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Effects of Bacillus subtilis KN-42 on Growth Performance, Diarrhea and Faecal Bacterial Flora of Weaned Piglets

  • Hu, Yuanliang;Dun, Yaohao;Li, Shenao;Zhao, Shumiao;Peng, Nan;Liang, Yunxiang
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.8
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    • pp.1131-1140
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    • 2014
  • This research focused on the effects of different doses of Bacillus subtilis KN-42 on the growth performance, diarrhea incidence, faecal bacterial flora, and the relative number of Lactobacillus and Escherichia coli in faeces of weaned piglets to determine whether the strain can serve as a candidate antimicrobial growth promoter. A total of 360 piglets (initial body weight $7.14{\pm}0.63$ kg) weaned at $26{\pm}2$ days of age were randomly allotted to 5 treatment groups (4 pens per treatment with 18 pigs per pen) for a 28-day trial. Dietary treatments were basal diet without any antimicrobial (negative control; NC), basal diet supplemented with 120 mg/kg feed of neomycin sulfate (positive control; PC) and basal diet supplemented with $2{\times}10^9$ (L), $4{\times}10^9$ (M) and $20{\times}10^9$ (H) CFU/kg feed of B. subtilis KN-42. During the overall period, average daily gain and feed efficiency of piglets were higher in groups PC, M, and H than those in group NC (p<0.05), and all probiotics and antibiotics groups had a lower diarrhea index than group NC (p<0.05). The 16S rDNA gene-based methods were used to analyze faecal bacterial flora on day 28 of experiment. The result of denaturing gradient gel electrophoresis analysis showed that supplementation of B. subtilis KN-42 to the diet changed the bacterial communities, with a higher bacterial diversity and band number in group M than in the other four groups. Real-time polymerase chain reaction analysis showed that the relative number of Lactobacillus were higher in groups PC and H than in group NC (p<0.05), and the supplemented B. subtilis KN-42 to the diet also reduced the relative number of E. coli (p<0.05). These results suggest that dietary addition of B. subtilis KN-42 can improve the growth performance and gastrointestinal health of piglets.

Recombinant Expression of Agarases: Origin, Optimal Condition, Secretory Signal, and Genome Analysis (한천분해효소의 재조합발현 : 기원, 활성조건, 분비신호와 게놈분석 등)

  • Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.30 no.3
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    • pp.304-312
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    • 2020
  • Agarase can be used in the field of basic science, as well as for production of agar-derived high-functional oligosaccharides and bioenergy production using algae. In 2012, we summarized the classification, origin, production, and applications of agar. In this paper, we briefly review the literature on the recombinant expression of agarases from 2012 to the present. Agarase genes originated from 19 genera, including Agarivorans, Flammeovirga, Pseudoalteromonas, Gayadomonas, Catenovulum, Microbulbifer, Cellulophaga, Saccharophagus, Simiduia, and Vibrio. Of the 47 recombinant agarases, there were only two α-agarases, while the rest were β-agarases. All α-agarases produced agarotetraose, while β-agarases yielded many neoagarooligosaccharides ranging from neoagarobiose to neoagarododecaose. The optimum temperature ranged between 25 and 60℃, and the optimum pH ranged from 3.0 to 8.5. There were 14 agarases with an optimum temperature of 50℃ or higher, where agar is in sol state after melting. Artificial mutations, including manipulation of carbohydrate-binding modules (CBM), increased thermostability and simultaneously raised the optimum temperature and activity. Many hosts and secretion signals or riboswitches have been used for recombinant expression. In addition to gene recombination based on the amino acid sequence after agarase purification, recombinant expression of the putative agarase genes after genome sequencing and metagenome-derived agarases have been studied. This study is expected to be actively used in the application fields of agarase and agarase itself.

Isolation and Expression of Dormancy-associated protein 1 (DRM1) in Poplar (Populus alba × P. glandulosa) (현사시나무에서 Dormancy-associated protein 1 (DRM1) 유전자의 분리와 발현특성 구명)

