• Korean Journal of Soil Science and Fertilizer
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• v.38 no.5
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• pp.287-293
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• 2005

In this work, to develop soil inoculant which maintains stable viable cells and normalized quality, studies on micro-encapsulation with bacteria and yeast cells were performed by investigating materials and methods for micro-encapsulation as well as variation and stability of encapsulated cells. Preparation of capsule was conducted by application of extrusion system using micro-nozzle and peristaltic pump. K-carragenan and Na-alginate were selected as best carrier for gelation among K-carageenan, Na-alginate, locust bean gum, cellulose acetate phthalate (CAP), chitosan and gelatin tested. Comparing the gels prepared with Bacillus sp. KSIA-9 and carriers of 1.5% concentration, although viable cell of K-carragenan and Na-alginate was six times higher than those of other, Na-alginate was finally selected as carrier for gelation because it is seven times cheaper than K-carragenan. The gel of 1.5% Na-alginate was also observed to have the best morphology with circular hardness polymatrix and highest viable cell. When investigating the stability of encapsulated cells and the stabilizer effect, free cells were almost dead within 30 or 40 days whereas encapsulated cells decreased in 10% after 30 days and 15-30% even after 120 days. As stabilizer for maintaining viable cell, both 1% starch and zeolite appeared to possess the level of 70-80% cell for bacteria and yeast until after 120 days.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.4
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• pp.297-306
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• 2020

The aims of the present study were to confirm that regulation of the PA and environment via TGF-β regulation of sperm by Percoll-separated in porcine uterine epithelial cells. And, it was performed to identify the cytokines (TGF-β1, 2 and 3, TGF-β receptor1 and 2; interleukin, IL-6, IL-8) and PA-related genes (urokinase-PA, uPA; tissue-PA, tPA; PA inhibitor, PAI; uPA-receptor, uPAR) by spermatozoa. The experiment used porcine uterus epithelial cells (pUECs) and uterine tissue epithelial cells, Boar sperm were separated by discontinuous Percoll density gradient (45/90%), and tissues were co-incubated with spermatozoa, followed by real-time PCR. PA activity was measured of sperm by discontinuous Percoll density gradient (45/90%) for 24 hours. To measure viability and acrosome damage of sperm double stained propidium iodide (PI) and SYBR-14 or FITC-PNA were used. In results, binding ratio of Percoll-separated sperm was found no differences, but sperms isolated from 90% Percoll layer reduced PA activity (p < 0.05). when co-cultured sperm selected Percoll in porcine uterus tissues epithelial cells, 90% layer sperm increased TGF-β R1, contrastively tPA and PAI-1 in comparison with control (p < 0.05). 45% sperm was decreased the expression of uPA (p < 0.05). TGF-β decreased PA activity in the supernatant collected from pUECs (p < 0.05). Especially, The group including uPA, PAI-1 were induce sperm intact, while it was reduced in sperm damage when compared to control (p < 0.05). Also, there was no significant difference group of tPA and tPA+I in the dead sperm and acrosome damage compared to control. The expression of tPA and PAI showed a common response. Percoll-separated spermatozoa in 90% layer reduced tPA and IL-related gene mRNA expression. Thus, Percoll-sparated sperm in 90% layer show that it can suppress inflammation through increased expression of TGF-β and downregulation of PA and IL in epithelial cells compared to 45% layer Percoll.

• Toxicological Research
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• v.15 no.1
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• pp.79-87
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• 1999

To know the stress response and antioxidative effect of sulfur containing compounds, we observed the expression of the stress protein (heat shock protein; inducible protein) from mouse tissues and evaluated the protective effects to hydroxyl radical in mouse brain cell culture. Cysteine, methionine or sodium sulfide was fed by oral administration of 1 ml/per 6hr/three times with 1 mM, 2mM or 3mM to mouse, respectively. After that, the stress proteins were extracted from mouse tissues and analyzed the features of expression. The stress proteins by sulfur containing compounds were showed different aspects in the kinds and concentrations of their compounds, and in the tissues of mouse. In the liver, the stress proteins were appeared at different time on the concentration of sulfur containing compounds and had less than 20 KDa as small molecules. In general, the molecular weights of stress protein in liver, the stress proteins were appeared at different time on the concentration of sulfur containing compounds and had less than 20 KDa as small molecules. In general, the molecular weights of stress protein in the spleen were evaluated from 32KDa to 50KDA, and the induced times were relatively late at high concentration of cysteine, early at low concentration of methionine or sodium sulfide. The stress proteins in mouse muscle were detected mostly between 24hr after treatment of sulfur containing compounds. Their molecular weights were 15~24KDa. In the antioxidative effects of sulfur containing compounds to hydroxyl radical, cell viabilities were measured by 63.2% at 10 $\mu\textrm{M}$, 65.5% at 50 $\mu\textrm{M}$, 68.6% at 100 $\mu\textrm{M}$, 78.3% at 150 $\mu\textrm{M}$, or 83.0% at 200 $\mu\textrm{M}$ of cysteine, respectively. At addition of methionine, the cell viabilities were assessed as 58.1% at 10 $\mu\textrm{M}$, 62.8% at 50 $\mu\textrm{M}$, 75.7% at 100 $\mu\textrm{M}$, 78.6% at 150 $\mu\textrm{M}$, and 79.2% at 200 $\mu\textrm{M}$ after 4hrs exposure with 20mU/ml glucose oxidase (GO) system, while the numbers of live cells to hydroxyl radicals in treatment of sodium sulfide were showed 48.6% at 10 $\mu\textrm{M}$, 54.8% at 100 $\mu\textrm{M}$, 51.8% at 150 $\mu\textrm{M}$, and 51.6% at 200 $\mu\textrm{M}$ in the neuronal cells. In the inhibitory effects on the proliferation of tumor cells, percentages of dead cells of the CT-26 or HeLa cell were generally less than 30% even 48hr after addition of sulfur containing compounds. Conclusively, the results of these experiments indicate that stress protein by sulfur containing compounds can be used as physiological indicator for animal nutrition and for environment, and also that cysteine and methionine can play critical roles as an antioxidant.

