• Journal of Life Science
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• v.21 no.10
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• pp.1377-1384
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• 2011

Astaxanthin (ATX) is a red-orange carotenoid pigment that occurs naturally in a wide variety of living organisms. In this study we investigated the inhibitory effects of ATX on the induction of inducible nitric oxide synthase (iNOS), nitric oxide (NO), proinflammatory cytokines, nuclear factor-kappa B(NF-${\kappa}B$) and reactive oxygen species (ROS) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. In addition, we tested the superoxide radical scavenging activity of ATX by scavenging assay. iNOS and NF-${\kappa}B$ expressions were determined by immunoblot analysis. Interleukin (IL)-6 and tumour necrosis factor-${\alpha}$ (TNF-${\alpha}$) were assayed by ELISA. NO production was monitored by measuring the amount of nitrite. ROS was examined by using the 2', 7'-Dichlorodihydrofluorescin diacetate (DCFH-DA) method. At a concentration of 100 ${\mu}M$, ATX inhibited the expression level of LPS-induced NF-${\kappa}B$, as well as the production of LPS-induced NO and proinflammatory cytokines (IL-6 and TNF-${\alpha}$), by suppressing iNOS expression. In particular, the maximal inhibition rate of IL-6 and TNF-${\alpha}$ production by ATX (100 ${\mu}M$) was 65.2----- and 21.2-----, respectively. In addition, ATX inhibited the LPS-induced transcriptional activity of NF-${\kappa}B$, and this was associated with suppressing the translocations of NF-${\kappa}B$ from the cytosol to the nucleus. Moreover, at various concentrations (25-100 ${\mu}M$), ATX inhibited the intracellular level of ROS. At a concentration of 5 mg/ml, the superoxide radical scavenging activity of ATX was 1.33 times higher than ${\alpha}$-tocopherol of the same concentration. These results showed that ATX inhibited the expression of iNOS and the production of NO and proinflammatory cytokines resulting from ROS production and NF-${\kappa}B$ activation in macrophages. Furthermore, ATX was found to be more effective in superoxide radical scavenging activities compared to ${\alpha}$-tocopherol. These findings are expected to strengthen the position of ATX as anti-inflammatory medicine and antioxidant.

• Applied Microscopy
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• v.30 no.4
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• pp.377-387
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• 2000

There are a few tanycytes between the general ependymal cells lining the ependymal layer of the brain ventricle. These cells are considered as modified ependymal cells which possess a long basal process. Tanycytes are known to have an ability to communicate by absorbing substances from cerebrospinal fluid and transporting them to the blood vessels and/or to the neurons in the CNS. The third and fourth ventricular tanycytes were mainly studied as subjects but it's rare to find reports about the tanycytes of the area postrema. Glial fibrillary acidic protein is an intermediate filament protein that is expressed especially in astrocytes of the CNS. But GFAP is also found in filament of the tanycytes and its process. Therefore this study was carried out for the examination of the GFAP immunoreactive tanycytes lining the area postrema of the bat, and we also examined the ultrastructure of tanycytes using electron microscope. GFAP immunoreactive tanycytes were located in the caudal portion of the fourth ventricle, and especially mainly in the transitional zone between the floor of the caudal fourth ventricle and ependymal layer lining the area postrema. A few GFAP immunoreactive tanycytes were also found in the ependymal layer lining the area postrema, and some groups of tanycytes were found in the ependymal layer of the area postrema near the floor of the caudal fourth ventricle , The processes of tanycytes were stained deeply with anti-GFAP antibody. Especially the GFAP immunoreactive tanycytes lining the area postrema had very long processes that cross the whole width of the area postrema. In the electron microscope, the cell body of ependymal tanycyte was located on the ependymal layer and had a long basal process. Intermediate filaments were observed around the nucleus and well developed in the process of tanycrte. Longitudinal oriented long mitochondria and a few lipid droplets were also found in this process. After immunocytocheical staining, the gold particles were found only in the intermediate filaments. We could not determine the function of the tanycytes in the area postrema. Thus, further investigation is required to determine the functional relationship between the tanycytes and the area postrema in hibernating animal, the bat.

