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Nanotechnology-Mediated Radiation Therapy
Published in D. Sakthi Kumar, Aswathy Ravindran Girija, Bionanotechnology in Cancer, 2023
repair mechanisms, and (iii) cell-cycle effects [118, 122]. PEGylated AuNPs functionalized with a known ROS sensor dihydro-rhodamine 123 (DHR-123) having a size of 20 nm were engineered to effectively monitor the ROS generation in tumor cells that led to apoptotic cell death following radiation exposure [123]. AuNPs with core sizes of 2 nm, 5 nm, and 19 nm when functionalized with prostate-specific membrane antigen-1 (PSMA-1) ligand reported the highest cellular uptake of the smallest size PSMA-1 targeted AuNPs in PSMA expressing tumors followed by improvement in radiation enhancement [124]. Over the years, significant progress has been made to exploit AuNPs not only as a versatile drug delivery platform and imaging modality, but also innovative radiosensitizers and obstacles in the form of in vivo efficacy/fate. However, for the full potential use of AuNPs, further study about safety concerns along with a comprehensive understanding of the radio sensitization mechanisms and radiation energy parameters needs to be conducted.
Models of Toxicity Screening Using Cultured Cells
Published in John J. Lemasters, Constance Oliver, Cell Biology of Trauma, 2020
Roberta L. Grant, Daniel Acosta
The effects of local anesthetics on mitochondrial function were investigated using a fluorescent dye, rhodamine 123 (Rho123). Rho123 is a lipophilic cationic dye that accumulates in the mitochondria because of its negative inner membrane potential. If the mitochondrial membrane potential is dissipated, the dye leaks into the cytoplasm and eventually out of the cell. Tetracaine caused a dissipation of mitochondrial membrane potential at concentrations equal to or greater than 2.5 mM after 15 min of treatment. A concentration of 5 mM caused an immediate dissipation of mitochondrial membrane potential. This concentration may occur clinically after a drop of tetracaine is applied to the eye. Proparacaine at the highest concentration of 5 mM occasionally dissipated mitochondrial membrane potential but this was not a consistent finding. Cocaine caused extensive cell rounding so the effect on mitochondrial membrane potential could not be evaluated accurately by this method.
Fluorescence in Histochemical Reactions
Published in Victoria Vladimirovna Roshchina, Fluorescence of Living Plant Cells for Phytomedicine Preparations, 2020
Victoria Vladimirovna Roshchina
Hydrogen peroxide detection may be carried out with 2,7-dichlorodihydrofluorescein (DCF)-based probes or dihydrorhodamine 123 (DHR)-based dyes. Probes based on DCFH-DA (dichlorodihydrofluorescein diacetate) cross the plasma membrane, and esterases cleave the acetate groups to produce DCFH, which is oxidized to DCF, an anthracene-like chemical structure that is capable of fluorescence. The same scheme can be applied to DHR-based probes; at the top of DCF molecules formulae are amino (-NH2) groups in the case of rhodamine 123. The third possibility is to use Amplex Red® dye, which undergoes oxidative deacetylation in the presence of cellular peroxidases, leading to the production of fluorescent resorufin.
Antioxidant activity of calcitriol reduces direct methamphetamine-induced mitochondrial dysfunction in isolated rat heart mitochondria
Published in Toxin Reviews, 2022
Ahmad Salimi, Morteza Minouei, Mohsen Niknejad, Elham Mojarad Aylar
The mitochondrial membrane potential (ΔΨm) was measured using rhodamine 123. Rhodamine 123 is a cationic, green-fluorescent dye used to detect ΔΨm. Therefore, the mitochondrial membrane potential (ΔΨm) can be measured indirectly by measurement of fluorescence intensity of rhodamine 123 (Mersa et al.2020). Briefly, isolated cardiac mitochondria (1000 µg/ml) were suspended in MMP buffer (68 mM D-mannitol, 220 mM sucrose, 5 mM KH2PO4, 10 mM KCl, 2 mM MgCl2, 50 μM EGTA, 10 mM HEPES, 5 mM sodium succinate, 1 µM of rhodamine123, 2 μM Rotenone at pH 7.4). To test the effect of calcitriol against methamphetamine-induced mitochondrial membrane potential collapse, the isolated cardiac mitochondria were treated with various concentrations of calcitriol (1, 2.5 and 5 µM) and 250 µM methamphetamine at 37 °C in 60 min. After 1 h of incubation the fluorescence intensity of rhodamine 123 was measured using a Cyflow Space-Partec flowcytometry.
Exenatide-loaded inside-porous poly(lactic-co-glycolic acid) microspheres as a long-acting drug delivery system with improved release characteristics
Published in Drug Delivery, 2020
Junqiu Zhai, Zhanlun Ou, Liuting Zhong, Yu-e Wang, Li-Ping Cao, Shixia Guan
Exenatide was purchased from Jianyuan Pharmaceutical Technology Co., Ltd. (Shenzhen China). Poly(vinyl alcohol) 210 (PVA), sodium azide, and sodium dodecyl sulfate (SDS) were obtained from Aladdin Bio-Chem Technology Co., Ltd. (Shanghai, China). PLGA (85/15) (lactic acid (LA)/glycolic acid (GA) = 85/15, Mw = 50,000), PLGA (75/25) (LA/GA= 75/25, Mw = 50,000), and PLGA (50/50) (LA/GA = 50/50, Mw = 50,000) were all purchased from Shandong Academy of Pharmaceutical Sciences (Pilot Plant; Jinan, China). Micro-BCA kit was supplied by Shenggong Biological Engineering Co. Ltd. (Shanghai, China). Rhodamine 123 (Rh123) was supplied by Shanghai Yuanye Bio-technology Co., Ltd. (Shanghai, China). Dichloromethane (DCM), NH4HCO3, and dimethyl carbonate (DMC) were all purchased from Damao Chemical Reagent Factory (Tianjin, China). All chemicals and reagents were analytical reagent grade.
trans-Anethole Abrogates Cell Proliferation and Induces Apoptosis through the Mitochondrial-Mediated Pathway in Human Osteosarcoma Cells
Published in Nutrition and Cancer, 2021
Kritika Pandit, Sandeep Kaur, Ajay Kumar, Renu Bhardwaj, Satwinderjeet Kaur
For estimating the functional status of mitochondria, the alterations in mitochondrial membrane potential were assessed with the fluorescent probe Rh-123 as described by (29). Rhodamine 123 is a cationic dye used to determine the changes in the mitochondrial membrane potential of MG-63 cells treated with trans-Anethole. In brief, MG-63 cells (4 × 105cells/well) were treated with trans-Anethole at different concentrations (25 µM, 50 µM, and 100 µM) for 24 h. After incubation, the cell culture was stained with Rh-123 (5 μg/ml) for 30 min in dark. Cells were collected and the pellet was washed with 1 ml of PBS and centrifuged at 1,500 rpm. The fluorescence intensity from 20,000 events was evaluated by FACS analysis using the BD Accuri C6 Flow cytometer.