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Lactobacillus casei Secreting ${\alpha}$-MSH Induces the Therapeutic Effect on DSS-Induced Acute Colitis in Balb/c Mice

  • Yoon, Sun-Woo;Lee, Chul-Ho;Kim, Jeong-Yoon;Kim, Jie-Youn;Sung, Moon-Hee;Poo, Har-Young
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1975-1983
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    • 2008
  • The neuropeptide ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) has anti-inflammatory property by down regulating the expressions of proinflammatory cytokines. Because ${\alpha}$-MSH elicits the anti-inflammatory effect in various inflammatory disease models, we examined the therapeutic effect of oral administration of recombinant Lactobacillus casei, which secretes ${\alpha}$-MSH (L. casei-${\alpha}$-MSH), on dextran sulfate sodium (DSS)-induced colitis in Balb/c mice. Thus, we constructed the ${\alpha}$-MSH-secreting Lactobacillus casei by the basic plasmid, pLUAT-ss, which was composed of a PldhUTLS promoter and ${\alpha}$-amylase signal sequence from Streptococcus bovis strain. Acute colitis was induced by oral administration of 5% DSS in drinking water for 7 days. To investigate the effect of L. casei-${\alpha}$-MSH on the colitis, L. casei or L. casei-${\alpha}$-MSH was orally administered for 7 days and their effects on body weight, mortality rate, cytokine production, and tissue myeloperoxidase (MPO) activity were observed. Administration of L. casei-${\alpha}$-MSH reduced the symptom of acute colitis as assessed by body weight loss (DSS alone: $14.45{\pm}0.2\;g$; L. casei-${\alpha}$-MSH: $18.2{\pm}0.12\;g$), colitis score (DSS alone: $3.6{\pm}0.4$; L. casei-${\alpha}$-MSH: $1.4{\pm}0.6$), MPO activity (DSS alone: $42.7{\pm}4.5\;U/g$; L. casei-${\alpha}$-MSH: $10.25{\pm}0.5\;U/g$), survival rate, and histological damage compared with the DSS alone mice. L. casei-${\alpha}$-MSH-administered entire colon showed reduced in vitro production of proinflammatory cytokines and $NF-{\kappa}B$ activation. The ${\alpha}$-MSH-secreting recombinant L. casei showed significant anti-inflammatory effects in the murine model of acute colitis and suggests a potential therapeutic role for this agent in clinical inflammatory bowel diseases.

Interaction of Pseudostellaria heterophylla with Quorum Sensing and Quorum Quenching Bacteria Mediated by Root Exudates in a Consecutive Monoculture System

  • Zhang, Liaoyuan;Guo, Zewang;Gao, Huifang;Peng, Xiaoqian;Li, Yongyu;Sun, Shujing;Lee, Jung-Kul;Lin, Wenxiong
    • Journal of Microbiology and Biotechnology
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    • v.26 no.12
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    • pp.2159-2170
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    • 2016
  • Many plant-pathogenic bacteria are dependent on quorum sensing (QS) to evoke disease. In this study, the population of QS and quorum quenching (QQ) bacteria was analyzed in a consecutive monoculture system of Pseudostellaria heterophylla. The isolated QS strains were identified as Serratia marcescens with SwrIR-type QS system and exhibited a significant increase over the years of monoculture. Only one QQ strain was isolated from newly planted soil sample and was identified as Bacillus thuringiensis, which secreted lactonase to degrade QS signal molecules. Inoculation of S. marcescens to P. heterophylla root could rapidly cause wilt disease, which was alleviated by B. thuringiensis. Furthermore, the expression of lactonase encoded by the aiiA gene in S. marcescens resulted in reduction of its pathogenicity, implying that the toxic effect of S. marcescens on the seedlings was QS-regulated. Meanwhile, excess lactonase in S. marcescens led to reduction in antibacterial substances, exoenzymes, and swarming motility, which might contribute to pathogensis on the seedlings. Root exudates and root tuber extracts of P. heterophylla significantly promoted the growth of S. marcescens, whereas a slight increase of B. thuringiensis was observed in both samples. These results demonstrated that QS-regulated behaviors in S. marcescens mediated by root exudates played an important role in replanting diseases of P. heterophylla.

