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Evaluation of Fermentation Ability of Microbes for Whole Crop Barley Silage Inoculant (보리 사일리지용 미생물의 발효능력 평가)

  • Kim, Jong-Geun;Ham, Jun-Sang;Chung, Eui-Soo;Park, Hyung-Soo;Lee, Joung-Kyong;Jung, Min-Woong;Choi, Ki-Choon;Cho, Nam-Chul;Seo, Sung
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.3
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    • pp.235-244
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    • 2009
  • This experiment was conducted to develop a new silage inoculant for barley at forage analysis laboratory, Grassland and Forages Division, National Institute of Animal Science, RDA from 2000 to 2002. Barley is very important crop in Korea. The great part of them is utilized as forage. Generally, it contains a lot of grains that are feed of animal, especially whole crop silage in ruminant. Efficient lactic acid bacteria were isolated from good barley silage by plating MRS agar containing 0.02% sodium azide, and assessed by growing and acid producing ability in MRS broth. Four lactic acid bacteria were selected, and were found to be Gram positive, rods and catalase negative and were identified to be Lactobacillus plantarum on the basis of the biochemical characteristics and utilization of substrates. Barley was ensiled at dough stage following treatment with four lactic acid bacteria, commercial inoculant, and no additive (control). After 2 months, B2-2 bacteria inoculated silage was lower pH and higher lactic acid content than others treatments. The Flieg's score and grade of B2-2 bacteria treated silage were higher than commercial inoculant. According to this experiment, Lactobacillus plantarum B2-2 (NLRI 201) was recommendable for good silage inoculant of whole crop barley silage.

Effects of β-Glucan from Paenibacillus polymyxa and L-theanine on Growth Performance and Immunomodulation in Weanling Piglets

  • Hwang, Y.H.;Park, B.K.;Lim, J.H.;Kim, M.S.;Song, I.B.;Park, S.C.;Jung, H.K.;Hong, J.H.;Yun, H.I.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.12
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    • pp.1753-1759
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    • 2008
  • Forty weanling piglets ($5.6{\pm}0.5kg$ and 26 to 30 d of age) were used in a 28-d experiment to determine the effects of ${\beta}$-glucan from Paenibacillus polymyxa and L-theanine on growth performance. Piglets were randomly allotted to four groups (n = 10, 2 animals per pen) provided with the basal feed (control), ${\beta}$-glucan 400 mg/kg feed, L-theanine 80 mg/kg feed or ${\beta}$-glucan plus l-theanine (combination of the above-mentioned concentrations). Body weight and feed consumption were recorded during four weeks. Subsequently, the immunomodulatory effects of ${\beta}$-glucan and L-theanine were investigated for lipopolysaccharide (LPS)-induced cytokine production in vitro and in vivo on day 28. Although there were no significant differences in the growth performances among the treatment groups, ${\beta}$-glucan plus L-theanine had 5.6% greater ADG (p = 0.074) on day 21 to 28. ${\beta}$-Glucan alone or plus L-theanine increased interleukin (IL)-10 levels and decreased interferon (IFN)-$\gamma$ and tumor necrosis factor (TNF)-${\alpha}$ levels in cultured medium by LPS treatment (p<0.05). Plasma IL-10 levels were also increased in the piglets fed with ${\beta}$-glucan alone or plus L-theanine after LPS challenge ($25{\mu}g/kg$, i.p.), whereas plasma IFN-$\gamma$ and TNF-${\alpha}$ levels were decreased (p<0.05). The levels of IFN$\gamma$ in piglets fed with ${\beta}$-glucan plus L-theanine showed the greatest inhibition after LPS challenges. In conclusion, treatment of ${\beta}$-glucan alone or plus L-theanine might lessen inflammatory responses against Gram-negative bacterial infection via the inhibition of pro-inflammatory cytokine production and enhancement of anti-inflammatory cytokine production. Further studies are needed to determine an optimal concentration of ${\beta}$-glucan and L-theanine for improved growth performance.

