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Expression of amino acid transport system L in the differentiation of periodontal ligament fibroblast cells (치주인대섬유모세포의 분화과정에서 아미노산 수송계 L의 발현)

  • Hwang, Kyu-Young;Kim, Do-Kyung;Kim, Chang-Hyun;Jang, Hyun-Seon;Park, Joo-Cheol;Choi, Seong-Mi;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.36 no.3
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    • pp.783-796
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    • 2006
  • The periodontium is a topographically complex organ consisting of epithelial tissue, soft and mineralized tissues. Structures comprising the periodontium include the gingiva, periodontal ligament (PDL) , cementum and the alveolar bone. The molecular mechanism of differentiation in PDL fibroblast cells remain unclear. Amino acid transporters play an important role in supplying nutrition to normal and cancer cells and for cell proliferation. Amino acid transport system L is a major nutrient transport system responsible for the Na+-independent transport of neutral amino acids including several essential amino acids. The system L is divided into two major subgroups, the L-type amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2). In this study, the expression pattern of amino acid transport system L was, therefore, investigated in the differentiation of PDL fibroblast cells. To determine the expression level of amino acid transport system L participating in intracellular transport of amino acids in the differentiation of PDL fibroblast cells, it was examined by RT-PCR, observation of cell morphology, Alizaline red-S staining and uptake analysis after inducing experimental differentiation in PDL fibroblast cells isolated from mouse molar teeth. The results are as follows. 1. The LAT1 mRNA was expressed in the early stage of PDL fibroblast cell differentiation. This expression level was gradually reduced by differentiation- inducing time and it was not observed after the late stage. 2. The expression level of LAT2 mRNA was increased in time-dependent manner during differentiation induction of PDL fibroblast cells. 3. There was no changes in. the expression level of 4F2hc mRNA, the cofactor of LAT1 and LAT2, during differentiation of PDL fibroblast cells. 4. The expression level of ALP mRNA was gradually increased and the expression level of Col I mRNA was decreased during differentiation of PDL fibroblast cells. 5. The L-leucine transport was reduced by time from the early stage to the late stage in PDL fibroblast cell differentiation. As the results, it is considered that among neutral ammo acid transport system L in differentiation of PDL fibroblast cells, the LATl has a key role in cell proliferation in the early stage of cell differentiation and the LAT2 has an important role in the late stage of cell differentiation for providing cells with neutral amino acids including several essential amino acids.

The Effects of Samul-tang-ga-dansam for Wound Healing (사물탕(四物湯) 가(加) 단참(丹參)의 상처 치료에 대한 효과)

  • Eun-Byeol Lee;Hyeon-Ji Kim;Chae-Young Kim;Ji-Su Choi;Chang-Hoon Woo;Young-Jun Kim;Hee-Duk An
    • Journal of Korean Medicine Rehabilitation
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    • v.33 no.2
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    • pp.1-18
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    • 2023
  • Objectives The purpose of this study was to evaluate the antioxidant, anti-inflammatory and wound healing effects of Samul-tang-ga-dansam water extract (SD) in wound-induced mice. Methods The mice were divided into five groups (n=7): the normal group, the control group, the positive control group, the low-dose SD group and the high-dose SD group. The normal group had no wounds and the other groups were wounded on the back with a leather punch. Distilled water was administered to the control group, 200 mg/kg of vitamin E was administered to the positive control group. In the low-dose SD group and the high-dose SD group, 1.23 g/kg and 2.47 g/kg of SD were administered, respectively. Antioxidant and anti-inflammatory protein levels were evaluated using western blot analysis. Skin tissue was analyzed by H&E, Masson's trichrome staining method. Results Oral administration of the SD significantly reduced the visible skin damage and decreased the reactive oxygen species and ONOO- activity of the serum. It significantly increased heme oxygenase-1, superoxide dismutase, catalase, GPx-1/2, Nrf2 and Keap-1 which are antioxidant-related factors in skin tissue and reduced NF-κB p65, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor α, interleukin (IL)-1β, IL-6 which are inflammation-related factors. Also, SD significantly decreased NOX2, p22phox and p47phox and increased α-smooth muscle actin and COL1A1 protein expression in fibroblasts involved in connective tissue repair. According to histological examination, the thickened epithelial layer was thinned and collagen fibers were increased to accelerate wound healing. Conclusions It is suggested that Samul-tang-ga-dansam has antioxidant and anti-inflammatory effects and promotes wound tissue repair.

