• Journal of Biomedical Engineering Research
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• v.40 no.5
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• pp.143-150
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• 2019

To evaluate the toxicity of ophthalmic drug, the Draize test and Bovine Corneal Opacity and Permeability (BCOP) test commonly used. In Draize test, experimental animals were under stress and pain due to long-term exposure of drug. In addition, regarding physiological functions, animal model is not perfectly reflected a human eye condition. Although some models such as $EpiOcular^{TM}$, HCE model, LabCyte Cornea-Model, and MCTT $HCE^{TM}$ were already presented advanced cornea ex-vivo model to replace animal test. In this sense, cornea tissue structure mimicked ex-vivo toxicity model was fabricated in this study. The corneal epithelial cells (CECs) and keratocytes (CKs) isolated from rabbit eyeball were seeded on non-patterned silk film (n-pSF) and patterned silk film (pSF) at $32,500cells/cm^2$ and $6,500cells/cm^2$. Sequentially, n-pSF and pSF were stacked to mimic a multi-layered stroma structure. The thickness of films was about $15.63{\mu}m$ and the distance of patterns was about $3{\mu}m$. H&E stain was performed to confirm the cell proliferation on silk film. F-actin of CKs was also stained with Phalloidin to observe the cytoskeletal alignment along with patterns of the pSF. In the results, CECs and CKs were shown the good cell attachment on the n-pSF and pSFs. Proliferated cells expressed the specific phenotype of cornea epithelium and stroma. In conclusion, we successfully established the ex-vivo cornea toxicity model to replace the eye irritation tests. In further study, we will set up the human ex-vivo cornea toxicity model and then will evaluate the drug screening efficacy.

• Food Science of Animal Resources
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• v.33 no.4
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• pp.522-530
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• 2013

The ability of probiotics to adhere to the intestinal epithelium likely plays an important role in their colonization of the gastrointestinal tract. Here, we performed high-throughput screening (HTS) for suitable characteristics of potential probiotic bacteria using attachment and colonization ability through a C. elegans surrogate in vivo model. A total of 100 strains of lactic acid bacteria (LAB) isolated from infant feces were subjected to the colonization assay using C. elegans intestine. Based on colonization ability, we showed that nine isolates have a high attachment ability during whole experimental periods (up to 168 h), compared to Lactobacillus rhamnosus strain GG as a control. Also, through the use of an in vitro cell attachment model, nine isolates revealed highly binding activity to the mucus layer. Next, the selected 9 isolates were assayed for their survival ability when exposed to acidic and bile conditions as well as cholesterol reduction and the utilization of prebiotic substrates. As a result, the isolated nine strains were determined to be highly resistant to acid and bile conditions. In addition, they have significant activity for the reduction of cholesterol and utilization of several prebiotic substrates as a carbon source. Finally, the selected nine strains were identified by either L. rhamnosus or L. plantarum (4 strains for L. rhamnosus and 5 strains for L. plantarum, respectively). Taken together, we propose that the direct colonization of probiotics using C. elegans may be applicable to the rapid screening of valuable probiotic strains in vivo.

• Journal of Nutrition and Health
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• v.49 no.5
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• pp.277-287
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• 2016

Purpose: This study was performed to compare total phenolic contents, in vitro antioxidant capacity, and reduction effect of Korean food groups on ex vivo DNA damage in human cells and analyze correlations between each indicator. Methods: Vegetable foods in the Korean diet based the results of the KNHANES V-2 (2011) were classified into 10 food groups: cereals, fruits, vegetables, nuts, kimchi, seaweeds, potatoes, mushrooms, legumes, and oils. Eighty-four foods constituted more than 1% of the total intake in each food group and finally designated as vegetable foods in the Korean diet. Total phenolic content of each food group was measured. Further, in vitro antioxidant capacity was measured based on DPPH radical scavenging assay, TEAC assay, and $ORAC_{ROO{\cdot}}$ assay. Ex vivo DNA damage in human lymphocytes was assessed using comet assay. Results: Total phenolic contents of food groups of the Korean diet increased in the order of mushrooms, fruits, vegetables, seaweeds, and kimchi. Meanwhile, antioxidant rankings of food groups as mean values from the three in vitro test methods increased in the order of mushrooms, seaweeds, vegetables, kimchi, and fruits. Protection against ex vivo DNA damage in human lymphocytes was highest in mushrooms, followed by vegetables, fruits, seaweeds, and kimchi. The rankings of the food groups for total phenolic content, in vitro DAC, and ex vivo DNA protection activity were similar, and correlations between each indicator were significantly high. Conclusion: Mushrooms, fruits, vegetables, and seaweeds among the tested food groups in the Korean diet showed high total phenolic contents, in vitro antioxidant capacities, and protection against DNA damage. Correlations between each indicator in terms of total phenolic content, in vitro antioxidant capacity, and ex vivo DNA protection between each food group were found to be particularly high.

• Restorative Dentistry and Endodontics
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• v.31 no.3
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• pp.192-202
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• 2006

The purpose of this study was to examine the viability of PDL cells in rat molars by using in vivo MTT assay, which was used to compare fast cryopreservation group by liquid nitrogen $(-196^{\circ}C)\;with\;4^{\circ}C$ cold preservation group. A total of 74 Sprague-Dawley white female rats of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted as atraumatically as possible under ketamine anesthesia. Ten teeth of each group were divided as six experimental groups depending upon the preservation. Cryopreservation groups were Group 1 (5% DMSO 6% HES in F medium) Group 2 (10% DMSO in F medium), Group 3 (5% DMSO 6% HES in $Viaspan^(R)$). Group 4 (10% DMSO in $Viaspan^(R)$) which were cryopreserved for 1 week and cold preservation groups were Group 5 (F medium) , Group 6 ($Viaspan^(R)$) at $4^{\circ}C$ for 1 week. Immediate extraction group was used as a control. After preservation and thawing, the in vivo MTT assay was processed. Two way ANOVA and Duncan's Multiple Range Test was performed at the 95 % level of confidence, Another 2 teeth of each group were treated as the same manner and frozen sections $10{\mu}m$ thick for microscopic observation. The value of optical density obtained after in vivo MTT analysis was divided by the value of eosin staining for tissue volume standardization. Group 1, 2 had significantly higher optical density than Group 3 and 4 which had the lowest OD value. Group 6 had higher OD value than in Group 5 (P<0.05). Histological findings of periodontal ligament cell, after being stained with MTT solution were consistent with the in vivo MTT assay results. In this study, the groups which were frozen with DMSO as a cryoprotectant and the groups with F medium showed the best results.

