• International Journal of Oral Biology
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• v.33 no.4
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• pp.205-211
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• 2008

Gingival overgrowth can cause dental occlusion and seriously interfere with mastication, speech, and dental hygiene. It is observed in 25 to 81% of renal transplant patients treated with cyclosporine A (CsA). CsA-induced gingival overgrowth (CIGO) is caused by quantitative alteration of the extracellular matrix components, particularly collagen. However, the molecular mechanisms involved in the pathogenesis of CIGO remain poorly understood, despite intense clinical and laboratory investigations. The aim of the present work is to identify differentially expressed genes closely associated with CIGO. Human gingival fibroblasts were isolated by primary explant culture of gingival tissues from five healthy subjects (HGFs) and two patients with the CIGO (CIGO-HGFs). The proliferative activity of CsA-treated HGFs and CIGO-HGFs was examined using the MTT assay. The identification of differentially expressed genes in CsA-treated CIGO-HGF was performed by differential display reverse transcriptase-polymerase chain reaction (RT-PCR) followed by DNA sequencing. CsA significantly increased the proliferation of two HGFs and two CIGO-HGFs, whereas three HGFs were not affected. Seven genes, including the beta subunit of prolyl 4-hydroxylase (P4HB) and testican 1, were upregulated by CsA in a highly proliferative CIGO-HGF. The increased P4HB and testican-1 mRNA levels were confirmed in CsA-treated CIGO-HGFs by semiquantitative RT-PCR. Furthermore, CsA increased type I collagen mRNA levels and suppressed MMP-2 mRNA levels, which are regulated by P4HB and testican-1, respectively. These results suggest that CsA may induce gingival overgrowth through the upregulation of P4HB and testican-1, resulting in the accumulation of extracellular matrix components.

• Korean Journal of Microbiology
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• v.50 no.1
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• pp.73-83
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• 2014

The objective of the this study was to investigate the binding capacity and removal ability of lactic acid bacterial strains obtained from Korean soybean paste for mutagenic heterocyclic amines (HCAs) formed during cooking of protein-rich food at high temperature. Among 19 strains identified by carbohydrate fermentation and 16S rRNA sequencing, the live cell or cell-free culture supernatant of Lactobacillus acidophilus D11, Enterococcus faecium D12, Pediococcus acidilactici D19, L. acidophilus D38, Lactobacillus sakei D44, Enterococcus faecalis D66, and Lactobacillus plantarum D70 inhibited the mutagenesis caused by either 3-amino-1,4-dimethyl-5H-pyrido[4,3-b] indole (Trp-P-1) or 3-amino-1-methyl-5H-pyrido[4,3-b] indole (Trp-P-2) in Salmonella typhimurium TA98 and TA100. The bacterial cells of the isolated strains showed greater binding activity than the pure cell wall, exopolysaccharide, and pepetidoglycan. The carbohydrate moieties of the cell wall or protein molecules on the cell surface have a significant role in binding Trp-P-1 and Trp-P-2, since protease, heating, sodium metaperiodate, or acidic pH treatments significantly (P<0.05) reduced the binding efficacy of the tested bacteria. Addition of metal ions or sodium dodecyl sulfate decreased the binding ability of E. faecium D12, L. acidophilus D38, and E. faecalis D66. Therefore, the binding mechanisms of these strains may consist of ion-exchange and hydrophobic bonds. Especially, the high mutagen binding by L. acidophilus D38 and L. plantarum D70 may reduce the accumulation or absorption of Trp-P-1 and Trp-P-2 in the small intestine via increased excretion of a mutagen-bacteria complex.

• Journal of Korean Society of Environmental Engineers
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• v.35 no.12
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• pp.973-977
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• 2013

These studies were attempted to investigate the change of microbial community of anode of microbial fuel cell using swine wastewater in the enrichment step with the lapse of time. Microbial fuel cells enriched by a 1 : 1 mixture of anaerobic digestive juices of the sewage treatment plant and livestock wastewater. Enrichment culture step was divided into three stages to indentify the microorganisms. It was separated by each lag phase, exponential phase, and stationary phase. These steps were determined by the change of the current value. The current after enrichment was generated about $0.84{\pm}0.06mA$. We were cut out the different 17 bands in the DGGE fingerprint gel to do sequencing. The bands which the concentration was increasing or newly appearing with the lapse of time were included for this study. In the lag and exponential phase, Clostridium, Rhodocyclaceae, Bacteriodetes, and Uncultured bacterium etc. were detected. There were in the stationary phase Geobacter sp., Rhodocyclaceae, Candidatus, Nitrospira, Flavobactriaceae and uncultured bacterium etc. Geobactor among microorganisms detected in this study is known as the Electrochemically active microorganisms. It may include electrochemically active microorganisms to be considered as electrical activity microorganisms.

