• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.2
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• pp.181-191
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• 2006

NFI-C null mice demonstrated aberrant odontoblast differentiation and thus abnormal dentin formation while other tissues/organs in the body, including ameloblasts, appear to be unaffected and normal. However little is known about the mechanism of NFI-C function in odontoblast differentiation and dentin formation. Odontoblasts are tall, highly polarized cells that are responsible for formation and maintenance of the predentin and dentin. An indication of their polarity is the acquisition of specialized intercellular junctions. As preodontoblasts differentiate into odontoblasts, they are Joined and attached at the apical end by well developed terminal webs of cytoskeletal actins, and associated tight as well as adherent njunctions. In this study, in order to investigate if disruption of the NFI-C gene interferes with formation of a specific or other structural proteins of the intercellular junctions, we examined morphological characteristic of the aberrant odontoblast in NFI-C null mice using light and electron microscope. In addition, we determined the expression of major structural proteins of intercellular junctions, ZO-1 and occludin, during the differentiation of odontoblasts using immunohitochemistry. The results were as follows : 1. In light microscopy, abnormal odontoblasts of incisors of the NFI-C null mice were round in shape, lost their polarity, and trapped in osteodentin-like mineralized tissue. Mutant molars have relatively normal crowns, but short and abnormal differentiating adontoblasts in root formation area. 2. Electron microscopy of abnormal odontoblasts revealed the dissociation of the round osteoblast-like cells, the loss of their cellular polarity, and the absence of an intercellular junctional complex known as the tight junctions. 3. A mutant incisor showed labeling for ZO-1 at the proximal and distal ends of secreting ameloblasts, while staining for ZO-1 was not observed in the abnormal odontoblasts. 4. A normal incisor showed immunoreactivity for occludin in the differentiating odontoblasts. However, staining for occludin was not observed in the abnormal odontoblasts of mutant incisor. These results suggest that NFI-C gene causes dissociation of odontoblast and thus abberant odontoblast differentiation and abnormal dentin formation by interfering with the formation of intercellular junctions.

• Journal of Life Science
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• v.24 no.9
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• pp.967-972
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• 2014

In the present study, we prepared eighty-five different kinds of lees extracts and their solvent fractions and investigated their anti-proliferative activities against human colorectal cancer HCT116 cells. HCT116 cells were treated with eighty-five solvent fractions of lees extracts and then cell viability was measured using MTS assay. Among the treated solvent fractions, three solvent fractions (KSD-E1-3, KSD-E2-3, and KSD-E4-3) were selected based on cell viability assay. In addition, we performed an oligo DNA microarray analysis to analyze the gene expression changes by treatment of KSD-E1-3 in HCT116 cells. Among the upregulated genes, we selected 4 genes (NAG-1, ATF3, p21, and DDIT3) and performed RT-PCR using gene-specific primers. Among the treated solvent fractions, KSD-E1-3 dramatically induced the expressions of the four selected genes. In addition, we investigated whether the upregulations of those genes were dependent on the transcription factor p53's presence using p53 null HCT116 cells. The results indicate that the upregulations of NAG-1, ATF3, and DDIT3 are not dependent on the p53 presence, whereas p21 is dependent on the p53 presence. These findings may help to understand the molecular mechanisms of the anti-proliferative activity mediated by rice wine lees in human colorectal cancer cells.

• Food Science and Preservation
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• v.18 no.3
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• pp.366-373
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• 2011

Recent studies suggested that Cheonnyuncho is a significant source of bioactive phenolic compounds, comparable to phytochemicals, including green tea and onion. In this study, the hot-water and 80% ethanolic extracts of Cheonnyuncho were assessed as to their total phenol content, total flavonoids content, antioxidant activity (DPPH radical-scavenging activity and reducing power), and anti-obesity activity. The results showed that the total phenol contents of the hot water extract and the 80% ethanolic extract were $16.52{\pm}3.87$ and $13.44{\pm}0.85$ mg GAE/g, respectively. The total flavonoids content was detected only in the 80% ethanolic extract, however, with a 778.08 ${\mu}g$ catechin equivalents/g content. The DPPH radical-scavenging activity and reducing power of the 80% ethanolic extract from Cheonnyuncho was significantly higher than those of the water extract (p < 0.05). During the adipocyte differentiation, the 80% ethanolic extract of Cheonnyuncho more significantly inhibited lipid accumulation and ROS production than the 3T3-L1 cells that were treated with hot water extract. Furthermore, the 80% ethanolic extract of Cheonnyuncho suppressed the mRNA abundance of the adipogenic transcription factor, $PPAR{\gamma}$ (peroxisome proliferator-activated receptor ${\gamma}$), and its target gene, aP2 (adipocyte protein 2). These results indicate that Cheonnyuncho extracts can inhibit adipogenesis through a mechanism that involves direct down regulation of $PPAR{\gamma}$ gene expression or via modulation of ROS production associated with radical-scavenging activities.

