• Clinical and Experimental Pediatrics
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• v.46 no.7
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• pp.702-709
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• 2003

Purpose : $IFN{\gamma}$ sentitizes many tumor cells to $TNF{\alpha}$ and FASL-mediated apoptosis by enhancing the expression of TNF or FAS/CD95 receptor and modulating the activation of caspase and Bcl-2 family. It has been reported that $IFN{\gamma}$ and $TNF{\alpha}$ synergistically caused differentiation and growth inhibition of neuroblastoma cells. Even though some neuroblastoma cell express FASR/FASL on the cell surface, they could not induce apoptosis by ligation of the FAS/CD95 receptor. But the treatment of $IFN{\gamma}$ is reported to induce apoptosis in some neuroblastoma cell lines through the CD95/CD95L autocrine circuit. In this study, we examined whether $IFN{\gamma}$ could affect $TNF{\alpha}$ and agonistic FAS/CD95 antibody(CH-11)-induced apoptosis against neuroblastoma cell lines that had shown diverse drug sensitivity and resistance. Methods : CHLA-15, CHLA-90 and LA-N-2 neuroblastoma cells were cultured in IMDM and treated with recombinant $IFN{\gamma}$, $TNF{\alpha}$ and CH-11 antibody. Cell viability was measured by DIMSCAN with a fluorescent calcein-AM. Apoptosis was analyzed through flow cytometry using Annexin V-PE and 7-ADD staining and confirmed by pancaspase and caspase-8 blocking experiments. The expression of TNF RI and FAS/CD95 receptor was evaluated by flow cytometry using the corresponding antibody and PE-conjugated secondary antibody. Results : $IFN{\gamma}$ or $TNF{\alpha}$ alone had no demonstrable cytotoxic effects, whereas both cytokines in combination induced apoptosis synergistically in CHLA-15 and CHLA-90 cells. Although there was no cytotoxicity with the ligation of CH-11 alone in CHLA-90 cells, pretreatment of $IFN{\gamma}$ increased the sensitivity of CH-11-mediated apoptosis. The expression of TNFRI and FAS/CD95R were non-specifically enhanced after treatment of $IFN{\gamma}$ without relation to sensitivity to $TNF{\alpha}$ and CH-11. This finding suggest up-regulation of both receptors may contribute to sensitization of $TNF{\alpha}$ and CH-11-mediated apoptosis by $IFN{\gamma}$ in only sensitive cell lines. Conclusion : $IFN{\gamma}$ induced sensitization of $TNF{\alpha}$ and agonistic FAS/CD95 antibody-mediated apoptosis on some neuroblastoma cells through up-regulation of TNFRI and FAS/CD95 receptor.

• Journal of Life Science
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• v.20 no.1
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• pp.1-8
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• 2010

We recently reported the distributions of AMPA ($\alpha$-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate) receptor subtypes glutamate receptors (GluR) 1 and GluR4 in the superior colliculi (SC) of hamsters with antibody immunocytochemistry and the effect of enucleation on these distributions. We also compared these labelings to those of calcium-binding proteins calbindin D28K, calretinin, parvalbumin, and GABA. In the present study, we investigated whether the GluR1- and GluR4-immunoreactive (IR) neurons are interneurons or projection neurons by injection of the retrograde tracer horseradish peroxidase (HRP) into one of each major ascending and descending pathways of the SC. HRP injections were made into a tecto-reticulospinal pathway (TRS) and dorsal lateral geniculate nucleus (dLGN). Animals were then allowed to recover and to survive for 48 hr before perfusion. Sections containing retrograde-labeled neurons were then treated for GluR-immunoreactivity. HRP injections proved that only a small population of the GluR1-IR cells project into TRS (1.4%) and dLGN (2.6%). However, a large subpopulation of GluR4-IR cells project into TRS (32.7%). The differential compositions of inter/projection neurons, along with our previous studies on the separate distribution of the GluR subunits, its differential co-localization with calcium-binding proteins and GABA, and differential reactions to enucleations, strongly imply the functional variety of the receptor subunits in visual behavior responses.

• Applied Microscopy
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• v.35 no.4
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• pp.10-15
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• 2005

Cultured neurons from Drosophila brain on a glass coverslip to understand the structural basis of synapse were prepared for TEM observations. Neurons on a coverslip were fixed, dehydrated and embedded in Epon without separating from coverslip. After polymerization, the block was placed in 49% hydrofluoric acid to remove the coverslip. The block was examined under a light microscope to select exact neurons, then trimmed and sectioned for TEM observation.