Function of Global Regulator CodY in Bacillus thuringiensis BMB171 by Comparative Proteomic Analysis

  • Qi, Mingxia;Mei, Fei;Wang, Hui;Sun, Ming;Wang, Gejiao;Yu, Ziniu;Je, Yeonho;Li, Mingshun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.152-161
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    • 2015
  • CodY is a highly conserved protein in low G+C gram-positive bacteria that regulates genes involved in sporulation and stationary-phase adaptation. Bacillus thuringiensis is a grampositive bacterium that forms spores and parasporal crystals during the stationary phase. To our knowledge, the regulatory mechanism of CodY in B. thuringiensis is unknown. To study the function of CodY protein in B. thuringiensis, BMB171codY- was constructed in a BMB171 strain. A shuttle vector containing the ORF of cry1Ac10 was transformed into BMB171 and BMB171codY-, named BMB171cry1Ac and BMB171codY-cry1Ac, respectively. Some morphological and physiological changes of codY mutant BMB171codY-cry1Ac were observed. A comparative proteomic analysis was conducted for both BMB171codY-cry1Ac and BMB171cry1Ac through two-dimensional gel electrophoresis and MALDI-TOF-MS/MS analysis. The results showed that the proteins regulated by CodY are involved in microbial metabolism, including branched-chain amino acid metabolism, carbohydrate metabolism, fatty acid metabolism, and energy metabolism. Furthermore, we found CodY to be involved in sporulation, biosynthesis of poly-β-hydroxybutyrate, growth, genetic competence, and translation. According to the analysis of differentially expressed proteins, and physiological characterization of the codY mutant, we performed bacterial one-hybrid and electrophoretic mobility shift assay experiments and confirmed the direct regulation of genes by CodY, specifically those involved in metabolism of branched-chain amino acids, ribosomal recycling factor FRR, and the late competence protein ComER. Our data establish the foundation for in-depth study of the regulation of CodY in B. thuringiensis, and also offer a potential biocatalyst for functions of CodY in other bacteria.

Biodistribution of [S-35] Labeled Antisense Oligodeoxynucleotides Increased Tumor Targeting With Microsphere Coinjection

  • Choe, Jae-Gol;Park, Gil-Hong;Claudio Nastruzzi;Yoon S. Cho-Chung;Kim, Meyoung-Kon
    • Environmental Mutagens and Carcinogens
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    • v.22 no.2
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    • pp.65-69
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    • 2002
  • To elucidate the effect of microsphere coinjection on the administration of oligodeoxynucleotides (ODN), we have investigated biodistribution of [S-35]-labeled antisense ODN targeted to cAMP-dependent protein kinase (PKA) RI-$\alpha$ subunit in nude mice xenografted with WiDr (human colon cancer, ATCC CCL218). The strategy of using microsphere has been proposed for cancer treatment as a carrier of therapeutic ODN so that it could offer an advantage with respect to maintaining constant ODN levels in blood and obtaining higher therapeutic ODN concentration at tumor sites. Comparative biodistribution studies were performed in nude mice (female, 20 g of body weight, n = 4-6) xenografted with WiDr cancer cells, when 0.1 $\mu$Ci (specific activity, 2.94 mCi/$\mu$mole) of [S-35]-labeled RI-$\alpha$ antisense ODN was injected alone or with microsphere (PLG-18, polylactic copolymer with cationic surfactant DDAB18). Peak tumor uptake of [S-35]-labeled ODN was significantly increased from 17.7% (at 6 h) of injected dose per gram of tissue (ID/g) to 42.5% (at 24 h) ID/g when microsphere was coinjected with ODN. The different biodistribution in the kidney accumulation (e.g., 100.2% ID/g for ODN alone and 54.9%/ID/g for microshpere coinjection) may contribute to higher blood concentration (e.g., 21.5%ID/$m\ell$ for ODN alone and 37.5%ID/$m\ell$ for microsphere coinjection) of radiolabeled ODN. Of importance is the fact that the whole body retention of radioactivity increased with microsphere coinjection from 50.8%ID/g to 68.0%ID/g after 24-h of injection. This decreased kidney accumulation and increased whole body retention of [S-35]-labeled ODN resulted in a significant improvement of ODN targeting to the tumor site. In conclusion, the coinjection of microsphere appears to be an important carrier system in vehiculation of antisense oligonucleotide to the tumor tissue in vivo.

