• The Korean Journal of Physiology and Pharmacology
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• v.3 no.1
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• pp.19-27
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• 1999

Apoptosis has been implicated in the pathophysiological mechanisms of various neurodegenerative diseases. In a variety of cell types, oxidative stress has been demonstrated to play an important role in the apoptotic cell death. However, the exact mechanism of oxidative stress-induced apoptosis in neuronal cells is not known. In this study, we induced oxidative stress in IMR-32 human neuroblastoma cells with tert- butylhydroperoxide (TBHP), which was confirmed by significantly reduced glutathione content and glutathione reductase activity, and increased glutathione peroxidase activity. TBHP induced decrease in cell viability and increase in DNA fragmentation, a hallmark of apoptosis, in a dose-dependent manner. TBHP also induced a sustained increase in intracellular $Ca^{2+}$ concentration, which was completely prevented either by EGTA, an extracellular $Ca^{2+}$ chelator or by flufenamic acid (FA), a non-selective cation channel (NSCC) blocker. These results indicate that the TBHP-induced intracellular $Ca^{2+}$ increase may be due to $Ca^{2+}$ influx through the activation of NSCCs. In addition, treatment with either an intracellular $Ca^{2+}$ chelator (BAPTA/AM) or FA significantly suppressed the TBHP-induced apoptosis. Moreover, TBHP increased the expression of p53 gene but decreased c-myc gene expression. Taken together, these results suggest that the oxidative stress-induced apoptosis in neuronal cells may be mediated through the activation of intracellular $Ca^{2+}$ signals and altered expression of p53 and c-myc.

• Archives of Pharmacal Research
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• v.22 no.4
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• pp.404-409
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• 1999

The inhibitory effects of the constituents of Gastrodia elata Bl. (GE) on glutamate-induced apoptosis in human neuronal cells were investigated using IMR32 human neuroblastoma cells. Glutamate (GLU) induced DNA fragmentation, a hallmark of apoptosis, in a dose-dependent manner. GLU also induced a slow and sustained increase in intracellular $Ca^{2+}$ concentration. Treatment with EGTA, an extracellular $Ca^{2+}$ chelator, in a nominal $Ca^{2+}$ -free buffer solution abolished the GLU-induced intracellular $Ca^{2+}$ increase, indicating that GLU stimulated Ca2+ influx pathway in the IMR32 cells. BAPTA, an intracellualr $Ca^{2+}$ chelator, significantly inhibited the GLU-induced apoptosis assessed by the flow cytometry measuring hypodiploid DNA content indicative of apoptosis, implying that intracellular $Ca^{2+}$ rise may mediate the apoptotic action of GLU. Vanillin (VAN) and p-hydroxybenzaldehyde(p-HB), known constituents of GE, significantly inhibited both intracellular $Ca^{2+}$ rise and apoptosis induced by GLU. These results suggest that the apoptosis-inhibitory actions of the constituents of GE may account, at least in part, for the basis of their antiepileptic activities. These results further suggest that intracelluarl $Ca^{2+}$ signaling pathway may be a molecular target of the constituents of GE.

• Advances in Traditional Medicine
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• v.10 no.3
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• pp.141-149
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• 2010

A concern of safety issue for traditional herbal medicine is the possibility of some herbs containing heavy metals responsible for several cases of metal poisoning. In our previous study, the physiological levels of heavy metals, such as Cd, Cu, Hg and Pb, were examined to evaluate a potential risk in patients taking herbal medicine. In this study, the levels of 12 heavy metals, including non-metallic element, Se, were analyzed in hair and compared to the results from the previous study. In the previous study, the levels of Cd, Cu, Hg and Pb in hair showed a significantly lower odds ratio than 1. In this study, however, all metals did not show any significant odds ratio higher or lower than 1 even if 5 of 12 metals showed lower odds ratio than 1. In addition, the levels of metal concentrations, especially for Cu, Fe, Pb and Zn, exceeding the WHO reference values were observed in hair. However, any evidence for metal accumulation in hair caused by taking herbal medicines for long duration was not observed in analysis of multiple regression and odds ratio from case-control study. This result would show another possibility for a role of herbs as a non-enzymatic chelator inhibiting the gastrointestinal absorption of heavy metals.

