• Asian-Australasian Journal of Animal Sciences
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• v.23 no.3
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• pp.292-301
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• 2010

Follicle-stimulating hormone (FSH) is a pituitary glycoprotein hormone that is encoded by separate alpha- and betasubunit genes. It plays a key role in stimulating and regulating ovarian follicular development and egg production in chicken. FSH signal transduction is mediated by the FSH receptor (FSHR) that exclusively interacts with the beta-subunit of FSH, but characterization of prokaryotic expression of the FSHb gene and its effect on the expression of the FSHR gene in local chickens have received very little attention. In the current study, the cDNA fragment of the FSHb gene from Dagu chicken was amplified using reverse transcription polymerase chain reaction (RT-PCR), and inserted into the pET-28a (+) vector to construct the pET-28a-FSHb plasmid. After expression of the plasmid in E. coli BL21 (DE3) under inducing conditions, the recombination protein, $FSH{\beta}$ subunit, was purified and injected into the experimental hens and the effect on the mRNA expression levels of the FSHR gene was investigated. Sequence comparison showed that the coding region of the FSHb gene in the local chicken shared 99%-100% homology to published nucleotides in chickens; only one synonymous nucleotide substitution was detected in the region. The encoded amino acids were completely identical with the reported sequence, which confirmed that the sequences of the chicken FSHb gene and the peptides of the $FSH{\beta}$ subunit are highly conserved. This may be due to the critical role of the normal function of the FSHb gene in hormonal specificity and regulation of reproduction. The results of gene expression revealed that a recombinant protein with a molecular weight of about 19 kDa was efficiently expressed and it was identified by Western blotting analysis. After administration of the purified $FSH{\beta}$ protein, significantly higher expression levels were demonstrated in uterus, ovary and oviduct samples (p<0.05). These observations suggested that the expressed $FSH{\beta}$ protein possesses biological activity, and has a potential role in regulation of reproductive physiology in chickens.

• IMMUNE NETWORK
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• v.2 no.3
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• pp.142-149
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• 2002

Background: Our previous study showed that purified protein derivative (PPD)-stimulated pleural mononuclear cells (PMC) from tuberculous pleurisy (Tbp) produced significantly more $IFN-{\gamma}$ (10- to 70-fold) after in vitro PPD stimulation than freshly isolated pleural cells from malignant pleurisy. The present study was designed to determine whether blocking the CD40-CD40 ligand (CD40L) interaction decreases $IFN-{\gamma}$ production by altering IL-12 levels. Methods: IL-12 and $IFN-{\gamma}$ production after neutralizing anti-CD40L antibody treatment was compared to the efficacy of anti-CD80, anti-CD86, and a combination of anti-CD80 and CD86 (CD80+86) monoclonal antibodies (mAb). These activities were measured by enzyme-linked immunosorbent assays (ELISAs) and reverse transcription-polymerase chain reaction (RT-PCR), after in vitro stimulation with PPO antigen (Ag). Results: Neutralization of CD80, CD86 and CD80+86 did not decrease $IFN-{\gamma}$ and IL-12 production in Tbp-PMC, whereas neutralization of CD40L significantly depressed IL-12 p40 and $IFN-{\gamma}$. In addition, neutralization of CD40L completely inhibited IL-12 p40 and $IFN-{\gamma}$ mRNA expression. Conclusion: The CD40-CD40L interaction might play a major role in IL-12 and $IFN-{\gamma}$ production in Tbp-PMC, thus contributing to protective immunity in human tuberculosis.

• IMMUNE NETWORK
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• v.9 no.6
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• pp.236-242
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• 2009

Background: Melanoma is the most fatal form of skin cancer due to its rapid metastasis. Recently, several studies reported that selenium can induce apoptosis in melanoma cells. However, the precise mechanism remains to be elucidated. In this study, we investigated the effect of selenium on cell proliferation in murine melanoma and on tumor growth and metastasis in C57BL/6 mice. Methods: Cell proliferation was measured by MTT assay in selenium-treated melanoma cells. Cell cycle distribution was analysized by staining DNA with propidum iodide (PI). mRNA and protein expression related to cell cycle arrest was measured by reverse transcription PCR and western blot. Tumor growth and metastasis was measured by in vivo model. Results: Selenium was suppressed the proliferation of melanoma cells in a dose dependent manner. The growth inhibition of melanoma by selenium was associated with an arrest of cell cycle distribution at G0/G1 stage. The mRNA and protein level of CDK2/CDK4 was suppressed by treatment with selenium in a time-dependent manner. In vivo, tumor growth was not suppressed by selenium; however tumor metastasis was suppressed by selenium in mouse model. Conclusion: These results suggest that selenium might be a potent agent to inhibit proliferative activity of melanoma cells.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.42 no.4
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• pp.377-385
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• 2016

