• Journal of fish pathology
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• v.29 no.1
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• pp.35-43
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• 2016

In comparison to the numbers of such studies of fish, few studies have been carried out on the immunity, physiology and ecology of abalone, while studies on abalone disease are also extremely rare. Moreover, mass mortality of cultured abalone due to pathogenic bacteria has not been reported in the southern coast of Korea. However, Vibrio-like bacteria have been isolated from dead abalone, which indicates that a review is required in order to determine the cause of abalone mortality. Use of an antimicrobial agent to minimize the damage caused by disease in abalone farms is common, but the therapeutic effects are insignificant. Demand for probiotics has increased, but research on the development of probiotics for use in abalone culture is very rare. Therefore, the present study isolated KC16-2 from fermented kimchi soup and investigated the characteristics of the isolate as a candidate probiotic bacterium in abalone. KC16-2 was identified as Bacillus amyloliquefaciens (B. amyloliquefaciens KC16-2) based on its biochemical properties and 16S rRNA gene sequence. B. amyloliquefaciens KC16-2 showed inhibitory effects against the growth of various vibrios in vitro, and kept the numbers constant until four days after inoculation in marine water at a temperature of $15{\sim}25^{\circ}C$, indicating the possible use of KC16-2 as a probiotic, except in the winter. The growth of KC16-2 was inhibited by bile salt, but the numbers increased over time suggesting the bacteria were still alive in the abalone's digestive tract. Abalone fed with a diet including KC16-2 for 12 weeks showed good growth, but showed no significant differences from the control group. However, the mortality of the abalone supplied the probiotic diet was reduced to half that of the control group in a challenge test with Vibrio tubiashii. Therefore, we suggest that B. amyloliquefaciens KC16-2 could be used as a probiotic bacterium for control of the mortality of abalone caused by opportunistic pathogenic vibrios.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.4
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• pp.500-508
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• 2011

This study was carried out to investigate the effects of jujube extracts on intestinal microflora, along with their antioxidant activities, according to extraction method. The antimicrobial activities of the extracts were measured using the agar diffusion method with a jujube extract concentration of 50 mg/mL. Neither the first nor second jujube extracts were inhibitory against the tested intestinal bacteria. However, water extracts of jujube significantly enhanced the growth of lactic acid bacteria, especially Bifidobacterium bifidum and Bifidobacterium adolescentis. Total phenol compounds and flavonoid compounds were higher in the 1st than in the 2nd water extracts. The EDA values of both water and ethanol extracts increased in proportion to the extract concentration. The 1st water extract showed the highest value among all the others, which was 85.60% at the concentration of 0.05 mg/mL. Furthermore, the 1st water extract showed stronger antioxidant activity than the other samples with an activity of 679.91 mg AA eq/g. These results support the potential use of jujube water extracts as a functional food component and a valuable resource for the development of nutraceutical foods, to increase the growth of Bifidobacterium spp. in the human intestine.

• Tuberculosis and Respiratory Diseases
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• v.82 no.1
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• pp.42-52
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• 2019

Background: Transforming growth factor ${\beta}$ (TGF-${\beta}$), retinoic acid (RA), p38 mitogen-activated protein kinase (MAPK), and MEK signaling play critical roles in cell differentiation, proliferation, and apoptosis. We investigated the effect of RA and the role of these signaling molecules on the phosphorylation of Smad2/3 (p-Smad2/3) induced by TGF-${\beta}1$. Methods: A549 epithelial cells and CCD-11Lu fibroblasts were incubated and stimulated with or without all-trans RA (ATRA) and TGF-${\beta}1$ and with MAPK or MEK inhibitors. The levels of p-Smad2/3 were analyzed by western blotting. For animal models, we studied three experimental mouse groups: control, bleomycin, and bleomycin+ATRA group. Changes in histopathology, lung injury score, and levels of TGF-${\beta}1$ and Smad3 were evaluated at 1 and 3 weeks. Results: When A549 cells were pre-stimulated with TGF-${\beta}1$ prior to RA treatment, RA completely inhibited the p-Smad2/3. However, when A549 cells were pre-treated with RA prior to TGF-${\beta}1$ stimulation, RA did not completely suppress the p-Smad2/3. When A549 cells were pre-treated with MAPK inhibitor, TGF-${\beta}1$ failed to phosphorylate Smad2/3. In fibroblasts, p38 MAPK inhibitor suppressed TGF-${\beta}1$-induced p-Smad2. In a bleomycin-induced lung injury mouse model, RA decreased the expression of TGF-${\beta}1$ and Smad3 at 1 and 3 weeks. Conclusion: RA had inhibitory effects on the phosphorylation of Smad induced by TGF-${\beta}1$ in vitro, and RA also decreased the expression of TGF-${\beta}1$ at 1 and 3 weeks in vivo. Furthermore, pre-treatment with a MAPK inhibitor showed a preventative effect on TGF-${\beta}1$/Smad phosphorylation in epithelial cells. As a result, a combination of RA and MAPK inhibitors may suppress the TGF-${\beta}1$-induced lung injury and fibrosis.

