Spectrofluorometric determination of intracellular levels of reactive oxygen species in drug-sensitive and drug-resistant cancer cells using the 2′,7′-dichlorofluorescein diacetate assay

Loetchutinat, Chatchanok; Kothan, S.; Dechsupa, Nathupakorn; Meesungnoen, Jintana; Jay‐Gerin, Jean‐Paul; Mankhetkorn, Samlee

doi:10.1016/j.radphyschem.2004.06.011

Cited by 217 publications

(96 citation statements)

References 28 publications

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“…Конеч-ная концентрация красителя в инкубационной среде составляла 10 мкМ/мл. ДХФ-ДА проявляет селективную чувствительность к Н 2 О 2 и суперок-сид-аниону О 2 -и, окисляясь в их присутствии внутри клетки, приобретает флуоресцентные свойства [22]. Оценку флуоресцентной активно-сти проводили на планшетном ридере Fluoroscan Ascent FL (Labsystems, США) при экстинкции 485 нм и эмиссии 538 нм, Scaling Factor: 1/1 и вы-ражали в единицах флуоресценции.…”

Section: т 20 №unclassified

COMPARATIVE STUDY OF ECHINOCHROME A, OXYGENATED CAROTENOIDS, GINSENOSIDE Rh2, LUTEOLIN DISULFATE AND METFORMIN AS A MEAN TO POTENTIATE ANTITUMOR EFFECT OF DOXORUBICIN

Tsybulsky¹,

Popov²,

Климович³

et al. 2018

Med. immunol.

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Section: т 20 №unclassified

COMPARATIVE STUDY OF ECHINOCHROME A, OXYGENATED CAROTENOIDS, GINSENOSIDE Rh2, LUTEOLIN DISULFATE AND METFORMIN AS A MEAN TO POTENTIATE ANTITUMOR EFFECT OF DOXORUBICIN

Tsybulsky¹,

Popov²,

Климович³

et al. 2018

Med. immunol.

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“…ROS levels were measured by a spectrofluorimetric method (LOETCHUTINAT et al, 2005), where the sperm was incubated in Tris-HCl in the presence of 2',7'-dichloro dihydrofluorescein diacetate (DCHFDA) for 60 min at 37 °C in the dark. This dye is a fluorogenic probe commonly used to detect cellular ROS production.…”

Section: Assessment Of Oxidative Stressmentioning

confidence: 99%

Use of a colloid to optimize centrifugation in the selection of bovine sperm for IVF

Gonçalves

Leivas

Míssio

et al. 2018

SCA

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This study aimed to evaluate the effect of the force and duration of centrifugation and the impact of cushioned centrifugation on sperm selection by Percoll gradient, on sperm quality and development kinetics of in vitro produced bovine embryos. Two experiments were performed. In Experiment I, a pool of semen was selected by Percoll gradients and the pellet was divided into four groups and distributed in a 2 × 2 factorial, with two forces (2200 × g or 9000 × g) and two durations (1 min or 3 min) of centrifugation. In Experiment II, semen was divided into two groups and selected by Percoll gradient with Cushion Fluid (CF) or without CF (Control) in the second centrifugation. The morphofunctionality, biochemical characteristics and fertilizing capacity of the selected sperms were evaluated. In addition, the development of the resulting bovine embryos was monitored for 48 h post-insemination. Duncan and Chi-square tests (P < 0.05) were used to compare the means. In Experiment I, there was a significant increase in sperm vigor (P < 0.05) after sperm selection in all treatments. The force and duration of centrifugation did not have any effect on sperm motility, vigor, and recovery rate among the different treatments (P > 0.05). In Experiment II, the recovery rate and reactive oxygen species (ROS) production in semen were similar among treatments (P > 0.05) although a higher ROS production was observed in the CF fertilization medium. Total fertilization rate was superior in the CF group (65.4 ± 5.3%) compared to that in Control (39.6 ± 4.9%). However, the normal fertilization and cleavage rate did not differ between the Control (94 ± 6.3% and 58.3 ± 8.3%) and CF (89 ± 7.1% and 75.0 ± 7.3%) groups. The reduction in the force and duration of centrifugation did not decrease the sperm recovery during selection by the Percoll gradient and the use of CF in the second centrifugation did not affect the normal fertilization and development of bovine IVF embryos up to 48 h. Key words: Bovine embryo. Cushion centrifugation. In vitro production. Percoll gradients. Sperm recovery. ResumoEsse estudo objetivou avaliar o efeito da força e duração da centrifugação, e o impacto da centrifugação amortecida na seleção espermática por gradientes de Percoll, na qualidade espermática e cinética do desenvolvimento de embriões bovinos produzidos in vitro. Dois experimentos foram realizados. No experimento I, um pool de sêmen foi selecionado por gradientes de Percoll e o pellet dividido em quatro grupos e distribuído em um fatorial 2 x 2, com duas forças (2200 e 9000 X g) e dois tempos (1 e 3 min) de centrifugação. No Experimento II, o sêmen foi dividido em dois grupos e selecionado com (CF) ou sem CushionFluid (Controle) na segunda centrifugação. A morfofuncionalidade, características bioquímicas e capacidade fecundante dos espermatozoides selecionados foram avaliadas. Além disso, o desenvolvimento dos embriões bovinos resultantes foi monitorado por 48 horas pós-inseminação. Os testes Duncan e Qui-quadrado (P<0,05) foram usados para ...