In order to investigate the possibility of Sesamum indicum L. (S. indicum) extract as an active ingredient for wrinkle-care cosmetics, we prepared 70% ethanolic extract of S. indicum and measured its elastase inhibitory activity and collagenase inhibitory activity. We also evaluated the effect of S. indicum extract on protein and mRNA expression of MMPs in fibroblast cell (CCD-986sk). For anti-wrinkle effects, elastase inhibition activities and collagenase inhibition activities were 37.8% and 45% at a dose of $1,000{\mu}g/mL$ of S. indicum 70% ethanol extract. For a cell viability test, measured on fibroblast cell by ethanol extract of S. indicum, results showed 96% with cell viability at $100{\mu}g/mL$ concentration. According to the results of western blot of ethanol extract from S. indicum the expression of the matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-3 (MMP-3) protein was decreased by 63%, 43%, 49% at $100{\mu}g/mL$ concentration. Reverse transcription-polymerase chain reaction (PCR) of ethanol extract from S. indicum showed that the expression of MMP-1, MMP-2, MMP-3 mRNA was decreased by 82%, 79%, 82% at $100{\mu}g/mL$ concentration. The findings suggest that 70% ethanol extract from S. indicum has potential as a cosmeceutical ingredient with anti-wrinkle effects.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.2
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• pp.174-180
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• 2016

In this study, the effects of fermented goat milk (F-GM) on immunological activity and physical strength were examined. Splenocytes obtained from mice administered with F-GM showed increased responsinveness to mitogens, concanavalin-A (ConA), and lipopolysaccharide (LPS). Treatment with F-GM also significantly augmented production of interleukin (IL)-2 and interferon (IFN)-${\gamma}$, but not IL-4 or IL-10 from ConA-stimulated splenocytes. The activity of F-GM administration to enhance production of IL-2 and IFN-${\gamma}$ was confirmed based on mRNA expression of these cytokines by reverse transcription polymerase chain reaction. After immunization with keyhole limpet hemocyanin (KLH, 20 mg/mouse), mice administered F-GM showed significantly higher antibody titers against KLH than those of phosphate-buffered saline-treated mice, and showed the highest titer 5 weeks after KLH immunization. Analysis for determining isotypes of antibodies revealed that administration of F-GM elicited KLH-specific antibody titers of IgG1, IgG2a, and IgM. In a delayed type hypersensitivity (DTH) test carried out 7 weeks after the primary immunization, F-GM-treated mice showed a higher DTH reaction than the control mice. Furthermore, the swimming test found that administration of F-GM significantly increased swimming time. These results suggest that administration of F-GM enhances not only immune responses against antigens but also physical strength.

• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.185-193
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• 2008

We evaluated the response to salt stress of two different ginseng lines, STG3134 and STG3159, which are sensitive and tolerant, respectively, to salt treatment. Plants were exposed to a 5 dS/m salt solution, and chlorophyll fluorescence was measured. STG3134 ginseng was more sensitive than STG3159 to salt stress. To characterize the cellular response to salt stress in the two different lines, changes in protein expression were investigated using a proteomic approach. Total protein was extracted from detached salt-treated leaves of STG3134 and STG3159 ginseng, and then separated by two-dimensional polyacrylamide gel electrophoresis(2-DE). Approximately 468 protein spots were detected by 2-DE and Coommassie brilliant blue staining. Twenty-two proteins were found to be reproducibly up- or down-regulated in response to salt stress. Among these proteins, twelve were identified using MALDI-TOF MS and ESI-Q-TOF and classified into several functional groups: photosynthesis-related proteins(oxygen-evolving enhancer proteins 1 and 2, rubisco and rubisco activase), detoxification proteins(polyphenol oxidase) and defense proteins($\beta$-1,3-glucanase, ribonuclease-like storage protein, and isoflavone reductase-like protein). The protein levels of ribonuclease-like storage protein, which was highly induced in STG3159 ginseng as compared to STG3134, correlated tightly with mRNA transcript levels, as assessed by reverse-transcription(RT)-PCR. Our results indicate that salinity induces changes in the expression levels of specific proteins in the leaves of ginseng plants. These changes may, in turn, playa role in plant adaptation to saline conditions.