• Microbiology and Biotechnology Letters
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• v.34 no.3
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• pp.235-243
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• 2006

The study was performed to identification of producing microbe Non-Cariogenicity Sugar (NCS; Fuc($1{\to}4$) gaINAc($2{\to}6$)NeuAc) with anti-caries activity, and to optimization of production condition. A typical strain which produced the NCS was identified alkalophilic Bacillus sp. S-1013 through the results of morphological, biochemical and chemotaxonomic characteristics and 16S rDNA sequencing. The optimal medium composition for the maximal production of the NCS from alkalophilic Bacillus sp. S-1013 was as follow: soluble starch 30 g, dextrin 15 g, yeast extract 5 g, peptone 10 g, $K_{2}HPO_4$ 2 g in a liter of distilled water. Optimal temperature and pH were 25 and 11.0, respectively. The highest production of NCS was shown 60 hrs cultivation using the optimal medium, and then NCS productivity and dry cell weight of culture broth increased 4.24 and 2.67 time than initial medium, respectively.

• Journal of Plant Biotechnology
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• v.37 no.2
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• pp.174-185
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• 2010

Full-length cDNAs are essential for the correct annotation of genomic sequences and for the functional analysis of genes and their products. To elucidate the functions of a large population of Chinese cabbage (Brassica rapa) genes and to search efficiently for agriculturally useful genes, we have been taking advantage of the full-length cDNA Over-eXpresser (FOX) gene hunting system. With oligo dT column it purify the each mRNA from the flower organs, leaf and stem tissue. And about 120,000 cDNAs from the library were transformed into $\lambda$-pFLCIII-F vector. Of which 115,000 cDNAs from the library were transformed into T-DNA binary vector, pBigs for transformation study. We used normalized full-length cDNA and introduced each cDNA into Arabidopsis by in planta transformation. Full-length Chinese cabbage cDNAs were expressed independently under the CaMV 35S promoter in Arabidopsis. Selfed seeds were harvested from transgenic Arabidopsis. We had selected 2,500 transgenic plants by hygromycin antibiotic tolerant test, and obtained a number of transgenic mutants. Each transgenic Arabidopsis was investigated in morphological changes, fertility and leaf colour. As a result, 285 possible morphological mutants were identified. Introduced cDNA was isolated by PCR amplification of the genomic DNA from the transgenic mutants. Sequencing result and BLAST analysis showed that most of the introduced cDNA were complete cDNAs and functional genes. Also, we examined the effect of Bromelain on enhancing resistance to soft rot in transgenic Chinese cabbage 'Osome'. The bromelain gene identified from FOX hunting system was transformed into Chinese cabbage using Agrobacterium methods. Transformants were screened by PCR, then RT-PCR and real time PCR were performed to analyze gene expression of cysteine protease in the T1 and T2 generations. The anti-bacterial activity of bromelain was tested in Chinese cabbages infected with soft rot bacteria. The results showed that the over-expressed bromelain gene from pineapple conferred enhanced resistance to soft rot in Chinese cabbage.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.18 no.1
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• pp.23-29
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• 2018

Carbamoyl phosphate synthetase 1 deficiency (CPS1D) is a rare autosomal recessive urea cycle disorder characterized by hyperammonemia. CPS1D is caused by mutations in the CPS1 gene on chromosome 2q35. Based on the age of onset, there are two phenotypes: the neonatal type and the delayed-onset type. The severity of clinical manifestation depends on the degree of CPS1 residual enzymatic activity, and can result in hyperammonemia and neurological dysfunction. We report a case of CPS1D in a neonate who developed vomiting, decreased consciousness and hyperammonemia at 25th day after birth. She showed excellent response to treatment including hydration, ammonia-lowering drugs and a low-protein diet without hemodialysis. Her growth, development and neurological outcomes were fair at the last follow-up at 17 months of age.

• Korean Journal of Microbiology
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• v.51 no.1
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• pp.48-60
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• 2015

612 strains isolated from Korean traditional fermented food in Sunchang and their investigated biochemical characterization and ability of biogenic amines non-producing. We selected the SCJ4 having various activity by measurement of extracellular enzyme, antioxidant and antimicrobial activities. Selected strain SCJ4 by 16S rRNA sequencing and biochemical characterization was named Bacillus subtilis SCJ4. And then, we investigated cell growth of SCJ4, and optimized of culture medium constituents using response surface methodology as statistically method. Response surface methodology used Plackett-Burman experimental design for screening of medium constituent. Tryptone, peptone and $MgSO_4$ as medium constituent improving cell growth selected. In order to find out optimal concentration on each constituent, we carried out central composite design. Consequently, optimized concentrations of tryptone, peptone and $MgSO_4$ were predicted to be 15.35 g/L, 12.235 g/L, and 3.5 g/L respectively. Through the model verification, we confirmed about 1.28-fold improvement of the dried cell weight from 0.8767 g/L to 1.1222 g/L when compared to basal medium.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.15 no.1
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• pp.44-48
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• 2015

