• Title/Summary/Keyword: Brain Korea 21 Program

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Synthesis of a PEGylated tracer for radioiodination and evaluation of potential in tumor targeting

  • Abhinav Bhise;Sushil K Dwivedi;Kiwoong Lee;Jeong Eun Lim;Subramani Rajkumar;Woonghee Lee;Seong Hwan Cho;Jeongsoo Yoo
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.7 no.2
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    • pp.79-84
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    • 2021
  • Radiopharmaceuticals are important for tumor diagnosis and therapy. To deliver a radiotracer at the desired target excluding non-targeted tissues is difficult The development of a targeted tracer that has a good clearance profile while maintaining high biostability and biocompatibility is key to optimizing its biodistribution and transport across biological barriers. Improving the hydrophilicity of radiotracers by PEGylation can reduce serum binding, allowing the tracer to circulate without retention and reducing its affinity for non-targeted tissues. In this study, we synthesized a new benzamido tracer (SnBz-PEG36) with the introduction of a low molecular weight polyethylene glycol unit (PEG36, ~2,100 Da). The tumor targeting efficiency and biodistribution of [131I]-Bz-PEG36 or radiotracer-loaded liposomes were evaluated after their administration to normal mice or mouse tumor models including CT26 (xenograft) and 4T1 (xenograft and orthotopic). Most of the radiotracer was cleared out rapidly (1-24 h post-administration) through the kidney and there was little tumor uptake.

Functional Analysis of the Stress-Inducible Soybean Calmodulin Isoform-4 (GmCaM-4) Promoter in Transgenic Tobacco Plants

  • Park, Hyeong Cheol;Kim, Man Lyang;Kang, Yun Hwan;Jeong, Jae Cheol;Cheong, Mi Sun;Choi, Wonkyun;Lee, Sang Yeol;Cho, Moo Je;Kim, Min Chul;Chung, Woo Sik;Yun, Dae-Jin
    • Molecules and Cells
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    • v.27 no.4
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    • pp.475-480
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    • 2009
  • The transcription of soybean (Glycine max) calmodulin isoform-4 (GmCaM-4) is dramatically induced within 0.5 h of exposure to pathogen or NaCl. Core cis-acting elements that regulate the expression of the GmCaM-4 gene in response to pathogen and salt stress were previously identified, between -1,207 and -1,128 bp, and between -858 and -728 bp, in the GmCaM-4 promoter. Here, we characterized the properties of the DNA-binding complexes that form at the two core cis-acting elements of the GmCaM-4 promoter in pathogen-treated nuclear extracts. We generated GUS reporter constructs harboring various deletions of approximately 1.3-kb GmCaM-4 promoter, and analyzed GUS expression in tobacco plants transformed with these constructs. The GUS expression analysis suggested that the two previously identified core regions are involved in inducing GmCaM-4 expression in the heterologous system. Finally, a transient expression assay of Arabidopsis protoplasts showed that the GmCaM-4 promoter produced greater levels of GUS activity than did the CaMV35S promoter after pathogen or NaCl treatments, suggesting that the GmCaM-4 promoter may be useful in the production of conditional gene expression systems.

Pathogen Inducible Voltage-Dependent Anion Channel (AtVDAC) Isoforms Are Localized to Mitochondria Membrane in Arabidopsis

  • Lee, Sang Min;Hoang, My Hanh Thi;Han, Hay Ju;Kim, Ho Soo;Lee, Kyunghee;Kim, Kyung Eun;Kim, Doh Hoon;Lee, Sang Yeol;Chung, Woo Sik
    • Molecules and Cells
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    • v.27 no.3
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    • pp.321-327
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    • 2009
  • Voltage-dependent anion channels (VDACs) are reported to be porin-type, ${\beta}$-barrel diffusion pores. They are prominently localized in the outer mitochondrial membrane and are involved in metabolite exchange between the organelle and the cytosol. In this study, we have investigated a family of VDAC isoforms in Arabidopsis thaliana (AtVDAC). We have shown that the heterologous expression of AtVDAC proteins can functionally complement a yeast mutant lacking the endogenous mitochondrial VDAC gene. AtVDACs tagged with GFP were localized to mitochondria in both yeast and plant cells. We also looked at the response of AtVDACs to biotic and abiotic stresses and found that four AtVDAC transcripts were rapidly up-regulated in response to a bacterial pathogen.

