• BMB Reports
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• 제51권5호
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• pp.207-208
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• 2018

Chitinase-Like Proteins (CLPs) are an evolutionarily conserved protein which lose their enzymatic activity for degrading chitin macromolecules. Chitinase-3-like-1 (Chi3l1) is a type of CLP that is highly expressed in epithelial cells, macrophages, etc., and is known to have correlations with type 2 inflammation and cancer. Although the increased level of Chi3l1 in the blood was reported in various disease patients, the function of Chi3l1 in adaptive immunity has been totally unknown. Recently, we found that Chi3l1 is expressed in T cells and has a negative regulatory role in T-cell activation and proliferation. A genetic ablation study of Chi3l1 in T cells showed hyperresponsiveness to TcR stimulation, which increased proliferation and Th1 differentiation. A significant increase of $IFN{\gamma}$ signaling in Chi3l1-deficient T cells synergistically increased Th1 and CTL functions against melanoma cells in vitro and in vivo. In addition, targeted knockdown by Chi3l1 siRNA complexed with the cell-penetrating peptide dNP2, which showed decreased pulmonary melanoma metastasis with increased infiltration of Th1 and CTL in the lung. This study first suggests that Chi3l1 is a novel regulator of Th1/CTL responses and could be a target for treating cancer to increase tumor immunity.

• 대한의생명과학회지
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• 제24권1호
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• pp.23-29
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• 2018

Abdominal obesity is considered as one of the most risky factors governing the development of metabolic diseases. Here we identify that human chitinase 3-like 1 (CHI3L1, also called YKL-40 in human) single nucleotide polymorphism (SNP), rs883125, is associated with abdominal obesity in Korean women. Korean women subjects with the rs883125 G/G or C/G genotype present higher waist-hip ratio than subjects with C/C genotype suggesting that human subjects who G nucleotide substitution at the rs883125 tended to more accumulate intra-abdominal fat at the abdominal cavity. In addition, Chi3l1 gene expression is increased in adipose tissue from obese mice and pro-inflammatory cytokine enhances Chi3l1 expression in adipocytes, indicating that Chi3l1 is greatly related with obesity and obesity-induced pro-inflammatory responses. Taken together, the minor allele of rs883125 is associated with a higher prevalence of abdominal obesity in Korean women. These findings suggest that genotype of rs883125 can be a biomarker of incident abdominal obesity and abdominal obesity-related metabolic diseases.

• IMMUNE NETWORK
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• 제21권3호
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• pp.22.1-22.17
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• 2021

Chitinase-3-like-1 (CHI3L1) is known to induce inflammation in the progression of allergic diseases. Previous our studies revealed that 2-({3-[2-(1-cyclohexen-1-yl)ethyl]-6,7-dimethoxy-4-oxo-3,4-dihydro-2-quinazolinyl}sulfanyl)-N-(4-ethylphenyl)butanamide (K284-6111; K284), the CHI3L1 inhibiting compound, has the anti-inflammatory effect on neuroinflammation. In this study, we investigated that K284 treatment could inhibit the development of atopic dermatitis (AD). To identify the effect of K284, we used phthalic anhydride (5% PA)-induced AD animal model and in vitro reconstructed human skin model. We analyzed the expression of AD-related cytokine mediators and NF-κB signaling by Western blotting, ELISA and quantitative real-time PCR. Histological analysis showed that K284 treatment suppressed PA-induced epidermal thickening and infiltration of mast cells. K284 treatment also reduced PA-induced release of inflammatory cytokines. In addition, K284 treatment inhibited the expression of NF-κB activity in PA-treated skin tissues and TNF-α and IFN-γ-treated HaCaT cells. Protein-association network analysis indicated that CHI3L1 is associated with lactoferrin (LTF). LTF was elevated in PA-treated skin tissues and TNF-α and IFN-γ-induced HaCaT cells. However, this expression was reduced by K284 treatment. Knockdown of LTF decreased the expression of inflammatory cytokines in TNF-α and IFN-γ-induced HaCaT cells. Moreover, anti-LTF antibody treatment alleviated AD development in PA-induced AD model. Our data demonstrate that CHI3L1 targeting K284 reduces AD-like skin inflammation and K284 could be a promising therapeutic agent for AD by inhibition of LTF expression.