Genotoxicity and Anti-Oxidative Effectiveness Study of Functional Food Additive Containing Astaxanthin (Astaxanthin 함유 기능성 식품소재의 유전독성 및 항산화능 검사)

  • Kim, Jun-Sung;Park, Jin-Hong;Jin, Hua;Cho, Hyun-Sun;Hwang, Soon-Kyung;Nah, Woon-Seong;Kang, Hwan-Goo;An, Gil-Hwan;Cho, Myung-Haing
    • Toxicological Research
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    • v.22 no.4
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    • pp.381-390
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    • 2006
  • Astaxanthine is a pigment that belongs to the family of the xanthophylls, the oxygenated derivatives of carotenoids whose synthesis in plants derives from lycopene. Astaxanthine is also a carotenoid widely used in salmonid and crustacean aquaculture to provide the pink color characteristic of that. Recent study reported that astaxanthine has the role as a detoxicant against the free radicals. On our study, we estimated the genotoxicity in ICR mice and possibility as antioxidant reagents of mutant Phaffia rhodozyma strain over expressing the astaxanthine by gamma-lay and carophyll pink including astaxanthine in apoE knock out mice, respectively. In our study, we administered Phaffia rhodozyma (2 mg and 3 mg) and carophyll pink for 4 and 8 week. The clinical sign and mortality were not detected compared with control groups. In the mutant frequency of hprt gene and chromosome aberration in splenic cells, there was not detected abnormality. There was not critical change in hematological and serum biochemical test compared to control. In expression level of repair enzyme, increase of catalase were detected and increase of expression level of Nrf-2 was detected in Phaffia rhodozyma (3 mg) and carophyll pink in 8 week treated group. In GSH level, the group of treated with Phaffia rhodozyma (3 mg) showed the increase of the GSH. In conclusion, mutant Phaffia rhodozyma and caphyll pink may be applied to the effective food additives to reduce the free radical.

Evaluation of at Rest Lateral Stress Coefficient Influenced by Particle Condition (입자의 조건에 따른 정지토압계수 평가)

  • Lee, Jung-Hwoon;Lee, Dong-Ryeol;Yun, Tae-Sup;Lee, Jun-Hwan
    • Journal of the Korean Geotechnical Society
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    • v.28 no.8
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    • pp.21-29
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    • 2012
  • At-rest lateral stress coefficient that is used for the evaluation of geotechnical structures such as foundations and retaining walls plays a significant role in the analysis and design, as a state variable of in-situ stress condition. In the widely applied Jaky's Ko equation stress condition can be inferred from the internal friction angle obtainable from the laboratory experimentation whereas the eguation mares it challenging to evaluate the influences and criteria of particle characteristics which is essential for the application of friction angles in practices. Thus, this study experimentally explored the behaviors of Ko depending on the relative density, particle shape, and surface roughness effect during a range of loading stages. The Ko values of Jumumjin sand, glass beads, and etched glass beads were measured using a customized Ko device housing strain gauges during loading-unloading-reloading steps, and the effect of dominant factors on Ko is analyzed. Results show that the high Ko prevails for both round and angular specimens with low relative density and the surface roughness has a nominal effect. The angular particles exhibit low Ko for specimens with similar relative density. The characteristics of relevance between Ko and friction angles with varying relative density are also investigated based on the experimental results using empirical correlations and previously reported values.

Isolation of Aeromonas sobria Containing Hemolysin Gene from Arowana (Scleropages formosus) (Arowana(Scleropages formosus)에서 Hemolysin Gene을 지닌 Aeromonas sobria 분리 및 특성)

  • Jun, Jin-Woo;Kim, Ji-Hyung;Casiano, Choresca Jr.;Dennis, K. Gomez;Shin, Sang-Phil;Han, Jee-Eun;Park, Se-Chang
    • Journal of Veterinary Clinics
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    • v.27 no.1
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    • pp.62-65
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    • 2010
  • Arowana (Scleropages formosus) is the most valuable group of ornamental fishes and very much in demand in the ornamental fish trade and commands high price ranging from hundreds to thousands of dollars per fish. In this paper, we described a case of mortality of arowana from a private aquarium in Korea. A bacterial pathogen from fish organs (brain, kidney, liver) was cultured, identified and confirmed using Vitek System 2, API 20E test, multiplex PCR and 16S rRNA gene sequencing. The morphological and biochemical properties of the bacterium isolated from the brain, kidney and liver of the fish were similar to Aeromonas sobria. Positive amplification products using the multiplex PCR assay for detection of A. sobria were obtained from these organs. The 16S rRNA gene of the isolates from fish was identical and exhibited 100% sequence similarity with A. sobria (AY987762.1) strain available from GenBank. This bacterium contained hemolysin gene, a virulence factor that plays an important role in outbreaks of disease and is pathogenic to humans as well as in fish. Although this opportunistic bacterium was isolated from a fish without any external symptoms, this pathogen may act as a reservoir and enhance chances of zoonosis to human such as during handling.

