• Korean Journal of Food Science and Technology
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• v.49 no.2
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• pp.222-227
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• 2017

Red ginseng prepared from fresh 6-year-old ginseng treated with colloidal gold nanoparticles was extracted using hot water to evaluate its total phenolic and flavonoid contents, antioxidant capacity, and neuroprotective effects. Water extract of red ginseng treated with gold nanoparticles (WERGGN) had total phenolic and total flavonoid contents of 212.2 mg gallic acid equivalents/$^{\circ}Bx$ and 3.5 mg catechin equivalents/$^{\circ}Bx$, respectively. The antioxidant capacities of WERGGN measured using ABTS, DPPH, and ORAC assays were 272.3, 141.2, and 868.4 mg vitamin C equivalents/$^{\circ}Bx$, respectively. The WERGGN showed protective effects on the viability of neuron-like PC-12 cells against oxidative stress induced by hydrogen peroxide in a dose-dependent manner, partly because of a reduction in intracellular oxidative stress. Acetylcholinesterase and butyrylcholinesterase, which degrade the neurotransmitter acetylcholine to terminate neurotransmission, were inhibited by treatment with WERGGN. These results suggest that WERGGN is useful as a functional material to decrease oxidative stress and neuronal damage.

• Journal of Veterinary Clinics
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• v.26 no.3
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• pp.279-281
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• 2009

Amyloid-producing odontogenic tumors were described in the gingival masses of two Maltese dogs. Gingival masses were surgically removed and submitted for diagnosis. On histopathology, the masses were poorly demarcated, infiltrative, and consisted of cuboidal and columnar epithelial cells with palisading pattern. Nodular deposit of congophilic amyloid-like material and mineralization were another features of the tumors. Immunohistochemically, the neoplasticd cells were positive to pancytokeratin and neuron-specific enolase but were negative to vimentin. The amorphous homogeneous eosinophilic materials were positive to Congo red stain and showed apple-green color under the polarized microscope. Based on these results, both cases were diagnosed as amyloid-producing odontogenic tumors in dogs.

• International Journal of Oral Biology
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• v.40 no.2
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• pp.103-109
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• 2015

Growing evidence suggests that mitochondrial reactive oxygen species (ROS) are involved in various pain states. This study was performed to investigate whether ROS-induced changes in neuronal excitability in trigeminal subnucleus caudalis are related to ROS generation in mitochondria. Confocal scanning laser microscopy was used to measure ROS-induced fluorescence intensity in live rat trigeminal caudalis slices. The ROS level increased during the perfusion of malate, a mitochondrial substrate, after loading of 2',7'-dichlorofluorescin diacetate ($H_2DCF-DA$), an indicator of the intracellular ROS; the ROS level recovered to the control condition after washout. When pre-treated with phenyl N-tert-butylnitrone (PBN) and 4-hydroxy-2,2,6,6-tetramethylpiperidene-1-oxyl (TEMPOL), malate-induced increase of ROS level was suppressed. To identify the direct relation between elevated ROS levels and mitochondria, we applied the malate after double-loading of $H_2DCF-DA$ and chloromethyl-X-rosamine (CMXRos; MitoTracker Red), which is a mitochondria-specific fluorescent probe. As a result, increase of both intracellular ROS and mitochondrial ROS were observed simultaneously. This study demonstrated that elevated ROS in trigeminal subnucleus caudalis neuron can be induced through mitochondrial-ROS pathway, primarily by the leakage of ROS from the mitochondrial electron transport chain.

• Biomolecules & Therapeutics
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• v.27 no.1
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• pp.78-84
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• 2019

Cell therapeutic agents for treating degenerative brain diseases using neural stem cells are actively being developed. However, few systems have been developed to monitor in real time whether the transplanted neural stem cells are actually differentiated into neurons. Therefore, it is necessary to develop a technology capable of specifically monitoring neuronal differentiation in vivo. In this study, we established a system that expresses cell membrane-targeting red fluorescent protein under control of the Synapsin promoter in order to specifically monitor differentiation from neural stem cells into neurons. In order to overcome the weak expression level of the tissue-specific promoter system, the partial 5' UTR sequence of Creb was added for efficient expression of the cell surface-specific antigen. This system was able to track functional neuronal differentiation of neural stem cells transplanted in vivo, which will help improve stem cell therapies.

• Biomolecules & Therapeutics
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• v.13 no.3
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• pp.165-173
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• 2005

The main aim of this study was to investigate stress related activities of ginsenosides and their action mechanism. Control group and ginsenoside supplemented groups were exposed to stress while no-stress group was not done. Animals of each group (n=$8\~10$) were orally administerd 100 mg red ginseng extract (R-G), or 10 mg ginsenosides/kg body weight once a day. Animals were given materials for 5 days without stress, and then were given supplements for 5 days with restraint and electroshock stress. Mice were given materials for 5 days for experiments on anti-fatigue effect. After loading final stress, stress-related behavioral changes of experimental animals were examined and plasma corticosterone levels were measured. R-G and ginsenoside $Rb_{1}$ supplementation partially blocked the stress effects on locomotion and elevated plus-maze test in rats and mice. They also partially blocked stress induced behavioral changes such as freezing, smelling, face-washing, rearing behavior in rats. R-G and $Rb_{1}$ decrease adrenal gland size and plasma corticosterone level, which were increased by stress in rats. R-G increased enduring time on the Rota rod, cold water and horizontal wire, but $Rb_{1}$ didn't. Effects of $Rb_{1}$ on plusmaze test were inhibited by administration of flumazenil. These results suggest that $Rb_{1}$ is the main antistress principle in ginseng and it's effect is modulated by GABAnergic nervous system.

• Journal of Korean Neurosurgical Society
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• v.49 no.4
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• pp.205-211
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• 2011

Objective : We investigated the neuroprotective effect of anthocyanin, oxygen radical scavenger extracted from raspberries, after traumatic spinal cord injury (SCI) in rats. Methods : The animals were divided into two groups : the vehicle-treated group (control group, n=20) received an oral administration of normal saline via stomach intubation immediately after SCI, and the anthocyanin-treated group (AT group, n=20) received 400 mg/kg of cyanidin 3-O-${\beta}$-glucoside (C3G) in the same way. We compared the neurological functions, superoxide expressions and lesion volumes in two groups. Results : At 14 days after SCI, the AT group showed significant improvement of the BBB score by $16.7{\pm}3.4%$, platform hang by $40.0{\pm}9.1%$ and hind foot bar grab by $30.8{\pm}8.4%$ (p<0.05 in all outcomes). The degree of superoxide expression, represented by the ratio of red fluorescence intensity, was significantly lower in the AT group ($0.98{\pm}0.38$) than the control group ($1.34{\pm}0.24$) (p<0.05). The lesion volume in lesion periphery was $32.1{\pm}2.4\;{\mu}L$ in the control and $24.5{\pm}2.3\;{\mu}L$ in the AT group, respectively (p<0.05), and the motor neuron cell number of the anterior horn in lesion periphery was $8.3{\pm}5.1$ cells/HPF in the control and $13.4{\pm}6.3$ cells/HPF in the AT group, respectively (p<0.05). Conclusion : Anthocyanin seemed to reduce lesion volume and neuronal loss by its antioxidant effect and these resulted in improved functional recovery.

• Journal of Life Science
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• v.17 no.2 s.82
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• pp.198-203
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• 2007

DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.