In situ analysis of apoptosis in Aspergillus nidulans with ethidium bromide and acridine orange

Leles, S M; Factori, R; Rocha, C.L.M.S.C.

doi:10.4238/2013.august.12.5

Cited by 4 publications

(6 citation statements)

References 28 publications

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“…In the current research both acridine orange and ethidium bromide could enter the apoptotic cells, and an orange color was produced that was visible by a fluorescence microscope. In a study Aspergillus colonies that were in the apoptosis phase appeared orange, while the healthy colonies appeared green, and the dead colonies were red (Leles & Factori, 2013).…”

Section: Annexin V-pimentioning

confidence: 99%

Anti-cancer effects of the methanolic extract of Persian Gulf brittle star

Farrokyar¹,

Baharara²,

Ramzani³

2018

nbr

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Nowadays natural compounds are increasingly used in various fields since they cause fewer adverse side effects than their chemical counterparts. In addition, preparation and production of natural compounds are comparatively simple. The present study aimed to evaluate the anti-cancer effects of brittle star methanol extract on MCF-7 cells. Anti-cancer effects of brittle star methanol extract were examined on the MCF-7 cell line. Moreover, the toxicity of this compound was assessed using cell apoptosis methods, ethidium bromide/acridine orange (EB/AO) staining and Annexin V Apoptosis Detection Kit. According to the MTT assay, the methanolic extract of brittle star could prevent proliferation in a time-and dose-dependent manner. The calculated inhibitory concentration that decreased the cell-viability by half (IC50) was estimated at 150 µg/ml. On the other hand, results of EB/AO staining were indicative of increased apoptotic cells in the treatment group. In addition, results of Annexin V/PI protocol demonstrated a significant enhancement in the percentage of late apoptotic cells (29.02%) in the treatment groups (150 µg/ml) and the percentage of necrotic cells were 2.77% for a period of 24 hours. According to the results, the methanolic extract of Persian Gulf brittle star induced apoptosis in the MCF-7 cell line. These effects could be attributed to the presence of certain active biological compounds, such as Saponins, Naphthoquinones and Carotenoids. Furthermore, our findings suggested that brittle star might have beneficial effects on cancer management. Therefore, this natural marine compound could be used as a complementary element in the prevention and treatment of cancer.

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Section: Annexin V-pimentioning

confidence: 99%

Anti-cancer effects of the methanolic extract of Persian Gulf brittle star

Farrokyar¹,

Baharara²,

Ramzani³

2018

nbr

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“…As a result, acridine orange can be used in apoptotic studies to stain apoptotic cells orange or red depending on the degree of loss of membrane integrity. This dye also has the capacity to label dead cells thus differentiating the apoptotic cells into early apoptosis (green) and late apoptosis (orange red) hence it offers superior accuracy than the older methods [10,11,38].…”

Section: Acridine Orangementioning

confidence: 99%

“…Therefore, dual staining of cells with acridine orange and propidium iodide/ ethidium bromide aids the detection of four main types of cells: (a) Viable cells with uniform green nuclei with organised chromatin structure (b) Necrotic cells with uniform orange red or red nuclei with organised structure due to loss of membrane integrity (c) Early apoptotic cells with irregular structured green nucleic but chromatin is condensed as apoptotic bodies or green patches/fragments (d) late apoptotic cells, with orange or red nuclei with extremely condensed or fragmented chromatin [10,11,38,[40][41][42]. Thus, since the cellular detail and nuclear outline are distinct, co-staining cells with acridine orange and ethidium bromide/propidium iodide offers a rapid, stable, sensitive, and easy-to-perform way to simultaneously visualise and identify all the possible nuclear stages with increased accuracy and ease of interpretation.…”

Section: Stromal Cell Line Hs-5 Cells Stained With Acridine Orange Wh...mentioning

confidence: 99%

“…Therefore, these cytotoxicity assays directly or indirectly assess cytotoxicity by providing information on cell membrane integrity, cell metabolism, cell machinery, cell mortality and cell proliferation [5][6][7][8]. Some of these sensitive and reproducible assays are used to obtain colorimetric, luminometric and fluorometric measurements hence entails the use of different dyes such as trypan blue exclusion dye, neutral red, acridine orange and propidium iodide [1,[9][10][11][12]. These dyes offer different advantages and disadvantages hence a combination of two or more dyes is usually employed in in vitro studies to avoid overestimation or underestimation of the toxicity of a chemical substance thereby increasing the reliability of the results obtained.…”

