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Development of ITS sequence based SCAR marker and multiplex-SCAR assay for the rapid authentication of Tetrapanacis Medulla and Akebiae Caulis (통초(通草), 목통(木通) 신속 감별용 ITS 염기서열 기반 SCAR 마커 및 Multiplex-SCAR 분석법 개발)

  • Noh, Pureum;Kim, Wook Jin;Park, Inkyu;Yang, Sungyu;Choi, Goya;Moon, Byeong Cheol
    • The Korea Journal of Herbology
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    • v.36 no.1
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    • pp.9-17
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    • 2021
  • Objectives : Tetrapanacis Medulla and Akebiae Caulis are one of the most frequently adulterated herbal medicines because of their confusability of terms in the ancient writings and the similarity of morphological features of dried herbal products. The major adulterant is Aristolochia manshuriensis (Guanmutong) which has a serious safety concern with its toxicity. To ensure the safety and quality of the two herbal medicines, it is necessary to discriminate the toxic adulterant from authentic species. The aim of this study is to develop SCAR markers and to establish the multiplex-SCAR assay for discrimination of four plant species related to Tetrapanacis Medulla and Akebiae Caulis. Methods : ITS regions of fifteen samples of four species (Tetrapanax papyrifer, Fatsia japonica, Aristolochia manshuriensis, and Akebia quinata) collected from different sites were amplified and sequenced. Fifteen obtained ITS sequences were aligned and analysed for the detection of species-specific sequence variations. The SCAR markers were designed based on the sequence alignments and then, multiplex-SCAR assay enhancing rapidity was optimized. Results : ITS sequences clearly distinguished the four species at the species level. The developed SCAR markers and multiplex-SCAR assay were successfully discriminated four species and detected the adulteration of commercial product samples by comparison of the amplified DNA fragment sizes. Conclusions : These SCAR markers and multiplex-SCAR assay are a rapid, simple, and reliable method to identify the authentic Tetrapanacis Medulla and Akebiae Caulis from adulterants. These genetic tools will be useful to ensure the safety and to standardize the quality of the two herbal medicines.

A Novel Anti-PD-L1 Antibody Exhibits Antitumor Effects on Multiple Myeloma in Murine Models via Antibody-Dependent Cellular Cytotoxicity

  • Ahn, Jae-Hee;Lee, Byung-Hyun;Kim, Seong-Eun;Kwon, Bo-Eun;Jeong, Hyunjin;Choi, Jong Rip;Kim, Min Jung;Park, Yong;Kim, Byung Soo;Kim, Dae Hee;Ko, Hyun-Jeong
    • Biomolecules & Therapeutics
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    • v.29 no.2
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    • pp.166-174
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    • 2021
  • Multiple myeloma is a malignant cancer of plasma cells. Despite recent progress with immunomodulatory drugs and proteasome inhibitors, it remains an incurable disease that requires other strategies to overcome its recurrence and non-response. Based on the high expression levels of programmed death-ligand 1 (PD-L1) in human multiple myeloma isolated from bone marrow and the murine myeloma cell lines, NS-1 and MOPC-315, we propose PD-L1 molecule as a target of anti-multiple myeloma therapy. We developed a novel anti-PD-L1 antibody containing a murine immunoglobulin G subclass 2a (IgG2a) fragment crystallizable (Fc) domain that can induce antibody-dependent cellular cytotoxicity. The newly developed anti-PD-L1 antibody showed significant antitumor effects against multiple myeloma in mice subcutaneously, intraperitoneally, or intravenously inoculated with NS-1 and MOPC-315 cells. The anti-PD-L1 effects on multiple myeloma may be related to a decrease in the immunosuppressive myeloid-derived suppressor cells (MDSCs), but there were no changes in the splenic MDSCs after combined treatment with lenalidomide and the anti-PD-L1 antibody. Interestingly, the newly developed anti-PD-L1 antibody can induce antibody-dependent cellular cytotoxicity in the myeloma cells, which differs from the existing anti-PD-L1 antibodies. Collectively, we have developed a new anti-PD-L1 antibody that binds to mouse and human PD-L1 and demonstrated the antitumor effects of the antibody in several syngeneic murine myeloma models. Thus, PD-L1 is a promising target to treat multiple myeloma, and the novel anti-PD-L1 antibody may be an effective anti-myeloma drug via antibody-dependent cellular cytotoxicity effects.