• Korean Journal of Plant Resources
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• v.20 no.1
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• pp.22-27
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• 2007

Effects of prunella vulgaris L., chrysanthemum indicum L. and pueraria Radix on plasma lipid composition and histological consideration were investigated in hyperlipidemic rats. Concentration of plasma ${\beta},-lipoprotein$ showed a tendence to decrease in prunella vulgaris L., chrysanthemum indicum L. and pueraria Radix treatment groups. However these values were showed not significantly different from control group. Concentration of plasma FFA in prunella vulgaris L., chrysanthemum indicum L. and pueraria Radix treatment groups showed a lower values compared to control group and concentration of plasma FFA of prunella vulgaris L. and pueraria Radix treatment group showed a lower values than prunella vulgaris L. and chrysanthemum indicum L. treatment group. Concentration of plasma glucose and triglyceride showed a tendence to decrease in prunella vulgaris L., chrysanthemum indicum L. and pueraria Radix treatment groups. However plasma glucose values showed not significantly different from control group. Plasma total cholesterol concentration showed a low and HDL-cholesterol concentration showed a high in prunella vulgaris L. and pueraria Radix treatment group. However LDL-cholesterol concentrations were not significantly different in treatment groups. Histological consideration of heart, liver and kidney in prunella vulgaris L. and pueraria Radix treatment groups showed a good features in fat accumulation condition than those of other treatment groups. However in the condition of high fat accumulation in tissues, heart, liver and kidney were showed a slight congestion and a bashed cell nucleus.

• Journal of Life Science
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• v.26 no.5
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• pp.555-562
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• 2016

In this study, we investigated the effects of Undaria pinnatifida (UP), Petalonia binghamiae (PB) and Punctaria latifolia (PL) extracts on the inhibition of proliferation and apoptosis in human gastric and breast cancer cells. AGS, MDA-MB-231 and SK-BR-3 cells were treated with 0, 50, 100, and 200 μg/ml concentrations of the extracts to determine their anti-proliferative effects, using the MTT assay. The UP, PB and PL extracts inhibited proliferation of AGS, MDA-MB-231 and SK-BR-3 cells in a dose-dependent manner, and the PL extract was found to be the most effective. DAPI staining was also performed to determine changes in the cell nucleus. Further, the AGS, MDA-MB-231 and SK-BR-3 cells were treated with 0, 50, 100, and 200 μg/ml of only the PL extract. DAPI staining showed increased chromatin condensation, which is indicative of apoptosis, in the 200 μg/ml group. The expression of the Bax, Bcl-2, and PARP proteins in AGS, MDA-MB-231 and SK-BR-3 cells treated with the PL extract was also determined by western blot analysis. The expression of Bax (a pro-apoptotic protein) and cleaved-PARP was increased, whereas the expression of Bcl-2 (an anti-apoptotic protein) was decreased compared with the control. These findings indicate that the PL extract may have potential as an alternative anticancer drug and nutraceutical.

• Journal of Korean Ophthalmic Optics Society
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• v.9 no.2
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• pp.333-343
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• 2004

Impression cytology refers to application of cellulose acetate filter material to the ocular surface to remove the superficial layers of the conjunctival epithelium. The technique is non-invasive, is easy to perform, causes minimal discomfort to the patient, and can be used to follow changes in the conjunctival ocular surface over time. With this method, the morphology of the conjunctival ocular surface can be studied and the degree of squmaous metaplasia assessed. This study was performed to evaluate the conjunctival surface by impression cytology in dry eye patients. A total of 70 students with no contact lens wearing history were recruited. Subjects were required to fill in a McMonnies dry eye symptom questionnaire. The non-invasive tear thinning time(TIT) test of each subject was measured, followed by Schirmer tear test(STI), tear film break-up time(TBUT) tests and Rose-bengal staining were performed as a baseline. Conjunctival epithelial cells from the inferior bulbar conjunctiva were harvested by the impression cytology technique. The specimens collected were labelled and stained with PAS(Periodic Acid Schift)-haematoxylin. The goblet cells and conjunctival epithelial cells were observed under a light microscope of 400x magnification. The specimens were classified according to the Nelson Grading scale which was based on the degree of squamous metaplasia such as changes of goblet cells density, size/form, N:C(nucleus : cytoplasm) ratio. Dry eye patients were observed morphological changes of the epithelial cells, different nuclear alterations, decrease of the goblet cells density. The degree of cytological changes was related to severity of dry eye conditions. When the epithelial cell morphology was graded according to the system described by Nelson, specimens from the control group revealed 91.43% of the eyes to be grade 0 and 8.57% to be grade 1, whereas of the dry eye patients, 20% were grade 0, 42.86% grade 1, 34.29% grade 2 and 2,86% grade 3. Impression cytology represents a non- or minimally invasive biopsy of the ocular surface epithelium with no side effects or contraindications. It has demonstrated to be a useful diagnostic aid for a wide variety of processes involving the ocular surface. This technique is a safe, simple method and may help increase understanding of various ocular surface alterations in dry eye patients.