Application of SYBR Green real-time PCR assay for the specific detection of Salmonella spp. (Salmonella spp. 특이적인 검출을 위한 SYBR Green real-time PCR 기법 적용)

  • Shin, Seung Won;Cha, Seung Bin;Lee, Won-Jung;Shin, Min-Kyoung;Jung, Myunghwan;Yoo, Anna;Jung, Byeng Yeal;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.53 no.1
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    • pp.25-28
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    • 2013
  • The aim of this study was to applicate and evaluate a SYBR Green real-time PCR for the specific detection of Salmonella spp. Specificity of the PCR method was confirmed with 48 Salmonella spp. and 5 non-Salmonella strains using invA gene primer. The average threshold cycle ($C_T$) of Salmonella spp. was $11.83{\pm}0.78$ while non-Salmonella spp. was $30.86{\pm}1.19$. Correlation coefficients of standard curves constructed using $C_T$ versus copy number of Salmonella Enteritidis ATCC 13076 showed good linearity ($R^2=0.993$; slope = 3.563). Minimum level of detection with the method was > $10^2$ colony forming units (CFU)/mL. These results suggested that the SYBR Green real-time PCR might be applicable for the specific detection of Salmonella spp. isolates.

Isolation of CONSTANS as a TGA4/OBF4 Interacting Protein

  • Song, Young Hun;Song, Na Young;Shin, Su Young;Kim, Hye Jin;Yun, Dae-Jin;Lim, Chae Oh;Lee, Sang Yeol;Kang, Kyu Young;Hong, Jong Chan
    • Molecules and Cells
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    • v.25 no.4
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    • pp.559-565
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    • 2008
  • Members of the TGA family of basic domain/leucine zipper transcription factors regulate defense genes through physical interaction with NON-EXPRESSOR OF PR1 (NPR1). Of the seven TGA family members, TGA4/octopine synthase (ocs)-element-binding factor 4 (OBF4) is the least understood. Here we present evidence for a novel function of OBF4 as a regulator of flowering. We identified CONSTANS (CO), a positive regulator of floral induction, as an OBF4-interacting protein, in a yeast two-hybrid library screen. OBF4 interacts with the B-box region of CO. The abundance of OBF4 mRNA cycles with a 24 h rhythm under both long-day (LD) and short-day (SD) conditions, with significantly higher levels during the night than during the day. Electrophoretic mobility shift assays revealed that OBF4 binds to the promoter of the FLOWERING LOCUS T (FT) gene, a direct target of CO. We also found that, like CO and FT, an OBF4:GUS construct was prominently expressed in the vascular tissues of leaf, indicating that OBF4 can regulate FT expression through the formation of a protein complex with CO. Taken together, our results suggest that OBF4 may act as a link between defense responses and flowering.

Identification and Characterization of Alternative Promoters of the Rice MAP Kinase Gene OsBWMK1

  • Koo, Sung Cheol;Choi, Man Soo;Chun, Hyun Jin;Park, Hyeong Cheol;Kang, Chang Ho;Shim, Sang In;Chung, Jong Il;Cheong, Yong Hwa;Lee, Sang Yeol;Yun, Dae-Jin;Chung, Woo Sik;Cho, Moo Je;Kim, Min Chul
    • Molecules and Cells
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    • v.27 no.4
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    • pp.467-473
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    • 2009
  • Our previous study suggested that OsBWMK1, a gene which encodes a member of the rice MAP kinase family, generates transcript variants which show distinct expression patterns in response to environmental stresses. The transcript variants are generated by alternative splicing and by use of alternative promoters. To test whether the two alternative promoters, pOsBWMK1L (promoter for the OsBWMK1L splice variant) and pOsBWMK1S (promoter for the OsBWMK1S splice variant), are biologically functional, we analyzed transgenic plants expressing GUS fusion constructs for each promoter. Both pOsBWMK1L and pOsBWMK1S are biologically active, although the activity of pOsBWMK1S is lower than that of pOsBWMK1L. Histochemical analysis revealed that pOsBWMK1L is constitutively active in most tissues at various developmental stages in rice and Arabidopsis, whereas pOsBWMK1S activity is spatially and temporally restricted. Furthermore, the expression of pOsBWMK1S::GUS was upregulated in response to hydrogen peroxide, a plant defense signaling molecule, in both plant species. These results suggest that the differential expression of OsBWMK1 splice variants is the result of alternative promoter usage and, moreover, that the mechanisms controlling OsBWMK1 gene expression are conserved in both monocot and dicot plants.

