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Substantial Protective Immunity Conferred by a Combination of Brucella abortus Recombinant Proteins against Brucella abortus 544 Infection in BALB/c Mice

  • Arayan, Lauren Togonon;Huy, Tran Xuan Ngoc;Reyes, Alisha Wehdnesday Bernardo;Hop, Huynh Tan;Son, Vu Hai;Min, WonGi;Lee, Hu Jang;Kim, Suk
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.330-338
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    • 2019
  • Chronic infection with intracellular Brucella abortus (B. abortus) in livestock remains as a major problem worldwide. Thus, the search for an ideal vaccine is still ongoing. In this study, we evaluated the protective efficacy of a combination of B. abortus recombinant proteins; superoxide dismutase (rSodC), riboflavin synthase subunit beta (rRibH), nucleoside diphosphate kinase (rNdk), 50S ribosomal protein (rL7/L12) and malate dehydrogenase (rMDH), cloned and expressed into a pMal vector system and $DH5{\alpha}$, respectively, and further purified and applied intraperitoneally into BALB/c mice. After first immunization and two boosters, mice were infected intraperitoneally (IP) with $5{\times}10^4CFU$ of virulent B. abortus 544. Spleens were harvested and bacterial loads were evaluated at two weeks post-infection. Results revealed that this combination showed significant reduction in bacterial colonization in the spleen with a log protection unit of 1.31, which is comparable to the average protection conferred by the widely used live attenuated vaccine RB51. Cytokine analysis exhibited enhancement of cell-mediated immune response as IFN-${\gamma}$ is significantly elevated while IL-10, which is considered beneficial to the pathogen's survival, was reduced compared to control group. Furthermore, both titers of IgG1 and IgG2a were significantly elevated at three and four-week time points from first immunization. In summary, our in vivo data revealed that vaccination with a combination of five different proteins conferred a heightened host response to Brucella infection through cell-mediated immunity which is desirable in the control of intracellular pathogens. Thus, this combination might be considered for further improvement as a potential candidate vaccine against Brucella infection.

Exocyclic GpC DNA methyltransferase from Celeribacter marinus IMCC12053 (Celeribacter marinus IMCC12053의 외향고리 GpC DNA 메틸트랜스퍼라아제)

  • Kim, Junghee;Oh, Hyun-Myung
    • Korean Journal of Microbiology
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    • v.55 no.2
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    • pp.103-111
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    • 2019
  • DNA methylation is involved in diverse processes in bacteria, including maintenance of genome integrity and regulation of gene expression. CcrM, the DNA methyltransferase conserved in Alphaproteobacterial species, carries out $N^6$-adenine or $N^4$-cytosine methyltransferase activities using S-adenosyl methionine as a co-substrate. Celeribacter marinus IMCC12053 from the Alphaproteobacterial group was isolated from a marine environment. Single molecule real-time sequencing method (SMRT) was used to detect the methylation patterns of C. marinus IMCC12053. Gibbs motif sampler program was used to observe the conversion of adenosine of 5'-GANTC-3' to $N^6$-methyladenosine and conversion of $N^4$-cytosine of 5'-GpC-3' to $N^4$-methylcytosine. Exocyclic DNA methyltransferase from the genome of strain IMCC12053 was chosen using phylogenetic analysis and $N^4$-cytosine methyltransferase was cloned. IPTG inducer was used to confirm the methylation activity of DNA methylase, and cloned into a pQE30 vector using dam-/dcm- E. coli as the expression host. The genomic DNA and the plasmid carrying methylase-encoding sequences were extracted and cleaved with restriction enzymes that were sensitive to methylation, to confirm the methylation activity. These methylases protected the restriction enzyme site once IPTG-induced methylases methylated the chromosome and plasmid, harboring the DNA methylase. In this study, cloned exocyclic DNA methylases were investigated for potential use as a novel type of GpC methylase for molecular biology and epigenetics.