• Journal of the Korean Applied Science and Technology
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• v.36 no.4
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• pp.1303-1311
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• 2019

This study was to investigate the antioxidant, anti-inflammatory and neuronal cell protective effects of Poria cocos Wolf and Corni fructus extracted by water and 70% ethanol. Total polyphenol content in water extract of Poria cocos Wolf was significantly higher than those of other extracts. DPPH and ABTS free radical scavenging activity in water extract of Corni fructus was higher than those of other extracts. DPPH and ABTS free radical scavenging activity were increased in a dose-dependent manners. In order to effectively extract total polyphenol contents and anti-oxidant components in Poria cocos Wolf and Corni fructus, hot water extraction method is more efficient than ethanol extraction method. Poria cocos extracts were found to be a superior NO production inhibitory effect compared to Corni fructus extracts. In a neuronal cell viability assay using MPP⁺, the water extract of Poria cocos Wolf protected against MPP⁺-induced neurotoxicity than those of Corni fructus extract. It is considered to be a potential functional material with antioxidant, anti-inflammatory and neuronal protective effect against to oxidative stress according to the extract methods of extracting Poria cocos Wolf and Corni fructus.

• Applied Microscopy
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• v.29 no.4
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• pp.459-470
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• 1999

To investigate the changes during the differentiation of the cerebral neurons of the embryogenic day (ED) 9 and 10, investigated the ultrastructural changes in the cerebral neurons by Electromicroscope, also cerebral protein, the activity of dehydronases (LDH, MDH and SDH) and changes of adenosine triphosphate concentration were analyzed, the result obtained are as follows. In the ultrastructural changes in the cerebral neurons, chromatin in 9 day-old chick embryos are comparatively distributed to even in neucleoplasm and could investigate very prominently that nuclear membrane is double-layer. Esperially, Rough endoplasmic reticulum (RER) and Golgi complex are developed well, also polysome is investigated and synaptic vesicles were scattered. In 10 day-old chick embryos, chromatin evenly spread and nuclear membrane could be differentiated prominently. Rough endoplasmic reticulum (RER) contain cytoplasm, mitochondria and Golgi complex are comparatively developed well. In 9 day-old cultural group of chick embryo cerebrum were separated 37 polypeptide bands and In 10 day-old cultural group of chick embryo cerebrum were separated 38 poly -peptide bands. The more culture time increase, the more the activity of dehydronases (LDH, MDH and SDH) increase. LDH activity was 11.07 (9th day) and 12.12 (10th day), MDH activity was 11.89 (9th day) and 13.44 (10th day) and SDH activity was 8.45 (9th day) and 10.52 (10th day) respectively. The ATP concentration degreesed 10 day-old cultural group than 9 day-old cultural group.

• The Transactions of the Korea Information Processing Society
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• v.3 no.3
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• pp.439-448
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• 1996

Extraction and selection of the informative features play a central role in pattern recognition. This paper describes a modified back-propagation algorithm that performs selection of the informative features and trains a neural network simultaneously. The algorithm is mainly composed of three repetitive steps : training, connection pruning, and input unit elimination. Afer initial training, the connections that have small magnitude are first pruned. Any unit that has a small number of connections to the hidden units is deleted,which is equivalent to excluding the feature corresponding to that unit.If the error increases,the network is retraned,again followed by connection pruning and input unit elimination.As a result,the algorithm selects the most im-portant features in the measurement space without a transformation to another space.Also,the selected features are the most-informative ones for the classification,because feature selection is tightly coupled with the classifi-cation performance.This algorithm helps avoid measurement of redundant or less informative features,which may be expensive.Furthermore,the final network does not include redundant parameters,i.e.,weights and biases,that may cause degradation of classification performance.In applications,the algorithm preserves the most informative features and significantly reduces the dimension of the feature vectors whiout performance degradation.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.02a
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• pp.599-599
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• 2013

최근의 원자간력현미경(AFM)은 생체물질을 대상으로 여러 구조적 형상뿐만 아니라 물리적 특성 측정이 가능하여 바이오분야에 다양이 활용되고 있다. 줄기세포의 신경세포로 분화 인지에 대한 연구와 관련하여 본 연구에서는 AFM의 한 기능인 Force-Distance curve 측정법을 활용하여 신경암세포주라 불리는 SH-SY5Y를 대상으로 분화 전과 후의 세포막의 stiffness 변화를 측정하였다. 세포막의 stiffness값은 시료표면과 맞닿은 AFM 탐침에 계속적으로 수직방향의 힘이 가해질 시 AFM 캔티레버의 구부러짐 정도로 측정된다. SH-SY5Y는 RA (retinoic acid) 처리에 의해 분화유도 되었으며, 생물학적 방법인 western blotting법을 통해 분화여부를 확인하였다. 측정영역은 AFM topography 이미지 상에서 roughness가 가장 낮은 분화 전과 후 SH-SY5Y의 핵 주변영역으로 선정하였다. 선정된 영역 내에 여러 부분의 분화 전후 세포막의 stiffness 값을 측정하여 통계화한 결과, 분화 전과 후 세포막의 stiffness 차이를 확인할 수 있었다. 분화 전 SH-SY5Y 세포막의 stiffness는 0.79445 N/m인 반면, 분화 후 SH-SY5Y 세포막의stiffness는 0.60324 N/m로 확인되었다. 이는 분화 전에 비하여 분화 후 SH-SY5Y 세포막의 stiffness가 약 24.07% 감소된 것으로 판단할 수 있다. 본 연구는 생물학적 복잡한 방법이 아닌 간단한 방법으로 세포의 stiffness의 변화 측정을 통한 세포의 분화를 판별할 수 있는 방법을 개발한 것으로 여러 줄기세포의 특정세포로 분화여부 판단에 활용할 수 있을 것으로 사료된다.

