• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.77-86
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• 2011

Alcohol dehydrogenase (E.C.1.1.1.1) is an enzyme present in higher plants involved in the anaerobic fermentation pathway that catalyzes the reduction of pyruvate to ethanol, resulting in continuous $NAD^+$ regeneration. It also plays an important role in many plant developments including tolerance to anoxia condition. Here, a cDNA clone encoding alcohol dehydrogenase (ADH) was isolated from Chinese cabbage (Brassica rapa) seedlings. The gene named Bradh1 had a total length of 1,326 bp that contains a single open reading frame of 1,140 bp. The predicted protein consists of 379 amino acid residues with a calculated molecular mass of 41.17 kDa. Expression pattern analysis revealed a tissue-specific expressing gene in different tissues and strongly expressed in the shoot, roots and seeds of Chinese cabbage. Agrobacterium transformation of full-length cDNA Bradh1 into rice Gopumbyeo showed high efficiency. Furthermore, induction of ADH in transgenic rice enhanced tolerance to anaerobiosis stresses and elevated mRNA transcripts. The overexpression of Bradh1 in rice increases germination under anaerobiosis stresses, implying the possibility of developing new varieties suited for direct seeding or flood-prone rice field.

• BMB Reports
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• v.41 no.8
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• pp.587-592
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• 2008

Cotesia vestalis is an endoparasitoid of Plutella xylostella larvae and injects a polydnavirus (CvBV) into its host during oviposition. In this report we characterize the gene, CvBV3307, and its products. CvBV3307 is located on segment S33 of the CvBV genome, is 517 bp, and encodes a putative protein of 122 amino acids, including a serine-rich region. The expression pattern of CvBV3307 in parasitized larvae and the subcellular localization of CvBV3307 only in granulocytes indicated that it might be involved in early protection of parasitoid eggs from host cellular encapsulation and in manipulating the hormone titer and developmental rhythm of host larvae. Western blot analysis showed that the size of the immunoreactive protein (about 55 kDa) in parasitized hosts at 48 hours post parasitization (h p.p.) is much larger than the predicted molecular weight of 13.6 kDa, which suggests that CvBV3307 undergoes extensive post-translational modification in hosts.

• The Korean Journal of Food And Nutrition
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• v.34 no.5
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• pp.441-448
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• 2021

This study was designed to investigate the intracellular signaling pathways and immunoenhancing effect of macrophage activation by crude polysaccharides (CPP) extracted from citrus peels. CPP did not affect the cytotoxicity of RAW264.7 cells, but showed dose-dependent effects on cell viability. Also, CPP showed high production of chemokine (nitric oxide (NO)) and cytokines (interleukin (IL)-6 and tumor necrosis factor (TNF)-α). CPP increased IL-6, TNF-α, and inducible nitric oxide synthase (iNOS) mRNA expression dose-dependently. CPP also strongly induced the phosphorylation of the ERK, p38, and IκBα pathways in RAW 264.7 cells. In anti-pattern recognition receptors (PRRs) experiments, the effect of CPP on NO production was strongly suppressed by neutralizing toll-like receptor (TLR)2, TLR4, and Dectin1 antibodies, whereas IL-6 and TNF-α production by CPP was mainly suppressed by mannose receptor (MR). Therefore, these results suggest that CPP treatment-induced NO production was regulated by the ERK, p38, and NF-κB pathways through TLR2, TLR4, and Dectin1 receptors, whereas IL-6 and TNF-α production was primarily regulated by the ERK, p38, and NF-κB pathways through MR receptors.

• Journal of dental hygiene science
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• v.23 no.4
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• pp.264-270
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• 2023

Background: Resin-based dental materials release residual monomers or other substances from incomplete polymerization into the oral cavity, thereby causing adverse biological effects on oral tissue. 10-Methacryloyloxydecyl dihydrogen phosphate (10-MDP), an acidic monomer containing dihydrogen phosphate and methacrylate groups, is the most commonly used component of resin-based dental materials, such as restorative composite resins, dentin adhesives, and resin cements. Although previous studies have reported the cytotoxicity and biocompatibility in various cultured cells, the effects of resin monomers on cellular aging have not been reported to date. Therefore, this study aimed to investigate the effects of the resin monomer 10-MDP on cellular senescence and inflamm-aging in vitro. Methods: After stimulation with 10-MDP, MC3T3-E1 osteoblast-like cells were examined for cell viability by WST-8 assay and reactive oxygen species (ROS) production by flow cytometry. The protein and mRNA levels of molecular markers of aging were determined by western blotting and RT-PCR analysis, respectively. Results: Treatment with 0.05 to 1 mM 10-MDP for 24 hours reduced the survival of MC3T3-E1 cells in a concentration-dependent manner. The intracellular ROS levels in the 10-MDP-treated experimental group were significantly higher than those in the control group. 10-MDP at a concentration of 0.1 mM increased p53, p16, and p21 protein levels. Additionally, an aging pattern was observed with blue staining due to intracellular senescence-associated beta-galactosidase activity. Treatment with 10-MDP increased the levels of tumor necrosis factor-α, interleukin (IL)-1β, IL-6 and IL-8, however their expression was decreased by mitogen-activated-protein-kinase (MAPK) inhibitors. Conclusion: Taken together, these results suggest that the exposure of osteoblast-like cells to the dental resin monomer 10-MDP, increases the level of cellular senescence and the inflammatory response is mediated by the MAPK pathway.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.3
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• pp.385-392
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• 2011