Genetic diversity and selection of Tibetan sheep breeds revealed by whole-genome resequencing

  • Dehong Tian;Buying Han;Xue Li;Dehui Liu;Baicheng Zhou;Chunchuan Zhao;Nan Zhang;Lei Wang;Quanbang Pei;Kai Zhao
    • Animal Bioscience
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    • v.36 no.7
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    • pp.991-1002
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    • 2023
  • Objective: This study aimed to elucidate the underlying gene regions responsible for productive, phenotypic or adaptive traits in different ecological types of Tibetan sheep and the discovery of important genes encoding valuable traits. Methods: We used whole-genome resequencing to explore the genetic relationships, phylogenetic tree, and population genetic structure analysis. In addition, we identified 28 representative Tibetan sheep single-nucleotide polymorphisms (SNPs) and genomic selective sweep regions with different traits in Tibetan sheep by fixation index (Fst) and the nucleotide diversity (θπ) ratio. Results: The genetic relationships analysis showed that each breed partitioned into its own clades and had close genetic relationships. We also identified many potential breed-specific selective sweep regions, including genes associated with hypoxic adaptability (MTOR, TRHDE, PDK1, PTPN9, TMTC2, SOX9, EPAS1, PDGFD, SOCS3, TGFBR3), coat color (MITF, MC1R, ERCC2, TCF25, ITCH, TYR, RALY, KIT), wool traits (COL4A2, ERC2, NOTCH2, ROCK1, FGF5, SOX9), and horn phenotypes (RXFP2). In particular, a horn-related gene, RXFP2, showed the four most significantly associated SNP loci (g. 29481646 A>G, g. 29469024 T>C, g. 29462010 C>T, g. 29461968 C>T) and haplotypes. Conclusion: This finding demonstrates the potential for genetic markers in future molecular breeding programs to improve selection for horn phenotypes. The results will facilitate the understanding of the genetic basis of production and adaptive unique traits in Chinese indigenous Tibetan sheep taxa and offer a reference for the molecular breeding of Tibetan sheep.

Physico-Chemical Properties and Antimicrobial Activity of Pyocyanine Produced by Pseudomonase aeruginosa KLP-2 (Pseudomonas aeruginosa KLP-2가 생산한 Pyocyanine의 항균활성 및 생리화학적 성상)

  • 박은희;이상준;차인호
    • Journal of Life Science
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    • v.11 no.5
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    • pp.483-488
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    • 2001
  • The antimicrobial substance produced by Pseudomonas aeruginosa KLP-2 strain was purified and identified. The substance was identified as a pyocyanine by the fast atom bombardment mass(FAB-MS). In physic-chemical properties, the pyocyanine was dark blue needles, and was soluble in various organic solvents such as chlorogorm, methanol, ethanol and ethyl acetae. The pyocyanine possessed a ultraviolet absorbance spectrum in methanol, 0.1 M HCl, and chlorogorm. The maximum absorption peak of the pyocyanine showed at 318 mm in methanol. The molecular formula of the pyocyanine was determined to the $C_{13}$ H$_{10}$ N$_{2}$O and protonate molecular ion species (M+H)$^{+}$ was observed at m/z 211 by FAB-MS. The pyocyanine showed antimicrobial against Bacillus cereus, Bacillus subtilis, Micrococcus luteus, Rodococcus equi, Staphylococcus aureus, Streptococcus faecalis, E. col, Legionella pneumophila, Shigella flexneri Shigella boydii, shgella sonnei, NAG Vibrio cholerae, Vibrio parahaemolyticus, Vibro vulnificus, Yersinia enterocolitica, and Saccharomyces cerevisiae. However, Salmonella spp. Shigela dysenteriae, 3 strains of Pseudomonas aeruginosa, Klebsiela pneumoniae, and Aspergillus niger were resistant to the pyocyanine. The pyocyanine showed the highest antimicrobial activity aganist Legionella pneumophila based on the size of inhibition zone by the disk contained 0.5 $\mu\textrm{g}$ of the pyocyanine.e.

