• Molecules and Cells
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• 제38권2호
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• pp.112-121
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• 2015

Ectopic expression of $14-3-3{\zeta}$ has been found in various malignancies, including lung cancer, liver cancer, head and neck squamous cell carcinoma (HNSCC), and so on. However, the effect of $14-3-3{\zeta}$ in the regulation of interactions between tumor cells and the immune system has not been previously reported. In this study, we aimed to investigate whether and how $14-3-3{\zeta}$ is implicated in tumor inflammation modulation and immune recognition evasion. In oral squamous cell carcinoma (OSCC) cell lines and cancer tissues, we found that $14-3-3{\zeta}$ is overexpressed. In OSCC cells, $14-3-3{\zeta}$ knockdown resulted in the up-regulated expression of inflammatory cytokines. In contrast, $14-3-3{\zeta}$ introduction attenuated cytokine expression in human normal keratinocytes and fibroblasts stimulated with interferon-${\gamma}$ (IFN-${\gamma}$) and lipopolysaccharide (LPS). Furthermore, supernatants from $14-3-3{\zeta}$ knockdown OSCC cells dramatically altered the response of peritoneal macrophages, dendritic cells and tumor-specific T cells. Interestingly, Stat3 was found to directly interact with $14-3-3{\zeta}$ and its disruption relieved the inhibition induced by $14-3-3{\zeta}$ in tumor inflammation. Taken together, our studies provide evidence that $14-3-3{\zeta}$ may regulate tumor inflammation and immune response through Stat3 signaling in OSCC.

• BMB Reports
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• 제56권6호
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• pp.341-346
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• 2023

Acute myocardial infarction (AMI) is a multifaceted syndrome influenced by the functions of various extrinsic and intrinsic pathways and pathological processes, which can be detected in circulation using biomarkers. In this study, we investigated the secretome protein profile of induced-hypertrophy cardiomyocytes to identify next-generation biomarkers for AMI diagnosis and management. Hypertrophy was successfully induced in immortalized human cardiomyocytes (T0445) by 200 nM ET-1 and 1 μM Ang II. The protein profiles of hypertrophied cardiomyocyte secretomes were analyzed by nano-liquid chromatography with tandem mass spectrometry and differentially expressed proteins that have been identified by Ingenuity Pathway Analysis. The levels of 32 proteins increased significantly (>1.4 fold), whereas 17 proteins (<0.5 fold) showed a rapid decrease in expression. Proteomic analysis showed significant upregulation of six 14-3-3 protein isoforms in hypertrophied cardiomyocytes compared to those in control cells. Multi-reaction monitoring results of human plasma samples showed that 14-3-3 protein-zeta levels were significantly elevated in patients with AMI compared to those of healthy controls. These findings elucidated the role of 14-3-3 protein-zeta in cardiac hypertrophy and cardiovascular disorders and demonstrated its potential as a novel biomarker and therapeutic strategy.

• 대한전자공학회논문지
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• 제24권2호
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• pp.242-247
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• 1987

Electrophysical phenomena at the silicon semiconductor-electrolyte solution interfaces were analyzed based on the zeta potential of the electrical double layer and microelectrophoresis. The suspensions were composed of the p or n-type silicon particles suspended in the KCI or pH buffer solutions. The approximate diameter of the prepared and sampled sioicon semiconductor pardticles was 1.5\ulcorner. The sign of the zeta poetntials of the p and n-type silicon particles in the KCl and pH buffer solution was positive. A range of electrophoretic mobilities of the p and n-type silicons in the KCl solutions was 5.5-8.9x10**-4 cm\ulcornerV-sec and 4.2-7.9x10**-4cm\ulcornerV-sec, respectively. The range of zeta potentials corresponding to the electrophoretic mobilities is 70.4-114.0mV nad 53.9-101.2mV, respectively. On the other hand, a range of electrophoretic mobilities of the p and n-type silicons in the pH buffer solutions was 1.1x10**-4-2.2x10**-3cm\ulcornerV-sec and 0-2.1x10**-3cm\ulcornerV-sec, respectively. The range of zeta potentials corresponding to the electrophoretic mobilities is 14.1-281.6mV and 0-268.8mV, respectively. The zeta potentials and electrical double layers of the doped silicon semiconductors are decisively influenced by the positively charged ions in the solutions. The maximum values of the zeta potentials in the KCl solutions appeared at a concentration of about 10-\ulcorner. The isoelectric point of the n-type silicon semiconductors appeared at about a pH 7. The effect of the space charge of the doped silicon semiconductors can be neglected compare with the effect of the surface charge.