Although monoculture methods have been remarkably useful due to their simplicity, they have serious limitation because of the different types of cells communication with each other in many physiological situations. We demonstrated levels of markers of endothelial dysfunction such as tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and interleukin-1$\beta$ (IL-1$\beta$) as well as stimulation of receptor of advanced glycation endproducts (AGEs) on monoand co-culture system such as only monocyte (THP-1) cultivation system, only endothelial cell (HUVEC) cultivation system, and co-cultivation system of THP-1 and HUVEC. The mRNA levels of TNF-$\alpha$ and IL-1$\beta$ on HUVEC increased by the co-culture with monocyte after 4 hr at 100 ${\mu}g/mL$ glyceraldehyde-AGE. The secreted protein contents into medium of TNF-$\alpha$ and IL-1$\beta$ increased after 8 hr approximately 2~2.5 times compared to mono-cultivation. In contrast, the mRNA level of receptor of AGE (RAGE) was relatively insensitive on the co-culture system. The mediators by which monocytes activate endothelial cell have not been fully elucidated. In this study we confirmed production of soluble cytokines such as TNF-$\alpha$ and IL-1$\beta$ by monocytes. Use of monocyte conditioned medium, which contains both cytokines, can activate endothelial cell.

• IMMUNE NETWORK
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• v.1 no.3
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• pp.230-235
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• 2001

Background: Finding of the regulation of various gene expression by cytokine including $IFN-{\gamma}$ in hematopoietic stem cell will light up the understanding of pathogenesis of aplastic anemia in various aspects. To study on aplastic anemia, however, we have to circumvent the difficulty of directly obtaining bone marrow stem cells from the patient. Therefore, we tried to find out a cell can replace the bone marrow stem cells for study on cell signaling pathway and regulation of gene expression by $IFN-{\gamma}$. Materials and Methods: HL-60 cells, of 20 ng/mL of $IFN-{\gamma}$. Total RNA was isolated from the cells and RT-PCR of the indoleamine 2,3-dioxygenase (IDO), $IFN-{\gamma}$, TNF-${\alpha}$, $MIP-1{\alpha}$, and $TGF-{\beta}2$ was carried out for the estimation of the gene expression. Results: $IFN-{\gamma}$ induced IDO gene expression of mononuclear cells from umbilical cord blood showed similar pattern as compared to that of bone marrow. Whether $INF-{\gamma}$ was treated or not, $TNF-{\alpha}$ was expressed in both mononuclear cells from umbilical cord blood and bone marrow. However, HL-60 cells showed different expression patterns. HL-60 cells would express neither IDO nor $TNF-{\alpha}$ even under the culture with 20ng/mL of $IFN-{\gamma}$. Conclusion: Our results showed bone marrow can be replaced with mononuclear cells from umbilical cord blood in the study on the relation between aplastic anemia and $IFN-{\gamma}$ including $IFN-{\gamma}$ cell signaling pathway.

• The Journal of Internal Korean Medicine
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• v.25 no.4
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• pp.289-299
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• 2004

Background : Although the pathophysiology of asthma has been reported, its mechanism has not been fully elucidated. The mast cell is an effector cells in allergic inflammation and secretes a number of chemokines. Chemokines are important for the recruitment of leukocytes to sites of infection, which is essential in host defense. Chemokines also contribute to the pathogenesis of several disorders such as asthma, chronic bronchitis, atopic dermatitis, allergic rhinitis, and rheumatoid arthritis. Objective : In this study, the aim was to identify the effect of Baekryunchihyo-tang(白蓮治哮湯) on expression of chemokines. This was examined by RT-PCR using the human mast cell line (HMC-l) Materials and Methods : HMC-l cells were used, which is known to secrete and express chemokines. In order to investigate the protective effect of Baekryunchihyo-tang(白蓮治哮湯), HMC-l cells were incubated with pretreatment of Baekryunchihyo-tang(白蓮治哮湯) for 24 hrs. RT-PCR analyses of chemokine genes of cells pretreated with Baekryunchihyo-tang(白蓮治哮湯) showed that expressions of IL-8, $MIP-l{\beta}$, and RANTES genes in these cells were lower and $MIP-l{\alpha}$ showed a similar pattern compared to the calcium ionophore-treated group. In addition, cell cytotoxicity concentration measurements were performed by MTT assay method. Results : After stimulation with 1 uM calcium ionophore A23178 for 2 hrs, IL-8, major one of CXC chemokines, was highly expressed, and expression of $MIP-l{\beta}$ and RANTES (CC chemokines) increased, while expression of $MIP-l{\alpha}$ did not change. The cell cytotoxicity of Baekryunchihyo-tang(白蓮治哮湯) with treatments at various concentrations and times was not observed, respectively. Conclusion : This study suggests that Baekryunchihyo-tang(白蓮治哮湯) has dose-dependent effects on mRNA expression of IL-8(CXC chemokines), $MIP-l{\beta}$ and RANTES(CC chemokines) in human mast cellline(HMC-l). So these herbal medicines may inhibit the inflammatory process of asthma. Advanced studies are required to investigate the mechanism of inhibition by herbal medicine in the asthma model. This study provides basic data on the possibility of the clinical treatment of Baekryunchihyo-tang(白蓮治哮湯) for allergic disorders.