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Investigation on Bacillus anthracis isolated from Kyong-Ju (경주에서 분리된 탄저균에 대한 연구)

  • 이준규;이은미;차우양;김정화;김영환;이양수;김우현;정종식
    • Korean Journal of Veterinary Service
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    • v.18 no.1
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    • pp.41-56
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    • 1995
  • The present study was conducted to investigate results of B. anthracis isolated from Anthrax in the Kyong-Ju of Feb. 12. 1994. 1. In biochemical feature, B. anthracis was a gram-positive rod, non-motility, sporulation, capsulation. It was positive in gelatinase, starch hydrolysis, glucose. But negative in urease, arabinose, mannitol, xylose. 2. B. anthracis grew well on B4 Br A TSA after incubation for 24 hours. The organisim grew well on BA, Br. A, NA, TSA after incubation for 72 hours. The media grew well on Br A instead of BA. 3. On 5% blood agar by laboratory animal, ${\beta}$ -hemolysis was produced from 36 hours to 48 hours incubation. There was perfect ${\beta}$-hemolysis after incubation for 48 hours. On the other side ${\beta}$-hemolysis was begun on 5% goat blood agar after incubation for 60 hours. 4. In the test of antimicrobial susceptibility, B. anthracis was very sensitive to AM, CF, TE, ENR, GM, AN, DFX, S, P, TYLO, N, KM, C, E, Lins+Sp, NN, CC, CFP, CB were sensitive one by one. B. anthracis was no-sensitive to L, XNL, TIA, CL, SXT 5. B. anthracis had never sensitivity to direct inoculation of rat and chicken, after subcutanous inj. It was very sensitive to mouse and goat, hamster, guinea pig, rabbit had a sensibility one by one. 6. The dead laboratory animal which had been inoculated with B. anthracis preserved at $37^{\circ}C$ incubation, B. anthracis didn't cultivate on non-dissected animal after 80 hours but cultivate on dissected animal after 360 hours. 7. The rapidly death could cause high concentration, died from 420 after S. C. 8. The blood smeared samples of hamster from inoculation with B. anthracis, spore germinated In 37$^{\circ}C$ after 5 hours, in $32^{\circ}C$ after 6 hours, in room temperature after 9 hours, in $-4^{\circ}C$ to $-20^{\circ}C$ after 10 hours. 9. B, anthracis inoculated to laboratory animal after SC or PO. Mice and rats feces didn't cultivated with B. anthracis after SC, but did cultivated with B. anthracis after PO. 10. In the test of disinfectant, B. anthracis was high effective to $HgC1_2$, formalin, effect phenol, cresol, but non-effect NaOH, ethanol.

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Changes in Renal Function by Nebulized Colistimethate Treatment (Colistimethate 분무요법 시행 환자에서 투여 전후 신기능의 변화)

  • Ahn, Hye Jin;Jung, Yoo Jin;Kim, Jae Song;Kim, Soo Hyun;Son, Eun Sun
    • Korean Journal of Clinical Pharmacy
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    • v.27 no.2
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    • pp.92-98
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    • 2017
  • Background: Nebulized colistimethate is increasingly used, because there are problems such as renal dysfunction and low distribution within the lungs when colistimethate is administered intravenously. This study was designed to compare and analyze the changes in renal function by of nebulized colistimethate treatment for its safe administration. Methods: This study retrospectively reviewed the electronic medical records of adult patients above 19 years old, receiving only the nebulized colistimethate at least 4 days in Yonsei university health system from Nov 2014 to Aug 2015. Acute kidney injury (AKI) was determined by using the RIFLE criteria (Risk, Injury, Failure, Loss and End-stage renal disease) according to serum creatinine (SCr) levels before and after use of nebulized colistimethate. Results: 48 patients were included our study and their SCr increased significantly after nebulized colistimethate treatment ($SCr_0$ vs. $SCr_1$; $0.85{\pm}0.80$ vs. $1.00{\pm}0.82mg/dL$, n=48, p<0.001), but the changes were in normal range according to the standards at Yonsei university health $system^a$. Among 48 patients, 38 patients were in the non-AKI group (79.2%), and 10 patients developed AKI (20.8%). Within the AKI group, 2 patients were in the Injury group (20%) and the other 8 in the Risk group (80%). Conclusion: There was no significant difference in age, dosage and duration of treatment between AKI group and non-AKI group (p>0.05). The study has a significance in that it reviewed the safety of nebulized colistimethate only treatment to national patients, analyzing its nephrotoxicity. It has confirmed that nebulized colistimethate is a safer method than intravenous injection, and requires to establish a guideline for the use of nebulized colistimethate in further studies with broader patient groups. $^a$ : SCr Male 0.68-1.19 mg/dL, Female 0.49-0.91 mg/dL.

