• Molecules and Cells
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• v.45 no.9
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• pp.660-672
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• 2022

The target of rapamycin complex (TORC) plays a key role in plant cell growth and survival by regulating the gene expression and metabolism according to environmental information. TORC activates transcription, mRNA translation, and anabolic processes under favorable conditions, thereby promoting plant growth and development. Tomato fruit ripening is a complex developmental process promoted by ethylene and specific transcription factors. TORC is known to modulate leaf senescence in tomato. In this study, we investigated the function of TORC in tomato fruit ripening using virus-induced gene silencing (VIGS) of the TORC genes, TOR, lethal with SEC13 protein 8 (LST8), and regulatory-associated protein of TOR (RAPTOR). Quantitative reverse transcription-polymerase chain reaction showed that the expression levels of tomato TORC genes were the highest in the orange stage during fruit development in Micro-Tom tomato. VIGS of these TORC genes using stage 2 tomato accelerated fruit ripening with premature orange/red coloring and decreased fruit growth, when control tobacco rattle virus 2 (TRV2)-myc fruits reached the mature green stage. TORC-deficient fruits showed early accumulation of carotenoid lycopene and reduced cellulose deposition in pericarp cell walls. The early ripening fruits had higher levels of transcripts related to fruit ripening transcription factors, ethylene biosynthesis, carotenoid synthesis, and cell wall modification. Finally, the early ripening phenotype in Micro-Tom tomato was reproduced in the commercial cultivar Moneymaker tomato by VIGS of the TORC genes. Collectively, these results demonstrate that TORC plays an important role in tomato fruit ripening by modulating the transcription of various ripening-related genes.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2001.11a
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• pp.43-56
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• 2001

Colostrum contains various kinds of cytokines including TGF-${\beta}$ which is known to be multifunctional in immune response and act as an anti-inflammatory agent. First, we measured the amount of TGF-${\beta}$ in bovine and human colostrum. Expression pattern of TGF-${\beta}$ isotypes was dramatically different between human and bovine colostrial samples. Bovine colostrum collected on day 1 post-delivery retained $41.79{\pm}16.96ng/ml$ of TGF-${\beta}$ 1 and $108.4{\pm}78.65ng/ml$ of TGF-${\beta}$ 2 while in human, $284{\pm}124.75ng/ml$ of TGF-${\beta}$ 1 and $29.75{\pm}6.73ng/ml$ of TGF-${\beta}$ 2. Thus, TGF-${\beta}$ is the predominant TGF-${\beta}$ isotype in bovine colostrum and vice versa in human colostrum. Both TGF-${\beta}$ isotypes diminished significantly in human and bovine colostrum with time. Next, biological activity of colostrial samples was examined in vitro. Both human and bovine colostrum increased IgA synthesis by LPS-activated mouse spleen B cells, which is a typical effect of TGF-${\beta}$ on the mouse B cell differentiation. Futhermore, we found that anti-proliferative activity in MV1LU cells by colostrum samples disappeared by addition of anti-TGF-${\beta}$ 1 and anti-TGF-${\beta}$ 2 antibody. In conclusion, there are substantial amounts of biologically active TGF-${\beta}$ 1 and TGF-${\beta}$ 2 in bovine and human colostrum. The results that the colostrum can increase IgA expression has important implications since IgA is the major Ig class produced in the gastrointestinal tract. We have previously shown that the stimulatory effect of Bifidobacteria bifidum on spllen B cells was quite similar to that of LPS which is a well-known polyclonal activator for murine B cells. In the present study, we further asked whether B. bifidum regulate the synthesis of IgA by mucosal lymphoid cells present in Peyers patches (PP) and mesenteric lymph nodes (MLN). B. bifidum alone, but not C. perfringens, significantly induced overall IgA and IgM synthesis by both MLN and PP cells. This observation indicates that B. bifidum possesses a modulatory effect on the mucosal antibody production in vivo. We, therefore, investigated the mucosal antibody prodduction following peroral administration of B. bifidum to mice. Ingested B. bifidum significantly increased the numbers of Ig (IgM, IgG, and IgA) secreting cells in the culture of both MLN and spleen cells, indicating that peroally introduced B. bifidum enhances mucosal and systemic antibody response. Importantly, however, B. bifidum itself does not induce the own specific antibody responses, implying that B. bifidum do not incite any unwanted immune reaction. Subsequently, it was found that excapsulation of B. bifidum further augments the total IgA production by increasing the number of IgA-secreting cells in the culture of both MLN and spleen cells. Finally, we found that the immuno-stimulating activity of B. bifidum is due to its cell wall components but not due to any actively secreting component(s) from bacteria. Thus our data reveal that peroral administration of B. bifidum can enhance intestinal IgA production and that encapsulation of B. bifidum further reinforces the IgA production.