• Mycobiology
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• 제45권4호
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• pp.385-391
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• 2017

The ability of Bacillus subtilis, strain ALICA to produce three mycolytic enzymes (chitinase, ${\beta}$-1,3-glucanase, and protease), was carried out by the chemical standard methods. Bacillus subtilis ALICA was screened based on their antifungal activity in dual plate assay and cell-free culture filtrate (25%) against five different phytopathogenic fungi Alternaria alternata, Macrophomina sp., Colletotrichum gloeosporioides, Botrytis cinerea, and Sclerotium rolfesii. The B. subtilis ALICA detected positive for chitinase, ${\beta}$-1,3-glucanase and protease enzymes. Fungal growth inhibition by both strain ALICA and its cell-free culture filtrate ranged from 51.36% to 86.3% and 38.43% to 68.6%, respectively. Moreover, hyphal morphological changes like damage, broken, swelling, distortions abnormal morphology were observed. Genes expression of protease, ${\beta}$-1,3-glucanase, and lipopeptides (subtilosin and subtilisin) were confirmed their presence in the supernatant of strain ALICA. Our findings indicated that strain ALICA provided a broad spectrum of antifungal activities against various phytopathogenic fungi and may be a potential effective alternative to chemical fungicides.

• 한국토양비료학회지
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• 제45권1호
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• pp.66-73
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• 2012

Plant growth promoting traits like production of indoleacetic acid (IAA), ammonia, hydrogen cyanide (HCN), siderophore, and like the enzyme activities of catalase, ACC deaminase, cellulase, chitinase and protease were assayed in vitro for twenty one phosphorus solubilizing bacteria isolated from soil isolates. Except SPP-5 and SPP-15 strains, all the other isolated strains produced IAA in various amounts of 10 to $23{\mu}g\;ml^{-1}$. All strains showed positive response for ammonia production and ACC deaminase activity implying that they are capable of growing in a N-free basal medium. Catalase activity was found to be superior in SPP-2, SPP-7, SPP-12 and SPP-17 compared to the other strains tested. HCN production was detected by 15 strains and among them SPP-9, SPP-15, SAph-11, and SAph-24 were found to be strong HCN producers. Except the isolates SPP-10, SPP-12, SPP-13 and SPP-14, all the other isolates produced more than 80% siderophore units. None of the strains showed cellulose and chitinase activity. SAph-8, SAPh-11, SAPh-24 and SPP-15 strains showed 35.84, 50.33, 56.64 and 34.78 U/ml protease activities, respectively. SPP-1, SPP-2, SPP-3, SPP-11, SPP-17, SPP-18, SAph-11 and SAph-24 strains showed positive response for all the tested plant growth promotion traits except cell wall degrading enzyme activities. According to the results, all the tested phosphorus solubilizing isolates could exhibit more than three or four plant growth promoting traits, which may promote plant growth directly or indirectly or synergistically. Therefore, these phosphorus solubilizing strains could be employed as bio-inoculants for agriculture soils.

• 한국작물학회지
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• 제43권3호
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• pp.172-178
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• 1998