Relationship between biofilm formation and the antimicrobial resistance in the Staphylococcus spp. isolated from animal and air

  • Seo, Yeon-Soo;Lee, Deog Young;Kang, Mi Lan;Lee, Won Jung;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.49 no.3
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    • pp.231-236
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    • 2009
  • Biofilm has been described as a barrier, which produced by microorganisms to survive and protect themselves against various environments, like antibiotic agents. Staphylococcus spp. is a common cause of nosocomial and environmental infection. Thirty-six and thirty-five Staphylococci were isolated from animals and air, respectively. Based on the biofilm forming ability of the bacterium reported in our previous report, relationship between biofilm formation and antibiotic-resistance was investigated in this study. Regarding antibiotics susceptibility, cefazolin was the most effective agent to the bacteria. Strong biofilm-forming Staphylococcus spp. isolates might have a higher antibiotic resistance than weak biofilm isolates regardless of the presence of antibiotic resistance genes (p < 0.05). This result suggested that the chemical complexity of the biofilm might increase the antibiotic resistance due to the decrease of antibiotic diffusion into cells through the extensive matrix.

Role of polyethylene glycol (PEG) linkers: trends in antibody conjugation and their pharmacokinetics

  • Kondapa Naidu Bobba;Abhinav Bhise;Subramani Rajkumar;Woonghee Lee;Jeongsoo Yoo
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.6 no.2
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    • pp.155-164
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    • 2020
  • Polyethylene glycol (PEG) has been the most commonly used polymer for the past few decades in the field of biomedical applications due to its gold standard stealth effect. PEGylation of antibody-drug conjugates, liposomes, peptides, nanoparticles, and proteins is done to improve their pharmaceutical efficacy and pharmacokinetic properties. PEGylation of antibodies with various PEG linkers improves targeting ability by increasing the blood circulation time and thus enhances the biodistribution profiles. It also assists in minimizing the immediate capture by the reticuloendothelial system. In this review, we summarize the effect of PEG linkers in an antibody conjugation and their pharmacokinetics in the field of biomedical imaging.

Development of Immunochromatography Strip-Test Using Nanocolloidal Gold-Antibody Probe for the Rapid Detection of Aflatoxin B1 in Grain and Feed Samples

  • Shim, Won-Bo;Yang, Zheng-You;Kim, Jung-Sook;Kim, Ji-Young;Kang, Sung-Jo;Woo, Gun-Jo;Chung, Young-Chul;Eremin, Sergei A.;Chung, Duck-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.17 no.10
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    • pp.1629-1637
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    • 2007
  • An immunochromatography (ICG) strip test using a nanocolloidal gold-antibody probe was developed and optimized for the rapid detection of aflatoxin B1 (AFB1). A monoclonal antibody specific to AFB1 was produced from the cloned hybridoma cell (AF78), coupled with nanocolloidal gold, and distributed on the conjugate pad of the ICG strip test. The visual detection limit of the ICG strip test was 0.5 ng/ml, and this method showed a cross-reaction to aflatoxin B2, G1, and G2. In total, 172 grain and feed samples were collected and analyzed by both the ICG strip test and HPLC. The results of the ICG strip test showed a good agreement with those obtained by HPLC. These results indicated that the ICG strip test has a potential use as a rapid and cost-effective screening tool for the determination of AFB1 in real samples and could be applied to the preliminary screening of mycotoxin in food and agricultural products, generating results within 15 min without complicated steps.

Identification of Antigenic Proteins in Trichomonas vaginalis

  • Lee, Hye-Yeon;Hyung, Su-Jin;Lee, Jong-Woong;Kim, Ju-Ri;Shin, Myeong-Heon;Ryu, Jae-Sook;Park, Soon-Jung
    • Parasites, Hosts and Diseases
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    • v.49 no.1
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    • pp.79-83
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    • 2011
  • Trichomoniasis is a sexually transmitted disease due to infection with Trichomonas vaginalis, and it can cause serious consequences for women's health. To study the virulence factors of this pathogen, T. vaginalis surface proteins were investigated using polyclonal antibodies specific to the membrane fractions of T. vaginalis. The T. vaginalis expression library was constructed by cloning the cDNA derived from mRNA of T. vaginalis into a phage ${\lambda}$ Uni-ZAP XR vector, and then used for immunoscreening with the anti-membrane proteins of T. vaginalis antibodies. The immunoreactive proteins identified included adhesion protein AP65-1, ${\alpha$-actinin, kinesin-associated protein, teneurin, and 2 independent hypothetical proteins. Immunofluorescence assays showed that AP65-1, one of the identified immunogenic clones, is prevalent in the whole body of T. vaginalis. This study led us to identify T. vaginalis proteins which may stimulate immune responses by human cells.