• YAKHAK HOEJI
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• v.57 no.6
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• pp.400-405
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• 2013

In this study, we measured the complex efficiency of a physiologically active antibody, a chelator and radiosiotopes using the ESI-TOF/Ms system for develop good radiopharmaceuticals. For a precise measurement, TLC is a low accuracy method. Loading of same amount of sample is difficult for each test, and work to quantify accurately the results obtained through TLC cannot be afforded compared to the use of other analytical instruments. The method of analysis using a mass spectrometer is capable of a mass analysis of proteins for quantitative analysis. The conjugates of the chelator (CHX-A- DTPA) and the antibody (IgG) were separated through MWCO, and were analyzed using ESI-TOF and MALDI-TOF mass spectrometry. The analysis using MALDI-TOF is roughly divided into measurements on mass spectrometry. When conjugating a small molecular weight of CHX-A-DTPA and a large molecular weight of IgG, distinguishing the peak of the conjugate and the peak of IgG was difficult. However, an ESI-TOF mass spectrometer system is capable of an analysis of mass by decentralizing the IgG. It is utilized as a technique for measuring the metabolic processes during conjugation and the stability evaluation of radiopharmaceuticals. When establishing this technique, the accuracy of the overall radiophar-maceutical analysis is expected to be able to be improved.

• Journal of Life Science
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• v.7 no.3
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• pp.228-233
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• 1997

A thiol protease has been isolated and partially purified from encysted brine shrimp Artemia franciscana using a four-step procedure(filtration, salting out, gel filtration and ion exchange chromatography). The optimum pH of the enzyme for caseinolytic activity was appeared to be 3.0, and the enzymematic activity was stable up to pH 6.0 but lost completely at the pH higher than 8.0. The optimal temperature of the enzyme was appeared to be 35$^{\circ}$C, and ninety percent of the enzyme activity was lost at 45$^{\circ}$C. Various metal ions, e.g., zinc, copper, iron, inhibited the enzyme activity; however, heavy metal chelator, e.g., EDTA, stimulated the enzyme activity. The protease was concluded to be a member of the thiol group protease, since it was inhibited by thiol protease inhibitors and iodoacetate. The protease was also concluded to be a acid protease based on optimum pH.

• Journal of Radiopharmaceuticals and Molecular Probes
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• v.8 no.1
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• pp.45-49
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• 2022

With the development of monoclonal antibodies, therapeutic or diagnostic radioisotope has been successfully delivered at tumor sites with high selectivity for antigens. Different approaches have been applied to improve the tumor-to-normal ratio by considering the in vivo stability of radioimmunoconjugates as a prerequisite. Various stable and inert antibody radiolabeling techniques for radioimmunoconjugate preparation have been extensively evaluated to enhance in vivo stability. Antibody radiolabeling techniques should be rapid and easy; they should not disrupt the immunoreactivity and in vivo behavior of antibodies, which are coupled with a bifunctional chelator (BFC) to stably coordinate with a radiometal. For the design of BFCs, radiometal coordination properties must be considered. However, various diagnostic radionuclides, such as ⁸⁹Zr, ⁶⁴Cu, ⁶⁸Ga, ¹¹¹ln, and ^99mTc, or therapeutic radionuclides, such as ¹⁷⁷Lu, ⁶⁷Cu, ⁹⁰Y, and ²²⁵Ac, have been increasingly used for antibody radiolabeling. In addition to useful radionuclides, ⁶⁴Cu and ¹⁷⁷Lu with the most accessible or the highest production rates in many countries should be considered. In this review, we mainly discussed antibody radiolabeling techniques and conditions that involve ⁶⁴Cu and ¹⁷⁷Lu radiometals.

