• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.296-301
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• 2015

Chronic kidney diseases are based on uncontrolled immunological and inflammatory responses to pathophysiological renal circumstances such as glomerulonephritis, which is caused by immunological mechanisms of glomerular inflammation with increased production of renal pro-inflammatory cytokines. Pentoxifylline (PTX) exhibits anti-inflammatory properties by inhibiting cytokine and chemokine production through aggregation of erythrocytes and thrombocytes. We synthesized a series of 1,7-substituted methylxanthine derivatives by the Traube purine reaction, and the formation of purine ring was completed through nitrosation, a reduction of the nitroso to the amine by catalytic hydrogenation as derivatives of PTX. Then we studied biological activities such as renal anti-inflammatory effects of the synthesized compounds in the production of cytokines such as nitric oxide (NO), interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) and of chemokines such as monocyte chemoattractant protein-1 and IL-8 in Raw 264.7 and HK-2 cells. Renal antiinflammatory activities of this novel series of N-1 and N-7-substituted methylxanthine showed that the N-7 methyl-group-substituted analogs (S7b) showed selective 61% and 77% inhibition of the production of NO and IL-8. The other replacement of the N-1-(CH₂)₄COCH₃ roup, as in the case of compound S6c, also showed an effective 50% and 77% inhibition of TNF-α and IL-8 production in LPS-stimulated Raw 264.7 and HK-2 cells.

• Journal of Plant Biotechnology
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• v.46 no.3
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• pp.165-171
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• 2019

Marker-assisted backcrossing (MABC) is useful for selecting an offspring with a highly recovered genetic background for a recurrent parent at early generation to various crops. Moreover, marker-assisted backcrossing (MABC) along with marker-assisted selection (MAS) contributes immensely to overcome the main limitation of the conventional breeding and it accelerates recurrent parent genome (RPG) recovery. In this study, we were employed to incorporate rin gene(s) from the donor parent T13-1084, into the genetic background of HK13-1151, a popular high-yielding tomato elite inbred line that is a pink color fruit, in order to develop a rin HK13-1084 improved line. The recurrent parent genome recovery was analyzed in early generations of backcrossing using SNP markers obtained from genotyping-by-sequencing analysis. From the $BC_1F_1$ and $BC_2F_1$ plants, 3,086 and 4868 polymorphic SNP markers were obtained via GBS analysis, respectively. These markers were present in all twelve chromosomes. The background analysis revealed that the extent of RPG recovery ranged from 56.7% to 84.5% and from 87.8% to 97.8% in $BC_1F_1$ and $BC_2F_1$ generations, respectively. In this study, No 5-1 with 97.8% RPG recovery rate among $BC_2F_1$ plants was similar to HK13-1151 strain in the fruit shape. Therefore, the selected plants were fixed in $BC_2F_2$ generation through selfing. MAS allowed identification of the plants that are more similar to the recurrent parent for the loci evaluated in the backcross generations. MABC can greatly reduce breeding time as compared to the conventional backcross breeding. For instance, MABC approach greatly shortened breeding time in tomato.

• Korean Journal of Microbiology
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• v.28 no.2
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• pp.145-150
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• 1990

The effects of environmental factors on degradation of Aroclor 1242 were investigated with four Gram-negative bacterial isolates. Their biodegradabilities of the Aroclor were well correlated to their growth rates on the Aroclor added as a sole carbon and energy source. The optimum concentration of the Aroclor for biodegradation of the substrate in MM2 medium was 0.5mg/ml in HK-100, HK-123, and MS-1003 strains, but 1 mg/ml in DJ-26 strain. The optimum temperature and pH were $30^{\circ}C$ and 7.0, respectively, for all the strains. On the basis of the results which the strain of DJ-26 showed the highest degradability of the Aroclor as well as the highest growth rate under the optimum environmental conditions, the bacterial isolate identified as Pseudomonas sp. was found to be a strain usable for treatment of the toxic and recalcitrant chemical pollutants, such as polychlorinated aromatic hydrocarbons.