Phylogenetic Analysis on Wild Cordyceps Collected from Miryang Region of South Korea (밀양근교에서 채집한 야생 동충하초 계통의 PCR 산물에 근거한 계통 유전학적 연구)

  • Park, Hyeancheal;Lee, Sangmong;Park, Namsook
    • Korean Journal of Plant Resources
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    • v.34 no.1
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    • pp.1-16
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    • 2021
  • The phylogenetic relationships among thirty-two strains (P1~P32; including Cordyceps sp., Paecilomyces sp., Beauveria sp., Aranthomyces sp., Isaria sp. and Himenostilbe sp.) in Miryang region located in the southern part of Korea, were investigated based on internal transcribed spacer (ITS) sequences of ribosomal DNA. A fragment of ITS region was amplified by polymerase chain reaction (PCR) using the specific primer pairs ITS1 and ITS4. After obtained same size of PCR products from various strains, we cloned them into a pGEM-T easy vector to determine their sequences. BLAST analyses of the nucleotide sequence ITS1, 5.8S and ITS2 gene fragments revealed the identity and their phylogenetic relationship. Among 32 strains isolated from Miryang region, Cordyceps militaris was shared 100% sequences with Genbank (AY49191, EU825999, AY491992), while some species are not shared perfectly with reported sequences. For example, strain P17 (P. tenuipes in Ulju-gun Gaji Mountain) has some differences among the other strains of P. tenuipes (Miryang-si Jocheon-eup, Miryang-si Gaji Mountain) and those of gene bank. We conclude that ITS analyses with strains in the suburbs of Miryang in this study can be effectively used as a tool for classification, evaluation and collection of the natural eco-type genetic resources.

A comparison study on shear bond strength of 3D printed resin and conventional heat-cured denture base resin to denture relining materials (3D-프린팅 의치상 레진과 열중합 의치상 레진에서의 의치 첨상 재료 간의 전단결합강도 비교 연구)

  • Cho, Sung-Yoon;Song, Young-Gyun
    • Journal of Dental Rehabilitation and Applied Science
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    • v.37 no.4
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    • pp.232-243
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    • 2021
  • Purpose: The purpose of this study was to evaluate the shear bond strength of various 3D printed denture base resins and the conventional denture base resin to various denture relining materials. Materials and Methods: For denture base materials, a heatcured (Vertex RS) and two types of 3D printed DENTCA Denture base II, NextDentTM Base) were used. And 4 types denture relining materials (Tokuyama Rebase II fast, Kooliner, Denture Liner, Denture Liner, Lang Jet Denture Repair Kit) with different components were used. It was classified into 12 groups. Adhesion was performed between the resin base and the relining materials in accordance with ISO/TS 11405 standard. The shear bonding strength was measured, and then the adhesion interface was observed with a stereoscopic microscope and a scanning electron microscope. The fracture pattern was investigated through the analysis of the fragment. Results: In the 3D printed denture resin group, the shear bonding strength with relining materials was significantly lower than that of the heat-cured resin group (P < 0.05). The group of polymethyl methacrylate -based relining materials, high shear bonding strength was shown regardless of the type of denture. As for the fracture pattern, adhesive fracture appeared in most groups, and cohesive, mixed fracture appeared in some groups. Conclusion: The polymethyl methacrylate -based denture relining materials showed high shear bonding strength values compared to other denture relining materials. But, for direct methods, it is considered advantageous in terms of shear bonding strength to use a isobutyl methacrylate-based denture relining materials.

Numerical analysis of melt migration and solidification behavior in LBR severe accident with MPS method

  • Wang, Jinshun;Cai, Qinghang;Chen, Ronghua;Xiao, Xinkun;Li, Yonglin;Tian, Wenxi;Qiu, Suizheng;Su, G.H.
    • Nuclear Engineering and Technology
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    • v.54 no.1
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    • pp.162-176
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    • 2022
  • In Lead-based reactor (LBR) severe accident, the meltdown and migration inside the reactor core will lead to fuel fragment concentration, which may further cause re-criticality and even core disintegration. Accurately predicting the migration and solidification behavior of melt in LBR severe accidents is of prime importance for safety analysis of LBR. In this study, the Moving Particle Semi-implicit (MPS) method is validated and used to simulate the migration and solidification behavior. Two main surface tension models are validated and compared. Meanwhile, the MPS method is validated by the L-plate solidification test. Based on the improved MPS method, the migration and solidification behavior of melt in LBR severe accident was studied furthermore. In the Pb-Bi coolant, the melt flows upward due to density difference. The migration and solidification behavior are greatly affected by the surface tension and viscous resistance varying with enthalpy. The whole movement process can be divided into three stages depending on the change in velocity. The heat transfer of core melt is determined jointly by two heat transfer modes: flow heat transfer and solid conductivity. Generally, the research results indicate that the MPS method has unique advantage in studying the migration and solidification behavior in LBR severe accident.

