• Title/Summary/Keyword: fAB

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Intracellular Expression of CTB in Vibrio cholerae Strains in Laboratory Culture Conditions

  • Hunseok Choi;Seonghyeon Son;Donghyun Lee;Jonghyun Bae;Eunyoung Seo;Dong Wook Kim;Eun Jin Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.736-744
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    • 2023
  • The introduction of the toxT-139F allele triggers the expression of TCP (toxin co-regulated pilus) and CT (cholera toxin) under simple laboratory culture conditions in most Vibrio cholerae strains. Such V. cholerae strains, especially strains that have been used in OCVs (oral cholera vaccines), can induce antibody responses against TCP in animal models. However, CT produced in these V. cholerae strains is secreted into the culture medium. In this study, V. cholerae strains that can express intracellular CTB under the control of the toxT-139F allele have been constructed for potential application in OCVs. First, we constructed a recombinant plasmid directly linking the ctxAB promoter to ctxB without ctxA and confirmed CTB expression from the plasmid in V. cholerae containing the toxT-139F allele. We constructed another recombinant plasmid to express NtrCTB, from which 14 internal amino acids-from the 7th to the 20th amino acid-of the leader peptide of CTB have been omitted, and we found that NtrCTB remained in the cells. Based on those results, we constructed V. cholerae strains in which chromosomal ctxAB is replaced by ntrctxB or ntrctxB-dimer. Both NtrCTB and NtrCTB-dimer remained in the bacterial cells, and 60% of the NtrCTB-dimer in the bacterial cells was maintained in a soluble form. To develop improved OCVs, these strains could be tested to see whether they induce immune responses against CTB in animal models.

Association between Genotypes of the Isocitrate Dehydrogenase 3, beta subunit (IDH3B) Gene and Carcass Traits in an F2 Crossbred Population of Landrace × Jeju (Korea) Black Pigs (제주흑돼지와 랜드레이스 교배 2세대의 도체형질과 IDH3B 유전자형의 상관관계)

  • Kang, Yong-Jun;Jeong, Dong Kee;Cho, In-Cheol;Han, Sang-Hyun
    • Journal of Life Science
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    • v.26 no.4
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    • pp.414-418
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    • 2016
  • This study tested the association between genetic polymorphisms of the isocitrate dehydrogenase 3, beta subunit (IDH3B) gene and economic traits in an F2 crossbred population of Landrace × Jeju (South Korea) Black pigs. A 304-bp insertion/deletion mutation in promoter region was screened for determining genotypes of the IDH3B gene in a total of 1,105 F2 pigs. Three genotypes (AA, AB, and BB) were identified in the founder, F1, and F2 populations. Association analysis showed significant differences in carcass weights (CW), backfat thicknesses in three positions of the body (4th-5th ribs, BF5; 11th-12th ribs, BF12; 13th rib-1st lumbar, BFL), and carcass lengths (CL) (p<0.05), but not in meat color (MC), eye muscle area (EMA), or marbling scores (MARB) (p>0.05). The F2 IDH3B BB homozygotes showed heavier CW (80.790±0.725 kg) and shorter CL (101.875±0.336 cm) than the other genotypes (p<0.05). In addition, the BF levels between the 4th - 5th and 11th - 12th vertebrae were thicker in the carcasses of pigs with the IDH3B BB genotype than with the other genotypes (p<0.05). These results suggested that genetic variations in the IDH3B gene may serve as molecular genetic markers for improving the Landrace × Jeju Black pig crossbreeding systems.

Genome Sequencing and Genome-Wide Identification of Carbohydrate-Active Enzymes (CAZymes) in the White Rot Fungus Flammulina fennae

  • Lee, Chang-Soo;Kong, Won-Sik;Park, Young-Jin
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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    • pp.300-312
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    • 2018
  • Whole-genome sequencing of the wood-rotting fungus, Flammulina fennae, was carried out to identify carbohydrate-active enzymes (CAZymes). De novo genome assembly (31 kmer) of short reads by next-generation sequencing revealed a total genome length of 32,423,623 base pairs (39% GC). A total of 11,591 gene models in the assembled genome sequence of F. fennae were predicted by ab initio gene prediction using the AUGUSTUS tool. In a genome-wide comparison, 6,715 orthologous groups shared at least one gene with F. fennae and 10,667 (92%) of 11,591 genes for F. fennae proteins had orthologs among the Dikarya. Additionally, F. fennae contained 23 species-specific genes, of which 16 were paralogous. CAZyme identification and annotation revealed 513 CAZymes, including 82 auxiliary activities, 220 glycoside hydrolases, 85 glycosyltransferases, 20 polysaccharide lyases, 57 carbohydrate esterases, and 45 carbohydrate binding-modules in the F. fennae genome. The genome information of F. fennae increases the understanding of this basidiomycete fungus. CAZyme gene information will be useful for detailed studies of lignocellulosic biomass degradation for biotechnological and industrial applications.

