{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T11:30:06Z","timestamp":1792063806876,"version":"4.4.0"},"reference-count":62,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2025,2,11]],"date-time":"2025-02-11T00:00:00Z","timestamp":1739232000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Research Foundation (NRF) innovation scholarship","award":["SFH180525334799"],"award-info":[{"award-number":["SFH180525334799"]}]},{"name":"National Research Foundation (NRF) innovation scholarship","award":["MND200617532846"],"award-info":[{"award-number":["MND200617532846"]}]},{"name":"National Research Foundation (NRF) innovation scholarship","award":["UID116354"],"award-info":[{"award-number":["UID116354"]}]},{"name":"HA Taylor Will Trust","award":["SFH180525334799"],"award-info":[{"award-number":["SFH180525334799"]}]},{"name":"HA Taylor Will Trust","award":["MND200617532846"],"award-info":[{"award-number":["MND200617532846"]}]},{"name":"HA Taylor Will Trust","award":["UID116354"],"award-info":[{"award-number":["UID116354"]}]},{"name":"NRF Development","award":["SFH180525334799"],"award-info":[{"award-number":["SFH180525334799"]}]},{"name":"NRF Development","award":["MND200617532846"],"award-info":[{"award-number":["MND200617532846"]}]},{"name":"NRF Development","award":["UID116354"],"award-info":[{"award-number":["UID116354"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Background\/Objectives: Tuberculosis remains a major public health crisis, and it is imperative to search for new antimycobacterial drugs. Natural products, including medicinal macrofungi, have been used as sources for the discovery of pharmaceuticals; however, research on their antimycobacterial activity remains limited. This study aimed to isolate and identify the bioactive compound responsible for antimycobacterial activity, thereby expanding on the limited knowledge regarding the antimicrobial activity and bioactive compounds present in Gymnopilus\u00a0junonius. Methods: Bioassay-guided fractionation using column chromatography and preparative thin-layer chromatography were employed to isolate the active compound. Antimycobacterial activity against Mycobacterium tuberculosis H37 was assessed using a resazurin microplate assay (REMA). The chemical structure was determined by 1H nuclear magnetic resonance (NMR) spectroscopy, heteronuclear single quantum coherence (HSQC) spectroscopy, heteronuclear multiple bond correlation (HMBC) spectroscopy, and high-resolution electrospray ionization mass (HR-ESI-MS) spectrometry. Transmission electron microscopy (TEM) was used to observe the ultrastructural changes in M. tuberculosis induced by the compound. Cytotoxicity was evaluated in African green monkey kidney cells (Vero), human liver cells (C3A), and zebrafish embryos\/larvae. Results: Bioassay-guided fractionation led to the isolation of gymnopilene, which showed inhibitory activity against M. tuberculosis (MIC: 31.25 \u00b5g\/mL). TEM analysis revealed that treatment with gymnopilene caused ultrastructural damage observed as the disruption and disintegration of the cell wall. While gymnopilene demonstrated cytotoxicity in Vero and C3A cells, no toxicity was observed in zebrafish embryos\/larvae for the crude extract. Conclusions: This study highlights that macrofungi, such as G. junonius, could be a valuable resource of bioactive compounds.<\/jats:p>","DOI":"10.3390\/antibiotics14020179","type":"journal-article","created":{"date-parts":[[2025,2,11]],"date-time":"2025-02-11T05:34:32Z","timestamp":1739252072000},"page":"179","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Exploring Antimycobacterial Potential: Safety Evaluation and Active Compound Isolation from Gymnopilus junonius"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7453-8119","authenticated-orcid":false,"given":"Jenske","family":"Didloff","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Microbiology, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gerhardt J.","family":"Boukes","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Microbiology, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South Africa"},{"name":"Afrigen Biologics (Pty) Ltd., c\/o South African Medical Research Council, P.O. Box 19070, Tygerberg 7505, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mutenta N.","family":"Nyambe","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmacy, Nutrition and Dietetics, Lusaka Apex Medical University, Lusaka 10101, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0223-2660","authenticated-orcid":false,"given":"Denzil R.","family":"Beukes","sequence":"additional","affiliation":[{"name":"School of Pharmacy, University of Western Cape, Bellville 7535, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mookho