• BMB Reports
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• 제50권6호
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• pp.335-340
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• 2017

Although doxorubicin (Dox)-induced oxidative stress is known to be associated with cytotoxicity, the precise mechanism remains unclear. Genotoxic stress not only generates free radicals, but also affects actin cytoskeleton stability. We showed that Dox-induced RhoA signaling stimulated actin cytoskeleton alterations, resulting in central stress fiber disruption at early time points and cell periphery cortical actin formation at a later stage, in HeLa cells. Interestingly, activation of a cofilin phosphatase, chronophin (CIN), was initially evoked by Dox-induced RhoA signaling, resulting in a rapid phosphorylated cofilin turnover leading to actin cytoskeleton remodeling. In addition, a novel interaction between CIN and $14-3-3{\zeta}$ was detected in the absence of Dox treatment. We demonstrated that CIN activity is quite contrary to $14-3-3{\zeta}$ binding, and the interaction leads to enhanced phosphorylated cofilin levels. Therefore, initial CIN activation regulation could be critical in Dox-induced actin cytoskeleton remodeling through RhoA/cofilin signaling.

• Asian-Australasian Journal of Animal Sciences
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• 제28권3호
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• pp.420-427
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• 2015

It is necessary to understand the cellular uptake and cytotoxicity of food-grade delivery systems, such as ${\beta}$-lactoglobulin (${\beta}$-lg) nanoparticles, for the application of bioactive compounds to functional foods. The objectives of this study were to investigate the relationships between the physicochemical properties of ${\beta}$-lg nanoparticles, such as particle size and zeta-potential value, and their cellular uptakes and cytotoxicity in Caco-2 cells. Physicochemical properties of ${\beta}$-lg nanoparticles were evaluated using particle size analyzer. Flow cytometry and confocal laser scanning microscopy were used to investigate cellular uptake and cytotoxicity of ${\beta}$-lg nanoparticles. The ${\beta}$-lg nanoparticles with various particle sizes (98 to 192 nm) and zeta-potential values (-14.8 to -17.6 mV) were successfully formed. A decrease in heating temperature from $70^{\circ}C$ to $60^{\circ}C$ resulted in a decrease in the particle size and an increase in the zeta-potential value of ${\beta}$-lg nanoparticles. Non-cytotoxicity was observed in Caco-2 cells treated with ${\beta}$-lg nanoparticles. There was an increase in cellular uptake of ${\beta}$-lg nanoparticles with a decrease in particle size and an increase in zeta-potential value. Cellular uptake ${\beta}$-lg nanoparticles was negatively correlated with particle size and positively correlated with zeta-potential value. Therefore, these results suggest that the particle size and zeta-potential value of ${\beta}$-lg nanoparticles play an important role in the cellular uptake. The ${\beta}$-lg nanoparticles can be used as a delivery system in foods due to its high cellular uptake and non-cytotoxicity.

