• Journal of Veterinary Clinics
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• v.21 no.2
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• pp.209-213
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• 2004

A 8 kg, 3-year-old male West Highland white terrier dog with a 1.5-year history of chronic, severely pruritic, seborrheic skin disorder was referred to the Veterinary Medical Teaching Hospital of the Tokyo University of Agriculture and Technology. On physical examination, lesions were observed on entire cutaneous surface, except for face, dorsum of body, and footpads. Skin lesions were characterized by diffuse erythema, erythematous papules, severe alopecia, hyperpigmentation, and lichenification. Tape strip tests of skin lesions revealed cocci and Malassezia infections. The intradermal allergy tests revealed positive reactions to Japanese cedar pollen, but the non-seasonal clinical signs were not compatible with atopic dermatitis caused by this pollen. Results of hematological examination, serum chemistry and thyroid gland profile were normal. Examination of skin biopsy exhibited hyperplastic superficial perivascular dermatosis with severe acanthosis, excessive keratinocyte mitoses, patchy or diffuse mild spongiosis, and lymphocytic exocytosis in epidermis. Perivascular to interstitial mononuclear cells infiltration was seen in the superficial dermis. Based on the results of examination described above, epidermal dysplasia was diagnosed. Treatments with administration of antibiotics, etretinate, and prednisolone orally combined with topical ketoconazole cream and antiseborreic shampoos had no good results. Following treatment with long-term oral itraconazole at 10 mg/kg daily and chlorhexidine shampoos was successful. However, when itraconazole therapy was stopped, the condition worsened twice within 2 or 3 months. Readministration of itraconazole produced improvement within 4 weeks. This dog has now been controlled periodical itraconazole therapy.

• Applied Microscopy
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• v.37 no.4
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• pp.209-217
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• 2007

The system of wound healing is very complex biological processing that includes inflammatory, reepithelialization, and matrix construction. For identification of the transitional pathway of the keratinocytes, we have employed immunohistochemical analysis using cytokeratin antibody after wounding. Epithelium in skin of the frog(Bombina orientalis) was examined with transmission electron microscopy. Cytokeratin was expressed in normal basal and gland cavity cells. At 3-hour basal layer cells were strong positive, however cells of the upper layer were negative reaction. Day1 and 2 after post-wounding, regenerating epithelial cell layer was positive reaction, especially basal layer cells were strong positive. At day 10 after wounding, the degree of positive reaction to basal cells of regenerating epithelial tissue was equal to day 7 wound tissue. At day of 19th, basal and spinous layer cells were strong positive reaction. Regenerating epithelial cells were positive but some basal cells were strong positive at day 27. From this result, we identified that the migration of the keratinocytes in amphibian skin wounds is initiated from basal layer fells and the keratinocytes migrate into basal and middle of the wound area.

• Journal of Physiology & Pathology in Korean Medicine
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• v.29 no.1
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• pp.58-65
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• 2015

This study is designed to investigate the protective effects of fermented Red Ginseng (FRG) against photoaging in vitro and in vivo. UVB was irradiated to the human keratinocyte HaCaT cell and dorsal skin of SKH-1 mice for the induction of photoaging. After treatment of non-fermented red ginseng (NRG), fermented red ginseng (FRG), and fortified fermented red ginseng (FFRG) to the UVB irradiated HaCaT cell, ROS production and activity of MMP-9 were examined by DCFC-DA assay and gelatin zymographic assay respectively. UVB irradiated SKH-1 mice were treated with NRG, FRG, and FFRG via oral(300 mg/Kg B.W./day) and topical($100{\mu}{\ell}/mouse/day$) route.All of NRG, FRG, and FFRG had significantly reduced the intracellular ROS production elicited by UVB, among them FRG slightly more reduced the ROS production than NRG and FFRG. FFRG had slightly more reduced the MMP-9 activity in UVB irradiated HaCaT cells than NRG and FFRG in high dose. Oral and topical treatment of NRG, FRG, and FFRG had decreased the expression of matrix metalloproteinase-2, -3, and -9 in dorsal skin of UVB irradiated mice. Among them, inhibitory effect of FRG on the expression of MMP-2 was apparent. We speculate that FRG has therapeutic potentials on the UVB irradiated photoaging.