  • Yoon, Seo-Kyung;Bae, Eun-Kyung;Choi, Hyunmo;Choi, Young-Im;Lee, Hyoshin
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.69-75
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    • 2017
  • Dormancy-associated protein (DRM) is involved in the dormancy physiology of plants and is conserved in almost all plant species. Recent studies found that DRM genes are involved in the abiotic stress response, and characterization studies of these genes have been conducted in several plants. However, few studies have focused on DRM genes in woody plants. Therefore, in this study, cDNA coding for DRM (PagDRM1) was isolated from poplar (Populus alba ${\times}$ P. glandulosa), and its structure and expression characteristics were investigated. PagDRM1 encodes a putative protein composed of 123 amino acids, and the protein contains two conserved domains (Domain I and Domain II). PagDRM1 is present as one or two copies in the poplar genome. Its expression level was highest in the stem, followed by mature leaves, roots, and flowers. During the growth of cultured cells in suspension, PagDRM1 was highly expressed from the late-exponential phase to the stationary phase. In addition, PagDRM1 expression increased in response to drought, salt stress, and treatment with plant hormones (e.g., abscisic acid and gibberellic acid). Therefore, we suggested that PagDRM1 not only plays an important role in the induction of dormancy, but also contributes to stress tolerance in plants.

Transduced Tat-Annexin protein suppresses inflammation-associated gene expression in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells

  • Lee, Sun-Hwa;Kim, Dae-Won;Back, Su-Sun;Hwang, Hyun-Sook;Park, Eun-Young;Kang, Tae-Cheon;Kwon, Oh-Shin;Park, Jong-Hoon;Cho, Sung-Woo;Han, Kyu-Hyung;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.44 no.7
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    • pp.484-489
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    • 2011
  • Annexin-1 (ANX1) is an anti-inflammatory protein as well as an important modulator in inflammation. However, the precise action of ANX1 remains unclear. To elucidate the protective effects of ANX1 on lipopolysaccharide (LPS)-induced murine macrophage Raw 264.7 cells, we constructed a cell-permeable Tat-ANX1 protein. The transduced Tat-ANX1 protein markedly inhibited the expression of cyclooxygenase-2, production of prostaglandin $E_2$, and generation of pro-inflammatory cytokines in the cells. Furthermore, transduced Tat-ANX1 protein caused a significant reduction in the activation of nuclear factor-kappa B (NF-${\kappa}B$) and mitogen-activated protein kinase (MAPK). The results indicate that Tat-ANX1 inhibits the production of inflammatory response cytokines and enzymes by blocking NF-${\kappa}B$ and MAPK. Therefore, Tat-ANX1 protein may be useful as a therapeutic agent against various inflammatory diseases.

Molecular and Morphologic Identification of Spirometra ranarum Found in the Stool of African Lion, Panthera leo in the Serengeti Plain of Tanzania

  • Eom, Keeseon S.;Park, Hansol;Lee, Dongmin;Choe, Seongjun;Kang, Yeseul;Bia, Mohammed Mebarek;Lee, Sang-Hwa;Keyyu, Julius;Fyumagwa, Robert;Jeon, Hyeong-Kyu
    • Parasites, Hosts and Diseases
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    • v.56 no.4
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    • pp.379-383
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    • 2018
  • The present study was performed with morphological and molecular analysis (cox1 and nad1 mitochondrial genes) to identify the proglottids of spirometrid tapeworm found in the stool of an African lion, Panthera leo, in the Serengeti plain of Tanzania. A strand of tapeworm strobila, about 75 cm in length, was obtained in the stool of a male African lion in the Serengeti National Park ($34^{\circ}$ 50' E, $02^{\circ}$ 30' S), Tanzania, in February 2012. The morphological features of the adult worm examined exhibited 3 uterine coils with a bow tie appearance and adopted a diagonal direction in the second turn. The posterior uterine coils are larger than terminal uterine ball and the feature of uteri are swirling rather than spirally coiling. The sequence difference between the Spirometra species (Tanzania origin) and S. erinaceieuropaei (GenBank no. KJ599680) was 9.4% while those of S. decipiens (GenBank no. KJ599679) differed by 2.1% in the cox1 and nad1 genes. Phylogenetic tree topologies generated using the 2 analytic methods were identical and presented high level of confidence values for the 3 major branches of the 3 Spirometra species in the cox1 gene. The morphological and molecular findings obtained in this study were nearly coincided with those of S. ranarum. Therefore, we can know for the first time that the African lion, Panthera leo, is to the definitive host of this tapeworm.