• Proceedings of the Materials Research Society of Korea Conference
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• 2004.05a
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• pp.67-67
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• 2004

• Preventive Nutrition and Food Science
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• v.21 no.1
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• pp.24-30
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• 2016

Fresh ginseng roots were aged in an oven at $80^{\circ}C$ for 14 d. The in vitro and in vivo antioxidant activities of this aged ginseng, in comparison with those of the white and red ginsengs, were evaluated. In in vitro antioxidant assays, the ethanolic extracts from aged ginseng showed significantly higher free radical scavenging activity and reducing power than those of the white and red ginsengs. In in vivo antioxidant assays, mice were fed a high fat diet supplemented with white, red, or aged ginseng powders. High fat feeding resulted in a significant increase in lipid peroxidation and a substantial decrease in antioxidant enzymes activities in the animals. However, diet supplementation of ginseng powders, particularly aged ginseng, markedly reduced lipid peroxidation and enhanced the antioxidant enzymes activities. The results illustrate that the aged ginseng has greater in vitro and in vivo antioxidant capacity than the white and red ginsengs. The aged ginseng also showed considerably higher total saponin, phenolic, and flavonoid contents, indicating that its antioxidant capacity may have been partly due to its high levels of antioxidant compounds. This new ginseng product may be useful as a functional food with strong antioxidant potential.

• Advances in Traditional Medicine
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• v.5 no.1
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• pp.29-36
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• 2005

Polysaccharide hydrogels constitute a structurally diverse class of biological macromolecules with a wide range of physicochemical properties. They also constitute important members of the family of industrial water-soluble polymers. They find application in Pharmacy as binders, disintegrants, suspending, emulsifying and sustaining agents. According to the International Pharmaceutical Excipients Council (IPEC), an excipient must have an established safety profile. Hence, in the present study, in vitro cytotoxicity on Vero and HEp-2 cell lines, and in vivo acute toxicity in rats were carried out to establish the safety of polysaccharide hydrogels from the seeds of Plantago ovata and Ocimum basilicum. The in vitro cytotoxicity was determined by MTT and SRB assays. In the in vivo acute toxicity, the effects of three different doses of hydrogels (100, 200 and 400 mg/kg body weight) on food and water intake, body weight, biochemical and hematological parameters were studied. The results of in vitro did not show any cytotoxicity on both the cell lines used. In the in vivo acute toxicity, the hydrogels did not show any toxic symptoms in all three dose levels. This establishes the safety of the selected hydrogels. Hence, they can be used as excipients in pharmaceutical dosage forms.

• Advances in Traditional Medicine
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• v.3 no.2
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• pp.72-79
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• 2003

The present investigation was undertaken to study whether tumor-associated macrophages of Daltons lymphoma (DL), a spontaneous transplantable T cell lymphoma can be activated to tumoricidal state by alcoholic extract of Tinospora cordifolia (ALTC). In vivo administration of ALTC (200 mg/kg body weight) in DL-bearing mice resulted in an enhanced RNI production and an augmented cytotoxic response of tumor-associated macrophages. Earlier we had reported that DL-bearing mice show a regression of thymus and an enlargement of spleen. In vivo administration of ALTC to DL-bearing hosts resulted in a decrease in the weight of spleen and counts of splenocytes along with an increase in the weight of thymus as compared to control DL-bearing mice. In vivo administration of ALTC in DL-bearing mice also resulted in an increase in the proliferation of splenocytes/thymocytes and BMC. The results of this study indicate that the ALTC upon in vivo administration in DL-bearing shows immuno-modulatory effects and thus may have clinical significance.

• Korean Journal of Animal Reproduction
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• v.21 no.1
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• pp.9-18
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• 1997

These studies were carried out to establish an effective and practical system for commercialization of embryo production techniques by analyzing several factors influencing in vivo embryo production in Korean native cattle. In vivo embryos were flushed 226 times from 128 donors. The results obtained in studies on the factors influencing in vivo embryo production were as follows : 1. There were no significant differences in the number of total recovered, fertilized, transferable and freezable embryos among the hormone doses(FSH-P, 28∼34mg; SUPER-OV, 75IU) used for superovulation. However, over 30mg doses of FSH-P showed a slightly higher effect than others. 2. There were slight decrease in the number of fertilized, transferable and freezable embryos in 3 times repeated superovulation. But there were no significant differences among 1, 2 and 3 times repeated superovulations. 3. Age of donors did not affect the number of transferable and freezable embryos, but the number of fertilzed embryos were highest in 2∼3 years old donors and were lowest in 8∼9 years old donors(P<0.05). 4. Season had a significant effect on the production of embryos(P<0.05). the embryo production, and the number of fertilized, transferable and freezable embryos were most effective in summer and follow by spring, autumn and winter. 5. The number of transferable and freezable embryos was highest in donors flushed at 7∼8 days after estrus(P<0.05).

• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2002.11a
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• pp.163-163
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• 2002