Section: Introductionmentioning

confidence: 99%

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Trypan Blue Exclusion Assay, Neutral Red, Acridine Orange and Propidium Iodide

Ude¹,

Afi-Leslie²,

Okeke³

et al. 2023

Cytotoxicity - Understanding Cellular Damage and Response

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Cytotoxicity and cell viability assessments are very important parameters that are widely used in fundamental research and drug development to determine the safety profile of toxic compounds. These assays measure the degree to which a substance can cause toxic damage to cells or cell death. There are different assays that have been employed to determine the cytotoxicity of substances. These assays either determine enzymatic function, cell viability, mitochondrial activity, lipid metabolism, cell proliferation and/or cell death. These assays entail use of different kinds of dyes such as trypan blue exclusion dye, neutral red, acridine orange and propidium iodide to stain the cells. Trypan blue dye permeates compromised cell membrane to stain necrotic cells. However, this can lead to false positive and false negative results as it does not provide information on sub-lethal injury. As a result, neutral red and acridine orange can be used as counterstains for trypan blue to stain the lysosome of live cells. Acridine orange can also be used to stain nucleic acids in living cells and is usually co-stained with propidium iodide or ethidium bromide. This is because propidium iodide and ethidium bromide permeate only compromised plasma membrane thus co-staining cells with these dyes can provide vital information that can be used to differentiate between live and dead cells.

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“…Nesse contexto, o emprego de Aspergillus nidulans como sistema biológico preliminar constitui uma alternativa eficaz por se tratar de célula eucariótica com mecanismos celulares e genéticos mais complexos que Salmonella thyphimurium (teste de Ames), fornecendo rápida triagem das atividades biológicas de agentes (31), como já realizados por outros pesquisadores (32)(33)(34)(35)(36)(37)(38). Além disso, segundo Fenech (39), a utilização de ensaios microbiológicos para avaliação de genotoxicidade é muito comum, uma vez que são sistemas rápidos, baratos e produzem resultados valiosos.…”

Section: Introductionunclassified

Bioatividade de frações de sementes e pericarpo de Annona squamosa (Annonaceae) em Aspergillus nidulans

Guidoti

Romero

et al. 2021

SaBios

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As plantas são amplamente utilizadas na medicina popular e apresentam os chamados constituintes bioativos, produzidos sob a forma de metabólitos secundários, capazes de desencadear efeitos farmacológico ou toxicológico no homem e nos animais. Annona squamosa, conhecida popularmente como fruta-do-conde, é empregada pela população na cura de doenças; possui amplo espectro nutricional e representa uma excelente fonte de diversas classes de constituintes bioativos, como alcaloides, terpenos, taninos, compostos fenólicos e acetogeninas. Apesar das atividades biológicas já descritas para essa planta, é necessária a investigação de extratos e/ou isolados a fim de verificar a genotoxicidade de extratos de frações obtidas a partir de sementes e pericarpo de A. squamosa na germinação de conídios de Aspergillus nidulans, utilizando como parâmetro a estimativa de mortos e malformados. Os resultados indicaram que a fração acetato de etila do pericarpo apresentou, em menores concentrações (2,5 e 25 μg.mL-1) efeitos pró-reparo, e antiapoptótico na maior concentração (250 μg.mL-1). A fração metanólica do extrato do pericarpo demonstrou efeito apoptótico, também em menores concentrações e efeito genotóxico na maior concentração. Quanto ao extrato das sementes, a fração acetato de etila também indicou ativação de apoptose em todas as concentrações, enquanto a fração metanólica apresentou efeito pró-reparo. A diversidade de bioatividades de frações dessa planta reflete a presença de inúmeras classes de metabólitos secundários que poderão atuar beneficamente na saúde humana. Portanto, o presente trabalho contribui na escolha de frações para futuro isolamento e identificação química de metabólitos com potencial medicinal.

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In situ analysis of apoptosis in Aspergillus nidulans with ethidium bromide and acridine orange

Cited by 4 publications

References 28 publications

Anti-cancer effects of the methanolic extract of Persian Gulf brittle star

Anti-cancer effects of the methanolic extract of Persian Gulf brittle star

Trypan Blue Exclusion Assay, Neutral Red, Acridine Orange and Propidium Iodide

Bioatividade de frações de sementes e pericarpo de Annona squamosa (Annonaceae) em Aspergillus nidulans

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