• Journal of Life Science
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• v.31 no.3
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• pp.356-365
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• 2021

Recently, we sequenced the entire genome of a freshwater agar-degrading bacterium Cellvibrio sp. KY-GH-1 (KCTC13629BP) to explore genetic information encoding agarases that hydrolyze agarose into monomers 3,6-anhydro-L-galactose (L-AHG) and D-galactose. The KY-GH-1 strain appeared to possess nine β-agarase genes and two α-neoagarobiose hydrolase (α-NABH) genes in a 77-kb agarase gene cluster. Based on these genetic information, the KY-GH-1 strain-caused agarose degradation into L-AHG and D-galactose was predicted to be initiated by both endolytic GH16 and GH86 β-agarases to generate NAOS (NA4/NA6/NA8), and further processed by exolytic GH50 β-agarases to generate NA2, and then terminated by GH117 α-NABHs which degrade NA2 into L-AHG and D-galactose. More recently, by employing E. coli expression system with pET-30a vector we obtained three recombinant His-tagged GH50 family β-agarases (GH50A, GH50B, and GH50C) derived from Cellvibrio sp. KY-GH-1 to compare their enzymatic properties. GH50A β-agarase turned out to have the highest exolytic β-agarase activity among the three GH50 isozymes, catalyzing efficient NA2 production from the substrate (agarose, NAOS or AOS). Additionally, we determined that GH117A α-NABH, but not GH117B α-NABH, could potently degrade NA2 into L-AHG and D-galactose. Sequentially, we examined the enzymatic characteristics of GH50A β-agarase and GH117A α-NABH, and assessed their efficiency for NA2 production from agarose and for production of L-AHG and D-galactose from NA2, respectively. In this review, we describe the benefits of recombinant GH50A β-agarase and GH117A α-NABH originated from Cellvibrio sp. KY-GH-1, which may be useful for the enzymatic hydrolysis of agarose for mass production of L-AHG and D-galactose.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.26 no.1
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• pp.32-39
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• 1981

In order to study the genetic system of the blast resistant varieties, the conidial suspension of mutant races of T-2$^{+t}$, N-2$^{+t}$, C-8$^{+t}$ was inoculated at 4-5 leaf stage by injector for F_2 seedlings from the crosses between seven resistant varieties and four maker lines easily detectable at seedling stage. The results are summarized as follows; 1. The fertility of cross between Semi-dwaf testers and Indica resistant varieties except Carreon was about 74 percents. 2. The segregation modes of resistance varied with varieties and blast races. However, the resistance was expressed as dominance in all cases. Tetep, Tadukan and Carreon showed more complicated segregation for resistance than that of the bred lines. 3. For blast races used, four segregation ratios such as 3:1, 9:7, 13:3 and 37:27 were found in the Tatukan, Tetep, and IR747, and three segregation ratios such as 3:1, 13:3 and 15:1 in the Carreon, and two segregation ratios of 3:1 and 13:3 with Suweon 287, Suweon 288, and Iri342. 4. In the segregation of the resistance to the each races, the ratios of 3:1, 13:3, 15:1 were fitted to T-2$^{+t}$, and the ratios of 3:1, 13:3, 9:7 and 37:27 to N-2$^{+t}$ and C-8$^{+t}$. 5. Suweon 287, Suweon 288 and Iri342 carried one simple dominant gene and inhibitor gene was considered in some cross combinations. Meanwhile Tadukan, Tetep and IR747 seemed to carry one to three resistant genes, and in some cross combinations, the expression of these genes were simple dominant, inhibiting, duplicating and complimentary action. 6. Resistance genes to blast races, T-2$^{+t}$, N-2$^{+t}$ and C-8$^{+t}$ in the Tadukan, Tetep, Carreon, Suweon 287, Suweon 288 and Iri342 were found to be independent with the linkage group of II(lg), VIII(la), XI(bc), and XII(gl).bc), and XII(gl).