• Development and Reproduction
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• v.13 no.4
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• pp.249-256
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• 2009

The estrogen receptor-related receptors (ERRs) are a group of nuclear receptor superfamily of transcription factors. ERRs and estrogen receptors (ERs) have overlapping affinities for coactivators and DNA binding sites, but differ markedly in ligand binding and activation. The three mammalian ERR genes have been implicated in diverse physiological processes ranging from placental development to maintenance of bone density, whereas the molecular diversity, function, and regulation of ERRs in non-mammalian species are not well understood. In the present study, to investigate the involvement of ERR in transcription and embryogenesis in marine invertebrates, a cDNA encoding ERR (SnERR) was cloned from the gonad in Strongylocentrotus nudus, by polymerase chain reaction (PCR). The amino acid sequence of SnERR showed high homology with that of S. purpuratus (91%). A phylogenetic tree clearly showed that SnERR is a member of the ERR family and clustered in echinodermata group as supported by a high bootstrap value. We examined gene expression of SnERR during embryonic development of S. nudus using real-time PCR. During the embryonic development, the mRNA of ERR was significantly high levels in early development stages (4~64 cell) and larval stages. The SnERR slightly activated transcription through the classical estrogen response elements (EREs) in the presence of genistein. In addition, peroxisome proliferator-activated receptor $\gamma$ coactivator (PGC)-$1\alpha$ knwon as a coactivator of ERR enhanced the snERR-mediated transactivation, suggesting that the PGC-$1\alpha$ is a coactivator of SnERR.

• Development and Reproduction
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• v.11 no.2
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• pp.85-96
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• 2007

Mesenchymal stem cells (MSC) are promising candidates for cell-based therapies. One major obstacle for their clinical use is the unsafety of fetal bovine serum (FBS), which is a crucial part of all media currently used for the culture of MSC. We investigated the effect of human cord serum (HCS) on the growth response, mRNA and protein expressions of human amnion-derived stem cells (HAM). HAM were isolated from the amnion after a Caesarean section and cultured in DMEM supplemented with 10% FBS, 5% HCS or 10% HCS. During culture, their biological characteristics at earlier and later passages were analyzed using RT-PCR and immunocytochemistry. Regardless of serum sources, HAM showed the prominent expression of Oct-4, Rex-1, SCF, FGF-5, BMP-4, nestin, GATA-4, NCAM and HLA ABC genes. The expression profile was observed even at later passages. Similarly, HAM cultured in either FBS or HCS exhibited the distinct protein expression of collagen I, II, III and XII, fibronectin, $\alpha$-smooth muscle actin, vimentin, CK18, CD54, FSP, TRA-1-60, SSEA-3, -4 and HLA ABC. However, desmin expression was only observed in HAM cultured in medium supplemented with FBS and vWF expression was only found in HAM cultured in medium supplemented with HCS. Overall pattern of gene and protein expression of HAM was typical of known adult stem cells such as bone marrow-derived MSC. In conclusion, HCS could be as effective as FBS for the culture of HAM.

• BMB Reports
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• v.38 no.6
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• pp.668-675
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• 2005

A full-length cDNA encoding taxadiene synthase (designated as TmTXS), which catalyzes the first committed step in the Taxol biosynthetic pathway, was isolated from young leaves of Taxus media by rapid amplification of cDNA ends (RACE). The full-length cDNA of TmTXS had a 2586 bp open reading frame (ORF) encoding a protein of 862 amino acid residues. The deduced protein had isoelectric point (pI) of 5.32 and a calculated molecular weight of about 98 kDa, similar to previously cloned diterpene cyclases from other Taxus species such as T. brevifolia and T. chinenisis. Sequence comparison analysis showed that TmTXS had high similarity with other members of terpene synthase family of plant origin. Tissue expression pattern analysis revealed that TmTXS expressed strongly in leaves, weak in stems and no expression could be detected in fruits. This is the first report on the mRNA expression profile of genes encoding key enzymes involved in Taxol biosynthetic pathway in different tissues of Taxus plants. Phylogenetic tree analysis showed that TmTXS had closest relationship with taxadiene synthase from T. baccata followed by those from T. chinenisis and T. brevifolia. Expression profiles revealed by RT-PCR under different chemical elicitor treatments such as methyl jasmonate (MJ), silver nitrate (SN) and ammonium ceric sulphate (ACS) were also compared for the first time, and the results revealed that expression of TmTXS was all induced by the tested three treatments and the induction effect by MJ was the strongest, implying that TmTXS was high elicitor responsive.