• Journal of Ginseng Research
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• 제45권4호
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• pp.465-472
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• 2021

Background: Ginseng can help regulate brain excitability, promote learning and memory, and resist cerebral ischemia in the central nervous system. Ginsenosides are the major effective compounds of Ginseng, but their protein targets in the brain have not been determined. Methods: We screened proteins that interact with the main components of ginseng (ginsenosides) by affinity chromatography and identified the 14-3-3 ζ protein as a potential target of ginsenosides in brain tissues. Results: Biolayer interferometry (BLI) analysis showed that 20(S)-protopanaxadiol (PPD), a ginseng saponin metabolite, exhibited the highest direct interaction to the 14-3-3 ζ protein. Subsequently, BLI kinetics analysis and isothermal titration calorimetry (ITC) assay showed that PPD specifically bound to the 14-3-3 ζ protein. The cocrystal structure of the 14-3-3 ζ protein-PPD complex showed that the main interactions occurred between the residues R56, R127, and Y128 of the 14-3-3 ζ protein and a portion of PPD. Moreover, mutating any of the above residues resulted in a significant decrease of affinity between PPD and the 14-3-3 ζ protein. Conclusion: Our results indicate the 14-3-3 ζ protein is the target of PPD, a ginsenoside metabolite. Crystallographic and mutagenesis studies suggest a direct interaction between PPD and the 14-3-3 ζ protein. This finding can help in the development of small-molecular compounds that bind to the 14-3-3 ζ protein on the basis of the structure of dammarane-type triterpenoid.

• 한국응용과학기술학회지
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• 제38권3호
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• pp.801-812
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• 2021

바이오틴을 피부에 전달하기 위해 고압균질기를 사용하여 바이오틴과 세라마이드를 모두 함유하는 에토좀에 대하여 연구하였다. 바이오틴이 주된 성분으로 사용되었으며, 세라마이드 NP는 인지질 이중층의 지지체로 활용되었다. 바이오틴은 수용성 내부에 포획되었고, 세라마이드 NP는 에토좀의 이중층에 흡착되었다. 세라마이드 NP를 함유한 에토좀의 물성을 살펴보면 소포체의 크기는 80~130 nm, 다분산지수는 0.09~0.16, 제타 전위는 -40~-49 mV로 측정되었다. 세라마이드 NP가 없는 소포체의 크기는 124.80±1.46 nm, 다분산지수와 제타전위는 각각 0.088±0.018과 -45.48±1.27 mV이었다. 따라서 세라마이드 NP가 함유된 에토좀은 세라마이드 NP가 없는 에토좀에 비해 소포체의 물리적 특성이 개선됨을 알 수 있었다. 세라마이드 NP를 함유한 에토좀의 피부흡수율은 12시간 후 6.13~14.98%이었으며, 반면에 세라마이드 NP가 없는 에토좀의 피부흡수율은 7.08%이었다. 결론적으로 세라마이드 NP를 함유한 에토좀은 피부흡수 효율을 향상시킬뿐만 아니라 소포체의 안정성에도 긍정적인 영향을 미쳤다.

• Journal of Ginseng Research
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• 제46권5호
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• pp.666-674
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• 2022

Background: Ginsenosides and their metabolites have antidepressant-like effects, but the underlying mechanisms remain unclear. We previously identified 14-3-3 ζ as one of the target proteins of 20 (S)-protopanaxadiol (PPD), a fully deglycosylated ginsenoside metabolite. Methods: Corticosterone (CORT) was administered repeatedly to induce the depression model, and PPD was given concurrently. The tail suspension test (TST) and the forced swimming test (FST) were used for behavioral evaluation. All mice were sacrificed. Golgi-cox staining, GSK 3β activity assay, and Western blot analysis were performed. In vitro, the kinetic binding analysis with the Biolayer Interferometry (BLI) was used to determine the molecular interactions. Results: TST and FST both revealed that PPD reversed CORT-induced behavioral deficits. PPD also ameliorated the CORT-induced expression alterations of hippocampal Ser9 phosphorylated glycogen synthase kinase 3β (p-Ser9 GSK 3β), Ser133 phosphorylated cAMP response element-binding protein (p-Ser133 CREB), and brain-derived neurotrophic factor (BDNF). Moreover, PPD attenuated the CORT-induced increase in GSK 3β activity and decrease in dendritic spine density in the hippocampus. In vitro, 14-3-3 ζ protein specifically bound to p-Ser9 GSK 3β polypeptide. PPD promoted the binding and subsequently decreased GSK 3β activity. Conclusion: These findings demonstrated the antidepressant-like effects of PPD on the CORT-induced mouse depression model and indicated a possible target-based mechanism. The combination of PPD with the 14-3-3 ζ protein may promote the binding of 14-3-3 ζ to p-GSK 3β (Ser9) and enhance the inhibition of Ser9 phosphorylation on GSK 3β kinase activity, thereby activating the plasticity-related CREBeBDNF signaling pathway.