• Journal of Nutrition and Health
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• v.50 no.5
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• pp.415-425
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• 2017

Purpose: Many studies have suggested that neuronal cells protect against oxidative stress-induced apoptotic cell death by polyphenolic compounds. We investigated the neuroprotective effects and the mechanism of action of Momordica charantia ethanol extract (MCE) against $H_2O_2-induced$ cell death of human neuroblastoma SK-N-MC cells. Methods: The antioxidant activity of MCE was measured by the quantity of total phenolic acid compounds (TPC), quantity of total flavonoid compounds (TFC), and 2,2-Diphenyl-1-pycrylhydrazyl (DPPH) radical scavenging activity. Cytotoxicity and cell viability were determined by CCK-8 assay. The formation of reactive oxygen species (ROS) was measured using 2,7-dichlorofluorescein diacetate (DCF-DA) assay. Antioxidant enzyme (SOD-1,2 and GPx-1) expression was determined by real-time PCR. Mitogen-activated protein kinases (MAPK) pathway and apoptosis signal expression was measured by Western blotting. Results: The TPC and TFC quantities of MCE were 28.51 mg gallic acid equivalents/extract g and 3.95 mg catechin equivalents/extract g, respectively. The $IC_{50}$ value for DPPH radical scavenging activity was $506.95{\mu}g/ml$ for MCE. Pre-treatment with MCE showed protective effects against $H_2O_2-induced$ cell death and inhibited ROS generation by oxidative stress. SOD-1,2 and GPx-1 mRNA expression was recovered by pre-treatment with MCE compared with the presence of $H_2O_2$. Pre-treatment with MCE inhibited phosphorylation of p38 and the JNK pathway and down-regulated cleaved caspase-3 and cleaved PARP by $H_2O_2$. Conclusion: The neuroprotective effects of MCE in terms of recovery of antioxidant enzyme gene expression, down-regulation of MAPK pathways, and inhibition apoptosis is associated with reduced oxidative stress in SK-N-MC cells.

• The Journal of Internal Korean Medicine
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• v.19 no.2
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• pp.381-391
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• 1998

The present study was done to investigate the effects of Bombusae concretio Salicea (BCS) on cultured astrocyte cell system and lipid peroxidation in $A{\beta}25-35$ treatment conditions. Cell killing was significantly enhanced by addition of increasing concentrations of $A{\beta}25-35$. Pretreatment of BCS attenuated in cell killing enhanced by increasing concentrations of $A{\beta}25-35$. MDA level induced by $A{\beta}25-35$ treatment was significantly increased and the level was slightly reduced by pretreatment of BCS. The present study showed that $A{\beta}25-35$ strongly increased MDA level and the level was enhanced by addition of increasing concentrations of In conclusion, it was shown that $A{\beta}25-35$ is not only potent lipid peroxide inducer, but also cause protection of neurodegeneration induced by $A{\beta}25-35$. It can be concluded that the activation of antioxidative enzymes may be related to the inhibition of lipid peroxidative reactions. We cannot fully explain to effects of BCS at present; however, the ability of BCS to reduce cell killing and MDA level induced by $A{\beta}25-35$ suggest that BCS may be a protective agent for free radical generating compounds such as $A{\beta}25-35$.

• The korean journal of orthodontics
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• v.34 no.6 s.107
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• pp.506-513
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• 2004

Osteoclast action is necessary for alveolar bone remodeling in orthodontic tooth movement. The nervous system has also been reported to be associated with bone remodeling. This study was aimed to investigate the changes of osteoclasts in the periodontal ligament (PDL) space after surgical resection of the inferior alveolar nerve (IAN). Experimental rats were divided into young and adult groups. A surgical resection procedure of the IAN was carried out in the left side of the mandible and a sham operation in the right side of the mandible. The number of osteoclasts on the bundle bone surface and the resorption activity of the osteoclasts were histomorphometrically measured. The changes in distribution of substance P (SP) immunoreactive (IR) nerve fiber were evaluated in the PDL and pulp. SP-IR nerve fiber was depleted in both the PDL and pulp of the IAN resection side in both groups, which confirmed the resection of IAN to be successfully conducted. The number of osteoclasts in the IAN resection side was significantly reduced in both the young and adult groups (p<0.01 and p<0.05), whereas the resorption activity of osteoclasts did not show any significant difference between the IAN resection side and the sham operation side in both groups (p>0.05 and p<0.05). The adult group showed that the number of osteoclasts reduced significantly (p<0.01) and the resorption activity didn't change in comparison with the young group (p>0.05). These results suggest that surgical resection of the IAN and aging reduce the population of the recruited osteoclasts within the PDL, but don't affect on the osteoclastic resorption activity.