S.","family":"Lerata","sequence":"additional","affiliation":[{"name":"School of Pharmacy, University of Western Cape, Bellville 7535, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9013-5579","authenticated-orcid":false,"given":"Velile","family":"Vilane","sequence":"additional","affiliation":[{"name":"Center for High-Resolution Transmission Electron Microscopy (HRTEM), Physics Department, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1528-1808","authenticated-orcid":false,"given":"Michael","family":"Lee","sequence":"additional","affiliation":[{"name":"Center for High-Resolution Transmission Electron Microscopy (HRTEM), Physics Department, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7117-2502","authenticated-orcid":false,"given":"Sharlene","family":"Govender","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Microbiology, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3816-1419","authenticated-orcid":false,"given":"Maryna","family":"van de Venter","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Microbiology, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.sajb.2017.07.010","article-title":"Cytotoxicity of thirteen South African macrofungal species against five cancer cell lines","volume":"113","author":"Boukes","year":"2017","journal-title":"S. Afr. J. Bot."},{"key":"ref_2","unstructured":"World Health Organization (2024). Global Tuberculosis Report 2024."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"193","DOI":"10.4103\/0970-2113.83977","article-title":"Tuberculosis chemotherapy in the 21st century: Back to the basics","volume":"28","author":"Joshi","year":"2011","journal-title":"Lung India"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1733","DOI":"10.1016\/j.biopha.2018.04.176","article-title":"Drug targets exploited in Mycobacterium tuberculosis: Pitfalls and promises on the horizon","volume":"103","author":"Bhat","year":"2018","journal-title":"Biomed. Pharmacother."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Alsayed, S.S.R., and Gunosewoyo, H. (2023). Tuberculosis: Pathogenesis, current treatment regimens and new drug targets. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24065202"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1038\/ja.2017.126","article-title":"New antituberculous drugs derived from natural products: Current perspectives and issues in antituberculous drug development","volume":"71","author":"Igarashi","year":"2018","journal-title":"J. Antibiot."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Intorasoot, S., Intorasoot, A., Tawteamwong, A., Butr-Indr, B., Phunpae, P., Tharinjaroen, C.S., Wattananandkul, U., Sangboonruang, S., and Khantipongse, J. (2022). In Vitro Antimycobacterial Activity of Human Lactoferrin-Derived Peptide, D-hLF 1-11, against Susceptible and Drug-Resistant Mycobacterium tuberculosis and Its Synergistic Effect with Rifampicin. Antibiotics, 11.","DOI":"10.3390\/antibiotics11121785"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Nowacka, N., Nowak, R., Drozd, M., Olech, M., Los, R., and Malm, A. (2015). Antibacterial, antiradical potential and phenolic compounds of thirty-one polish mushrooms. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0140355"},{"key":"ref_9","first-page":"196","article-title":"Anti-Mycobacterium tuberculosis Active Metabolites from an Endophytic Streptomyces sp. YIM65484","volume":"9","author":"Zhou","year":"2015","journal-title":"Rec. Nat. Prod."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/17460441.2017.1255604","article-title":"Current challenges in drug discovery for tuberculosis","volume":"12","author":"Kumar","year":"2017","journal-title":"Expert Opin. Drug Discov."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.phytol.2019.11.016","article-title":"Drimane sesquiterpenoids from cultures of the basidiomycete Gymnopilus sp. BCC 19384","volume":"35","author":"Isaka","year":"2020","journal-title":"Phytochem. Lett."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Caldas, L.A., Santos, P.D., Carbonero, E.R., Ionta, M., Miyazawa, M., Caixeta, E.S., Fregnan, A.M., N\u00f3brega, B.B., Di Medeiros, M.C.B., and Menolli, N. (2022). Immunomodulatory effect of polysaccharides from the mushroom-forming basidiomycete Gymnopilus imperialis (Agaricomycetes, Basidiomycota). Pharmaceuticals, 15.","DOI":"10.3390\/ph15101179"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1007\/s12272-020-01213-6","article-title":"Trichothecene and tremulane sesquiterpenes from a hallucinogenic mushroom Gymnopilus junonius and their cytotoxicity","volume":"43","author":"Lee","year":"2020","journal-title":"Arch. Pharm. Res."},{"key":"ref_14","unstructured":"Branch, M. (2001). Sasol Mushrooms of Southern Africa: A First Field Guide, Penguin Random House South Africa."