• Journal of Plant Biotechnology
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• v.47 no.3
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• pp.227-234
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• 2020

To produce the miraculin protein in suspension cultures, rice (Oryza sativa L.) was transformed with Agrobacterium tumefacience EHA105 containing the miraculin AB512278 gene. The cell suspension cultures were established using cell lines selected from transgenic rice callus. The integration of the miraculin gene into the rice chromosome was confirmed using genomic PCR analysis. In addition, RT-PCR analysis indicated that the miraculin gene is expressed in the selected suspension cell lines. Thus, the recombinant miraculin was expressed in the transgenic suspension cell line, HK-2. Therefore, we have successfully developed a HK-2 line that produces miraculin. These results demonstrate that transformed cell suspension cultures can be used to produce a taste-modifying protein such as miraculin.

• Biomolecules & Therapeutics
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• v.21 no.3
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• pp.190-195
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• 2013

Cisplatin is a member of platinum-containing anti-cancer drugs that causes cross-linking of DNA and ultimately cancer cell apoptosis. The therapeutic function of cisplatin on various types of cancers has been widely reported but the side effects have been discovered together and nephrotoxicity has been regarded as major side effect of cisplatin. To select candidates for new sensitive nephrotoxicity biomarker, we performed proteomic analysis using 2-DE/MALDI-TOF-MS followed by cisplatin treatment in human kidney cell line, HK-2 cells, and compared the results to the gene profile from microarray composed of genes changed in expression by cisplatin from formerly reported article. Annexin A5 has been selected to be the most potential candidate and it has been identified using Western blot, RT-PCR and cell viability assay whether annexin A5 is available to be a sensitive nephrotoxic biomarker. Treatment with cisplatin on HK-2 cells caused the increase of annexin A5 expression in protein and mRNA levels. Over-expression of annexin A5 blocked HK-2 cell proliferation, indicating correlation between annexin A5 and renal cell toxicity. Taken together, these results suggest the possibility of annexin A5 as a new biomarker for cisplatin-mediated nephrotoxicity.

• Microbiology and Biotechnology Letters
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• v.45 no.1
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• pp.35-42
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• 2017

This study aimed to investigate the effects of probiotic fermentation by comparing the skin antioxidant and anti-inflammatory properties of non-fermented (ARE) and fermented (ARE-F) hot water extracts of Agastache rugosa leaves. ARE-F was obtained via ARE fermentation using Lactobacillus rhamnosus HK-9. In vitro, anti-inflammatory properties were evaluated by analyzing the levels of nitric oxide (NO), reactive oxygen species (ROS), and inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-stimulated HaCaT keratinocytes. In vitro antiradical activity was measured using 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay. Attenuation of LPS-stimulated NO (p < 0.01), ROS (p < 0.001) and iNOS (p < 0.05) levels by ARE-F was significantly stronger than that by ARE in HaCaT keratinocytes. However, no differences were observed between the DPPH radical scavenging activities of ARE and ARE-F. ARE-F possesses enhanced skin antioxidant and anti-inflammatory properties, suggesting that probiotic bacterial fermentation can be considered an effective tool for augmenting some pharmacological properties of A. rugosa leaves. In brief, the skin antioxidant and anti-inflammatory potentials of A. rugosa leaf extract are augmented by the fermentation with L. rhamnosus HK-9, a probiotic bacterium.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.48 no.1
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• pp.77-85
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• 2022

Retinaldehyde (RA), vitamin A derivative, is an intermediate between retinol and retinoic acid and has an excellent wrinkle improving effect. In this study, Drug-in-cyclodextrin-in-liposome (DCL) was used to enhance the stability and skin penetration of RA. The complex of RA and hydroxypropyl-beta-cyclodextrin (HP-β-CD) was prepared by the freeze-drying method, and the presence or absence of inclusion of retinal was confirmed by UV-Vis spectrometer, FT-IR and SEM images. RA was captured in HP-β-CD about 95.6% on 1 : 15 (w/w). The retinal-HP-β-CD complex was encapsulated in liposomes using a homomixer and microfluidizer, with an average particle size of 215 ± 4.2 nm and a zeta potential of -31.2 ± 0.5 mv. In the evaluation of the degradation stability of RA, degradation rate of RA-HP-β-CD-liposomes in water was 1.8% higher than RA-liposome (5.8%), RA-HP-β-CD complex (9.7%) and RA alone (37.6%). RA cream (0.05% RA) including RA-HP-β-CD-liposomes was prepared for clinical test with wrinkle-improving efficacy and skin dermis denseness evaluated for 2 or 4 weeks. RA cream showed a significant wrinkle improving effect without skin irritation. In conclusion, it was confirmed that the double stabilization technology using the DCL system contribu tes to the effect of improving skin wrinkles by increasing the stabilization of retinal.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.117-127
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• 2012