Application of AFLPs to Phylogenetic Analysis of Aegilops (AFLPs에 의한 Aegilops의 계통발생학적 재평가)

  • Park, Yong-Jin;Shim, Jae-Wook
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.6
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    • pp.790-799
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    • 1997
  • Aegilops genus is known to include the donor species of the Band D genome of the bread wheat(ABD). An effort to establish a better strategy for phylogenetic relationships about Aegilops polyploids by AFLPs(Amplified Fragment Length Polymorphisms) was conducted using the 19 Aegilops sPP. and T. aestivum. The 207 polymorphic bands from the amplified products on the 6% acrylamide denaturing sequencing gels were obtained with the 7 AFLP primer combinations, and used to account for the genetic similarities and cluster analysis using NTSYS program. According to the genome analysis, the $M^h$-genome of Ae. heldreichii was estimated as an intermediate genome between the M-genome of Ae. comosa and N-genome of Ae. uniaristata and supposed to be incorporated in the establishing process of UM-genome as a possible diploid donor. And Ae. ventricosa(DN) was more close to Ae. umbellulata(U) than Ae. squarrosa(D). The close relationship between Ae. squarrosa and T. aestivum was perceived as a diploid donor of D-genome. As for the polyploid species, hexaploid Ae. triaristata was more closely related to Ae. columnaris rather than tetraploid Ae. triaristata. The clustered groups were, basically same to the previous Gihara's sections based on phenotypes and pairing analysis of interspecific hybrids. AFLP was evaluated as an efficient and powerful method in the genome evaluation of closely related species.

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Developing system of forest habitat quality assessment for endangered species (멸종위기 야생생물 산림 서식지 질적 평가 체계 개발)

  • Kwang Bae Yoon;Sunryoung Kim;Seokwan Cheong;Jinhong Lee;Jae Hwa Tho;Seung Hyun Han
    • Korean Journal of Environmental Biology
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    • v.40 no.3
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    • pp.307-315
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    • 2022
  • In terms of habitat conservation, it is essential to develop a habitat assessment system that can evaluate not only the suitability of the current habitat, but also the health and stability of the habitat. This study aimed to develop a methodology of habitat quality assessment for endangered species by analyzing various existing habitat assessment methods. The habitat quality assessment consisted of selecting targeted species, planning of assessment, selecting targeted sites, assessing performance, calculating grade, and expert verification. Target sites were selected separately from core and potential habitats using a species distribution model or habitat suitability index. Habitat assessment factors were classified into ecological characteristic, landscape characteristic, and species-habitat characteristic. Ecological characteristic consisted of thirteen factors related to health of tree, vegetation, and soil. Landscape characteristic consisted of five factors related to fragment and connectivity of habitat. Species-habitat characteristic consisted of factors for evaluating habitat suitability depending on target species. Since meanings are different depending on characteristics, habitat quality assessment of this study could be used by classifying results for each characteristic according to various assessment purposes, such as designation of alternative habitats, assessment of restoration project, and protected area valuation for endangered species. Forest habitat quality assessment is expected to play an important role in conservation acts of endangered species in the future through continuous supplementation of this system in regard to quantitative assessment criteria and weighting for each factor with an influence.

Stephen Sondheim's Experiments and Strategies for Musical Fusion Style (스티븐 손드하임의 뮤지컬 융합 양식을 향한 실험과 전략)

  • Lee, Eun-Hye
    • Journal of Korea Entertainment Industry Association
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    • v.13 no.7
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    • pp.15-23
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    • 2019
  • This paper aims to identify the characteristics implied in the process of change of Stephen Sondheim's musical style and to derive the structural communication tools that connect the creator and audience and the implications. The strategies that appear in Sondheim's musical works are the following. First, Sondheim's early musical works follow the principles of book musical and emphasize the integration faithful to the basic elements such as the rhythm and meaning of the lyrics. Second, in the era of early postmodernism, Sondheim attempts the principle of extreme deconstruction through the concept musical Company and pursues rational thought through fragmented dramatic flow. Third, while maintaining dramatic linearity to overcome the lack of immersion caused by extreme deconstruction, Sondheim realizes a more elaborate and formalized fusion style of the principles of integration and deconstruction through the choruses and the songs of the characters in Little Night Music and Sweeney Todd. Finally, Sondheim attempts multiple experiments to break down the boundaries of act and scene and to express new subject matters through his works such as Sunday Park with George and Into the Woods. In conclusion, Sondheim's fusion experiments and strategies are the process of finding a framework and a medium of communication to induce dramatic immersion of audience and at the same time create rational thinking about social problems to deliver the message of the creative work with the leading aesthetics of the modern musical's box-office success.