• The Korean Journal of Pharmacology
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• v.26 no.2
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• pp.193-207
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• 1990

Transforming growth factor ${\beta}(TGF-{\beta})$ is a multifunctional polypeptide with diverse effects on the proliferation, differentiation and other functions in many cell types. $TGF-{\beta}$ is highly abundant in bone matrix and induces divergent responses in many aspects of bone cell metabolism . Several lines of investigation indicate that matrix-associated $TGF-{\beta}$ is the products of bone cells themselves. However, exact bone cell type reponsible for the production of $TGF-{\beta}$ is still in controversy, The present study was undertaken to determine the cellular origin of matrix-associated $TGF-{\beta}$ and to assess how different bone cells respond to $TGF-{\beta}$. As a prerequisite for this, 5 bone cell populations of distinct phenotype were isolated from fetal calvaria with sequential enzyme digestion protocol and biochemical characterization. Calvarial cell populations released in early stage showed fibroblastic features whereas populations relesed later was enriched with osteoblast-like cell as judged by their acid and alkaline phosphatase activities, cAMP responsiveness to parathyroid hormone, calcitonin and prostaglandin $E_2$ and collagen synthesis rate. By polyacylamide gel and immunoblot analysis of bone and calvarial cell extracts, presence of $TGF-{\beta}$ in bone tissues and production of $TGF-{\beta}$ by bone cells were confirmed again. Subsequent analysis of calvarial cell extracts prepared as individual population revealed that all calvarial cell populations synthesize $TGF-{\beta}$. Exogenously added $TGF-{\beta}$ induced biphasic response upon bone cell proliferation under serum-free condition. In osteoblastic cell populations, it was stimulatory whereas inhibitory in fibroblastic cell populations. In contrast, collagen and noncollagen protein synthesis of all calvarial cell populations were stimulated by $TGF-{\beta}$. Enhancement of protein synthesis was found to be more general rather than specific for collagen synthesis. In addition, effects of $TGF-{\beta}$ on protein synthesis were independent to its effects on cell proliferation. In summary, production of $TGF-{\beta}$ by bone cells and differential actions on various cell populations observed in this study suggest that $TGF-{\beta}$ may play an important role in the regulation of bone metabolism by modulating the specific cellular functions in autocrine and paracrine fashion.

• Journal of Life Science
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• v.20 no.10
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• pp.1519-1524
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• 2010

Apigenin (4', 5, 7-trihydroxyflavone), a common dietary flavonoid abundantly present in fruits and vegetables, has shown remarkable anti-proliferative effects against various malignant cell lines. To observe the anti-proliferative effects, oral cavity cancer cell lines, $6{\times}10^3$ cells/well (96 well plate) of KB oral cavity tumor cells were plated and 24 hr later treated with apigenin for one day, after which MTT assay was performed. Apigenin induced cell death in a dose-dependent manner after incubation. Cell viability was significantly decreased in the group treated with 100 ${\mu}M$ apigenin for 24 hr (p<0.05) compared to the control group. To assess apoptosis, the nuclei of KB cells were stained with DAPI. The presence of chromatin condensation in the apigenin treated cells was detected on a fluorescent microscope (${\times}200$). We investigated the in vivo growth inhibitory effects of apigenin on oral cavity cancer KB tumor xenograft subcutaneously implanted in male nude mice. Apigenin was administered to mice by gavage at doses of 25 and 50 mg/kg/day in 0.2ml of PBS. Tumor volume was significantly decreased in 25 and 50 mg/kg apigenin-administration groups compared to the control group. For apoptosis analysis, TUNEL staining was performed. A significant increase in TUNEL positive cells was found in the 25 mg/kg apigenin administration group compared to the non- apigenin administration group. Histopathological changes were not observed. These results indicate that apigenin inhibits oral cavity cancer cell growth through the induction of apoptosis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.3
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• pp.487-493
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• 2004