Diversity and Phylogenetic Analysis of Culturable Marine Bacteria Isolated from Rhizosphere Soils of Suaeda japonica Makino in Suncheon Bay (순천만 칠면초의 근권으로부터 분리된 해양세균의 다양성 및 계통학적 분석)

  • You, Young-Hyun;Park, Jong Myong;Nam, Yoon-Jong;Kim, Hyun;Lee, Myung-Chul;Kim, Jong-Guk
    • Journal of Life Science
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    • v.25 no.2
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    • pp.189-196
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    • 2015
  • Bacterial diversity was studied in the rhizosphere of Suaeda japonica Makino, which is native to Suncheon Bay in South Korea. Soil samples from several sites were diluted serially, and pure isolation was performed by subculture using marine agar and tryptic soy agar media. Genomic DNA was extracted from 29 pure, isolated bacterial strains, after which their 16S rDNA sequences were amplified and analyzed. Phylogenetic analysis was performed to confirm their genetic relationship. The 29 bacterial strains were classified into five groups: phylum Firmicutes (44.8%), Gamma proteobacteria group (27.6%), Alpha proteobacteria group (10.3%), phylum Bacteriodetes (10.3%), and phylum Actinobacteria (6.8%). The most widely distributed genera were Bacillus (phylum Firmicutes), and Marinobacterium, Halomonas, and Vibrio (Gamma proteobacteria group). To confirm the bacterial diversity in rhizospheres of S. japonica, the diversity index was used at the genus level. The results show that bacterial diversity differed at each of the sampling sites. These 29 bacterial strains are thought to play a major role in material cycling at Suncheon Bay, in overcoming the sea/mud flat-specific environmental stress. Furthermore, some strains are assumed to be involved in a positive interaction with the halophyte S. japonica, as rhizospheric flora, with induction of growth promotion and plant defense mechanism.

Antifungal Activity of Bacillus Subtilis HK2 against Trichothecium Roseum Causing Pink Rot of Melon and White Stain Symptom on Grape (멜론 분홍빛썩음병과 포도 흰얼룩병의 원인균인 Trichothecium Roseum에 대한 Bacillus Subtilis HK2의 항균활성)

  • Oh, Soh-Young;Lee, En-Young;Nam, Ki-Woong;Yoon, Deok-Hoon
    • Korean Journal of Plant Resources
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    • v.29 no.1
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    • pp.39-45
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    • 2016
  • Pink Rot on melon and White Stain Symptom on grape are caused by Trichothecium roseum, one of the most important diseases of grape and melon. These diseases have been occurred in national-wide in Korea and causes irreversible damage on the grape and the melon at harvest season. This research presents the evaluation of the capacity of Bacillus subtillis HK2 to protect both melon and grape against T. reseum and establishes its role as a biocontrol agent. In this study, we isolated a Bacillus strain HK2 from rhizosphere soil, identified it as Bacillus subtillis by 16S rRNA analysis and demonstrated its antifungal activity against T. roseum. Under I-plate assay it was observed that the effect of hyphal growth inhibition was not due to production of volatile compounds. The optimum culture condition of HK2 was found at 30℃ and initial pH of 7.0. Application of HK2 culture suspension reduced 90.2% of white stain symptom on grape as compared to control, resulting in greater protection to grape against T. roseum infestation. Butanol extract of HK2 culture purified using flash column chromatography. The antifungal material was a polar substance as it showed antifungal activity in polar elute. Therefore, our results indicated a clear potential of B. subtilis HK2 to be used for biocontrol of Pink rot in melon and white stain symptom on grape caused by T. roseum.