},{"key":"ref_15","unstructured":"Gryzenhout, M. (2010). Pocket Guide to Mushrooms of South Africa, Penguin Random House South Africa."},{"key":"ref_16","first-page":"650","article-title":"Antibacterial activiity, In vitro antioxidant potential and GC-MS characterization of methanolic extract of Gymnopilus junonius, a wild mushroom from Southern Western Ghats, India","volume":"5","author":"Ragupathi","year":"2018","journal-title":"European J. Biomed. Pharm. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1038\/ja.2011.122","article-title":"Gymnopilin K: A new cytotoxic gymnopilin from Gymnopilus spectabilis","volume":"65","author":"Kim","year":"2012","journal-title":"J. Antibiot."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"55","DOI":"10.4489\/MYCO.2008.36.1.055","article-title":"Chemical constituents of Gymnopilus spectabilis and their antioxidant activity","volume":"36","author":"Lee","year":"2008","journal-title":"Mycobiology"},{"key":"ref_19","unstructured":"Boukes, G.J., Didloff, J., van de Venter, M., and Govender, S. (2018). Antitubercular and HIV-1 Enzyme Inhibitory Activities of South African Macrofungi, The 43rd FEBS Congress."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1016\/j.sajb.2024.01.021","article-title":"Investigation of the inhibition of respiratory bacterial pathogens and HIV-1 enzymes by twenty-one South African mushroom species","volume":"166","author":"Didloff","year":"2024","journal-title":"S. Afr. J. Bot."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1479","DOI":"10.1055\/s-0030-1250027","article-title":"Potential of Cameroonian plants and derived products against microbial infections: A review","volume":"76","author":"Kuete","year":"2010","journal-title":"Planta Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"7507549","DOI":"10.1155\/2019\/7507549","article-title":"Antibacterial and antibiotic modifying potential of crude extracts, fractions, and compounds from Acacia polyacantha Willd. against MDR Gram-negative bacteria","volume":"2019","author":"Mambe","year":"2019","journal-title":"Evid. Based Complement. Alternat. Med."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Aro, A.O., Dzoyem, J.P., Awouafack, M.D., Selepe, M.A., Eloff, J.N., and McGaw, L.J. (2019). Fractions and isolated compounds from Oxyanthus speciosus subsp. stenocarpus (Rubiaceae) have promising antimycobacterial and intracellular activity. BMC Complement. Altern. Med., 19.","DOI":"10.1186\/s12906-019-2520-x"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1021\/np50073a017","article-title":"Minor constituents of Gymnopilus spectabilis","volume":"54","author":"Findlay","year":"1991","journal-title":"J. Nat. Prod."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"114596","DOI":"10.1016\/j.jep.2021.114596","article-title":"Exploring four South African Croton species for potential anti-inflammatory properties: In vitro activity and toxicity risk assessment","volume":"282","author":"Rampa","year":"2022","journal-title":"J. Ethnopharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"745","DOI":"10.1016\/j.tiv.2011.01.005","article-title":"Relative embryotoxicity of two classes of chemicals in a modified zebrafish embryotoxicity test and comparison with their in vivo potencies","volume":"25","author":"Hermsen","year":"2011","journal-title":"Toxicol. Vitr."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.reprotox.2015.06.050","article-title":"From cutting edge to guideline: A first step in harmonization of the zebrafish embryotoxicity test (ZET) by describing the most optimal test conditions and morphology scoring system","volume":"56","author":"Beekhuijzen","year":"2015","journal-title":"Reprod. Toxicol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1707","DOI":"10.1055\/s-0032-1315370","article-title":"A review on antimicrobial activity of mushroom (Basidiomycetes) extracts and isolated compounds","volume":"78","author":"Alves","year":"2012","journal-title":"Planta Med."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Matija\u0161evi\u0107, D., Panti\u0107, M., Ra\u0161kovi\u0107, B., Pavlovi\u0107, V., Duvnjak, D., Sknepnek, A., and Nik\u0161i\u0107, M. (2016). The antibacterial activity of Coriolus versicolor methanol extract and its effect on ultrastructural changes of Staphylococcus aureus and Salmonella Enteritidis. Front. Microbiol., 7.","DOI":"10.3389\/fmicb.2016.01226"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"485","DOI":"10.22270\/jddt.v9i4.3083","article-title":"Investigation of bioactive compounds of Capsicum Frutescence and Annona Muricata by chromatographic techniques","volume":"9","author":"Maya","year":"2019","journal-title":"J. Drug Deliv. Ther."