System for Repeated Integration of Various Gene Expression Cassettes in the Yeast Chromosome (효모염색체내에 다양한 유전자발현 cassette의 반복적 integration을 위한 system 구축)

  • Kim, Yeon-Hee
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1277-1284
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    • 2018
  • In this study, a repeated yeast integrative plasmid (R-YIp) harboring Cre/loxP system was constructed to integrate various gene expression cassettes into the yeast chromosome. The R-YIp system contains a reusable selective marker (CgTRP1), loxP sequence, and target sequence for integration. Therefore, many gene expression cassettes can be integrated into the same position of the same yeast chromosome. In the present study, several model enzymes involving xylan/xylose metabolism were examined, including endoxylanase (XYLP), ${\beta}$-xylosidase (XYLB), xylose reductase (GRE3) and xylitol dehydrogenase (XYL2). Efficient expression of these genes was obtained using two promoters (GAL10p and ADH1p) and various plasmids (pGMF-GENE and pAMF-GENE plasmids) were constructed. The XYLP, XYLB, GRE3, and XYL2 genes were efficiently expressed under the control of the GAL10 promoter. Subsequently, R-YIps containing the GAL10p-GENE-GAL7t cassette were constructed, resulting in pRS-XylP, pRS-XylB, pRS-Gre3, and pRS-Xyl2 plasmids. These plasmids were sequentially integrated into chromosome VII of a Saccharomyces cerevisiae strain by repeated gene integration and selective marker rescue. These genes were integrated by the R-YIp system and were stably expressed in the yeast transformants to produce active recombinant enzymes. Therefore, we expect that the R-YIp system will be able to overcome current limitations of the host cells and allow selective marker selection for the integration of various genes into the yeast chromosome.

Spatio-Temporal Variation of Polymetallic Mineralization in the Wooseok Deposit (우석광상 다금속 광화작용의 시공간적 특성변화)

  • Im, Heonkyung;Shin, Dongbok;Jeong, Junyeong;Lee, Moontaek
    • Economic and Environmental Geology
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    • v.51 no.6
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    • pp.493-507
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    • 2018
  • The Wooseok deposit in Jecheon belongs to the Hwanggangri Mineralized Distict of the northeastern Ogcheon Metamorphic Belt. Its geology consists mostly of limestone of the Choseon Supergroup and the Cretaceous Muamsa granite intruded at the eastern area of the deposit. The deposit shows vertical occurrence of skarn and hydrothermal vein ores with W-Mo-Fe and Cu-Pb-Zn mineralization and skarn is developed only at lower levels of the deposit. Skarn minerals are replaced or cut by ore minerals in paragenetic sequence of magnetite-hematite, molybdenite-scheelite-wollframite, and higher abundances of pyrrhotite-chalcopyrite-pyrite-sphalerite-galena. Garnet has chemical compositions of $Ad_{65.9-97.8}Gr_{0.3-32.0}Pyr_{0.9-3.0}$, corresponding to andradite series, and pyroxene compositions are $Hd_{4.5-49.7}Di_{42.3-93.9}Jo_{0.5-7.9}$, prevailing in diopside compositions, both of which suggest oxidized conditions of skarnization. On the FeS-MnS-CdS ternary diagram, FeS contents of sphalerite in vein ores decrease with increasing MnS contents from bottom to top levels, possibly relating to W mineralization in deep and Pb-Zn mineralization in shallow level. Sulfur isotope values of sulfide minerals range from 5.1 to 6.8‰, reflecting magmatic sulfur affected by host rocks. W-Mo skarn and Pb-Zn vein mineralization in the Wooseok deposit were established by spatio-temporal variation of decreasing temperature and oxygen fugacity with increasing sulfur fugacity from bottom to top levels.

Conservation Treatment Based on Material Characteristics, Provenance Presumption and Deterioration Diagnosis of the Seven-Storied Jungwon Tappyeongri Stone Pagoda, Chungju, Korea (중원탑평리칠층석탑의 재질특성과 산지추정 및 손상도 진단을 통한 보존처리)

  • Lee, Chan Hee;Kim, Moo Yeon;Jo, Young Hoon;Lee, Myeong Seong
    • Korean Journal of Heritage: History & Science
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    • v.43 no.3
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    • pp.4-25
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    • 2010
  • This study was carried out on scientific conservation treatment based on material characteristics, provenance interpretation, and deterioration diagnosis for seven-storied Jungwon Tappyeongri stone pagoda in Chungju. As a result, main rock of the pagoda is biotite granite with magnetite-series (average $5.86{\times}10^{-3}$ SI unit), containing partly basic xenolith, pegmatite veinlet and feldspar phenocryst. As a result of the provenance presumption of the host rock, a rock around the Songgang stream was identified the same origin. Therefore the rock is appropriate for materials of the pagoda restoration. The deterioration assessment suggested that the pagoda was seriously exfoliated (2.7 to 5.5%), discolored (39.8 to 58.9), and contaminated with repair materials (3.5 to 9.4%), and bioorganisms (19.3 to 24.4%). Accordingly, conservation treatment was carried out based on preliminary investigation for stable conservation of the pagoda. Overall processes were sequentially proceeded by restoration of the replacement stone, cleaning, joining and consolidation. This study sets up an integrated conservation system from preliminary investigation to conservation treatment of the pagoda. Also, the study will contribute for establishing the future-oriented customized conservation treatment.