Freezing-resistant plants can survive subzero temperatures by withstanding extracellular ice formation. During cold acclimation, their leaves accumulate antifreeze proteins (AFPs) that are secreted into the apoplast and have the ability to modify the normal growth of ice crystals. Three barley, two wheat and two rye cultivars were grown under two different temperature regimes (20/16$^{\circ}C$ and 5/2$^{\circ}C$, day/night). Apoplastic proteins from winter cereals were separated by SDS-PAGE and detected with antisera to AFPs from winter rye. Apoplastic proteins accumulated to much higher levels in cold-acclimated (CA) leaves compared with nonacclimated (NA) ones in winter cereals. After cold acclimation, the protein concentration of apoplastic extracts increased significantly from 0.088 $mgmL^{-1}$ to 0.448 $mgmL^{-1}$, with about 5-fold increment. Also, the apoplastic protein content per gram leaf fresh weight in CA leaves ranged from 31 $\mu\textrm{g}$ $(gFW)^{-1}$ to 120 $\mu\textrm{g}$ $(gFW)^{-1}$ with an averaged value of 77 $\mu\textrm{g}$ $(gFW)^{-1}$, and coefficients of variation of 54.9%. The CA leaves in Musketeer (a Canadian winter rye cultivar) showed the greatest AFPs and antifreeze activity followed by 'Geurumil' (a Korean winter wheat cultivar), and 'Dongbori l' (Korean facultative barley cultivar). The proteins secreted into the wheat leaf apoplast at CA condition were more numerous than those observed in winter rye, where two $\beta$-1,3-glucanase-like proteins (GLPs), two chitinase-like proteins (CLPs) and two thaumatin-like proteins (TLPs) accumulated during cold acclimation. The proteins in barley leaf apoplast at CA conditions were a little different from those in wheat leaves. The AFPs were various among and within species. More freezing-resistant cultivars had more clear and numerous bands than less freezing-resistant ones. The high determination coefficient ($R^2$ =91 %) between freezing resistance and AFPs per gram leaf fresh weight indicated that the amount of AFPs was highly related to freezing resistance in winter cereal crops.

• 원예과학기술지
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• 제31권3호
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• pp.275-281
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• 2013

Low temperature is one of the major environmental factors that affect productivity including reduced growth and budding of vines, and changes of metabolic processes in grape (Vitis spp.). To screen the specific expression of abiotic stress-related genes against cold treatment in 'Kyoho' and 'Campbell Early' grapevines, expression of various defense-related genes was investigated by RT-PCR and real-time PCR. Among the 67 genes analyzed by RT-PCR and real-time PCR, 17 and 16 types of cDNA were up-regulated, while 5 and 6 types were down-regulated in cold-treated 'Kyoho' and 'Campbell Early' grapevines, respectively. Genes encoding carotene (Cart3564 and Cart4472), chalcone isomerase (CHI), cytochrome P450 (CYP), flavonol synthase (FLS), endo-${\beta}$-glucanase precursor (Glu), glutathione peroxidase (GPX), glutathione-S-transferase (GST), leucine-rich repeats (LRR), manganese superoxide dismutase (Mn-SOD), phenylalanine ammonia lyase (PAL), polygalacturonase-inhibiting protein (PGIP), proline rich protein 2 (PRP2), small heat shock protein (sHSP), temperature induced lipocalin (TIL), and thaumatin-like protein (TLP) were up-regulated, while those encoding CBF like transcription factor (CBF1), chitinase-like protein (CLP), cold induced protein (CIP), glycerol-3-phosphate acyltransferase (GPAT), and mitogen-activated protein kinase (MAPK) were down-regulated by low temperature treatment in both in 'Kyoho' and 'Campbell Early'.

• Journal of Plant Biotechnology
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• 제30권4호
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• pp.315-321
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• 2003

고비사막 접경지역의 식물인 Artermisia adamsii의 내건성 및 내동성 증진을 위해 조직배양과 CLP및 Dhn5유전자의 형질전환을 수행하였다. 다양한 호르몬 농도 조건에서의 성장을 확인한 결과, 0.05mg/L의 NAA와 0.5mg/L의 BAP조건과 0.1mg/L의 NAA와 0.5mg/L의 BAP가 포함된 배지의 암조건 하에서 최적의 캘러스 생장을 확인하였으며 유전자 도입 및 유전자의 발현이 확인된 캘러스가 세포 수준에서도 내건성 및 내동성이 증진되었음을 확인하였다. 그러나 Atremisia속 다른 식물과 다르게 조직절편에서 직접 기관분화를 유도하거나 캘러스를 통한 식물체 재생에 어려움이 있어 식물체 수준에서 형질전환에 따른 환경스트레스 내성의 증진을 확인하지 못하고 있다. 앞으로 진행될 A. adamsii의 식물체 재생에 대한 연구를 통해 내동성 및 내건성이 증진된 식물체를 육성하여 고비사막 지역적응력을 조사할 예정이다.