The beneficial effects of adipose-derived stem cell conditioned media (ADSC-CM) for skin regeneration have previously been reported, despite the precise mechanism of how ADSC-CM promotes skin regeneration remaining unclear. ADSC-CM contains various secretomes and this may be a factor in it being a good resource for the treatment of skin conditions. It is also known that ADSC-CM produced in hypoxia conditions, in other words Advanced Adipose-Derived Stem cell Protein Extract (AAPE), has excellent skin regenerative properties. In this study, a human primary skin cell was devised to examine how AAPE affects human dermal fibroblast (HDF) and human keratinocyte (HK), which both play fundamental roles in skin regeneration. The promotion of collagen formation by HDFs was observed at 0.32 mg/ml of AAPE. AAPE treatment significantly stimulated stress fiber formation. DNA gene chips demonstrated that AAPE in HKs (p<0.05) affected the expression of 133 identifiable transcripts, which were associated with cell proliferation, migration, cell adhesion, and response to wounding. Twenty five identified proteins, including MMP, growth factor and cytokines such as CD54, FGF-2, GM-CSF, IL-4, IL-6, VEGF, TGF-${\beta}2$, TGF-${\beta}3$, MMP-1, MMP-10, and MMP-19, were contained in AAPE via antibody arrays. Thus, AAPE might activate the HK biological function and induce the collagen synthesis of HDF. These results demonstrate that AAPE has the potential to be used for clinic applications aimed at skin regeneration.

• Journal of Microbiology and Biotechnology
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• v.25 no.1
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• pp.89-97
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• 2015

Bacillus subtilis HK176 with high fibrinolytic activity was isolated from cheonggukjang, a Korean fermented soyfood. A gene, aprE176, encoding the major fibrinolytic enzyme was cloned from B. subtilis HK176 and overexpressed in E. coli BL21(DE3) using plasmid pET26b(+). The specific activity of purified AprE176 was 216.8 ± 5.4 plasmin unit/mg protein and the optimum pH and temperature were pH 8.0 and 40℃, respectively. Error-prone PCR was performed for aprE176, and the PCR products were introduced into E. coli BL21(DE3) after ligation with pET26b(+). Mutants showing enhanced fibrinolytic activities were screened first using skim-milk plates and then fibrin plates. Among the mutants, M179 showed the highest activity on a fibrin plate and it had one amino acid substitution (A176T). The specific activity of M179 was 2.2-fold higher than that of the wild-type enzyme, but the catalytic efficiency (k_cat/K_m) of M179 was not different from the wild-type enzyme owing to reduced substrate affinity. Interestingly, M179 showed increased thermostability. M179 retained 36% of activity after 5 h at 45℃, whereas AprE176 retained only 11%. Molecular modeling analysis suggested that the 176^th residue of M179, threonine, was located near the cation-binding site compared with the wild type. This probably caused tight binding of M179 with Ca²⁺, whichincreased the thermostability of M179.

• IMMUNE NETWORK
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• v.13 no.2
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• pp.75-79
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• 2013

Evidence for immunoregulatory roles of prostaglandins (PGs) is accumulating. Since our observation of PG production by human follicular dendritic cells (FDCs), we investigated the regulatory mechanism of PG production in FDC and attempted to understand the functions of released PGs in the responses of adjacent lymphocytes. Here, using FDC-like cells, HK cells, we analyzed protein expression alterations in cyclooxygenase-2 (COX-2) in the presence of IL-4 or histone deacetylase (HDAC) inhibitors. Both IL-4 and HDAC inhibitors suppressed COX-2 expression in dose-dependent manners. Their effect was specific to COX-2 and did not reach to COX-1 expression. Interestingly, HDAC inhibitors gave rise to an opposing effect on COX-2 expression in peripheral blood monocytes. Our results suggest that IL-4 may regulate COX-2 expression in FDCs by affecting chromatin remodeling and provide insight into the role of cellular interactions between T cells and FDC during the GC reaction. Given the growing interests in wide-spectrum HDAC inhibitors, the differential results on COX-2 expression in HK cells and monocytes raise cautions on their clinical use.