• Journal of Life Science
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• v.22 no.11
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• pp.1500-1506
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• 2012

Transforming growth factor (TGF)-${\beta}$-dependent apoptosis is important in the elimination of damaged or abnormal cells from normal tissues, especially in liver, in vivo. To investigate which gene expressions are critical for TGF-${\beta}$-induced apoptosis in hepatocytes, gene expression profiling experiments were performed with TGF-${\beta}$-treated and non-treated mouse hepatocytes AML12 cells. Findings showed that serum and glucocorticoid-inducible protein kinase1 (SGK1) expression is markedly downregulated during TGF-${\beta}$-induced apoptosis. Findings confirmed that expression of SGK1 protein, as well as mRNA, is also markedly decreased with TGF-${\beta}$ treatment. Infection of adenoviral vector encoding constitutively active SGK1 (CA-SGK1), but not kinase dead SGK1 (KD-SGK1), attenuated TGF-${\beta}$-induced apoptosis. All of these results suggest that downregulation of SGK1 expression is critical for TGF-${\beta}$-induced apoptosis in AML12 cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.12
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• pp.1827-1834
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• 2014

Colon cancer is the third highest cause of death in Korea. Known dietary causes of colon cancer include a diet rich in fat and red meat as well as inadequate intake of dietary fiber, fruits, and vegetables. Therefore, recent research has focused on the anticancer effects of natural products. Opuntia humifusa is a type of prickly pear that is known to contain biologically active compounds that can be used in the treatment of diabetes mellitus, arteriosclerosis, and hyperglycemia. The aim of this study was to determine whether or not O. humifusa extract affects proliferation, cell death, and DNA fragmentation in human carcinoma HT-29 cells. O. humifusa is rich in carbohydrates, minerals (Mg, K, and Ca), and total phenolics. HT-29 cells were treated with extracts of O. humifusa at concentrations of 0, 0.25, 0.5, 1, and 2 mg/mL for 24 or 48 hours. O. humifusa extracts inhibited HT-29 cell growth in a dose-dependent manner. Hoechst 33342/PI double staining and Comet assay were performed to observe changes in nuclei of cancer cells undergoing cell death. The results of both tests showed that O. humifusa extract induced cell shrinkage, DNA fragmentation, and chromatin condensation dose-dependently in HT-29 cells. The results of this study suggest that O. humifusa extract inhibits the growth of HT-29 via induction of DNA fragmentation and chromatin condensation.

• Korean Journal of Food Science and Technology
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• v.38 no.2
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• pp.290-294
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• 2006

Inactivation rates of Salmonella enteritidis in vitro and in vivo were assessed using confocal microscopy and flow cytometry. S. enteritidis was inactivated with 1% (w/v) trisodium phosphate (TSP) and live cells, and inactive cells were distinguished by staining with fluorescent probe, LIVE/DEAD BacLight Bacteria Viability stain. After TSP treatment for 1 min, most of Salmonella cells changed from green (live cells) fluorescence to red (inactive cells) fluorescence, indication of effective sanitizing. Inactivation efficiency and contamination sites of S. enteritidis on chicken skin by TSP treatment were assessed using confocal laser microscopy. Precise flow cytometry histograms for viability changes of S. enteritidis. after TSP treatments were obtained. Efficiency of various sanitizer treatments on foodborne pathogens could be assessed using this method.