Mucopolysaccharidosis (MPS) IIIA is a lysosomal storage disorder caused by abnormalities of the enzyme Heparan N-sulfatase that is required for degradation of heparan sulfate. The patient in this study was a 5 year-old boy who presented with macrocephaly and developmental delay. Urinary excretion of glycosaminoglycan was increased (26 g/moL creatinine, reference range: <7 g/moL creatinine) and a distinct band of heparan sulfate was shown in electrophoresis. Heparan N-sulfatase activity was significantly decreased in skin fibroblasts (0.2 pmoL/min/mg protein, reference range: 9-64 pmoL/min/mg protein). PCR and direct sequencing analysis of the SGSH gene showed compound heterozygous mutations: c.1040C>T (p.S347F) and c.703G>A (p.D235N). This is the first report for a Korean patient with MPS IIIA who was confirmed by biochemical investigation and molecular genetic analyses.

• Microbiology and Biotechnology Letters
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• v.31 no.2
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• pp.103-110
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• 2003

The regulation of pyrimidine nucleotide synthesis has been proved to be controlled by a regulatory protein PyrR-mediated attenuation in the Gram-positive bacteria. After several bacterial genome sequencing projects, we have discovered the PyrR orthologues in the databases for Haemophilus influenzae and Synechocystis and sp. PCC6803 genome sequences. To investigate whether these PyrR orthologue proteins regulate pyrimidine nucleotide synthesis as well as the cases of Bacillus, the PyrR regions of each strains were amplified by PCR and cloned with pUC19 or T-vector in Escherichia coli and with a shuttle vector pHPS9 for E. coli and B. subtilis. For the regulation test of the PyrR orthologues, the aspartate-transcarbamylase (ATCase) assay was carried out. From the results of the ATCase assay, it was confirmed that Synechocystis sp. PCC6803 could not restore by pyrimidines to a B. subtilis, PyrR but H. influenzae PyrR could. For Purification of PyrR orthologue proteins, PyrR orthologue genes were cloned into the expression vector (pET14b). Over-expressed product of PyrR orthologue genes was purified and analyzed by the SDS-PACE. The purified PyrR orthologue proteins from H. influenzae and Synechocystis sp. PCC6803 turned out to be molecular mass of 18 kDa and 21 kDa, respectively. The result of uracil phosphoribosyl transferase (UPRTase) assay with purified PyrR orthologue proteins showed that H. influenzae PyrR protein only has UPRTase activity. In addition, we could predict several regulatory mechanisms that PyrR orthologue proteins regulate pyrimidine de novo synthesis in bacteria, through phylogenetic analysis for PyrR orthologue protein sequences.

• Journal of Life Science
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• v.26 no.10
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• pp.1163-1168
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• 2016

Plant growth-promoting rhizobacteria (PGPR) have gained worldwide importance and acceptance due to their agricultural benefits. These microorganisms are potential tools for sustainable agriculture, with effects on plant growth, biofertilization, induced systemic resistance, and biocontrol of plant pathogens. In this study, four different Pantoea species were isolated from field soil, and their plant growth-promoting characteristics were studied. Based on 16S rDNA gene sequencing analyses, the se were grouped into Pantoea ananatis, Pantoea citrea, Pantoea dispersa, Pantoea vagans and named as Pa1, Pc1, Pd1, Pv1, respectively. All of these strains have their ability for solubilization of insoluble phosphate depending on pH decrease at the range around pH 5 at 1days after inoculation and production of plant hormone indole acetic acid (IAA) with 85.3±16.3 μg/ml of Pa1, 183.9±16.8 μg/ml of Pc1, 28.8±17.3 μg/ml of Pd1 and 114.1±16.5 μg/ml of Pv1, respectively. Pa1, Pc1 and Pd1 also have high activity for production of gibberellin (GA₃) hormone with 331.1±19.2 μg/ml of Pa1, 288.5±16.8 μg/ml of Pc1, 309.2±18.2 μg/ml of Pd1, but Pv1 does not. Furthermore, all these species have significantly promoted the growth of the lettuce seedling plants at the range around 32~37% for fresh weight and 10~15% for shoot length enhancement, so that these microbe could be used as a potential bio-fertilizer agents.