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"307","DOI":"10.11113\/mjfas.v15n2-1.1542","article-title":"Cytotoxicity, antioxidant and phytochemical screening of propolis extracts from four different Malaysian stingless bee species","volume":"15","author":"Nafi","year":"2019","journal-title":"Mal. J. Fund. Appl. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"59","DOI":"10.22271\/phyto.2020.v9.i5a.12664","article-title":"Phytochemical constituents of dichloromethane fraction and essential oil of Napoleonaea imperialis rind","volume":"9","author":"Jack","year":"2020","journal-title":"J. Pharmacogn. Phytochem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"35851","DOI":"10.1021\/acsomega.2c04195","article-title":"Isolation and identification of flavones responsible for the antibacterial activities of Tillandsia bergeri extracts","volume":"7","author":"Lo","year":"2022","journal-title":"ACS Omega"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"012041","DOI":"10.1088\/1757-899X\/1011\/1\/012041","article-title":"Metabolite detection and antibacterial activity of fungal endophytic extracts isolated from Brotowali (Tinospora crispa) plants using TLC-bioautography assay","volume":"1011","author":"Fathoni","year":"2021","journal-title":"IOP Conf. Ser. Mater. Sci. Eng."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Dasgupta, A., and Acharya, K. (2019). Mushrooms: An emerging resource for therapeutic terpenoids. 3 Biotech, 9.","DOI":"10.1007\/s13205-019-1906-2"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"111982","DOI":"10.1016\/j.foodres.2022.111982","article-title":"New insights into antimicrobial and antibiofilm effects of edible mushrooms","volume":"162","author":"Moussa","year":"2022","journal-title":"Food Res. Int."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2077","DOI":"10.1021\/np800577p","article-title":"Lanostane-type triterpenes from the mushroom Astraeus pteridis with antituberculosis activity","volume":"71","author":"Stanikunaite","year":"2008","journal-title":"J. Nat. Prod."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"9834","DOI":"10.1021\/jf302433r","article-title":"Astraodoric acids A-D: New lanostane triterpenes from edible mushroom Astraeus odoratus and their anti-Mycobacterium tuberculosis H37Ra and cytotoxic activity","volume":"60","author":"Arpha","year":"2012","journal-title":"J. Agric. Food Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/j.phytochem.2012.11.022","article-title":"Lanostane triterpenes from cultures of the Basidiomycete Ganoderma orbiforme BCC 22324","volume":"87","author":"Isaka","year":"2013","journal-title":"Phytochemistry"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1731","DOI":"10.1016\/S0040-4039(00)81756-6","article-title":"Isolation and structure of gymnoprenols, a novel type of polyisoprenepolyols from Gymopilus spectabilis","volume":"24","author":"Nozoe","year":"1983","journal-title":"Tetrahedron Lett."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"156","DOI":"10.3184\/174751916X14546877525997","article-title":"A new monoacylglycerol from the fruiting bodies of Gymnopilus spectabilis","volume":"40","author":"Suh","year":"2016","journal-title":"J. Chem. Res."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Yamada, H., Yamaguchi, M., Chikamatsu, K., Aono, A., and Mitarai, S. (2015). Structome analysis of virulent Mycobacterium tuberculosis, which survives with only 700 ribosomes per 0.1 fl of cytoplasm. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0117109"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1016\/j.jiac.2015.11.014","article-title":"Antimycobacterial activity of methanolic plant extract of Artemisia capillaris containing ursolic acid and hydroquinone against Mycobacterium tuberculosis","volume":"22","author":"Jyoti","year":"2016","journal-title":"J. Infect. Chemother."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"12642","DOI":"10.1073\/pnas.1812334116","article-title":"1,4-Benzoquinone antimicrobial agents against Staphylococcus aureus and Mycobacterium tuberculosis derived from scorpion venom","volume":"116","author":"Sathyamoorthi","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1758","DOI":"10.1016\/j.tetlet.2017.03.066","article-title":"Lanostane triterpenoids from fruiting bodies of the bracket fungus Fomitopsis feei","volume":"58","author":"Isaka","year":"2017","journal-title":"Tetrahedron Lett."},{"key":"ref_46","first-page":"1934578X20903555","article-title":"Advances in pharmacological activities of terpenoids","volume":"15","author":"Yang","year":"2020","journal-title":"Nat. Prod. Commun."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Huang, W., Wang, Y., Tian, W., Cui, X., Tu, P., Li, J., Shi, S., and Liu, X. (2022). Biosynthesis investigations of terpenoid, alkaloid, and flavonoid antimicrobial agents derived from medicinal plants. Antibiotics, 11.","DOI":"10.3390\/antibiotics11101380"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Dulay, R.M., Valdez, B.C., Li, Y., Chakrabarti, S., Dhillon, B., Kalaw, S.P., Reyes, R.G., and Cabrera, E.C. (2021). Cytotoxicity of Gymnopilus purpureosquamulosus extracts on hematologic malignant cells through activation of the SAPK\/JNK signaling pathway. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0252541"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Serras, A.S., Rodrigues, J.S., Cipriano, M., Rodrigues, A.V., Oliveira, N.G., and Miranda, J.P. (2021). A critical perspective on 3D liver models for drug metabolism and toxicology studies. Front. Cell Dev. Biol., 9.","DOI":"10.3389\/fcell.2021.626805"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.etap.2016.06.030","article-title":"Non-cytotoxic concentrations of acetaminophen induced mitochondrial biogenesis and antioxidant response in HepG2 cells","volume":"46","author":"Zhang","year":"2016","journal-title":"Environ. Toxicol. Pharmacol."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Zhong, X., Silveira, E.S.R.d.C.d., and Zhong, C. (2017). Mitochondrial biogenesis in response to chromium (VI) toxicity in human liver cells. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18091877"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1016\/j.sajb.2015.08.016","article-title":"Isoalantolactone, a sesquiterpene lactone from Artemisia afra Jacq. ex Willd and its in vitro mechanism of induced cell death in HeLa cells","volume":"103","author":"Venables","year":"2016","journal-title":"S. Afr. J. Bot."},{"key":"ref_53","first-page":"197","article-title":"Column chromatography and HPLC analysis of phenolic compounds in the fractions of Salvinia molesta mitchell","volume":"5","author":"Gini","year":"2019","journal-title":"Egypt J. Basic and Appl. Sci."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"362","DOI":"10.1128\/JCM.36.2.362-366.1998","article-title":"Rapid, low-technology MIC determination with clinical Mycobacterium tuberculosis isolates by using the microplate Alamar Blue assay","volume":"36","author":"Franzblau","year":"1998","journal-title":"J. Clin. Microbiol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1055\/s-2005-837826","article-title":"Anti-tuberculosis constituents from the stem bark of Micromelum hirsutum","volume":"71","author":"Ma","year":"2005","journal-title":"Planta Med."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1083\/jcb.25.2.407","article-title":"A simplified lead citrate stain for use in electron microscopy J","volume":"25","author":"Venable","year":"1965","journal-title":"Cell Biol."},{"key":"ref_57","unstructured":"Freshney, R.I. (2015). Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications, John Wiley & Sons. [6th ed.]."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"A.3B.1","DOI":"10.1002\/0471142735.ima03bs21","article-title":"Trypan blue exclusion test of cell viability","volume":"21","author":"Strober","year":"1997","journal-title":"Curr. Protoc. Immunol."},{"key":"ref_59","doi-asserted-by":"crossref","unstructured":"Swanepoel, B., Nitulescu, G.M., Olaru, O.T., Venables, L., and van de Venter, M. (2019). Anti-Cancer Activity of a 5-Aminopyrazole Derivative Lead Compound (BC-7) and Potential Synergistic Cytotoxicity with Cisplatin against Human Cervical Cancer Cells. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20225559"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1089\/zeb.2017.1545","article-title":"ZebraPace: An Open-Source Method for Cardiac-Rhythm Estimation in Untethered Zebrafish Larvae","volume":"15","author":"Gaur","year":"2018","journal-title":"Zebrafish"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1148","DOI":"10.1016\/j.toxrep.2019.10.020","article-title":"Toxicity study on Clinacanthus nutans leaf hexane fraction using Danio rerio embryos","volume":"6","author":"Murugesu","year":"2019","journal-title":"Toxicol. Rep."},{"key":"ref_62","first-page":"9152757","article-title":"Evaluation of the Effect of Aqueous Momordica charantia Linn","volume":"2019","author":"Thiagarajan","year":"2019","journal-title":"Extract on Zebrafish Embryo Model through Acute Toxicity Assay Assessment. Evid. Based Complement. Alternat. Med."}],"container-title":["Antibiotics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6382\/14\/2\/179\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:31:01Z","timestamp":1760027461000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6382\/14\/2\/179"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,11]]},"references-count":62,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["antibiotics14020179"],"URL":"https:\/\/doi.org\/10.3390\/antibiotics14020179","relation":{},"ISSN":["2079-6382"],"issn-type":[{"value":"2079-6382","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,11]]}}}