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Comparisons of physical Fitness, Self Efficacy, Instrumental Activities of Daily Living, and Quality of Life between Institutionalized and Noninstitutionalized Elderly (가정노인과 양로원노인의 체력, 자기효능, 일상생활활동능력 및 삶의 질에 관한 연구)

  • 노유자;김춘길
    • Journal of Korean Academy of Nursing
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    • v.25 no.2
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    • pp.259-278
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    • 1995
  • This study was done to investigate levels of physical fitness, self efficacy(SE), instrumental activities of daily living(IADL), and quality of life (QL), and their relationships among the elderly Data were collected from 47 noninstitutionalized elderly dwell-ing in their own homes and 43 institutionalized elderly living in homes for older people. The ages of the subjects were 65 years and over. The data were col lected from January 20th to February 20th, 1995. Physical fitness was measured with T.K. K dyna mometer, grip dynamometer, stop watch, and Purdue pegboard. Structured questionnaires developed by Kim by Lawton & Brody, and by Ro were adopted to measure SE, IADL, and QL, respectively. The data were analyzed using t-test, ANOVA, and Pearson correlation coefficients. The results were as follows : 1. The noninstitutionalized elderly had higher scores in such measures of physical fitness as body weight, skinfold thickness, flexibility and coordination as compared to the institutionalized elderly. 2. On the following measures of physical fitness, men showed higher levels than women ; in height when standing, height when sitting, body weight, and muscle strength. Women had more trunk flexibility than men. 3. Muscle strength was positively correlated with height when standing, height when sitting, and body weight. The following measures of physical fitness, muscle strength, flexibility and coordi nation, were negatively correlated with "up and go”. 4. The noninstitutionalized elderly had higher scores in SE(t=2.28, p<0.05), IADL(t=2.24, p<0.05), and QL(t=2.41, p<0.05) as compared to the institutionalized elderly. 5. SE was positively correlated with both IADL(r=0.41, p<0.001) and QL(r=0.54, p<0.001), and the latter two variables were a positively correlated (r=0.30, p<0.001). 6. SE was positively correlated with the physical fitness measures of height when standing, body weight, strength of leg muscles, strength of back muscles, grip strength, and level of touching the floor, but negatively correlated with "up and go”. 7. Positive correlations were revealed between IADL and the physical fitness measures of height, strength of leg muscles, level of trunk muscle extension, level of touching the floor, and coordination. The IADL was negatively correlated with “up and go”. 8. QL was positively correlated with body weight (r=0.28, p<0.01) and skinfold thickness (r=0.26, p<0.05). 9. Age was negatively correlated with the physical fitness measures of height when standing, height when sitting, “up and go”, strength of leg muscles, strength of back muscles, grip strength, level of trunk muscle extension, and coordination. 10. Age was also negatively correlated with SE (r=-0.24, p<0.05) and IADL(r=-0.22, p<0.05). The above results suggest that caring elderly in their own homes were more effective and that nursing interventions to enhance physical fitness, SE, IADL, and QL especially for the institutionalized elderly are stressed.

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Cloning and Characterization of a Cellulase Gene from a Plant Growth Promoting Rhizobacterium, Bacillus subtilis AH18 against Phytophthora Blight Disease in Red-Pepper (고추역병을 방제하는 PGPR균주 Bacillus subtilis AH18의 항진균성 Cellulase 유전자의 Cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Jung, Hee-Kyoung;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.311-317
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    • 2006
  • Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

The Effect of Trans-cinnamaldehyde on the Gene Expression of Lipopolysaccharide-stimulated BV-2 Cells Using Microarray Analysis (Trans-Cinnamaldehyde가 Lipopolysaccharide로 처리된 BV-2 cell에 미치는 항염증 기전 연구: Microarray 분석)

  • Sun, Young-Jae;Choi, Yeong-Gon;Jeong, Mi-Young;Hwang, Se-Hee;Lee, Je-Hyun;Cho, Jung-Hee;Lim, Sabina
    • The Journal of Korean Medicine
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    • v.30 no.4
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    • pp.13-27
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    • 2009
  • Objectives: Trans-cinnamaldehyde (TCA) is the main component of Cinnamomi Ramulus and it has been reported that TCA inhibits inflammatory responses in various cell types. Inflammation-mediated neurological disorders induce the activation of macrophages such as microglia in brain, and these activated macrophages release various inflammation-related molecules, which can be neurotoxic if overproduced. In this study, we evaluated gene expression profiles using gene chip microarrays in lipopolysaccharide (LPS)-stimulated BV-2 cells to investigate the antiinflammatory effect of TCA on inflammatory responses in brain microglia. Methods: A negative control group was cultured in normal medium and a positive control group was stimulated with $1{\mu}g/ml$ in the absence of TCA. TCA group was pretreated with $10{\mu}g/ml$ before $1{\mu}g/ml$ LPS stimulation. The oligonucleotide microarray analysis was performed to obtain the expression profiles of 28,853 genes using gene chip mouse gene 1.0 ST array in this study. Results: In positive control group, 1522 probe sets were up-regulated in the condition of the cutoff value of 1.5-fold change and 341 genes with Unigene ID were retrieved. In TCA group, 590 probe sets were down-regulated from among 1522 probe sets and 33 genes with Unigene ID were retrieved, which included 6 inflammation-related genes. We found out that Id3 gene is associated with transforming growth factor-${\beta}$ (TGF-${\beta}$) signaling pathway and Klra8 gene is related to natural killer cell-mediated cytotoxicity pathway. Conclusions: The results mean that TCA inhibits inflammatory responses through down-regulating the expressions of inflammation-related genes in LPS-stimulated BV-2 cells.