AB INITIO CALCULATIONS OF STRONGLY CORRELATED ELECTRONS: ANTIFERROMAGNETIC GROUND STATE OF $UO_2$

  • YUN YOUNSUK;KIM HANCHUL;KIM HEEMOON;PARK KWANGHEON
    • Nuclear Engineering and Technology
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    • v.37 no.3
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    • pp.293-298
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    • 2005
  • We have performed the density functional theory calculations of $UO_2$ using the spin-polarized generalized gradient approximation (SP-GGA) and the SP-GGA+U approach. The SP-GGA+U approach correctly predicts the insulating electronic structure with antiferromagnetic ordering, but the SP-GGA calculations predict metallic behavior. The cohesive properties obtained from the SP-GGA+U calculations are in good agreement with the available experimental results and previous calculations. The spin-polarized local density of states shows that the antiferromagnetic ordering of $UO_2$ is governed by 5f orbitals of uranium ion. Our calculations demonstrate that the strong correlation of U 5f electrons should be taken into account for a reliable description of $UO_2$ physics.

Multiple Monoclonal Antibodies Produced in a Single Transgenic Plant (형질전환 식물체에서의 복합 단일 항체 단백질 생산)

  • Ahn, Mi-Hyun;Oh, Eun-Yi;Song, Mi-Ra;Lu, Zhe;Kim, Hyun-Soon;Joung, Hyouk;Ko, Ki-Sung
    • Journal of Life Science
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    • v.19 no.1
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    • pp.123-128
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    • 2009
  • Production of highly valuable immunotherapeutic proteins such as monoclonal antibodies and vaccines using plant biotechnology and genetic engineering has been studied as a popular research field. Plant expression system for mass production of such useful recombinant therapeutic proteins has several advantages over other existing expression systems with economical and safety issues. Immunotherapy of multiple monoclonal antibodies, which can recognize multiple targeting including specific proteins and their glycans highly expressed on the surface of cancer cells, can be an efficient treatment compared to a single targeting immunotherapy using a single antibody. In this study, we have established plant production system to express two different targeting monoclonal antibodies in a single transgenic plant through crossing fertilization between two different transgenic plants expressing anti-colorectal cancer mAbCO17-1A and anti-breast cancer mAbBR55, respectively. The F1 seedlings were obtained cross fertilization between the two transgenic parental plants. The presence, transcription, and protein expression of heavy chain (HC) and light chain (LC) genes of both mAbs in the seedlings were investigated by PCR, RT-PCR, and immunoblot analyses, respectively. Among all the seedlings, some seedlings did not carry or transcribe the HC and LC genes of both mAbs. Thus, the seedlings with presence and transcription of HC and LC genes of both mAbs were selected, and the selected seedlings were confirmed to have relatively stronger density of HC and LC protein bands compared to the transgenic plant expressing only each mAb. These results indicate that the F1 seedling plant with carrying both mAb genes was established. Taken together, plant crossing fertilization can be applied to generate an efficient production system expressing multiple monoclonal antibodies for immunotherapy in a single plant.

Detection of Tuberculous Lesion by Immunoscintigraphy Using Radiolabeled Specific Polyclonal Antibody Against M. bovis BCG in Rabbit: A Preliminary Result (가토에서 방사면역 신티그래피를 이용한 결핵병변의 진단 : 예비보고)