Identification of QTLs for Some Agronomic Traits in Rice Using an Introgression Line from Oryza minuta

  • Rahman, Md Lutfor;Chu, Sang Ho;Choi, Min-Sun;Qiao, Yong Li;Jiang, Wenzhu;Piao, Rihua;Khanam, Sakina;Cho, Young-Il;Jeung, Ji-Ung;Jena, Kshirod K.;Koh, Hee-Jong
    • Molecules and Cells
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    • v.24 no.1
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    • pp.16-26
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    • 2007
  • Wild progenitor species provide potential gene sources for complex traits such as yield and multiple resistances to biotic and abiotic stresses, and thus are expected to contribute to sustainable food supplies. An introgression line 'IR71033-121-15' was derived from a wild species Oryza minuta (2n = 48, BBCC, Acc No. 101141) at IRRI. Introgression analysis using 530 SSR and STS markers revealed that at least 14 chromosomal segments distributed over 12 chromosomes had been introgressed from O. minuta. An $F_{2:3}$ population from the cross between IR71033 and Junambyeo (a Korean japonica cultivar) consisting of 146 lines was used for quantitative trait loci (QTL) analysis of 16 agronomic traits. A total of 36 single-locus QTLs (S-QTLs) and 45 digenic epistasis (E-QTLs) were identified. In spite of it's inferiority of O. minuta for most of the traits studied, its alleles contributed positively to 57% of the QTLs. The other QTLs originated from either parent, IR71033 or Junambyeo. QTLs for phenotypically correlated traits were mostly detected on introgressed segments. Fourteen QTLs corresponded to QTLs reported earlier, indicating that these QTLs are stable across genetic backgrounds. Twenty-two QTLs controlling yield and its components had not been detected in previous QTL studies. Of these, thirteen consisted of potentially novel alleles from O. minuta. QTLs from O. minuta introgression could be new sources of natural variation for the genetic improvement of rice.

Monitoring and Analysis of Genetically Modified Ingredients of Imported Foods by PCR (PCR에 의한 수입식품의 유전자재조합 원료 분석 및 모니터링)

  • Kim, Hee-Yun;Park, Yong-Chjun;Ro, Hye-Lim;Jo, Jun-Il;Kim, Eun-Jung;Nam, Hae-Sun;Lee, Jin-Kyung;Lee, Jin-Ha;Kang, Yoon-Sook;Lee, Jong-Ook
    • Korean Journal of Food Science and Technology
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    • v.38 no.5
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    • pp.605-608
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    • 2006
  • Genetically modified (GM) ingredients found in imported raw materials and processed foods were monitored in the province Gyeongin in Korea. The analysis was performed according to "Testing methods for genetically modified foods of food standards and specifications" established in Korea. We received 120 items from the Gyeongin Regional KFDA. Only two of the 120 items analyzed in the samples, were contaminated with GM ingredients. However, we could not analyze the internal standard gene from 12 processed foods. We found that the extracted total DNA of the above 12 samples were extracted and found to be degraded. The total DNA contained a very small fragment of less than 300 base pair. Therefore, it seems that the total DNA is not large enough to serve as the template DNA for PCR analysis.

The anti-imflammatory effect and the mechanism of Formica yessensis extraction (홍의 추출물의 항염작용 및 그 기전 연구)

  • Kim, Jong-Min;Kim, Seung-Hyung;Yang, Won-Kyung;Jung, Taek-Geun;Kim, Se-Ran;Hwang, Sung-Joon;Yoo, Hwa-Seung
    • Journal of Haehwa Medicine
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    • v.25 no.1
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    • pp.71-86
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    • 2016
  • Objective : Hongyi (Formica yessensis) is the dried insect of fomicidae. In previous studies, it appeared possibilities on anti-thrombosis, preventing atherosclerosis, treating rheumatoid disease, and inhibiting hela cell. In this study, we investigated anti-inflammatory effects and mechanism of Hongyi. Methods : Hongyi A was extracted by water and made dried powder. Hongyi B was extracted by ethanol and made dried powder. We measured Nitric Oxide (NO) production on the mouse macrophages (RAW 264.7), mouse vascular endothelial cell (MOVAS) and human vascular endothelial cell (HUVEC) for anti-inflammatory effect. In addition, we conducted reverse transcription reaction (RT-PCR) for investigating the mechanism. Results : In RAW 264.7 macrophages stimulated by LPS, Hongyi A ($100{\mu}g/m{\ell}$) decreased NO production compared with LPS $2{\mu}g/ml$ control group with statistical significance (p<0.05). Hongyi A (50, $100{\mu}g/m{\ell}$) also decreased NO production compared with LPS $4{\mu}g/ml$ control group with statistical significance (p<0.01). Hongyi B (50, $100{\mu}g/m{\ell}$) decreased NO production compared with LPS $2{\mu}g/ml$ control group with statistical significance (p<0.01). Hongyi B (10, 50, $100{\mu}g/m{\ell}$) also decreased NO production compared with LPS $4{\mu}g/ml$ control group with statistical significance (p<0.01, p<0.001, p<0.001). In the MOVAS, Hongyi A and B increased NO production compared with control group. In the HUVEC, Hongyi B increased NO production compared with control group. The expression of NF-${\kappa}B$ in 12-hours MOVAS culture was decreased by Hongyi A and B (10, $50{\mu}g/ml$) compared with control group, but expression of $I{\kappa}B$ was increased. In the 24-hours MOVAS culture, expression of $I{\kappa}B$ was significantly increased. The expression of NF-${\kappa}B$ in 12-hours HUVEC culture was decreased by Hongyi A and B compared with control group, but expression of $I{\kappa}B$ was increased. Hongyi B also increased eNOS mRNA gene expression. Conclusions : Hongyi A and B showed anti-inflammatory effect in mouse macrophages with the activation of vascular endothelial cell through NO production in MOVAS and HUVEC repectively. Honyi B showed superior effect than Hongyi A, but additonal mechanism study should be conducted.