• The korean journal of orthodontics
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• v.39 no.4
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• pp.248-256
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• 2009

Objective: The aim of this study was to determine if human PDL cells can produce osteoclastogenic mRNA and examine how compressive stress affects the expression of osteoclastogenic mRNA in human PDL cells. Methods: Human PDL cells were obtained from biscupids extracted for orthodontic treatment. The compressive force was adjusted by increasing the number of cover glasses. PDL cells were subjected to a compressive force of 0.5, 1.0, 2.0, 3.0 or $4.0\;g/cm^2$ for 0.5, 1.5, 6, 24 or 48 hours. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed to examine levels of M-CSF, IL-$1{\beta}$, RANKL, OPG mRNA expression. Results: Human PDL cells could produce M-CSF mRNA. Human PDL cells under compressive stress showed increased M-CSF, IL-$1{\beta}$ and RANKL mRNAs expression in a force (up to $2\;g/cm^2$) and time-dependent manner. However, OPG mRNA expression was constant regardless of the level and duration of stress. Conclusions: Continuous compressive stress induced the mRNA expression of osteoclastogenic cytokines including M-CSF, RANKL, IL-$1{\beta}$ in PDL cells. Together with an unchanged OPG mRNA level, these results suggest that compressive stress-induced osteoclastogenesis in vivo is partly controlled by M-CSF, RANKL and IL-$1{\beta}$ expression in PDL cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.5
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• pp.664-670
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• 2016

Cheonggukjang (CKJ) is a Korean traditional food made of fermented soybeans. In comparison to normal intake of soybeans, Cheonggukjang has high digestibility with bioactive, antioxidant substances, and thrombolytic enzymes. Recent studies have reported anti-oxidant, anti-cancer, anti-inflammatory, anti-obesity activities as well as inhibitory activities against osteoporosis for CKJ. In this study, we identified the effects of CKJ on osteoblast differentiation by increasing the polyglutamic acid (PGA) content of CKJ. Alkaline phosphatase (ALP) activity and mineralization significantly increased in response to treatment with both natural CKJ (CKJ A) and PGA-increased CKJ (CKJ B). However, CKJ B exhibited higher ALP activity and mineralization than CKJ A. Real-time reverse transcription PCR demonstrated that mRNA expression of osteoblastic-associated genes such as type I collagen, alkaline phosphatase, osteocalcin, and osteopontin in C2C12 cells was significantly up-regulated by CKJ A or B treatment. These results indicate that treatment with CKJ has an anabolic effect on bone by increasing osteoblastic differentiation and ALP activity. Increasing PGA content in CKJ had a greater effect than CKJ A on up-regulation of osteoblastic gene expression in osteoblast cells.

• Korean Journal of Food Science and Technology
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• v.52 no.5
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• pp.441-449
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• 2020

The aim of this study was to compare the biological activities of whole-plant (WAE), bulb (BAE), and leaf (LAE) extracts of Allium macrostemon. The antioxidant activities, total polyphenol contents, and anti-adipogenic activities of WAE and LAE were superior to those of BAE, whereas the biological effects of WAE and LAE were similar. Therefore, the effect of LAE on adipogenesis was further investigated. Treatment of preadipocytes with LAE at 100 g/mL resulted in the inhibition of intracellular lipid accumulation by 49.64%. Consistent with this result, quantitative reverse transcription-PCR showed that LAE treatment decreased the gene expressions of CCAAT/enhancer-binding protein beta (C/EBPβ), peroxisome proliferator-activated receptor gamma (PPARγ), C/EBPα and stearoyl-CoA desaturase 1 (SCD1). Thus, LAE attenuates the adipogenesis of preadipocytes by suppressing the expression of adipogenic and lipogenic genes. These results suggest that LAE can be potentially useful as a functional ingredient to prevent obesity in the food industry.

• Journal of Life Science
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• v.28 no.6
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• pp.733-737
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• 2018

Cucurbitacin-I, a natural triterpenoid derived from Cucurbitaceae family plants, exhibits a number of potentially useful pharmacological and biological activities. Indeed, the previous study demonstrated that cucurbitacin-I reduced the proliferation of colon cancer cells by enhancing apoptosis and causing cell cycle arrest at the G2/M phase. CD44, a type I transmembrane protein with the function of adhering to cells, mediates between the extracellular matrix and other cells through hyaluronic acid. Recent studies have demonstrated that an overexpression of the CD44 membrane receptor results in tumor initiation and growth, specific behaviors of cancer stem cells, the development of drug resistance, and metastasis. The aim was to examine the effect of cucurbitacin-I on CD44 expression human ovarian cancer cells because the effect of cucurbitacin-I on CD44 expression has not been reported. The expressions of CD44 mRNA and protein were detected using a quantitative real-time reverse-transcription polymerase chain reaction and a Western blot analysis, respectively. Treatment with cucurbitacin-I inhibited the expression of CD44 mRNA and protein. A subsequent analysis revealed that cucurbitacin-I blocked the phosphorylation of activator protein-1 (AP-1) and nuclear factor kappa-B ($NF-{\kappa}B$), which are key regulators of CD44 expression. Taken together, the data demonstrate that cucurbitacin-I regulates the AP-1 and $NF-{\kappa}B$ signaling pathways, leading to decreased CD44 expression.