• Journal of Dairy Science and Biotechnology
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• v.33 no.3
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• pp.197-202
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• 2015

While various foodborne pathogenic bacteria can be detected more rapidly via polymerase chain reaction than via conventional plating methods, it is impossible to distinguish between viable and dead cells in DNA-based assays. Hence, propidium monoazide (PMA) treatment has been introduced to detect living cells. The purpose of this study is to evaluate the applicability of the PMA treatment and real-time qPCR method for the detection of Cronobacter sakazakii and to compare it to that of plate counting. Based on our positive results, we suggest the use of PMA treatment and real-time qPCR for the detection of viable Cronobacter sakazakii in various food sources and an update of the Korean Food Code.

• Journal of Microbiology and Biotechnology
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• v.28 no.2
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• pp.218-226
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• 2018

Nanometric Lactobacillus plantarum nF1 (nLp-nF1) is a biogenics consisting of dead L. plantarum cells pretreated with heat and a nanodispersion process. In this study, we investigated the immune-enhancing effects of nLp-nF1 in vivo and in vitro. To evaluate the immunostimulatory effects of nLp-nF1, mice immunosuppressed by cyclophosphamide (CPP) treatment were administered with nLp-nF1. As expected, CPP restricted the immune response of mice, whereas oral administration of nLp-nF1 significantly increased the total IgG in the serum, and cytokine production (interleukin-12 (IL-12) and tumor necrosis factor alpha (TNF-${\alpha}$)) in bone marrow cells. Furthermore, nLp-nF1 enhanced the production of splenic cytokines such as IL-12, TNF-${\alpha}$, and interferon gamma (IFN-${\gamma}$). In vitro, nLp-nF1 stimulated the immune response by enhancing the production of cytokines such as IL-12, TNF-${\alpha}$, and IFN-${\gamma}$. Moreover, nLp-nF1 given a food additive enhanced the immune responses when combined with various food materials in vitro. These results suggest that nLp-nF1 could be used to strengthen the immune system and recover normal immunity in people with a weak immune system, such as children, the elderly, and patients.

• Food Science and Preservation
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• v.5 no.1
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• pp.89-96
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• 1998

The antibacterial activity of clove (Eugenia caryophyllata Thumb) in culture troth against S. aureus was tested at 35, 5, -20, 50, 55 and 60$^{\circ}C$. Tryptic soy broth(TSB) containing 0∼0.5%(w/v) of clove was inoculated with 105∼107 CFU/ml of S. aureus and incubated at each temperature. The growth of S. aureus occured at 0.1% clove after a prolonged lag period while viable cells of S. aureus decreased more than 2 log cycles at 0.3 and 0.4% clove during 12 hours storage at 35$^{\circ}C$. During 32 days of refrigerated storage at 5$^{\circ}C$, survivors of S. aureus were decreased with the progress of time and increasing clove concentration. At the presence of 0.2, 0.3 and 0.4% clove, bacterial cells were dead after 32, 20 and 16 days of refrigerated storage, respectively. During 32 days of frozen storage at -20$^{\circ}C$, survivors of S. aureus were decreased less than 0.5 log cycle at 0% clove. At the presence of 0.1∼0.4% clove, survivors were decreased 2.5∼3.0 log cycles after 1 day and then decreased 0.4∼0.8 log cycles through the frozen storage. There were small changes in populations of S. aureus in TSB between different concentrations of clove during frozen storage. The D-values of S. aureus at clove 0, 0,2, 0.4% were 28.53, 15.14, 8.9 min at 50$^{\circ}C$, 18.43, 10.32, 6.74 min at 55$^{\circ}C$ and 12.78, 9.88, 5.72 min at 60$^{\circ}C$, respectively. The D-values for S. aureus were decreased with the increasing temperature and clove concentration.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.564-570
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• 2005

In the present study, the protective effects of Bcl-2 over-expression in a suspension culture (without any adaptation) and spent medium (low nutrient and high toxic metabolite conditions) were investigated. In the suspension culture without prior adaptation, the viability of the control cell line fall to 0% by day 7, whereas the Bcl-2 cell line had a viability of 65%. The difference in the viability and viable cell density between the Bcl-2 and control cell lines was more apparent in the suspension culture than the static culture, and became even more apparent on day 6. Fluorescence microscopic counting revealed that the major mechanism of cell death in the control cell line in both the static and suspension cultures was apoptosis. For the Bcl-2 cell lines, necrosis was the major mode of cell death in the static culture, but apoptosis became equally important in the suspension culture. When the NS0 6A1 cell line was cultured in spent medium taken from a 14 day batch culture, the control cell line almost completely lost its viability by day 5, whereas, the Bcl-2 still had a viability of 73%. The viable cell density and viability of the Bcl-2 cell line cultivated in fresh medium were 2.2 and 2.7 fold higher, respectively, than those of the control cultures. However, the viable cell density and viability of the Bcl-2 cultivated in the spent medium were 8.7 and 7.8 fold higher, respectively, than those of the control cultures. Most of the dead cells in the control cell line were apoptotic; whereas, the major cell death mechanisms in the Bcl-2 cell line were necrotic.