• 한국수산과학회지
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• 제48권4호
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• pp.439-446
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• 2015

Inflammation is a protective response to infection or injury. However, prolonged inflammation can contribute to the pathogenesis of many diseases, such as cancer, diabetes, arthritis, atherosclerosis, and Alzheimer's disease. Recent studies have shown that activated macrophages, inflammatory effector cells, can react to tissue insults in a polarized manner, in which their phenotypes are polarized into two major subtypes, categorized as M1 or M2. Classical M1 activation involves the production of pro-inflammatory cytokines, such as interleukin (IL)-6 and tumor necrosis factor (TNF)-${\alpha}$, and free radicals, while M2 or alternative activation is an anti-inflammatory phenotype involved in homeostatic processes, such as wound healing, debris scavenging, and the dampening of inflammation via the production of very low levels of pro-inflammatory cytokines and high levels of anti-inflammatory mediators, including IL-10. As part of our ongoing effort to isolate anti-inflammatory compounds from seaweeds, we investigated the effects of phlorotannins isolated from the brown alga Ecklonia stolonifera on macrophage polarization. Mouse peritoneal macrophages were treated with various concentrations of the extracts, and real-time RT-PCR analyses were performed to examine the expression of polarization markers: IL-$1{\beta}$, IL-6, and TNF-${\alpha}$ for M1 and arginase-1, peroxisome proliferator-activated receptor (PPAR)-${\gamma}$, found inflammatory zone-1 (Fizz-1), chitinase 3-like 3 (Ym1), and$Kr{\ddot{u}}ppel$-like factor 4 (Klf-4) for M2. The pretreatment of cells with eckol, dieckol, and phlorofucofuroeckol-A (PFF-A), isolated from the ethyl acetate fraction of E. stolonifera ethanolic extract, potentiated the anti-inflammatory M2 phenotype of the macrophages. These results indicate that phlorotannins derived from E. stolonifera can be used to enrich macrophages with markers of the M2 anti-inflammatory state.

• The Plant Pathology Journal
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• 제34권6호
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• pp.555-566
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• 2018

Two rhizobacteria Bacillus aryabhattai H26-2 and B. siamensis H30-3 were evaluated whether they are involved in stress tolerance against drought and high temperature as well as fungal infections in Chinese cabbage plants. Chinese cabbage seedlings cv. Ryeokgwang (spring cultivar) has shown better growth compared to cv. Buram-3-ho (autumn cultivar) under high temperature conditions in a greenhouse, whilst there was no difference in drought stress tolerance of the two cultivars. In vitro growth of B. aryabhattai H26-2 and B. siamensis H30-3 were differentially regulated under PEG 6000-induced drought stress at different growing temperatures (30, 40 and $50^{\circ}C$). Pretreatment with B. aryabhattai H26-2 and B. siamensis H30-3 enhanced the tolerance of Chinese cabbage seedlings to high temperature, but not to drought stress. It turns out that only B. siamensis H30-3 showed in vitro antifungal activities and in planta crop protection against two fungal pathogens Alternaria brassicicola and Colletotrichum higginsianum causing black spots and anthracnose on Chinese cabbage plants cv. Ryeokgwang, respectively. B. siamensis H30-3 brings several genes involved in production of cyclic lipopeptides in its genome and secreted hydrolytic enzymes like chitinase, protease and cellulase. B. siamensis H30-3 was found to produce siderophore, a high affinity iron-chelating compound. Expressions of BrChi1 and BrGST1 genes were up-regulated in Chinese cabbage leaves by B. siamensis H30-3. These findings suggest that integration of B. aryabhattai H26-2 and B. siamensis H30-3 in Chinese cabbage production system may increase productivity through improved plant growth under high temperature and crop protection against fungal pathogens.