• 한국재료학회지
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• 제14권9호
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• pp.634-641
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• 2004

The photocatalytic performance of $TiO_2$ thin films coated on porous alumina balls using various aqueous $TiOCl_2$ solutions as starting precursors, to which 1.0 $mol\%$ transition metal ($Ni^{2+},\;Cr^{3+},\;Fe^{3+},\;Nb^{3+},\;and\;V^{5+}$) chlorides had been already added, has been investigated, together with characterizations for $TiO_2$ sols synthesized simultaneously in the same autoclave through hydrothermal method. The synthesized $TiO_2$ sols were all formed with an anatase phase, and their particle size was between several nm and 30 nm showing ${\zeta}-potential$ of $-25{\sim}-35$ mV, being maintained stable for over 6 months. However, the $TiO_2$ sol added with Cr had a much lower value of -potential and larger particle sizes. The coated $TiO_2$ thin films had almost the same shape and size as those of the sol. The pure $TiO_2$ sol showed the highest optical absorption in the ultraviolet light region, and other $TiO_2$ sols containing $Cr^{3+},\;Fe^{3+}\;and\;Ni^{2+}$ showed higher optical absorption than pure sol in the visible light region. According to the experiments for removal of a gas-phase benzene, the pure $TiO_2$ film showed the highest photo dissociation rate in the ultraviolet light region, but in artificial sunlight the photo dissociation rate of $TiO_2$ coated films containing $Cr^{3+},\;Fe^{3+}\;and\;Ni^{2+}$ was measured higher together with the increase of optical absorption by doping.

• Asian Pacific Journal of Cancer Prevention
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• 제17권4호
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• pp.2277-2284
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• 2016

Ribosome-inactivating protein (RIP) from Mirabilis jalapa L. leaves has cytotoxic effects on breast cancer cell lines but is less toxic towards normal cells. However, it can easily be degraded after administration so it needs to be formulated into nanoparticles to increase its resistance to enzymatic degradation. The objectives of this study were to develop a protein extract of M. jalapa L. leaves (RIP-MJ) incorporated into nanoparticles conjugated with Anti-EpCAM antibodies, and to determine its cytotoxicity and selectivity in the T47D breast cancer cell line. RIP-MJ was extracted from red-flowered M. jalapa L. leaves. Nanoparticles were formulated based on polyelectrolyte complexation using low viscosity chitosan and alginate, then chemically conjugated with anti-EpCAM antibody using EDAC based on carbodiimide reaction. RIP-MJ nanoparticles were characterised for the particle size, polydispersity index, zeta potential, particle morphology, and entrapment efficiency. The cytotoxicity of RIP-MJ nanoparticles against T47D and Vero cells was then determined with MTT assay. The optimal formula of RIP-MJ nanoparticles was obtained at the concentration of RIP-MJ, low viscosity chitosan and alginate respectively 0.05%, 1%, and 0.4% (m/v). RIP-MJ nanoparticles are hexagonal with high entrapment efficiency of 98.6%, average size of 130.7 nm, polydispersity index of 0.380 and zeta potential +26.33 mV. The $IC_{50}$ values of both anti-EpCAM-conjugated and non-conjugated RIP-MJ nanoparticles for T47D cells (13.3 and $14.9{\mu}g/mL$) were lower than for Vero cells (27.8 and $33.6{\mu}g/mL$). The $IC_{50}$ values of conjugated and non-conjugated RIP-MJ for both cells were much lower than $IC_{50}$ values of non-formulated RIP-MJ (>$500{\mu}g/mL$).