This study has been undertaken to increase availability of native camellia in Jeonnam as a medicinal resource and to isolate the effective components from them. Fresh leaf and flower of camellia, single camellia tea and camellia tea mixed with green tea, herbs were screened for cytotoxicity on MCF -7 (human breast adenocarcinoma pleual effusion), Calu-6 (human pulmonary carcinoma), SNU-601 (human gastric carcinoma) cells. Also their multidrug-resistance reversing activity were evaluated using drug sensitive AML-2/WT and multidrug-resistant AML-2/D100 cells. Among the camellia extracts, young leaf and camellia tea mixed with green tea had strong growth inhibitory effects in below 100 $\mu\textrm{g}$/mL against human cancer cells. In result, young leaf showed the strongest inhibitory effects on MCF -7 ($IC_{50}$/ = 100 $\mu\textrm{g}$/mL ↑), Calu-6 ($IC_{50}$/ = 79 $\mu\textrm{g}$/mL), and SNU -601 ($IC_{50}$/ = 39 $\mu\textrm{g}$/mL), and AML-2/WT ($IC_{50}$/ = 64 $\mu\textrm{g}$/mL). Chemosensitizing effect was the extracts of mature leaf ($IC_{50}$/ = 97 $\mu\textrm{g}$/mL, RF=3.0), roasted tea ($IC_{50}$/ = 76 $\mu\textrm{g}$/mL, RF = 2.6 ↑) and steam tea ($IC_{50}$/ = 70 $\mu\textrm{g}$/mL, RF=2.8 ↑) strongly potentiate vincristine cytotoxicity in AML-2/D100 cells. But their cytotoxicities to both sensitive AML-2/WT and resistant AML-2/D100 cells were in the same order of magnitude. This results indicate that crude extracts of camellia mature leaves would contain some principles which have chemosensitizing activity.

• Korean Journal of Medicinal Crop Science
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• v.13 no.4
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• pp.154-160
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• 2005

This study was performed to compare anticancer and immune activities between natural Artemisia capillaris Thunb. extract and tissue cultured plant extract (hairy root, in vitro culture, callus). The inhibitory effect of cancer cell growth, human B cell growth and productivity of cytokines were examined. Furthermore, HPLC analysis was performed to confirm the components. The anticancer activities increased by more than 55% with the cultured callus of Artemisia capillaris T. for four cancer cell lines(Lung carcunoma, Stomach adenocarcinoma, Hepatocillular carcinoma, Breast adenocarcinoma), showing higher effect than natural Artemisia capillaris T. The extracts from hairy root and in vitro culture of Artemisia capillaris T. significantly increased the immune B cell growth. The immune B cell growth effect of natural Artemisia capillaris T. was higher than that of the tissue culture plants such as hairy root, in vitro culture and callus. Both natural and tissue cultured plants showed similar effects on cytokine secretion. The similar peak size was observed between natural Artemisia capillaris T. and cultured callus in HPLC analysis. As a results, the biological activities were not observed the difference between natural Artemisia capillaris T. and cultured callus. Thus, the cultured callus will be altered natural Artemisia capillaris T. in the environmental side and the resources preservative side

• Korean Journal of Microbiology
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• v.50 no.4
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• pp.345-350
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• 2014

The aim of this study was to investigate the effect of high temperature on the viability of probiotic organisms (Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae) mixed with animal feed under controlled conditions by simulating a farm feed bin in the summer. Following inoculation of probiotics into the feed, the pH and probiotic viability were monitored during an 8-day incubation at room temperature. Sterile and non-sterile feeds displayed different patterns of pH changes, with increased pH in non-sterile feed at 2 days, but a pattern of decreasing pH at 4 days. The viabilities of S. cerevisiae and B. subtilis after mono/co-inoculation were maintained without substantial changes during the incubation, whereas L. plantarum viability tended to decline. In both non-sterile and sterile feeds, the probiotics were maintained or grew without any antagonistic effects. Probiotic viability was also tested upon a shift to high temperature ($60^{\circ}C$). There was no distinct change in pH between sterile and non-sterile feeds after the temperature shift. L. plantarum and S. cerevisiae could not survive at the high temperature, whereas B. subtilis displayed normal growth, and it inhibited the growth of contaminant microbes. Fungal growth was not observed in non-sterile feed 2 days after supplementation with B. subtilis. Therefore, heat resistant B. subtilis could be safely used in feed bins to inhibit microbial contamination, even at high temperatures. The prevention of elevated temperature in feed bins is necessary for the utilization of L. plantarum and S. cerevisiae during the summer season.