• Journal of Biomedical Engineering Research
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• v.42 no.6
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• pp.251-258
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• 2021

Hyaluronic acid (HA) microspheres (MSs) crosslinked with polyethylene glycol diglycidyl ether (PEGDE) are prepared using a simple fluidic device (SFD) to investigate the optimized parameters. A solution mixture of PEGDE in 2-methyl-1-propanol was prepared as a continuous phase in SFD. HA solutions of 1 wt% concentration were introduced into SFD as a discontinuous phase. The HA solution prepared by stirring for more than 48 h exhibited spherical MSs at the needle tip inside the ring cap. As the flow rate of the continuous phase increased from 0.7 to 1.9 mL/min, the diameter of the MS decreased from 173±36 ㎛ to 129±13 ㎛. Although the PEGDE concentration in the range of 0.2 to 1.8 vol% did not affect the diameter of the MS, the microstructure of MS, consisting of inner hollow void and wall, was changed. The inner void and wall size decreased and increased from 79.5 ㎛ to 57.2 ㎛ and from 10.3 ㎛ to 21.4 ㎛, respectively, with increasing PEGDE concentration from 0.2 vol% to 1.8 vol%. FT-IR peaks located around 2867 cm^-1 and 1088 cm^-1 indicated that the HA MS prepared at different PEGDE concentrations were chemically crosslinked. The HA MSs containing different PEGDE concentrations exhibited quantitative cell viability of more than 98%. L-929 cells adhered well to the HA MSs and proliferated continuously with increasing culture time to 48 h regardless of PEGDE concentration, implying that the HA MSs are clinically safe and effective.

• Microbiology and Biotechnology Letters
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• v.47 no.3
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• pp.465-472
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• 2019

Candida albicans is an opportunistic human pathogen that causes infections. Candidiasis is often related to antifungal resistance because the pathogen has the ability to form biofilms. In a previous study, we found that the Salvia miltiorriza ethanol extract demonstrated anticandidal activity by altering membrane permeability and inhibiting the cell wall synthesis in C. albicans. Our results here demonstrate that $78{\mu}g/ml$ of the S. miltiorriza extract significantly diminished the early stage biofilms formed by 10 clinical C. albicans isolates by 51.3%; this was analyzed by 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt (XTT) reduction assay. The effect of the S. miltiorrhiza extract on the adhesion of C. albicans cells to polystyrene plates and germ tube formation was examined via microscopic investigation. Although the density of the adhered cells was remarkably reduced up on incubation with $39{\mu}g/ml$ S. miltiorrhiza extract, germ tube formation by C. albicans was rarely affected. Quantitative real-time PCR analysis showed that the S. miltiorrhiza extract downregulated the expression of C. albicans hypha-specific genes, EAP1 by 34.7% (p < 0.001), ALS1 by 45.0% (p < 0.001), ALS3 by 48.1% (p < 0.001), and ECE1 by 21.3% (p = 0.006), respectively. Our data suggest that the S. miltiorrhiza ethanol extract significantly inhibited the early stage of biofilm formation by C. albicans by interfering with cell adhesion, by downregulating EAP1, ALS1 and ALS3, and presumably by modifying the cell wall and membrane structure.