Screening of Biologically Active Compounds from Weeds I (잡초(雜草)에 함유(含有)된 생리활성물질(生理活性物質) 탐색(探索) I)

  • Kim, C.J.;Kang, B.H.;Lee, I.K.;Ryoo, I.J.;Park, D.J.;Lee, K.H.;Lee, H.S.;Yoo, I.D.
    • Korean Journal of Weed Science
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    • v.14 no.1
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    • pp.16-22
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    • 1994
  • Ninty three species of domestic weeds were collected and screened for antimicrobial, antitumor, antioxidant and herbicidal activities. Among them, few showed antifungal activities. Cuscuta japonica showed inhibitory activity against Alternaria mali, Ambrosia artemisiifolia and Geranium sibiricum against Phytophthora capsici, Aster yomema and Aster pilosus against Phytophthora parasitica. Ambrosia artemisiifolia, Artemisia princeps, Artemisia capillaris, Ludwigia prostrata, Chrysanthemum zawadskii, Bidens frondosa, and Geranium sibiricum showed broad antibacterial activities. Carex chordorhiza, Artemisia capillaris, Persicaria nodosa, Senecio koreanus, Pariicum bisulcatum, Geranium sibiricum showed antiblebbing activity on human chronic leukemia K562 cell, among them, Persicaria nodosa was the strongist. Angelica decursiva, Equisetum arvense, Cimicifuga heracleifolia, Persicaria nodosa, Geranium sibiricum, Oenothera odorata, Cyperus sanguinolentus showed antioxidant activities. Ludwigia prostrata and Peucedanum terebinthaceum showed strong herbicidal activities.

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Molecular cloning and characterization of β-mannanase B from Cellulosimicrobium sp. YB-43 (Cellulosimicrobium sp. YB-43의 mannanase B 유전자 클로닝과 특성 분석)

  • Yoon, Ki-Hong
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.336-343
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    • 2016
  • A mannanase gene was cloned into Escherichia coli from Cellulosimicrobium sp. YB-43, which had been found to produce two kinds of mannanase, and sequenced completely. This mannanase gene, designated manB, consisted of 1,284 nucleotides encoding a polypeptide of 427 amino acid residues. Based on the deduced amino acid sequence, the ManB was identified to be a modular enzyme including two carbohydrate binding domains besides the catalytic domain, which was highly homologous to mannanases belonging to the glycosyl hydrolase family 5. The N-terminal amino acid sequence of ManB, purified from a cell-free extract of the recombinant E. coli carrying a Cellulosimicrobium sp. YB-43 manB gene, has been determined as QGASAASDG, which was correctly corresponding to signal peptide predicted by SignalP4.1 server for Gram-negative bacteria. The purified ManB had a pH optimum for its activity at pH 6.5~7.0 and a temperature optimum at $55^{\circ}C$. The enzyme was active on locust bean gum (LBG), konjac and guar gum, while it did not exhibit activity towards carboxymethylcellulose, xylan, starch, and para-nitrophenyl-${\beta}$-mannopyranoside. The activity of enzyme was inhibited very slightly by $Mg^{2+}$, $K^+$, and $Na^+$, and significantly inhibited by $Cu^{2+}$, $Zn^{2+}$, $Mn^{2+}$, and SDS. The enzyme could hydrolyze mannooligosaccharides larger than mannobiose, which was the most predominant product resulting from the ManB hydrolysis for mannooligosaccharides and LBG.