• Journal of Life Science
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• v.19 no.6
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• pp.770-780
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• 2009

A new photosensitizer, 9-Hydroxypheophorbide-a (9-HpbD-a), was derived from Spirulina platensis. We conducted a series of experiments, in vitro and in vivo, to evaluate the anticancer effect and mechanism of photodynamic therapy using 9-HpbD-a and 660 nm diode lasers on a squamous carcinoma cell line. We studied the cytotoxic effects of pheophytin-a, 9-HpbD-a, 9-HpbD-a red and 660 nm diode lasers in a human head and neck cancer cell line (SNU-1041). Cell growth inhibition was determined by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. The effects of 9-HpbD was higher than those of 9-HpbD-a red or pheophytin-a in PDT. We then tested the cytotoxic effects of 9-hydroxypheophorbide-a (9-HpbD-a) in vitro. The cultured SNU-I041 cells were treated with serial concentrations of 9-HpbD-a followed by various energy doses (0, 0.1, 0.5, 3.2 J/$cm^{2}$) and by various interval times (0, 3, 6, 9, 12 hr) until laser irradiation, then MTT assay was applied to measure the relative inhibitory effects of photodynamic therapy (PDT). Optimal laser irradiation time was 30 minutes and the cytotoxic effects according to incubation time after 9-HpbD-a treatment increased until 6 hours, after which it then showed no increase. To observe the cell death mechanism after PDT, SUN-I041 cells were stained by Hoechst 33342 and propidium iodide after PDT, and observed under transmission electron microscopy (TEM). The principal mechanism of PDT at a low dose of 9-HpbD-a was apoptosis, and at a high dose of 9-HpbD-a it was necrosis. PDT effects were also observed in a xenografted nude mouse model. Group I (no 9-HpbD-a, no laser irradiation) and Group II (9-HpbD-a injection only) showed no response (4/4, 100%), and Group III (laser irradiation only) showed recurrence (1/4,25%) or no response (3/4, 75 %). Group IV (9-HpbD-a + laser irradiation) showed complete response (10/16, 62.5%), recurrence (4/16, 25%) or no response (2/16, 12.5%). Group IV showed a significant remission rate compared to other groups (p<0.05). These results suggest that 9-HpbD-a is a promising photosensitizer for the future and that further studies on biodistribution, toxicity and mechanism of action would be needed to use 9-HpbD-a as a photosensitizer in the clinical setting.

• Journal of Life Science
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• v.28 no.10
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• pp.1201-1211
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• 2018

The study compared the anti-aging, anti-adipogenesis, and anti-tumor effects of epigallocatechin-3- gallate (EGCG) in various cancer cell lines (SNU-601, MKN74, AGS, MCF-7, U87-MG, and A-549) and normal cell lines (MRC-5 fibroblasts, dental tissue-derived mesenchymal stem cells [DSC], and 3T3-L1 pro-adipocytes). Half inhibitory concentration ($IC_{50}$) values were significantly (p<0.05) higher in normal cell lines (~50 uM), when compared to that in cancer cell lines (~10 uM). For anti-aging effects, MRC-5 and DSC were exposed to 10 uM EGCG for up to five passages that did not display any growth arrest. Population doubling time and senescence-related ${\beta}-galactosidase$ ($SA-{\beta}-gal$) activity in treated cells were similar to untreated cells. For anti-adipogenic effects, mouse 3T3-L1 pre-adipocytes were induced to adipocytes in an adipogenic differentiation medium containing 10 uM EGCG, but adipogenesis in 3T3-L1 cells was not inhibited by EGCG treatment. For anti-tumor effects, the cancer cell lines were treated with 10 uM EGCG. PDT was significantly (p<0.05) increased in EGCG-treated SNU-601, AGS, MCF-7, and U87-MG cancer cell lines, except in MKN74 and A-549. The level of telomerase activity and cell migration capacity were significantly (p<0.05) reduced, while $SA-{\beta}-gal$ activity was highly up-regulated in EGCG treated-cancer cell lines, when compared to that in untreated cancer cell lines. Our results have demonstrated that EGCG treatment induces anti-tumor effects more efficiently as noted by decreased cell proliferation, cell migration, telomerase activity, and increased $SA-{\beta}-gal$ activity than inducing anti-aging and anti-adipogenesis. Therefore, EGCG at a specific concentration can be considered for a potential anti-tumor drug.