  • Lee, Jong-Doo;Shin, Kyoo-Ho;Cho, Sang-Nae;Shin, Jeon-Soo;Lee, Min-Geol;Yang, Woo-Ick;Park, Chang-Yoon;Yoo, Hyung-Sik;Lee, Jong-Tae;Awh, Ok-Doo;Park, Kyung-Bae;Kim, Jae-Rok
    • The Korean Journal of Nuclear Medicine
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    • v.25 no.2
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    • pp.245-251
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    • 1991
  • 결핵성 병변의 단순 x-ray 촬영이나 CT, MRI 소견은 매우 다양하며, 결핵과 전이암 혹은 원발성 암과 감별이 어려운 경우가 있어 결핵으로 확진하기 위하여서 조직 생검이나 수술 등 침습적인 진단 방법을 이용하여야 하였다. 그러므로 이러한 결핵 병변을 비 침습적인 방법으로 정확히 감별할 수 있는 방법을 연구하던 바, 결핵균에 대한 항체를 동위원소에 부착시켜 신티그래피로 진단할 수 있는지의 가능성을 동물실험을 통하여 알아보고자 하였다. 15마리의 가토에서 M.tuberculosis H37Rv를 슬관절에 주입시켜 결핵병변을 유발시키고, 대조군으로 2마리의 가토의 고환에 T.pallidum을 주입하여 매독병변을 유발시킨 후 M.bovis BCG에 대한 특이항체 (specific polyclonal antibody)와 정상 가토의 immunoglobulin을 I-131에 부착시켜 각각의 가토에 주입하여 preset time 10분간 감마카메라로 주사한 결과 다음과 같은 결과를 얻었다. (1) 8마리의 결핵에 감염된 가토에 M.bovis BCG에 대한 $F(ab')_2$를 1 mCi의 I -131 labeling 시킨후 주사한 결과 모두에서 주사후 2시간 부터 72시간까지 병소가 hot uptake으로 보였으며 주사후 24 시간에 가장 높은 target/background ratio를 보였다. (2) 2마리의 매독에 감염된 가토에서 anti-BCG $F(ab')_2$를 주사한 결과 2시간에서는 병소에 hot activity를 보였으나 24시간부터 급격히 activity가 감소하였다. (3) $F(ab')_2$ 대신에 intact antibody를 결핵에 감염된 가토에 투여한 결과 specific polyclonal antibody나 정상가토의 immunoglobulin 모두 결핵병소에 96시간까지 hot uptake를 보였다. 그러므로 결핵균에 대한 specific antibody fragment를 이용하면 방사면역 신티그램으로 진단이 가능하리라 사료되었고, intact antibody를 사용할 경우 sensitivity는 높으나 specificity는 적을 것으로 사료되었다.

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Prospective Observation Study on Hyperthyroidism Patients Treated with Korean Medicine (한의치료를 받은 갑상선 기능 항진증 환자에 대한 전향적 관찰연구)

  • Choi, Yu-jin;Shin, Seon-mi;Han, Yang-hee;Ahn, Se-young;Cho, Chung-sik
    • The Journal of Internal Korean Medicine
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    • v.39 no.1
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    • pp.84-96
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    • 2018
  • Objective: This is aprospective clinical case study that includes the administration of Ahnjeonbaekho-tang (AJBHT) to patients who have suffered from Graves' disease for the 8 weeks of the clinical research. Methods: Without administration of an antithyroid drug, AJBHT was administered 3 times a day for 8 weeks. In Case 1, AJBHT without modification was administered and in Case 2, in accordance with the patients' symptoms, a dose of Gypsum Fibrosum was increased by 4g from the beginning of week 2, and further increased by 16 g, for a total of 20 g between week 6 and week 8. Body mass index (BMI), thyroid function test (TFT), thyroid autoantibody test, complete blood cell count (CBC), and blood chemistry (BC) tests were conducted at an interval of 4 weeks, for a total of 3 times during the study. The clinical manifestations, a spectrum of symptoms of the patients, was observed by Wayne's Index, Euro-QOL-5 Dimension (EQ-5D), and Perceived Stress Scale (PSS). Results: In both cases, an acceptable significance of reduction of value in T3, fT4, Thyroid stimulating antibody (TSAb) and Thyroid stimulating hormone receptor antibody (TSH-R-Ab) was observed. After treatment, in TFT, T3 decreased by 31.66%, fT4 decreased by 32.82% in Case 1, and T3 decreased by 43.42%, and fT4 decreased by 37.32% in Case 2. In the thyroid autoantibody test, TSAb decreased by 7.59%, and TSH-R-Ab decreased by 53.19% in Case 1, and TSAb decreased by 33.45%, TSH-R-Ab decreased by at least 7.75% in Case 2. Besides this, there was a decreasing trend of Wayne's index, and a loss of typical symptoms of hyperthyroidism also declared the efficacy. Conclusion: From these results, AJBHT is very effective in the regulation of TFT and improving the symptoms of hyperthyroidism, and is also expected to be an effective alternative to antithyroid drugs for patients who have side effects or drug intolerance.

The eigensolutions of wave propagation for repetitive structures

  • Zhong, Wanxie;Williams, F.W.
    • Structural Engineering and Mechanics
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    • v.1 no.1
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    • pp.47-60
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    • 1993
  • The eigen-equation of a wave traveling over repetitive structure is derived directly form the stiffness matrix formulation, in a form which can be used for the case of the cross stiffness submatrix $K_{ab}$ being singular. The weighted adjoint symplectic orthonormality relation is proved first. Then the general method of solution is derived, which can be used either to find all the eigensolutions, or to find the main eigensolutions for large scale problems.