• Restorative Dentistry and Endodontics
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• v.45 no.4
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• pp.44.1-44.9
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• 2020

Objectives: Phytic acid (IP6), a naturally occurring agent, has been previously reported as a potential alternative to ethylenediaminetetraacetic acid (EDTA). However, its effect on adhesion to sodium hypochlorite (NaOCl)-treated dentin and its interactions with NaOCl have not been previously reported. Thus, in this study, the effects of IP6 on resin adhesion to NaOCl-treated dentin and the failure mode were investigated and the interactions between the used agents were analyzed. Materials and Methods: Micro-tensile bond strength (µTBS) testing was performed until failure on dentin treated with either distilled water (control), 5% NaOCl, or 5% NaOCl followed with chelators: 17% EDTA for 1 minute or 1% IP6 for 30 seconds or 1 minute. The failed specimens were assessed under a scanning electron microscope. The reaction of NaOCl with EDTA or IP6 was analyzed in terms of temperature, pH, effervescence, and chlorine odor, and the effects of the resulting mixtures on the color of a stained paper were recorded. Results: The µTBS values of the control and NaOCl with chelator groups were not significantly different, but were all significantly higher than that of the group treated with NaOCl only. In the failure analysis, a distinctive feature was the presence of resin tags in samples conditioned with IP6 after treatment with NaOCl. The reaction of 1% IP6 with 5% NaOCl was less aggressive than the reaction of the latter with 17% EDTA. Conclusions: IP6 reversed the adverse effects of NaOCl on resin-dentin adhesion without the chlorine-depleting effect of EDTA.

• Journal of Microbiology and Biotechnology
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• v.7 no.5
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• pp.352-355
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• 1997

The promoter region of the pupA gene of Pseudomonas putida WCS358 was fused with the structural genes for bioluminescence (luxCDABE) from Vibrio fischeri, and the resulting fusion plasmid harbored by the WCS358 host. The pup-lux fusion gene was then used for quantitative analysis of the iron-dependence of pupA promoter activity. Factors affecting bioluminescence produced by the pup-lux bioreporter were found to be cell activity, iron-chelator concentrations, Fe(III) concentrations, and nutrient components. Light production rates of the pup-lux bioreporter were inversely dependent upon iron molecules when $FeCl_3$ concentrations were between $10^{-2}$ and 1 ${\mu}M$ in nutrient-poor minimal media, and between 0.1 and 10 mM in nutrient-rich complex media.

• Journal of Radiopharmaceuticals and Molecular Probes
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• v.5 no.2
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• pp.129-134
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• 2019

Over a few decades, copper radioisotopes and their chelation chemistry for radiopharmaceuticals have played crucial role in the radiopharmaceutical science area. A variety of chelators have been required for their stable targeting ability in physiological conditions. For radiolabeling with copper-64 into biomolecules, thermodynamic stability, kinetic inertness, pH stability, and redox stability should be considered. In this regard, many researchers have attempted to develop the chelators that can bind with copper more tightly, rapidly and stably for copper radiolabeling. This review discusses the chemistry of copper, its suitable chelators and characteristics, while elucidating the evaluations of each chelator for radiolabeling.

• Korean Journal of Pharmacognosy
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• v.27 no.4
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• pp.350-353
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• 1996

The effects of acid hydrolysis product of Panax ginseng and MeOH extract of Euphorbia humifusa on the growth of human brain tumor cells were evaluated using U-373 MG human astrocytoma and SK-N-MC human neuroblastoma cells as model cellular systems. These plant extracts induced cytotoxicity in both cells in a dose-dependent manner. These cytotoxic effects were significantly inhibited by GSH, an antioxidant, in both cells. BAPTA/AM, an intracellular $Ca^{2+}$ chelator, significantly blocked the cytotoxic effects of these extracts in U-373 cells, but enhanced these effects in SK-N-MC cells. These results suggest that the plant extracts may be a valuable choice for the studies on the treatment of human brain tumors.