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Construction and validation of a synthetic phage-displayed nanobody library

  • Minju Kim;Xuelian Bai;Hyewon Im;Jisoo Yang;Youngju Kim;Minjoo MJ Kim;Yeonji Oh;Yuna Jeon;Hayoung Kwon;Seunghyun Lee;Chang-Han Lee
    • The Korean Journal of Physiology and Pharmacology
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    • v.28 no.5
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    • pp.457-467
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    • 2024
  • Nanobodies derived from camelids and sharks offer unique advantages in therapeutic applications due to their ability to bind to epitopes that were previously inaccessible. Traditional methods of nanobody development face challenges such as ethical concerns and antigen toxicity. Our study presents a synthetic, phage-displayed nanobody library using trinucleotide-directed mutagenesis technology, which allows precise amino acid composition in complementarity-determining regions (CDRs), with a focus on CDR3 diversity. This approach avoids common problems such as frameshift mutations and stop codon insertions associated with other synthetic antibody library construction methods. By analyzing FDA-approved nanobodies and Protein Data Bank sequences, we designed sub-libraries with different CDR3 lengths and introduced amino acid substitutions to improve solubility. The validation of our library through the successful isolation of nanobodies against targets such as PD-1, ATXN1 and STAT3 demonstrates a versatile and ethical platform for the development of high specificity and affinity nanobodies and represents a significant advance in biotechnology.

A retrospective study on related factors affecting the survival rate of dental implants

  • Jang, Hee-Won;Kang, Jeong-Kyung;Lee, Ki;Lee, Yong-Sang;Park, Pil-Kyoo
    • The Journal of Advanced Prosthodontics
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    • v.3 no.4
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    • pp.204-215
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    • 2011
  • PURPOSE. The aim of this retrospective study is to analyze the relationship between local factors and survival rate of dental implant which had been installed and restored in Seoul Veterans Hospital dental center for past 10 years. And when the relationship is found out, it could be helpful to predict the prognosis of dental implants. MATERIALS AND METHODS. A retrospective study of patients receiving root-shaped screw-type dental implants placed from January 2000 to December 2009 was conducted. 6385 implants were placed in 3755 patients. The following data were collected from the dental records and radiographs: patient's age, gender, implant type and surface, length, diameter, location of implant placement, bone quality, prosthesis type. The correlations between these data and survival rate were analyzed. Statistical analysis was performed with the use of Kaplan-Meier analysis, Chi-square test and odds ratio. RESULTS. In all, 6385 implants were placed in 3755 patients (3120 male, 635 female; mean age $65{\pm}10.58$ years). 108 implants failed and the cumulative survival rate was 96.33%. There were significant differences in age, implant type and surface, length, location and prosthesis type (P<.05). No significant differences were found in relation to the following factors: gender, diameter and bone quality (P>.05). CONCLUSION. Related factors such as age, implant type, length, location and prosthesis type had a significant effect on the implant survival.

The Importance of Filter Integrity Test to Ensure Sterility of Radiophamaceuticals for Using PET Image

  • Cho, Yong-Hyun;Park, Jun-Hyung;Hwang, Ki-Young;Kim, Hyung-Woo;Lee, Hong-Jae;Kim, Hyun-Ju
    • The Korean Journal of Nuclear Medicine Technology
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    • v.12 no.1
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    • pp.74-77
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    • 2008
  • The radiopharmaceuticals are routinely injected to blood vessel for acquiring PET image. For this reason, It is imperative that they undergo strict quality control measures. Especially, Sterility test is more important than any other quality control procedures. According to the FDA guideline, It requires filter integrity test used in the processing of sterile solutions. Among several methods, we can decide to use bubble point test. We usually use vented GS-filters (Millipore co., USA) which are sterilizinggrade (0.22 um pore size) and are placed upper site on product vial. After the synthesis of $^{18}F$-FDG, solutions wet the membrane in filter and then go into the product vial. By all synthesis steps have finished, we can observe the presence of the bubbles in the product vial. Since we have started this study, we have never found any bubbles in the product vial. Because the maximum pressure intensity of the filter which has set by manufacturer is up to 5 bars, but helium gas pressure is up to 1 bar in our module system. So, we can make 5 bars pressure using helium gas bombe and increase pressure up to 5 bars step by step. However, it does not happen to anything in vial.

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Detection of L-Xylosone and its Physiological Effects in Saccharomyces cerevisiae

  • Seok, Yeong-Jae;Yang, Kap-Seok;Kang, Ju-Gyeong;Kim, Seong-Tae;Huh, Won-Ki;Kang, Sa-Ouk
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.192-197
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    • 1996
  • L-Xylosone was detected as its quinoxaline derivative in the degradation solution of dehydro-L-ascorbic acid. The production rate of L-xylosone was much faster in aerated phosphate-cirate buffer (pH 5. 6) than in pure water. L-Xylosone and dehydro-L-ascorbic acid were identified in the crude extracts of Saccharomyces cerevisiae. The concentration of L-xylosone in the crude cell extracts was calculated to be about 0.2 nmol $(mg protein)^{-1}$. When L-xylosone was added to asynchronous culture of S. cerevisiae, it inhibited primarily the synthesis of protein and RNA. Examination of the effect of L- xylosone on synchronous culture of the yeast indicated that L-xylosone inhibited the initiation of DNA replication and that the cells were arrested at $G_1$, stage of cell division cycle. These results suggested a possibility that L-xylosone can act as an inhibitor of cell growth.