• Journal of Veterinary Clinics
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• v.31 no.4
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• pp.267-271
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• 2014

Accurate determination of in vivo oocyte maturation is particularly critical for dog cloning compared to other assisted reproductive technologies because oocytes in metaphase II stage have to be recovered in order to undergo somatic cell nuclear transfer right after its recovery. The aim of present study was to evaluate the reliability and to set a reference range of a chemiluminescence enzyme immunoassay (CLEIA) compared to radioimmunoassay (RIA) method to retrieve in vivo matured oocytes. Serum progesterone concentration during proestrus and estrus was analyzed by RIA and CLEIA to determine ovulation day (Day 0). On Day 3, in vivo oocytes were recovered surgically and evaluated microscopically maturation status after staining nucleus with bisbenzimidazole dye. Mean progesterone concentration by CLEIA ($7.64{\pm}0.06ng/ml$) was significantly higher than by RIA ($6.46{\pm}0.04ng/ml$, P < 0.0001). It was not different between CLEIA ($10.01{\pm}0.34ng/ml$) and RIA values ($7.91{\pm}0.14ng/ml$, P < 0.05) on Day 0, but significantly higher CLEIA level on Day -1 and Day 1 ($6.41{\pm}0.15$ and $14.25{\pm}0.44ng/ml$) was assessed compared to RIA ($4.95{\pm}0.10$ and $11.29{\pm}0.34ng/ml$). However, with both methods, progesterone level was significantly increased from Day -1 to Day 2. To determine oocyte maturation with CLEIA method, a wider and higher reference range has to be considered.

• Radiation Oncology Journal
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• v.17 no.4
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• pp.299-306
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• 1999

Purpose : The human genetic disorder ataxia-telangiectasia (AT) is a multisystem disease characterized by extreme radiosensitivity. The recent identification of the gene mutated in AT, ATM, and the demonstration that it encodes a homologous domain of phosphatidylinositol 3-kinase (PI3-K), the catalytic subunit of an enzyme involved in transmitting signals from the cell surface to the nucleus, provide support for a role of this gene in signal transduction. Although ionizing radiation was known to induce c-fos transcription, nothing is known about how ATM or PKCI mediated signal transduction pathway modulates the c-fos gene transcription and gene expression. Here we have studied the effect of PKCI on radiation sensitivity and c-fos transcription in normal and AT cells. Materials and Methods: Normal (LM217) and AT (AT5BIVA) cells were transfected with PKCI expression plasmid and the overexpression and integration of PKCI was evaluated by northern blotting and polymerase chain reaction, respectively. 5 Gy of radiation was exposed to LM and AT cells transfected with PKCI expression plasmid and cells were harvested 48 hours after radiation and investigated apoptosis with TUNEL method. The c-fos transcription activity was studied by performing CAT assay of reporter gene after transfection of c-fos CAT plasmid into AT and LM cells. Results: Our results demonstrate for the first time a role of PKCI on the radiation sensitivity and c-fos expression in LM and AT cells. PKCI increased radiation induced apoptosis in LM cells but reduced apoptosis in AT cells. The basal c-fos transcription activity is 70 times lower in AT cells than that in LM cells. The c-fos transcription activity was repressed by overexpression of PKCI in LM cells but not in AT cells. After induction of c-fos by Ras protein, overexpression of PKCI repressed c-fos transcription in LM cells but not in AT cells Conclusion: Overexpression of PKCI increased radiation sensitivity and repressed c-fos transcription in LM cells but not in AT cells. The results may be a. reason of increased radiation sensitivity of AT cells. PKCI may be involved in an ionizing radiation induced signal transduction pathway responsible for radiation sensitivity and c-fos transcription. The data also provided evidence for novel transcriptional difference between LM and AT cells.

• Clinical and Experimental Pediatrics
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• v.45 no.9
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• pp.1106-1113
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• 2002

Purpose : Nuclear ($factor-{\kappa}BNF-{\kappa}B$) is now recognized as playing a potential role in programmed cell death and the adaptive response to various stress. Cellular hypoxia is a primary manifestation of many cardiovascular diseases. It seems that vascular endothelial growth factor (VEGF) and insulin like growth factor-I(IGF-I) have a function as a protective molecule in the heart against several stress including hypoxia. In this study, the role of $NF-{\kappa}B$ to the cellular response and regulation of protective molecules against the acute hypoxia in the heart was studied. Methods : To cause acute hypoxic stress to the heart, Sprague Dawley rats were exposed to hypoxic chamer($N_2$ 92% and $O_2$ 8%). After the hypoxic exposure, nuclear proteins, total proteins and mRNA were isolated from heart. Translocation of the transcription factors $NF-{\kappa}B$, NF-ATc, AP-1 and NKX-2.5 were evaluated by electrophoretic mobility shift assay(EMSA). The expression of IGF-I and VEGF were studied before and after the hypoxic stress by competitive-PCR, Northern hybridization and Western hybridization. To confirm the role of the $NF-{\kappa}B$ in the heart, the rats also were pretreated with diethyl-dithiocarbamic acid(DDTC) into peritoneal cavity to block $NF-{\kappa}B$ translocation into nucleus. Results : The expression of $NF-{\kappa}B$, AP-1 and NF-ATc were increased by the hypoxic stress. Increased expression of the VEGF and IGF-I were also observed by the hypoxic stress. However, the blocking of the $NF-{\kappa}B$ translocation reduced those expressions of VEGF and IGF-I. Conclusion : These results suggest that $NF-{\kappa}B$ has a protective role against the acute hypoxia through several gene expression, especially VEGF and IGF-I in heart muscle.