Studies on the Characteristics of Spatial Components and Conversion of Sandan Area in the Rear Garden of Changdeokgung Palace (창덕궁 후원 산단(山壇) 권역 구성요소의 특질과 변용)

  • Jung, Woo-jin;Liu, Gi-suk;Sim, Woo-kyung
    • Korean Journal of Heritage: History & Science
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    • v.46 no.4
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    • pp.24-47
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    • 2013
  • This study focused on the constructional concepts, landscape design techniques and changes made of the Sandan(山壇) area in the rear garden of Changdeokgung Palace based on the spatial organization of the zone. The results of this study are as below. Constructional layers that have been maintained throughout the centuries from King Injo(仁祖), King Sukjong(肅宗), King Sunjo(純祖), the Japanese colonial period to the present, were found in the Sandan area. The Sandan area, which was developed with the establishment of Chuiseungjeong(取勝亭) during the ruling of King Injo(仁祖), was created as a resting place for the King, and its usage continued until King Sukjong(肅宗) built Nakminjeong(樂民亭). However, the whole area was reorganized to host ritual ceremonies, where ancestral rites were performed for the mountain gods, after Sandan and Baekunsa(白雲社) were built during King Gojong's reign, before the drawing of 'Painting of Eastern Palaces(東闕圖).' The architectural component used then appears to have been designed to serve ancestral ritual formalities for the god of land, or to satisfy the religious desires of the royal family not fulfilled by Confucian courtesy. These are characteristics that define the Sandan area. Meanwhile, the analysis of the ground plan of the Changdeokgung Palace revealed that the Sandan had been maintained until the Japanese colonial era, but was removed sometime after liberation. The area underwent extensive reform in the early 1970s, when the whole area was developed into a tourist destination called Bingcheon(氷泉). Then, a new road that runs through Bingcheon was laid, and the present condition reflects the construction result of the time. An interview with a person who had taken part in the repair work in the 1970s confirmed that the construction work at the time had the goal of establishing two drainage systems for drinking water and copied the Okryuchon(玉流川) well.

Precise Deterioration Diagnosis and Restoration Stone Suggestion of Jungdong and Banjukdong Stone Aquariums in Gongju, Korea (공주 중동 및 반죽동 석조의 정밀 손상도 진단과 복원석재 제안)

  • Jo, Young Hoon;Lee, Myeong Seong;Choi, Gi Eun;Lee, Chan Hee
    • Korean Journal of Heritage: History & Science
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    • v.44 no.3
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    • pp.92-111
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    • 2011
  • This study focus on the restoration stone selection of break-out part based on material characteristics analysis and the conservational safety diagnosis using various nondestructive techniques for Jungdong and Banjukdong Stone Auariums. As a result, the original rocks of the stone aquariums body are porphyritic granodiorite with magnetite-series having igneous lineation, microcline phenocryst, veinlet and basic xenolith. As a result of the provenance presumption of the host rock, a rock around Gamgokri area in Nonsan City was identified the genetically same rock. Therefore, the rock is appropriate for restoration materials of the break-out part. The deterioration assessment showed that the stone aquariums were highly serious scaling, scale off and blackening. Particularly, the front face of Banjukdong stone aquarium needs reinforcement of structural crack (760mm) caused from igneous lineation of biotite. Blackening contaminants on the stone aquariums surface occurred by combining iron oxide, manganese oxide and clay mineral. Also, major factors of efflorescence contaminants were identified as calcite (Jungdong stone aquariums) and gypsum (Banjukdong stone aquariums). The physical characteristics of stone aquariums appeared that the original and new stone is third (moderately weathered) and second grade (slightly weathered), respectively. This study sets up an integrated conservation system from material analysis to restoration stone selection and conservational safety diagnosis of Jungdong and Banjukdong stone aquariums.

