• Title/Summary/Keyword: Fluorometry

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Nobiletin attenuates neurotoxic mitochondrial calcium overload through K+ influx and ∆Ψm across mitochondrial inner membrane

  • Lee, Ji Hyung;Amarsanaa, Khulan;Wu, Jinji;Jeon, Sang-Chan;Cui, Yanji;Jung, Sung-Cherl;Park, Deok-Bae;Kim, Se-Jae;Han, Sang-Heon;Kim, Hyun-Wook;Rhyu, Im Joo;Eun, Su-Yong
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.3
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    • pp.311-319
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    • 2018
  • Mitochondrial calcium overload is a crucial event in determining the fate of neuronal cell survival and death, implicated in pathogenesis of neurodegenerative diseases. One of the driving forces of calcium influx into mitochondria is mitochondria membrane potential (${\Delta}{\psi}_m$). Therefore, pharmacological manipulation of ${\Delta}{\psi}_m$ can be a promising strategy to prevent neuronal cell death against brain insults. Based on these issues, we investigated here whether nobiletin, a Citrus polymethoxylated flavone, prevents neurotoxic neuronal calcium overload and cell death via regulating basal ${\Delta}{\psi}_m$ against neuronal insult in primary cortical neurons and pure brain mitochondria isolated from rat cortices. Results demonstrated that nobiletin treatment significantly increased cell viability against glutamate toxicity ($100{\mu}M$, 20 min) in primary cortical neurons. Real-time imaging-based fluorometry data reveal that nobiletin evokes partial mitochondrial depolarization in these neurons. Nobiletin markedly attenuated mitochondrial calcium overload and reactive oxygen species (ROS) generation in glutamate ($100{\mu}M$)-stimulated cortical neurons and isolated pure mitochondria exposed to high concentration of $Ca^{2+}$ ($5{\mu}M$). Nobiletin-induced partial mitochondrial depolarization in intact neurons was confirmed in isolated brain mitochondria using a fluorescence microplate reader. Nobiletin effects on basal ${\Delta}{\psi}_m$ were completely abolished in $K^+-free$ medium on pure isolated mitochondria. Taken together, results demonstrate that $K^+$ influx into mitochondria is critically involved in partial mitochondrial depolarization-related neuroprotective effect of nobiletin. Nobiletin-induced mitochondrial $K^+$ influx is probably mediated, at least in part, by activation of mitochondrial $K^+$ channels. However, further detailed studies should be conducted to determine exact molecular targets of nobiletin in mitochondria.

A Study on the Blue Fluorescence Characteristics of Silica Nanoparticles with Different Particle Size (실리카 나노 입자의 크기에 따른 청색 형광 특성 연구)

  • Yoon, Ji-Hui;Kim, Ki-Chul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.5
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    • pp.1-6
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    • 2019
  • Organic dye-doped silica nanoparticles are used as a promising nanomaterials for bio-labeling, bio-imaging and bio-sensing. Fluorescent silica nanoparticles(NPs) have been synthesized by the modified $St{\ddot{o}}ber$ method. In this study, dye-free fluorescent silica NPs of various sized were synthesized by Sol-Gel process as the modified $St{\ddot{o}}ber$ method. The functional material of APTES((3-aminopropyl)triethoxysilane) was added as an additive during the Sol-Gel process. The as-synthesized silica NPs were calcined at $400^{\circ}C$ for 2 hours. The surface morphology and particle size of the as-synthesized silica NPs were characterized by field-emission scanning electron microscopy. The fluorescent characteristics of the as-synthesized silica NPs was confirmed by UV lamp irradiation of 365 nm wavelength. The photoluminescence (PL) of the as-synthesized silica NPs with different size was analyzed by fluorometry. As the results, the as-synthesized silica NPs exhibits same blue fluorescent characteristics for different NPs size. Especially, as increased of the silica NPs size, the intensity of PL was decreased. The blue fluorescence of dye-free silica NPs was attributed to linkage of $NH_2$ groups of the APTES layer and oxygen-related defects in the silica matrix skeleton.

Comparison of Dissolved Ammonium Analytical Method in Seawater: Spetrophotometry and Fluorometry (해수 중 용존 암모늄 분석방법 비교: 분광광도법과 형광법)

  • SON, PURENA;PARK, JOONSEONG;RHO, TAEKEUN
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.25 no.4
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    • pp.81-96
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    • 2020
  • Berthlot's reaction spectrophotometric method is generally used for the analysis of dissolved ammonium in seawater, but in recent years, a fluorescence method using an orthophthaldialdehyde-sulfite (OPA) fluorescent reagent is actively used internationally. In this study, we investigated the effects of the detection limit between the analysis methods, the reagent refractive index inherent in the spectrophotometric method, and the use of different calibration curves to understand the cause of the difference in dissolved ammonium concentration (about 0.31 𝜇M) observed in the seawater samples and a nutrient reference material between two institutions (KIOST (spectrophotometric method, one-order linear regression gradient only), Australia CSIRO (fluorescence method, quadratic formula)) conducted onboard the Australian R/V Investigator in 2017. The method detection limit (0.063 𝜇M) and the reagent refractive index background value (0.054 𝜇M) of the spectrophotometric method measured in this study could explain the difference in dissolved ammonium concentration values of the two institutes about 20% and 17%, respectively. However, when the concentration of the calibration curve of the spectrophotometric method was calculated using the same quadratic as the fluorescence method or the slope and intercept of linear regression, the difference in the dissolved ammonium concentration between the two institutions was reduced to less than the detection limit of the spectrophotometric method. Therefore, the difference in the concentration of dissolved ammonium between the two institutions, found in the nutrient reference materials and the seawater field sample during the international onboard nutrient inter-comparison experiment, may be attributed to be the effect of the different calibration curves used in the two methods rather than the effect of the difference in two analytical methods. When comparing the dissolved ammonium data from seawater samples in the future, it is recommended to pay attention to the information on the baseline, number of standard solutions, and calibration curve used in the analysis.