Cloning and Expression of K11 Phage RNA Polymerase (K11 RNA 중합효소의 Cloning 및 발현)

  • Lee, Sang-Soo
    • The Journal of Natural Sciences
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    • v.9 no.1
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    • pp.19-24
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    • 1997
  • Using the PCR(polymerase chain reaction method), gone 1 of phage K11 coding for K11 phage RNA polymerase has been cloned and expressed under the control of lac promoter. K11 phage RNA polymerase was conventionally purified through the DEAE-sephacel and Affigel blue column chromatographies. The 0.2-0.3 M $NH_4Cl$ fractions of DAEA-sephacel column chromatography showed K11 phage RNA polymerase activity and further purification with Affigel blue column chromatography showed nearly single protein band on SDS-polyacryl amide gel. K11 phage RNA polymerase, which is one of the T7 group phage RNA polymerase (E. coil phage T7, T3 and Salmonella tyhimurium phage SP6 RNA polymerase), shares high degrees of homology with the other T7 group phage RNA polymerase. Previously we constructed T7 and SP6 promoter variants and revealed promoter specificity of T7 and SP6 RNA polymerase (Lee and Kang, 1993). To investigate the promoter specificity of K11 RNA polymerase in vitro K11 promoter activity was measured with SP6 promoter variants. The SP6 promoter variant share highest degrees of sequence homology with K11 promoter sequence show strongest promoter activity.

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Research article Black ginseng activates Akt signaling, thereby enhancing myoblast differentiation and myotube growth

  • Lee, Soo-Yeon;Go, Ga-Yeon;Vuong, Tuan Anh;Kim, Jee Won;Lee, Sullim;Jo, Ayoung;An, Jun Min;Kim, Su-Nam;Seo, Dong-Wan;Kim, Jin-Seok;Kim, Yong Kee;Kang, Jong-Sun;Lee, Sang-Jin;Bae, Gyu-Un
    • Journal of Ginseng Research
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    • v.42 no.1
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    • pp.116-121
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    • 2018
  • Background: Black ginseng (BG) has greatly enhanced pharmacological activities relative to white or red ginseng. However, the effect and molecular mechanism of BG on muscle growth has not yet been examined. In this study, we investigated whether BG could regulate myoblast differentiation and myotube hypertrophy. Methods: BG-treated C2C12 myoblasts were differentiated, followed by immunoblotting for myogenic regulators, immunostaining for a muscle marker, myosin heavy chain or immunoprecipitation analysis for myogenic transcription factors. Results: BG treatment of C2C12 cells resulted in the activation of Akt, thereby enhancing hetero-dimerization of MyoD and E proteins, which in turn promoted muscle-specific gene expression and myoblast differentiation. BG-treated myoblasts formed larger multinucleated myotubes with increased diameter and thickness, accompanied by enhanced Akt/mTOR/p70S6K activation. Furthermore, the BG treatment of human rhabdomyosarcoma cells restored myogenic differentiation. Conclusion: BG enhances myoblast differentiation and myotube hypertrophy by activating Akt/mTOR/p70S6k axis. Thus, our study demonstrates that BG has promising potential to treat or prevent muscle loss related to aging or other pathological conditions, such as diabetes.