• Journal of Life Science
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• v.31 no.2
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• pp.109-114
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• 2021

Oryzalin is a dinitroaniline herbicide that has been known to disrupt microtubules. Microtubules and microfilaments are components of cytoskeletons that are implicated in plant cell growth, which requires the synthesis of cellulose when cell walls elongate. In addition, microtubules are also involved in the sedimentation of statoliths, which regulate the perception of gravity in the columella cells of root tips. In this study, we investigated the effect of oryzalin on the growth and gravitropic response of Arabidopsis roots. The role of ethylene in oryzalin's effect was also examined using these roots. Treatment of oryzalin at a concentration of 10-4 M completely inhibited the roots' growth and gravitropic response. At a concentration of 10-6 M oryzalin, root growth was inhibited by 47% at 8 hr when compared to control. Gravitropic response was inhibited by about 38% compared to control in roots treated with 10-6 M oryzalin for 4 hr. To understand the role of oryzalin in the regulation of root growth and gravitropic response, we measured ethylene production in root segments treated with oryzalin. It was found that the addition of oryzalin stimulated ethylene production through the activation of ACC oxidase and ACC synthase genes, which are key components in the synthesis of ethylene. From these findings, it can be inferred that oryzalin inhibits the growth and gravitropic response of Arabidopsis roots by stimulating ethylene production. The increased ethylene alters the arrangement of the microtubules, which eventually interferes with the growth of the cell wall.

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.54-54
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• 2015

Crops lack genetic resistance to most necrotrophic soil-borne pathogens and parasitic nematodes that are ubiquitous in agroecosystems worldwide. To overcome this disadvantage, plants recruit and nurture specific group of antagonistic microorganisms from the soil microbiome to defend their roots against pathogens and other pests. The best example of this microbe-based defense of roots is observed in disease-suppressive soils in which the suppressiveness is induced by continuously growing crops that are susceptible to a pathogen. Suppressive soils occur globally yet the microbial basis of most is still poorly described. Fusarium wilt, caused by Fusarium oxysporum f. sp. fragariae is a major disease of strawberry and is naturally suppressed in Korean fields that have undergone continuous strawberry monoculture. Here we show that members of the genus Streptomyces are the specific bacterial components of the microbiome responsible for the suppressiveness that controls Fusarium wilt of strawberry. Furthermore, genome sequencing revealed that Streptomyces griseus, which produces a novel thiopetide antibiotic, is the principal species involved in the suppressiveness. Finally, chemical-genetic studies demonstrated that S. griseus antagonizes F. oxysporum by interfering with fungal cell wall synthesis. An attack by F. oxysporum initiates a defensive "cry for help" by strawberry root and the mustering of microbial defenses led by Streptomyces. These results provide a model for future studies to elucidate the basis of microbially-based defense systems and soil suppressiveness from the field to the molecular level.

• BMB Reports
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• v.38 no.5
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• pp.507-516
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• 2005

Antimicrobial peptides (AMPs) have been isolated and characterized from tissues and organisms representing virtually every kingdom and phylum. Their amino acid composition, amphipathicity, cationic charge, and size allow them to attach to and insert into membrane bilayers to form pores by 'barrel-stave', 'carpet' or 'toroidal-pore' mechanisms. Although these models are helpful for defining mechanisms of AMP activity, their relevance to resolving how peptides damage and kill microorganisms still needs to be clarified. Moreover, many AMPs employ sophisticated and dynamic mechanisms of action to carry out their likely roles in antimicrobial host defense. Recently, it has been speculated that transmembrane pore formation is not the only mechanism of microbial killing by AMPs. In fact, several observations suggest that translocated AMPs can alter cytoplasmic membrane septum formation, reduce cell-wall, nucleic acid, and protein synthesis, and inhibit enzymatic activity. In this review, we present the structures of several AMPs as well as models of how AMPs induce pore formation. AMPs have received special attention as a possible alternative way to combat antibiotic-resistant bacterial strains. It may be possible to design synthetic AMPs with enhanced activity for microbial cells, especially those with antibiotic resistance, as well as synergistic effects with conventional antibiotic agents that lack cytotoxic or hemolytic activity.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.475-481
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• 1999

