• Bulletin of the Korean Chemical Society
• /
• 제32권5호
• /
• pp.1599-1603
• /
• 2011

Novel chemosensors based on 2-(2'-hydroxyphenyl)quinoline were prepared and evaluated for sensing metal cations. The photophysical properties of chemosensors 1-3 were examined and their ion-selectivity was evaluated by measuring their fluorescent emission responses to alkali, alkaline earth, and transition metal ions. Chemosensors 1, 2 and 3 show ratiometric and enhanced fluorescence changes with transitional metals that are efficient fluorescence quenchers, especially 3 has a high binding constant with $Hg^{+2}$ in $CH_3CN$.

• Bulletin of the Korean Chemical Society
• /
• 제29권3호
• /
• pp.567-570
• /
• 2008

A new pyrene appended diaza-18-crown-6 ether derivative 1 has been prepared and its fluoroionophoric properties toward transition metal ions were investigated. Compound 1 exhibited a high Hg2+-selectivity over other transition metal ions as well as alkali and alkaline earth metal ions in aqueous acetonitrile solution. The ratiometric analysis of the monomer and excimer emissions of pyrene successfully signals the presence of Hg2+ ions. The detection limit for Hg2+ ions was found to be 3.1 ´ 10-6 M in 50% aqueous acetonitrile solution at pH 8.1. Competition experiments also suggest that the compound could be utilized as a selective and sensitive fluorescent chemosensor for the analysis of micromolar Hg2+ ions in physiological and environmental samples.

• Bulletin of the Korean Chemical Society
• /
• 제30권5호
• /
• pp.1093-1096
• /
• 2009

Two new chemosensors 1 and 2 derived from 5,12- and 5,7-dioxocyclams were prepared and their signaling behaviors toward transition metal ions were investigated. Chemosensors 1 and 2 showed very efficient responses toward $Cu_{2+}$ and $Hg_{2+}$ ions. Ratiometric analysis of the fluorescence changes in pyrene monomer and excimer emissions clearly demonstrated the $Cu_{2+}$- and $Hg_{2+}$-selective signaling behavior. The signaling mechanism of the chemosensors is due to conformation changes upon complexation with metal ions and the inherent quenching nature of the complexed $Cu_{2+}$ and $Hg_{2+}$ ions themselves.

• BMB Reports
• /
• 제46권12호
• /
• pp.600-605
• /
• 2013

Interplay between calcium ions ($Ca^{2+}$) and reactive oxygen species (ROS) delicately controls diverse pathophysiological functions of vascular smooth muscle cells (VSMCs). However, details of the $Ca^{2+}$ and ROS signaling network have been hindered by the absence of a method for dual measurement of $Ca^{2+}$ and ROS. Here, a real-time monitoring system for $Ca^{2+}$ and ROS was established using a genetically encoded hydrogen peroxide indicator, HyPer, and a ratiometric $Ca^{2+}$ indicator, fura-2. For the simultaneous detection of fura-2 and HyPer signals, 540 nm emission filter and 500 nm~ dichroic beamsplitter were combined with conventional exciters. The wide excitation spectrum of HyPer resulted in marginal cross-contamination with fura-2 signal. However, physiological $Ca^{2+}$ transient and hydrogen peroxide were practically measurable in HyPer-expressing, fura-2-loaded VSMCs. Indeed, distinct $Ca^{2+}$ and ROS signals could be successfully detected in serotonin-stimulated VSMCs. The system established in this study is applicable to studies of crosstalk between $Ca^{2+}$ and ROS.

• Journal of Microbiology and Biotechnology
• /
• 제34권4호
• /
• pp.783-794
• /
• 2024

The antifungal activity of fisetin against Candida albicans is explored, elucidating a mechanism centered on membrane permeabilization and ensuing disruption of pH homeostasis. The Minimum Inhibitory Concentration (MIC) of fisetin, indicative of its interaction with the fungal membrane, increases in the presence of ergosterol. Hoechst 33342 and propidium-iodide staining reveal substantial propidium-iodide accumulation in fisetin-treated C. albicans cells at their MIC, with crystal violet uptake assays confirming fisetin-induced membrane permeabilization. Leakage analysis demonstrates a significant release of DNA and proteins in fisetin-treated cells compared to controls, underscoring the antifungal effect through membrane disruption. Green fluorescence, evident in both the cytoplasm and vacuoles of fisetin-treated cells under BCECF, AM staining, stands in contrast to controls where only acidic vacuoles exhibit staining. Ratiometric pH measurements using BCECF, AM reveal a noteworthy reduction in intracellular pH in fisetin-treated cells, emphasizing its impact on pH homeostasis. DiBAC4(3) uptake assays demonstrate membrane hyperpolarization in fisetintreated cells, suggesting potential disruptions in ion flux and cellular homeostasis. These results provide comprehensive insights into the antifungal mechanisms of fisetin, positioning it as a promising therapeutic agent against Candida infections.