• Korean Chemical Engineering Research
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• v.43 no.1
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• pp.103-109
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• 2005

Isolation and primary culture technique of rabbit oral keratinocytes, and the study for effect of environmental factors on the cell growth were carried out in T75-flask. $1.92{\pm}0.59{\times}10^6$ viable cells were isolated by trypsin enzymatic digestion method from $0.25cm^2$ biopsy of rabbit oral mucosa. Primary culture with 10 mL of K-SFM containing 50 mg/L BPE, $5.0{\mu}g/L$ EGF and 0.15 mM $Ca^{2+}$ showed confluence after 8 days and doubling time was 2.54 days. Effect of medium types, medium volume and supplement types on the cell growth was investigated after the cultured keratinocytes had been harvested from primary confluence. Serum addition showed adverse effect and the increase of serum concentration didn't have an effect on the cell growth. The increase of medium volume decreased the cell growth. The increase of calcium concentration increased the cell growth and 2.0 mM was optimum value. In conclusion, when rabbit oral keratinocytes was cultured in T75-flask, the most effective conditions was to use 10 mL of K-SFM containing 50 mg/L BPE, $5.0{\mu}g/L$ EGF and 2.0 mM $Ca^{2+}$, and doubling time was 1.32 days. This study can provide the useful informations to develop a process and design a bioreactor for the culture of keratinocytes in human body like skin and cornea, as well as mucosa.

• International Journal of Oral Biology
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• v.32 no.1
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• pp.35-43
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• 2007

Tooth loss in elderly is mainly caused by alveolar bone loss via severe periodontitis. Although the severity of periodontitis is known to be affected by age, the aging process or the genetic changes during the aging of periodontal tissue cells are not well characterized. In this study, we investigated the effect of in vitro aging on the change of gene expression pattern in periodontal fibroblasts. Gingival fibroblasts (GF) and periodontal ligament fibroblasts (PDL) were obtained from two young patients and replicative senescence was induced by sequential subcultivation. When more than 90% cells were positively stained with senescence-associated ${\beta},-galactosidase$, those cells were regarded as aged cells. In aged GF and PDL, the level of phosphorylated retinoblastoma (RB) and $p16^{INK4A}$ protein was significantly decreased and increased, respectively. However, the protein level of p53 and p21, well known senescence-inducing genes, did not increase in aged GF and PDL. Although $p27^{Kip1}$ and $p15^{INK4B}$, another cyclin-dependent kinase inhibitors, were reported to be involved in replicative senescence of human cells, they were decreased in aged GF and PDL. Because senescent cells showed flattened and enlarged cell shape and are known to have increased focal adhesion, we examined the protein level of several integrins. Aged GF and PDL showed increased protein level of integrin ${\alpha}2$, ${\alpha}v$, and ${\beta}1$. When the gene expression profiles of actively proliferating young cells and aged cells were compared by cDNA microarray of 3,063 genes and were confirmed by reverse transcription-polymerase chain reaction, 7 genes and 15 genes were significantly and commonly increased and decreased, respectively, in aged GF and PDL. Among them, included are the genes that were known to be involved in the regulation of cell cycle, gene transcription, or integrin signaling. The change of gene expression pattern in GF and PDL was minimally similar to that of oral keratinocyte. These results suggest that $p16^{INK4A}/RB$ might be involved in replicative senescence of periodontal fibroblasts and the change of gene expression profile during aging process is cell type specific.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.222-222
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• 2004

Superoxide dismutases are key antioxidant enzymes in metabolism of reactive oxygen species. Three different isoforms of SOD exist in mammals. The extracellular SOD (EC-SOD) is the most recently discovered SOD family member. This isoform is a copper- and zinc-containing enzyme like Cu/Zn-SOD and a homotetrameric glycoprotein with a molecular weight of about 165 kDa in mouse. (omitted)

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.120-120
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• 2019

Allergic inflammatory disease has been increased by abnormal lifestyle and food habits. Especially, prevalence of atopic dermatitis (AD) has been elevated and treatment of AD has not been unclear. Red sweet pepper (RSP), named as Capsicum annuum L, has been known as having pharmacological effects such as antioxidant, detoxification and antibacterial effects. However, the beneficial effect of ethanolic extract of RSP on AD has not been partly examined yet. Therefore, the aim of this study was to investigate anti-inflammatory effects of RSP on AD in vitro and in vivo models. The treatment of RSP inhibited the secretion of inflammatory cytokine such as interleukin (IL)-6 and IL-8 in tumor necrosis factor (TNF)-${\alpha}$ and interferon (IFN)-${\gamma}$-stimulated human keratinocyte (HaCaT cell). Also, RSP extract regulated 2,4-dinitroflorobenzene (DNFB)-induced AD-like skin lesions in BALB/c mice. Oral administration of RSP ameliorated DNFB-induced AD-like symptoms. In presented results indicated that RSP inhibited inflammatory cytokines in HaCaT cell and ameliorated AD-like skin lesion through suppression of symptom of DNFB-induced skin inflammation. Thus, RSP might be a potential therapeutic agent for AD.

