  
Isabelle Chantret ◽  
Thibaut Léger ◽  
Layla Jamal ◽  
...  
Keyword(s):  
Solid Phase ◽  
Extraction Procedure ◽  
Lipid Ii ◽  
Liver Membrane ◽  
Solid Phase Extraction Procedure ◽  
Bacterial Lipid ◽  
First Time ◽  
Layer Chromatography ◽  
Hydrolysis Of

Mammalian protein N-glycosylation requires the transfer of an oligosaccharide containing 2 residues of N-acetylglucosamine, 9 residues of mannose and 3 residues of glucose (Glc3Man9 GlcNAc2) from Glc3Man9GlcNAc2-diphospho (PP)-dolichol (DLO) onto proteins in the endoplasmic reticulum (ER). Under some pathophysiological conditions, DLO biosynthesis is perturbed, and truncated DLO is hydrolyzed to yield oligosaccharyl phosphates (OSP) via unidentified mechanisms. DLO diphosphatase activity (DLODP) was described in vitro, but its characterization is hampered by a lack of convenient non-radioactive substrates. Our objective was to develop a fluorescence-based assay for DLO hydrolysis. Using a vancomycin-based solid-phase extraction procedure coupled with thin layer chromatography (TLC) and mass spectrometry, we demonstrate that mouse liver membrane extracts hydrolyze fluorescent bacterial lipid II (LII: GlcNAc-MurNAc(dansyl-pentapeptide)-PP-undecaprenol) to yield GlcNAc-MurNAc(dansyl-pentapeptide)-P (GM5P). GM5P production by solubilized liver microsomal proteins shows similar biochemical characteristics to those reported for human hepatocellular carcinoma HepG2 cell DLODP activity. To conclude, we show, for the first time, hydrolysis of lipid II by a eukaryotic enzyme. As LII and DLO are hydrolyzed by the same, or closely related, enzymes, fluorescent lipid II analogs are convenient non-radioactive substrates for investigating DLODP and DLODP-like activities.

scholarly journals Isolation of Cycloeucalenol from Boophone Disticha and Evaluation of its Cytotoxicity

2013 ◽  
Vol 8 (9) ◽  
pp. 1934578X1300800 ◽  
Author(s):  
Emmanuel Adekanmi Adewusi ◽  
Paul Steenkamp ◽  
Gerda Fouche ◽  
Vanessa Steenkamp
Keyword(s):  
Preparative Thin Layer Chromatography ◽  
2D Nmr ◽  
Neutral Red ◽  
Literature Report ◽  
Human Neuroblastoma ◽  
H Nmr ◽  
Low Toxicity ◽  
Dose Dependent Decrease ◽  
First Time ◽  
Layer Chromatography

Boophone disticha (Amaryllidaceae) is widely used in traditional medicine in southern Africa. Several alkaloids, volatile oils and fatty acids have been isolated from the plant. However, there has been no literature report of a triterpene from B. disticha. Cycloeucalenol, a cycloartane triterpene, together with its regio-isomer, was isolated from the ethyl acetate extract of the bulbs using column chromatography and preparative thin layer chromatography. Structural elucidation was carried out using 1D and 2D NMR and mass spectroscopy. The MTT and neutral red assays were used to assess the cytotoxicity of the compound in human neuroblastoma (SH-SY5Y) cells. The compound was obtained as a mixture of two regio-isomers, which were separated for the first time by chromatographic optimization. Integration of the 1H NMR spectrum showed that cycloeucalenol and its regio-isomer were present in a ratio of 1.04:1. A dose-dependent decrease in cell viability was observed using both cytotoxicity assays. IC50 values of 173.0 ± 5.1 μM and 223.0 ± 6.4 μM were obtained for the MTT and neutral red assays, respectively, indicative of the low toxicity of the compound. This work describes for the first time, the presence of triterpene compounds from the genus Boophone.

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