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“…The ROS levels were determined by a spectrofluorimetric method [28], using 2', 7'-dihydrodichlorofluorescein diacetate (DCHF-DA) assay. Sample aliquot (50 µL) was incubated with 5 µL of DCHF-DA (1 mM).…”

Section: Reactive Oxygen Species Levelsmentioning

confidence: 99%

Justicia insularis Improves the in vitro Survival and Development of Ovine Preantral Follicles Enclosed in Ovarian Tissue

Mbemya¹,

Guerreiro²,

Donfack³

et al. 2017

JPP

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Objectives: Evaluating the addition effect of J. insularis extract and FSH on the survival, activation and ROS production after in vitro culture of ovine preantral follicles enclosed in ovarian tissue. Methods: In the first experiment, ovarian fragments were fixed (non-cultured control) or in vitro cultured in α-MEM + (cultured control), α-MEM + supplemented with FSH 50 ng/mL, or in α-MEM + supplemented with J. insularis (JUS0.3; 1.25 or 5 mg/mL) for 1 or 7 days, at 39˚C, 5% CO 2 . In the second experiment, fragments were fixed or cultured in α-MEM + supplemented with anethole 300 µg/mL + FSH 50 ng/mL or in α-MEM + supplemented with anethole 300 µg/mL + 0.3 mg/mL JUS. Key findings: JUS0.3 was the only treatment that maintained the percentage of morphologically normal follicles similar to non-cultured control even after 7 days of culture. After 7 days of culture, a higher (p < 0.05) percentage of developing follicles was observed in JUS5 treatment compared with the other treatments except JUS1.25. In the second experiment, FSH maintained the percentage of normal follicles and promoted activation of primordial follicles. A reduction (p < 0.05) of stromal cell density was observed in MEM + +ANE supplemented with JUS or FSH. Conclusions: J. insularis in a concentration-dependent manner maintained the levels of ROS and improved in vitro follicular survival and activation of ovine primordial follicles.

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Spectrofluorometric determination of intracellular levels of reactive oxygen species in drug-sensitive and drug-resistant cancer cells using the 2′,7′-dichlorofluorescein diacetate assay

Cited by 217 publications

References 28 publications

COMPARATIVE STUDY OF ECHINOCHROME A, OXYGENATED CAROTENOIDS, GINSENOSIDE Rh2, LUTEOLIN DISULFATE AND METFORMIN AS A MEAN TO POTENTIATE ANTITUMOR EFFECT OF DOXORUBICIN

COMPARATIVE STUDY OF ECHINOCHROME A, OXYGENATED CAROTENOIDS, GINSENOSIDE Rh2, LUTEOLIN DISULFATE AND METFORMIN AS A MEAN TO POTENTIATE ANTITUMOR EFFECT OF DOXORUBICIN

Use of a colloid to optimize centrifugation in the selection of bovine sperm for IVF

Justicia insularis Improves the in vitro Survival and Development of Ovine Preantral Follicles Enclosed in Ovarian Tissue

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