Rapid in vivo Colonization Screening of Probiotic Bacteria Isolated from Human Infants using Caenorhabditis elegans Surrogate Host (Caenorhabditis elegans 생체대체모델을 이용한 한국 영유아분변 유래 프로바이오틱스 균주의 in vivo 장 우점능 검토)

  • Park, Miri;Jeong, Eun-Seon;Oh, Sangnam;Song, Min-Ho;Doo, Jae-Kyun;Jeong, Yong-Seob;Moon, Yong-Il;Kim, Younghoon
    • Food Science of Animal Resources
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    • v.33 no.4
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    • pp.522-530
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    • 2013
  • The ability of probiotics to adhere to the intestinal epithelium likely plays an important role in their colonization of the gastrointestinal tract. Here, we performed high-throughput screening (HTS) for suitable characteristics of potential probiotic bacteria using attachment and colonization ability through a C. elegans surrogate in vivo model. A total of 100 strains of lactic acid bacteria (LAB) isolated from infant feces were subjected to the colonization assay using C. elegans intestine. Based on colonization ability, we showed that nine isolates have a high attachment ability during whole experimental periods (up to 168 h), compared to Lactobacillus rhamnosus strain GG as a control. Also, through the use of an in vitro cell attachment model, nine isolates revealed highly binding activity to the mucus layer. Next, the selected 9 isolates were assayed for their survival ability when exposed to acidic and bile conditions as well as cholesterol reduction and the utilization of prebiotic substrates. As a result, the isolated nine strains were determined to be highly resistant to acid and bile conditions. In addition, they have significant activity for the reduction of cholesterol and utilization of several prebiotic substrates as a carbon source. Finally, the selected nine strains were identified by either L. rhamnosus or L. plantarum (4 strains for L. rhamnosus and 5 strains for L. plantarum, respectively). Taken together, we propose that the direct colonization of probiotics using C. elegans may be applicable to the rapid screening of valuable probiotic strains in vivo.

A Basic Study on Micro-Electric Potential accompanied with Specimen Failure during Uniaxial Compressive Test (일축 압축에 의한 시료 파괴 시 수반되는 미소 전위에 대한 기초 연구)

  • Kim, Jong-Wook;Park, Sam-Gyu;Song, Young-Soo;Sung, Nak-Hun;Kim, Jung-Ho;Cho, Seong-Jun
    • Geophysics and Geophysical Exploration
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    • v.10 no.3
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    • pp.203-210
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    • 2007
  • As a part of basic studies on monitoring of landslides and slope stability using SP measurements, micro-electric potentials of rock samples were measured accompanied with the rock failure by a uniaxial loading test were measured. The measurement system consists of a 8 channel A/D converter with 24 bit resolution, uniaxial loading tester, strain gages and 4 sets of electrode attached to a rock sample. Rock samples of granite, limestone, and sandstone were tested. Also, mortar samples were tested in order to monitor electric-potentials of a uniform sample. Micro-electric potentials were detected in all saturated samples and the strength of them increased as the loading force increased. Sandstone samples showed the largest strength of micro-electric potential and it followed limestone and granite samples, which indicates a positive relationship with porosity of rocks. The mechanism generating these micro-electric potential can be explained in terms of electro-kinetics. In case of dry samples, micro-electric potential could be observed only in sandstone samples, where piezoelectric effect played main role due to high contents of quartz in sandstone samples. We found that biggest micro-electric potentials were observed at the electrodes near the crack surface of rock samples. This is very encouraging result that SP monitoring can be applied to predicting landsliding or to estimate collapsing position combining with monitoring of acoustic emissions.

Technique for the Measurement of Crack Widths at Notched / Unnotched Regions and Local Strains (콘크리트의 노치 및 비노치 구역에서의 균열폭 및 국부 변형률 정밀 측정기법)

  • Choi, Sok-Hwan;Lim, Bub-Mook;Oh, Chang-Kook;Joh, Chang-Bin
    • Journal of the Korea Concrete Institute
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    • v.24 no.2
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    • pp.205-214
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    • 2012
  • Crack widths play an important role in the serviceability limit state. When crack widths are controlled sufficiently, the reinforcement corrosion can be reduced using only existing concrete cover thickness due to low permeability in the region of finely distributed hair-cracks. Thus, the knowledge about the tensile crack opening is essential in designing more durable concrete structures. Therefore, numerous researches related to the topic have been performed. Nevertheless accurate measurement of a crack width is not a simple task due to several reasons such as unknown potential crack formation location and crack opening damaging strain gages. In order to overcome these difficulties and measure precise crack widths, a displacement measurement system was developed using digital image correlation. Accuracy calibration tests gave an average measurement error of 0.069 pixels and a standard deviation of 0.050 pixels. Direct tensile test was performed using ultra high performance concrete specimens. Crack widths at both notched and unnotched locations were measured and compared with clip-in gages at various loading steps to obtain crack opening profile. Tensile deformation characteristics of concrete were well visualized using displacement vectors and full-field displacement contour maps. The proposed technique made it possible to measure crack widths at arbitrary locations, which is difficult with conventional gages such as clip-in gages or displacement transducers.