• 대한기계학회논문집B
• /
• 제36권5호
• /
• pp.533-538
• /
• 2012

본 논문에서는 산소 민감 발광 염료를 이용하여 마이크로 채널 내에서 세포가 배양되고 있을 때 산소농도를 측정하였다. 현재까지 알려진 여러 산소 민감 발광 염료 중 본 논문에서는 물에 잘 녹으며 장 시간 동안 사용하여도 독성이 없는 것으로 알려진 $[Ru(bpy)_3]^{2+}$를 사용하였으며 이와 더불어 산소 민감 염료 측정법의 단점을 보완하기 위하여 칼세인 염료를 이용하여 두 염료의 밝기 비율을 구하여 농도를 측정하였다. SCOMS 카메라와 마이크로 채널을 이용하여 캘리브레이션을 실시하고 농도와 밝기와의 관계를 구하였으며 이 관계를 이용하여 세포가 배양되는 조건에서의 배양액의 산소농도를 측정하였다. 실험 결과 채널입구에서 점점 멀어질수록 마이크로 채널 내에서 산소농도는 점점 낮아진다는 것을 관찰 할 수 있었다.

• International Journal of Oral Biology
• /
• 제46권4호
• /
• pp.176-183
• /
• 2021

Oral squamous cell carcinoma (OSCC) metastasis is characterized by distant metastasis and local recurrence. Combined chemotherapy with cisplatin and 5-fluorouracil is routinely used to treat patients with OSCC, and the combined use of gefitinib with cytotoxic drugs has been reported to enhance the sensitivity of cancer cells in vitro. However, the development of drug resistance because of prolonged chemotherapy is inevitable, leading to a poor prognosis. Therefore, understanding alterations in signaling pathways and gene expression is crucial for overcoming the development of drug resistance. However, the altered characterization of Ca²⁺ signaling in drug-resistant OSCC cells remains unclear. In this study, we investigated alterations in intracellular Ca²⁺ ([Ca²⁺]_i) mobilization upon the development of gefitinib resistance in human tongue squamous carcinoma cell line (HSC)-3 and HSC-4 using ratiometric analysis. This study demonstrated the presence of altered epidermal growth factor- and purinergic agonist-mediated [Ca²⁺]_i mobilization in gefitinib-resistant OSCC cells. Moreover, Ca²⁺ content in the endoplasmic reticulum, store-operated calcium entry, and lysosomal Ca²⁺ release through the transient receptor potential mucolipin 1, were confirmed to be significantly reduced upon the development of apoptosis resistance. Consistent with [Ca²⁺]_i mobilization, we identified modified expression levels of Ca²⁺ signaling-related genes in gefitinib-resistant cells. Taken together, we propose that the regulation of [Ca²⁺]_i mobilization and related gene expression can be a new strategy to overcome drug resistance in patients with cancer.

• Tuberculosis and Respiratory Diseases
• /
• 제85권2호
• /
• pp.147-154
• /
• 2022

Background: The expression of calcium signaling pathway molecules is altered in various carcinomas, which are related to the proliferation and altered characteristics of cancer cells. However, changes in calcium signaling in anti-cancer drug-resistant cells (bearing a T790M mutation in epidermal growth factor receptor [EGFR]) remain unclear. Methods: Afatinib-mediated changes in the level of store-operated Ca²⁺ entry (SOCE)-related proteins and intracellular Ca²⁺ level in non-small cell lung cancer cells with T790M mutation in the EGFR gene were analyzed using western blot and ratiometric assays, respectively. Afatinib-mediated autophagic flux was evaluated by measuring the cleavage of LC3B-II. Flow cytometry and cell proliferation assays were conducted to assess cell apoptosis and proliferation. Results: The levels of SOCE-mediating proteins (ORAI calcium release-activated calcium modulator 1 [ORAI1], stromal interaction molecule 1 [STIM1], and sarco/endoplasmic reticulum Ca²⁺ ATPase [SERCA2]) decreased after afatinib treatment in non-small cell lung cancer cells, whereas the levels of SOCE-related proteins did not change in gefitinib-resistant non-small cell lung cancer cells (PC-9/GR; bearing a T790M mutation in EGFR). Notably, the expression level of SOCE-related proteins in PC-9/GR cells was reduced also responding to afatinib in the absence of extracellular Ca²⁺. Moreover, extracellular Ca²⁺ influx through the SOCE was significantly reduced in PC-9 cells pre-treated with afatinib than in the control group. Additionally, afatinib was found to decrease the level of SOCE-related proteins through autophagic degradation, and the proliferation of PC-9GR cells was significantly inhibited by a lack of extracellular Ca²⁺. Conclusion: Extracellular Ca²⁺ plays important role in afatinib-mediated autophagic degradation of SOCE-related proteins in cells with T790M mutation in the EGFR gene and extracellular Ca²⁺ is essential for determining anti-cancer drug efficacy.