Inactivation of trypsin inhibitor and inhibitory activity of soybean(Glycine max) cultivars (대두(Glycine max) trypsin 억제제의 불활성화 및 품종별 억제활성)

  • Ryu, Byung-Woo;Han, Kang-Wan
    • Applied Biological Chemistry
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    • v.33 no.2
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    • pp.109-115
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    • 1990
  • This study was carried out to investigate the effect of L-cysteine and sodium sulfite on heat inactivation of soybean trypsin inhibitor(STI) and to determine cultivar difference in the inhibitory activity of STI. Effect of L-cysteine and sodium sulfite at different concentrations, pH's, and lengths of treatment on inactivation of STI were studied. The inactivation of STI was spectrophotometrically determined by measuring the rate of production of p-nitroaniline from synthetic substrate, N-benzoyl-DL-arginine-p-nitroanilide. Addition of L-cysteine and sodium sulfite increased magnitude of heat inactivation and greatly inhibited the re-activation of STI. There was no difference STI inactivation in among soybean cultivars employed. The trypsin inhibitory activity of STI of the soybean cultivars ranged from 64.7 to 86.4 TIU(trypsin inhibitor unit) per gram soyflour and the decreasing order of the TIU was Jangback>Hill>Jangyeab, Kwangkyo> Danyeab>Dangkyung>Paldal, Saeal, Duckyu>Hwangkeum. Inhibitory activity of STI was correlated with cysteine $content(r=0.6568^*)$ and with $digestivility(r=-0.7695^{**})$, but there was no correlation between the protein content and the inhibitory activity of STI.

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Quorum-Sensing Mechanisms in Bacterial Communities and Their Potential Applications (세균의 의사 소통(Quorum-Sensing) 기구와 그 잠재적 응용성)

  • Yoon, Sung-Sik
    • Food Science of Animal Resources
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    • v.26 no.3
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    • pp.402-409
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    • 2006
  • Although microorganisms are, in fact, the most diverse and abundant type of organism on Earth, the ecological functions of microbial populations remains poorly understood. A variety of bacteria including marine Vibrios encounter numerous ecological challenges, such as UV light, predation, competition, and seasonal variations in seawater including pH, salinity, nutrient levels, temperature and so forth. In order to survive and proliferate under variable conditions, they have to develop elaborate means of communication to meet the challenges to which they are exposed. In bacteria, a range of biological functions have recently been found to be regulated by a population density-dependent cell-cell signaling mechanism known as quorum-sensing (QS). In other words, bacterial cells sense population density by monitoring the presence of self-produced extracellular autoinducers (AI). N-acylhomoserine lactone (AHL)-dependent quorum-sensing was first discovered in two luminescent marine bacteria, Vibrio fischeri and Vibrio harveyi. The LuxI/R system of V. fischeriis the paradigm of Gram-negative quorum-sensing systems. At high population density, the accumulated signalstrigger the expression of target genes and thereby initiate a new set of biological activities. Several QS systems have been identified so far. Among them, an AHL-dependent QS system has been found to control biofilm formation in several bacterial species, including Pseudomonas aeruginosa, Aeromonas hydrophila, Burkholderia cepacia, and Serratia liquefaciens. Bacterial biofilm is a structured community of bacterial cells enclosed in a self-produced polymeric matrix that adheres to an inert or living surface. Extracellular signal molecules have been implicated in biofilm formation. Agrobacterium tumefaciens strain NT1(traR, tra::lacZ749) and Chromobacterium violaceum strain CV026 are used as biosensors to detect AHL signals. Quorum sensing in lactic acid bacteria involves peptides that are directly sensed by membrane-located histidine kinases, after which the signal is transmitted to an intracellular regulator. In the nisin autoregulation process in Lactococcus lactis, the NisK protein acts as the sensor for nisin, and NisR protein as the response regulator activatingthe transcription of target genes. For control over growth and survival in bacterial communities, various strategies need to be developed by which receptors of the signal molecules are interfered with or the synthesis and release of the molecules is controlled. However, much is still unknown about the metabolic processes involved in such signal transduction and whether or not various foods and food ingredients may affect communication between spoilage or pathogenic bacteria. In five to ten years, we will be able to discover new signal molecules, some of which may have applications in food preservation to inhibit the growth of pathogens on foods.