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Optimization of 1D 1H Quantitative NMR (Nuclear Magnetic Resonance) Conditions for Polar Metabolites in Meat

  • Kim, Hyun Cheol;Ko, Yoon-Joo;Kim, Minsu;Choe, Juhui;Yong, Hae In;Jo, Cheorun
    • Food Science of Animal Resources
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    • v.39 no.1
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    • pp.1-12
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    • 2019
  • The objective of this study was to establish an optimized 1D $^1H$ quantitative nuclear magnetic resonance (qNMR) analytical method for analyzing polar metabolites in meat. Three extraction solutions [0.6 M perchloric acid, 10 mM phosphate buffer, water/methanol (1:1)], three reconstitution buffers [20 mM 3-morpholinopropane-1-sulfonic acid, 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid, phosphate buffer], and two pulse programs (zg30, noesypr1d) were evaluated. Extraction with 0.6 M perchloric acid and 20 mM phosphate resulted in a stable baseline and no additional overlap for quantifying polar metabolites in chicken breast. In qNMR analysis, zg30 pulse program (without water-suppression) showed smaller relative standard deviation (RSD) and faster running time than noesypr1d (water-suppression). High-performance liquid chromatography was compared with qNMR analyses to validate accuracy. The zg30 pulse program showed good accuracy and lower RSD. The optimized qNMR method was able to apply for beef and pork samples. Thus, an optimized 1D $^1H$ qNMR method for meat metabolomics was established.

Low driving voltage and high stability organic light-emitting diodes with rhenium oxide-doped hole transporting layer

  • Leem, Dong-Seok;Park, Hyung-Dol;Kang, Jae-Wook;Lee, Se-Hyung;Kim, Jang-Joo
    • 한국정보디스플레이학회:학술대회논문집
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    • 2007.08b
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    • pp.1650-1653
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    • 2007
  • We demonstrate fluorescent green organic lightemitting diodes employing a rhenium oxide ($ReO_3$)-doped N,N-diphenyl-N,N'-bis(1,1'-biphenyl)-4,4'-diamine (NPB) hole transporting layer (HTL). The devices exhibit significantly reduced driving voltages as well as prolonged lifetime. Details of $ReO_3$ doping effects are described in terms of charge transfer complex and stabilization of HTL morphology.

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A Case of Posterior Antebrachial Cutaneous Neuropathy After Injection Therapy (주사 치료 후 발생한 후부 전박 피부 신경병증 1예)

  • Bae, Jong Seok;Jang, Min Uk;Kim, Min Ky;Yoon, Gyong Jae;Kim, Byoung Joon
    • Annals of Clinical Neurophysiology
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    • v.7 no.2
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    • pp.130-132
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    • 2005
  • Isolated posterior antebrachial cutaneous (PABC) neuropathy is rare. A 62 year-old man presented with hypesthesia over the skin region of PABC nerve territory after an injection at the proximal to the lateral epicondyle. Antidromic sensory nerve conduction studies of PABC nerve was not evoked on the affected side. Our case showed that sensory nerve conduction of PABC nerve can be performed without much difficulty and that this test is useful in confirming PABC neuropathy.

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Enhanced Growth Inhibition by Combined Gene Transfer of p53 and $p16^{INK4a}$ in Adenoviral Vectors to Lung Cancer Cell Lines (폐암세포주에 대한 p53 및 $p16^{INK4a}$의 복합종양억제유전자요법의 효과)

  • Choi, Seung -Ho;Park, Kyung-Ho;Seol, Ja-Young;Yoo, Chul-Gyu;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.1
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    • pp.67-75
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    • 2001
  • Background : Two tumor suppressor genes, p53 and p16, which have different roles in controlling the cell cycle and inducing apoptosis, are frequently inactivated during carcinogenesis including lung cancer. Single tumor suppressor gene therapies using either with p53 or p16 have been studied extensively. However, there is a paucity of reports regarding a combined gene therapy using these two genes. Methods : The combined effect of p53 and p16 gene transfer by the adenoviral vector on the growth of lung cancer cell lines and its interactive mechanism was investigated. Results : An isobologram showed that the co-transduction of p53 and p16 exhibited a synergistic growth in hibitory effect on NCI H358 and an additive effect on NCI H23. Cell cycle analysis demonstrated the induction of a synergistic G1/S arrest by a combined p53 and p16 transfer. This synergistic interaction was again confirmed in a soft agar confirmed in a soft agar clonogenic assay. Conclusion : These observations suggest the potential of a p53 and p16 combination gene therapy as another potent strategy in cancer gene therapy.

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