The study of lignin, a major component of secondary cell wall, has been partly focused on its removal from the woody part in the kraft pulping industry. Cinnamyl alcohol dehydrogenase (CAD; EC 1.1.l95) catalyzes the synthesis of cinnamyl alcohols from corresponding cinnamaldehydes. A cDNA clone, MdCADl, encoding putative CAD from apples (Malus domestica Borkh. cv Fuji) was characterized in this study. The clone contains an open reading frame of 325 amino acid residues, which shows a greater than 80％ identity with Eucalyptus CADl. MdCADl mRNA was detectable in vegetative tissues and was strongly expressed in the fruit. The expression pattern of MdCADl mRNA in the fruit peel after light exposure was also examined. The mRNA was rapidly increased until 1 day after light exposure and remained stable thereafter, suggesting that MdCADl is light inducible. The inducibility of the MdCADl gene was examined using several environmental stresses. Mechanical wounding of leaves increased the MdCADl mRNA level and the induction was further increased by salicylic acid. Southern blot hybridization showed that there is either one or a few copies of CAD genes in apples. To our knowledge, it is believed that MdCADl is the first CAD clone expressed predominantly in fruit.

• Genes and Genomics
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• v.40 no.11
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• pp.1237-1248
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• 2018

Introduction Cellulose microfibril is a major cell wall polymer that plays an important role in the growth and development of plants. The gene cellulose synthase A (CesA), encoding cellulose synthases, is involved in the synthesis of cellulose microfibrils. However, the regulatory mechanism of CesA gene expression is not well understood, especially during the early developmental stages. Objective To identify factor(s) that regulate the expression of CesA genes and ultimately control seedling growth and development. Methods The presence of cis-elements in the promoter region of the eight CesA genes identified in flax (Linum usitatissimum L. 'Nike') seedlings was verified, and three kinds of ethylene-responsive cis-elements were identified in the promoters. Therefore, the effect of ethylene on the expression of four selected CesA genes classified into Clades 1 and 6 after treatment with $10^{-4}$ and $10^{-3}M$ 1-aminocyclopropane-1-carboxylic acid (ACC) was examined in the hypocotyl of 4-6-day-old flax seedlings. Results ACC-induced ethylene either up- or down-regulated the expression of the CesA genes depending on the clade to which these genes belonged, age of seedlings, part of the hypocotyl, and concentration of ACC. Conclusion Ethylene might be one of the factors regulating the expression of CesA genes in flax seedlings.

• Biomolecules & Therapeutics
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• v.31 no.2
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• pp.141-147
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• 2023

Antibiotic resistance has emerged as a global threat to modern healthcare systems and has nullified many commonly used antibiotics. β-Lactam antibiotics are among the most successful and occupy approximately two-thirds of the prescription antibiotic market. They inhibit the synthesis of the peptidoglycan layer in the bacterial cell wall by mimicking the D-Ala-D-Ala in the pentapeptide crosslinking neighboring glycan chains. To date, various β-lactam antibiotics have been developed to increase the spectrum of activity and evade drug resistance. This review emphasizes the three-dimensional structural characteristics of β-lactam antibiotics regarding the overall scaffold, working mechanism, chemical diversity, and hydrolysis mechanism by β-lactamases. The structural insight into various β-lactams will provide an in-depth understanding of the antibacterial efficacy and susceptibility to drug resistance in multidrug-resistant bacteria and help to develop better β-lactam antibiotics and inhibitors.