• Journal of the korean academy of Pediatric Dentistry
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• v.27 no.2
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• pp.318-332
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• 2000

The purpose of this study was to investigate the effects of demineralized freeze-dried bone (DFDB) on mechanically exposed pulp of dog by evaluating the pulpal inflammation and healing process, formation of dental hard tissue, and structural changes of fibroblasts of the remaining pulp tissue. Teeth of 4 dogs, weighing 10kg, were used in this study. Class V cavities were prepared followed by exposed the pulp tissue mechanically by sterilized round bur. In control group, exposed pulps were capped with calcium hydroxide paste followed by sealed with IRM. In experimental groups, the exposed pulps of one group were capped with the collagen and those of the other group were capped with DFDB. All cavities were sealed with same manor as control group. The animals were sacrificed at the intervals of 3, 7, 14, and 28 days for histopathlogic evaluation. The specimens were observed by the light microscope and trans-electron microscope. The results were as follows: 1. Pulp necrosis was not observed in all groups. Inflammatory response was disappeared from 1 week in control group and group 2. But it was not disappeared until 2 weeks and also irregular arrangement of odontoblasts was showed at the lateral walls of root canal just beneath the amputated site of the pulp in group 1. 2. Dentinal bridge was formed incompletely at 2 weeks but it was formed completely at 4 weeks in control group. Odontoid tissue was also found in control group at 4 weeks from treatment. Amputated site of pulp was encapsulated with fibrous tissue and odontoblast and dentinal bridge was not found in group 1. Preodontoid tissue and reparative dentin which were formed by odontoblast differentiated around DFDB were found, but dentinal bridge was not found in group 2. 3. Cell with large basophillic-stained nuclei infiltrated to amputated site and DFDB at 1 week from treatment in control group and group 2. They were found more in group 2 than in control group. Odontoblasts arranged more regularly and reparative dentin was found more as time elapsed. 4. Dentin-formative odontoblasts which showed ultramicrostructure of cytoplasm with polarized nucleus, rEM, Golgi complex, secretory granules, secretion of organic matrix in control of group and group 2. In regards to above results, the demineralized freeze-dried bone(DFDB) induce odontoblastic differentiation and further come up to the dentin formation in amputated pulp.

• The Korean Journal of Mycology
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• v.21 no.1
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• pp.16-22
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• 1993

Morphological and cultural characteristics of fungi causing rice sclerotial diseases were examined. Hyphal widths of R. solani and R. oryzae were same and ranged $6.0-12.0\;{\mu}m$ with average $9.0\;{\mu}m$, the widest among those of the sclerotial fungi examined. Hyphal width of R. oryzae sativae ranged $6.0-9.0{\mu}m$ with average $7.4{\mu}m$. Hyphal width of R. cerealis was the narrowest among those of Rhizoctonia species examined, and the same was hyphal width of S. oryzae among those of Sclerotium species. Nuclear staining by HCL-Giemsa method showed that R. solani and R. oryzae had many nuclei within one hyphal cell, S. oryzae one nucleus, and the other sclerotial fungi mostly two nuclei. The nuclear number of R. solani was the largest, which ranged 2-17 with average 6.3. Average size of sclerotia of the sclerotial fungi except S. hydrophilum and S. oryzae produced in lesions ranged 1.0-2.0mm. Average size of sclerotia of S. hydrophilum and S. oryzae was 0.5mm and 0.24mm, respectively. Sclerotia of R. solani and R. oryzae produced in culture were more variable in size and larger than those produced in lesions. However, the sclerotial sizes of the other sclerotial fungi produced in culture were almost the same as those produced in lesions. Sclerotial colors of sclerotial fungi produced in lesions were similar to those produced in culture, but sclerotial shapes of some sclerotial fungi exhibited somewhat difference between the sclerotia produced in lesions and in culture. Optimum temperature for mycelial growth of R. cerealis was $23^{\circ}C$, and that of the other sclerotial fungi ranged from $27\;to\;33^{\circ}C$. Maximum temperature for mycelial growth of some sclerotial fungi was as high as $41^{\circ}C$, while that of R. cerealis was as low as $31^{\circ}C$. Minimum temperature for mycelial growth of R. cerealis was $2^{\circ}C$, and that of the other sclerotial fungi ranged from $6\;to\;10^{\circ}C$.