• Journal of the Korean Applied Science and Technology
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• v.38 no.1
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• pp.29-36
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• 2021

Among natural products, the shells of black walnut(Juglans nigra), which are not used commercially and mostly discarded, were examined to investigate the physiological activity and the efficacy for the cosmetic application. DPPH radical scavenging activity of hot water extract of black walnut shells was 76.06% at 300 ㎍/mL. ABTS radical scavenging activity of the extract was 61% at 1000 ㎍/mL, showing excellent antioxidant activity. When the black walnut shell extract was applied to HaCaT cells, a skin keratinocyte, the viability of the cells was 92.6% at 250 ㎍/mL, showing a remarkably low effect on cell viability. At the concentration of 500 ㎍/mL, 67.35% of nitric oxide(NO) production was inhibited. It also showed an inhibitory effect on Hyaluronidase 31 times higher than that of Vitamin C at 100 ㎍/mL concentration. In conclusion, the black walnut shell extract showed high potentials for the cosmeceutical applications, suggesting the possibility of using it as a high value-added natural material in various industries such as food, fragrance, healthcare, and pharmaceuticals.

• Biomolecules & Therapeutics
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• v.31 no.6
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• pp.640-647
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• 2023

The skin, the largest organ in the body, undergoes age-related changes influenced by both intrinsic and extrinsic factors. The primary external factor is photoaging which causes hyperpigmentation, uneven skin surface, deep wrinkles, and markedly enlarged capillaries. In the human dermis, it decreases fibroblast function, resulting in a lack of collagen structure and also decreases keratinocyte function, which compromises the strength of the protective barrier. In this study, we found that treatment with γ-aminobutyric acid (GABA) had no toxicity to skin fibroblasts and GABA enhanced their migration ability, which can accelerate skin wound healing. UVB radiation was found to significantly induce the production of matrix metalloproteinase 1 (MMP-1), but treatment with GABA resulted in the inhibition of MMP-1 production. We also investigated the enhancement of filaggrin and aquaporin 3 in keratinocytes after treatment with GABA, showing that GABA can effectively improve skin moisturization. In vivo experiments showed that oral administration of GABA significantly improved skin wrinkles and epidermal thickness. After the intake of GABA, there was a significant decrease observed in the increase of skin thickness measured by calipers and erythema. Additionally, the decrease in skin moisture and elasticity in hairless mice exposed to UVB radiation was also significantly restored. Overall, this study demonstrates the potential of GABA as functional food material for improving skin aging and moisturizing.

• Journal of Korean society of Dental Hygiene
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• v.16 no.3
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• pp.471-477
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• 2016

Objectives: Fluoride is widely used in the prevention and control of dental caries. The purpose of this study is to examine the biological effects of Sodium fluoride on the proliferation of oral normal cell in vitro(MDPC-23, HaCaT, HGF-1 cells). Methods: The proliferation of normal cells and the cyto-skeletal change of normal cells were assessed by WST-1 assay and F-actin stain assay. The statistical significances of the resulting data were analyzed using SPSS(Window 12.0). Results: The sodium fluoride(0-12 mM) treatment decreased the cell viability in a dose and time dependent manner: HaCaT(6 h): $100{\pm}0$, $98{\pm}0.39$, $82{\pm}2.68$, $75{\pm}0.83$, $69{\pm}1$, $67{\pm}1.42%$(p<0.005); HaCaT(24 h): $100{\pm}0$, $98{\pm}1.85$, $54{\pm}0.64$, $43{\pm}0.4$, $38{\pm}0.32$, $36{\pm}0.13%$(p<0.006), MDPC-23(6 h): $100{\pm}0$, $93{\pm}1.48$, $85{\pm}0.28$, $82{\pm}1.58$, $79{\pm}1.48$, $76{\pm}1.93%$(p<0.009); MDPC-23(24 h): $100{\pm}0$, $91{\pm}1.26$, $58{\pm}0.65$, $49{\pm}1$, $44{\pm}0.74$, $2{\pm}0.05%$(p<0.005), HGF-1(6 h): $100{\pm}0$, $97{\pm}2.93$, $89{\pm}5$, $71{\pm}5.42$, $58{\pm}4.82$, $43{\pm}3.47%$(p<0.009); HGF-1(24 h): $100{\pm}0$, $97{\pm}2.05$, $73{\pm}1.73$, $22{\pm}1.61$, $14{\pm}1.73$, $7{\pm}0.85%$(p<0.005). Thus, changes in cell morphology and disruption of filamentous(F)-actin organization were observed in higher concentration. Conclusions: These results suggest that higher concentrations of fluoride lead to a reduce the number of cells and morphology change of normal cell.