A Study on the Luminescence Properties of Eu3+ Ions Doped Vanadate (Eu3+ 이온이 첨가된 바나듐산염의 형광특성 연구)

  • Kang, Yeonhee;Yoon, Changyong
    • Journal of the Korean Society of Radiology
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    • v.13 no.3
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    • pp.445-451
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    • 2019
  • The fluorescence intensity and fluorescence lifetime of $Ba_2GdV_3O_{11}$, a vanadate compound based on $Ba^{2+}$ ion, were investigated by adding $Eu^{3+}$ as a rare earth ion which is an alkaline earth metal, which is distributed around active ions and has a large influence on fluorescent properties when used as a host in a phosphor. $Ba_2GdV_3O_{11}:Eu^{3+}$ phosphor was synthesized by solid state method and the crystallinity of the phosphor was confirmed by X - ray diffraction analysis. The fluorescence properties of the $Ba_2GdV_3O_{11}:Eu^{3+}$ phosphor were measured using optical and laser. The energy transfer and diffusion of the $Ba_2GdV_3O_{11}:Eu^{3+}$ phosphor are highly dependent on the concentration of $Eu^{3+}$. When the concentration of $Eu^{3+}$ is low, it shows strong fluorescence to the CT band. However, as the concentration of $Eu^{3+}$ increases, the fluorescence due to 4f - 4f transition is strong. The concentration of $Eu^{3+}$ ion increased and the energy between ions was diffused, and the lifetime of fluorescence decreased. Energy transfer occurs between two $Eu^{3+}$ ions at low $Eu^{3+}$ concentration and energy diffusion occurs at high $Eu^{3+}$ concentration.

Luminescence Properties of Europium-doped NaSr(PO3)3 Phosphor (Europium이 첨가된 NaSr(PO3)3형광체의 형광특성)

  • Yoon, Changyong;Park, Cheolwoo
    • Journal of the Korean Society of Radiology
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    • v.13 no.4
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    • pp.645-652
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    • 2019
  • Phosphor with phosphorus doped with rare earth ions was investigated by searching Sr and Eu phosphors suitable for substitution of Eu ions with similar ionic radius to polyphosphate host. The $NaSr(PO_3)_3$ phosphor was synthesized by the solid state method and the $NaSr(PO_3)_3:Eu^{2+}$ phosphor was prepared by the carbon thermal reduction method. Both of the phosphors were identified by X - ray diffraction. The excitation and emission spectra of $NaSr(PO_3)_3:Eu^{3+}$ increased fluorescence intensity and intensity quenching with increasing $Eu^{3+}$ concentration. The higher the $Eu^{3+}$ concentration in the emission spectrum, the higher the local symmetry of $Eu^{3+}$ environment. The mechanism of concentration quenching, in which fluorescence decreases due to the energy transfer between $Eu^{2+}$ ions with the closest critical distance between $Eu^{2+}$ ions with increasing $Eu^{2+}$ ion concentration, was confirmed in the emission spectrum of $NaSr(PO_3)_3:Eu^{2+}$ concentration. It is possible to change the fluorescent region through the post-processing of single rare earth ion added phosphors, and it is possible to change the fluorescence by applying the energy transfer and concentration quenching mechanism according to the local symmetry of $Eu^{3+}$ will be used for high phosphor development.

Recent advances on next-generation probiotics linked to the gut microbiome (장내 마이크로바이옴과 차세대 프로바이오틱스 연구 현황)

  • Choi, Hak-Jong
    • Food Science and Industry
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    • v.52 no.3
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    • pp.261-271
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    • 2019
  • Gut microbiome have recently provided evidence that the gut microbiota are capable of greatly influencing all aspects of physiology and immunology. Although a number of recent studies have shown that probiotics can modulate gut microbiota structure, the mechanism underlying this effect remains to be elucidated. In a disease state, the relative abundances of beneficial gut bacteria are generally reduced, which is restored by constant probiotic supplementation. Oral administration of probiotics improved the disease state by (1) inducing differentiation and function of regulatory T cells, (2) reducing inflammatory response, (3) modulating the gut environment, and (4) increasing the proportions of short-chain fatty acid- or beneficial metabolite-producing gut microbiota including the genera Bifidobacterium, Faecalibacterium, Akkermansia, etc. In this review, current knowledge on how probiotics can influence host's health by altering gut microbiota structure and on how probiotics and beneficial gut bacteria can be applied as next-generation probiotics will be discussed.