• Journal of Life Science
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• v.20 no.12
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• pp.1772-1776
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• 2010

Transcription factors Nrf2 and NF-${\kappa}B$ are important regulators of the innate immune response, and their cross-talks in inflammation have been reported. Previously, we demonstrated that gold(I)-compound auranofin, an inhibitor of NF-${\kappa}B$ signal, induced Nrf2 activation in human synovial cells and monocytic cells. To investigate whether the Nrf2 activation is involved in the mechanism of the auranofin-attenuated NF-${\kappa}B$ signaling, we examined the effects of Nrf2 knockdown on NF-${\kappa}B$ activation using rheumatic synovial cells. When the cells were transfected with a specific siRNA for Nrf2, the gene expression was perfectly blocked. However, the Nrf2 knockdown did not cancel the suppressive effect of auranofin on TNF-$\alpha$-induced $I{\kappa}B-{\alpha}$ degradation. Treatment with a specific siRNA for HO-1, which is a target of Nrf2 and plays a role in anti-inflammation, also did not affect the blocking activity of auranofin on $I{\kappa}B-{\alpha}$ degradation. In addition, auranofin-inhibited ICAM-1 expression was not restored by Nrf2 knockdown. These findings indicate that the activated Nrf2 and HO-1 are not associated with the suppressive action of auranofin on the pro-inflammatory cytokines-stimulated NF-${\kappa}B$ activation. This suggests that Nrf2/HO-1 and NF-${\kappa}B$ signals, which are regulated by auranofin, participate in the anti-inflammatory action of auranofin via independent pathways in rheumatic synovial cells.

• Journal of Preventive Medicine and Public Health
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• v.37 no.2
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• pp.157-165
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• 2004

Objectives : This study aimed to investigate the toxic effects of chromium (VI) on the placental function and reproduction in rats. For the study, the placental prolactin-growth hormone (PRL-GH) gene expression, placental trophoblast cell differentiation and reproductive data were analyzed. Methods : The pregnancies of F344 Fisher rats were checked by the presence of a copulatory plug or sperm in the vaginal smear, which was defined as day 0 of the pregnancy. Pregnant rats were divided into the three groups. The control group was given tap water (chromium level < 0.001 ppm) and the remaining groups were given 250 or 750 ppm of chromium (VI) [as potassium dichromate], from day 7 to 19 of the pregnancy. Rats were sacrificed at days 11 and 20 of pregnancy. The mRNA levels of PRL-GH and Pit-1a and b isotype genes were analyzed by Northern blot hybridization and reverse transcription-polymerase chain reaction (RT-PCR). The hormonal concentration was analyzed by radioimmunoassay, and the differentiation of placental trophoblast cells were observed by histochemical studies. Reproductive data, such as placental and fetal weights, pregnancy period, and litter size, were surveyed at day 20 of pregnancy and after birth. A statistical analysis was carried out using the SAS program (version 8.1). Results : The mRNA levels of the prolactin-growth hormone (PRL-GH) family of genes were dose dependently reduced by chromium exposure. The mRNA levels of Pit-1a and b isotype genes that induce the expression of the PRL-GH family of genes were also reduced by chromium exposure. The PRL-GH hormonal concentration in the rat placenta, fetus and maternal blood were decreased by chromium exposure. In the middle stage of pregnancy (day 11), a high dose of chromium suppressed the differentiation of spongiotrophoblast cells that secret the PRLGH hormones. In the last stage of pregnancy (day 20), a high dose of chromium induced apoptosis of placental cells. Reproductive data, such as placental and fetal weights, litter size, were reduced, but the pregnancy period was extended in the group exposed to chromium compared with the controls. Conclusion : Chromium (VI) disrupts the ordered functions of the placenta, which leads to reproductive disorders in rats.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.3
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• pp.171-178
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• 2006

Objective: Human embryonic stem (ES) cells have a great potential in regenerative medicine and tissue engineering. The human ES cells could be differentiated into specific cell types by treatments of growth factors and alterations of gene expressions. However, the efficacy of guided differentiation and isolation of specific cells are still low. In this study, we characterized isolated cells from differentiated human ES cells by magnetic activated cell sorting (MACS) system using specific antibodies to cell surface markers. Methods: The undifferentiated hES cells (Miz-hESC4) were sub-cultured by mechanical isolation of colonies and embryoid bodies were spontaneously differentiated with DMEM containing 10% FBS for 2 weeks. The differentiated cells were isolated to positive and negative cells with MACS system using CD34, human epithelial antigen (HEA) and human fibroblast (HFB) antibodies, respectively. Observation of morphological changes and analysis of marker genes expression were performed during further culture of MACS isolated cells for 4 weeks. Results: Morphology of the CD34 positive cells was firstly round, and then it was changed to small polygonal shape after further culture. The HEA positive cells showed large polygonal, and the HFB positive spindle shape. In RT-PCR analysis of marker genes, the CD34 and HFB positive cells expressed endodermal and mesodermal genes, and HEA positive cells expressed ectodermal genes such as NESTIN and NF68KD. The marker genes expression pattern of CD34 positive cells changed during the extension of culture time. Conclusion: Our results showed the possibility of successful isolation of specific cells by MACS system from undirected differentiated human ES cells. Thus, MACS system and marker antibodies for specific cell types might be useful for guided differentiation and isolation of specific cells from human ES cells.