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Dormancy - breaking Conditions of Bulrush(Scirpus juncoides Roxb.) (올챙고랭이(Scirpus juncoides Roxb.) 종실(種實)의 휴면타파조건(休眠打破條件)에 관한 연구(硏究))

  • Huh, S.M.;Guh, J.O.;Son, P.K.
    • Korean Journal of Weed Science
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    • v.6 no.2
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    • pp.102-109
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    • 1986
  • To know the ecological pattern of bulrush (Scirpus juncoides) seeds in dormancy-breaking responses as affected by different ripening processes, storage conditions, germination conditions, and some of known chemicals concerned, the study was conducted. Among other conditions detected, the burial in 2 cm depth paddy soil, $5^{\circ}C$ storage temperature, pre-maturing process (Green color), high concentration of chemicals used, and flooding paddy surfaces were the most efficient conditions for bulrush seeds to break dormancy and germinate, respectively.

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Recovery of Intracellular Enzyme Using Phage Lysogen and Expanded Bed Adsorption (Phage Lysogen과 Expanded Bed Adsorption을 이용한 세포내 효소의 회수)

  • 장우진;윤현식구윤모
    • KSBB Journal
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    • v.11 no.6
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    • pp.663-668
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    • 1996
  • In the production of recombinant protein using E. coli, phage lysogen system can be usefully applied for simultaneously achieving protein production at high cell concentration and recovery by cell disruption in the same bioreactor. A major drawback of this system is that the intracellular product and complex broth components are mixed together in culture broth and hence purification efficiency is reduced. With the E. coli double-lysogen system, the expanded bed adsorption is very useful because the pretreatment processes in a routine bioseparation process can be done in a single column operation, and therefore may contribute towards lowering the operating cost of overall recovery/purification process. In the operation of EBA, it has been observed that the change in broth feed volume does not influence much the protein recovery in a tested range. The amount of protein adsorption per mL of resin was increased from $3.44{\times}106unit to 5.28{\times}106unit$ by doubling the column length. By two-fold increase of the column diameter, the ratio of protein concentration in eluent to that in feed was increased from 0.8 to 2.1. It is concluded from the present investigation that the increase of column length and diameter is necessary to enhance the protein adsorption amount per volume of resin and protein concentration in the eluent. The development of resins with various physical properties will be necessary for more extensive application of EBA.

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Evaluation of Syringaresinol Content and MC3T3E1 Osteoblast Differentiation of Fermented Extracts of Eleutherococcus senticosus Using Lactobacillus ssp. with High 𝛽-Glucosidase Activity (𝛽-Glucosidase 활성이 높은 유산균을 이용한 한국가시오갈피 발효 추출물의 Syringaresinol의 함량 및 MC3T3E1조골세포 분화 평가)

  • Minji Kang;Minkyoung Kang;Sangnam Oh
    • Journal of Dairy Science and Biotechnology
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    • v.42 no.2
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    • pp.48-63
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    • 2024
  • This study investigated the 𝛽-glucosidase activity of lactic acid bacteria and specifically eleutheroside E and B from Acanthopanax senticosus after bioconverting them into syringaresinol (SYR). Out of 125 lactic acid bacteria strains isolated from kimchi and other sources, 46 exhibiting both extracellular and internal 𝛽-glucosidase activity were identified. Notably, strains LFFR 20-011 (Lactobacillus curvatus) and LFFR 20-043/LFR20-050 (Levilactobacillus brevis) enhanced SYR production by more than two-fold during Acanthopanax senticosus fermentation. Further investigation revealed that SYR significantly promoted osteoblast differentiation, as evidenced by the increased mRNA expression levels of early and mature osteoblast markers, including Runx2, type I collagen, and osteocalcin. These findings suggested that the enhanced presence of SYR through bioconversion by Acanthopanax senticosus may improve bone health. These results provide foundational data supporting the development of lactic-acid-bacteria-fermented Acanthopanax senticosus as a functional food aimed at promoting skeletal health in older adults.