Association study and expression analysis of olfactomedin like 3 gene related to meat quality, carcass characteristics, retail meat cut, and fatty acid composition in sheep

  • Listyarini, Kasita;Sumantri, Cece;Rahayu, Sri;Uddin, Muhammad Jasim;Gunawan, Asep
    • Animal Bioscience
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    • v.35 no.10
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    • pp.1489-1498
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    • 2022
  • Objective: The objective of this study was to identify polymorphism in olfactomedin like 3 (OLFML3) gene, and association analysis with meat quality, carcass characteristics, retail meat cut, and fatty acid composition in sheep, and expression quantification of OLFML3 gene in phenotypically divergent sheep. Methods: A total of 328 rams at the age of 10 to 12 months with an average body weight of 26.13 kg were used. A novel polymorphism was identified using high-throughput sequencing in sheep and genotyping of OLFML3 polymorphism was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Among 328 rams, 100 rams representing various sheep genotypes were used for association study and proc general linear model was used to analyse association between genotypes and phenotypic traits. Quantitative real-time polymerase chain reaction (qRT-PCR) was used for the expression analysis of OLFML3 mRNA in phenotypically divergent sheep population. Results: The findings revealed a novel polymorphism in the OLFML3 gene (g.90317673 C>T). The OLFML3 gene revealed three genotypes: CC, CT, and TT. The single nucleotide polymorphism (SNP) was found to be significantly (p<0.05) associated with meat quality traits such as tenderness and cooking loss; carcass characteristics such as carcass length; retail meat cut such as pelvic fat in leg, intramuscular fat in loin and tenderloin, muscle in flank and shank; fatty acids composition such as tridecanoic acid (C13:0), palmitoleic acid (C16:1), heptadecanoic acid (C17:0), ginkgolic acid (C17:1), linolenic acid (C18:3n3), arachidic acid (C20:0), eicosenoic acid (C20:1), arachidonic acid (C20:4n6), heneicosylic acid (C21:0), and nervonic acid (C24:1). The TT genotype was associated with higher level of meat quality, carcass characteristics, retail meat cut, and some fatty acids composition. However, the mRNA expression analysis was not different among genotypes. Conclusion: The OLFML3 gene could be a potential putative candidate for selecting higher quality sheep meat, carcass characteristics, retail meat cuts, and fatty acid composition in sheep.

Anaerobic digestion for food wastewater using HADS Pilot Plant and analysis of microbial community in the digester (HADS Pilot Plant를 이용한 음폐수의 혐기성 소화 및 미생물 군집 변화 분석)

  • Ju, Dong-Hun;Lee, Jung-Min;Park, Seong-Bum;Sung, Hyun-Je;Bae, Jae-Sang;Sang, Byoung-In
    • Journal of the Korea Organic Resources Recycling Association
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    • v.18 no.2
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    • pp.76-83
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    • 2010
  • We(Hansol EME Co. Ltd.) proceeded anaerobic digestion test for domestic food wastewater applying to two operating method for increasing of OLR. The methods are as follows. One was the rapidity operating method which was increasing the OLR continuously and rapidly and the other was the terraced operating method which was increasing the OLR having adaptation period for each step. As a result of this tests, the ratio of VFA/Alkalinity of the process was very unstable under the rapidity operating method then the volume of produced biogas was dramatically decreased. However the process was shown stable performance under the terraced operating method maintaining the ratio of VFA/Alkalinity less than 0.4. Also, the process was performing the biogas recovery of $0.8Nm^3/kgVS_{rem}/d$ and the VS removal ratio of 85%. T-RFLP analysis about the community of bacteria and methanogen is also conducted to check the change of the microbial community according to the methods of OLR increasing operation. The microbial community was changed by the methods of OLR increasing operation according to the result of T-RFLP analysis. Although the anaerobic digestion test was executed by same pilot plant, the reactivity and the tolerance of microbial community for surrounding environment could be considerably changed by the operating method for the process.