• Development and Reproduction
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• v.14 no.4
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• pp.243-251
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• 2010

ESRRB (Estrogen related receptor $\beta$) is an orphan receptor, and have a role on maintaining the undifferentiated state and self-renewal of pluripotent stem cell as a transcription factor which regulates the expression of OCT4 and NANOG genes. Also, Feng et al. (2009) reported that Esrrb, Oct4 and Sox2 could induce pluripotent stem cell from somatic cells. The aim of the present study was to develop the direct delivery system of human ESRRB protein into human amniotic fluid-derived stem cells (AFSCs) and to analyze the effect of ESRRB on the regulation of pluripotency-related genes. Human ESRRB has three isoforms arisen by alternative splicing. We cloned short-form ESRRB and made a fusion protein of ESRRB and R7 for an efficient protein transfer to cell. R7 as cell-penetrating peptide(CPP) can help to transfer ESRRB into cells. R7-ESRRB-His6 protein was observed in the cytoplasm and nuclei within 5 hours after treatment. Also, we could observe R7-ESRRB-His6 protein only in the nuclei within 24 hours. Realtime PCR showed that ESRRB increased expression of OCT4 and NANOG as well as SOX2 gene. Therefore, we demonstrated that R7-ESRRB-His6 proteins were efficiently transferred into the nuclei of AFSCs and work well as a possible transcription factor.

• Korean Journal of Microbiology
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• v.52 no.4
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• pp.428-436
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• 2016

The emergence of antibiotic-resistant bacteria has been increased and become a public health concern worldwide. Many bacterial infections can be sequentially treated with different types of antibiotics. Thus, this study was designed to evaluate the changes in survival, antibiotic susceptibility, mutant frequency, ${\beta}$-lactamase activity, biofilm formation, and gene expression in Klebsiella pneumoniae after exposure to sequential antibiotic treatments of ciprofloxacin and meropenem. Treatments include control (CON; no addition), 1/2 MIC ciprofloxacin addition (1/2CIP), 2 MIC ciprofloxacin addition (2CIP), initial 1/2 MIC ciprofloxacin addition followed by 1/2 MIC meropenem (8 h-incubation) and 2 MIC ciprofloxacin (16 h-incubation) (1/2CIP-1/2MER-2CIP), initial 1/2 MIC ciprofloxacin addition followed by 1/2 MIC meropenem (8 h-incubation) and 2 MIC meropenem (16 h-incubation) (1/2CIP-1/2MER-2MER), and initial 1/2 MIC ciprofloxacin addition followed by 2 MIC ciprofloxacin(8 h-incubation) and 2 MIC meropenem(16 h-incubation) (1/2CIP-2CIP-2MER). No growth of K. pneumoniae was observed for the 2CIP throughout the incubation period. The numbers of planktonic cells varied with the treatments (7~10 log CFU/ml), while those of biofilm cells were not significantly different among treatments after 24-h incubation, showing approximately 7 log CFU/ml. Among the sequential treatments, the least mutant frequency was observed at the 1/2CIP-1/2MER-2CIP (14%). Compared to the CON, 1/2CIP-2CIP-2MER decreased the sensitivity of K. pneumoniae to piperacillin, cefotaxime, and nalidixic acid. The highest ${\beta}$-lactamase activity was 22 nmol/min/ml for 1/2CIP-1/2MER-2CIP, while the least ${\beta}$-lactamase activity was 6 nmol/min/ml for 1/2CIP-2CIP-2MER. The relative expression levels of multidrug efflux pump-related genes (acrA, acrB, and ramA) were increased more than 2-fold in K. pneumoniae exposed to 1/2CIP-1/2MER-2MER and 1/2CIP-2CIP-2MER. The results suggest that the sequential antibiotic treatments could change the antibiotic resistance profiles in K. pneumoniae.