• Title/Summary/Keyword: Dual specificity

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Dual positional substrate specificity of rice allene oxide synthase-1: insight into mechanism of inhibition by type II ligand imidazole

  • Yoeun, Sereyvath;Rakwal, Randeep;Han, Oksoo
    • BMB Reports
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    • v.46 no.3
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    • pp.151-156
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    • 2013
  • Phylogenetic and amino acid sequence analysis indicated that rice allene oxide synthase-1 (OsAOS1) is CYP74, and is clearly distinct from CYP74B, C and D subfamilies. Regio- and stereo-chemical analysis revealed the dual substrate specificity of OsAOS1 for (cis,trans)-configurational isomers of 13(S)- and 9(S)-hydroperoxyoctadecadienoic acid. GC-MS analysis showed that OsAOS1 converts 13(S)- and 9(S)-hydroperoxyoctadecadi(tri)enoic acid into their corresponding allene oxide. UV-Visible spectral analysis of native OsAOS1 revealed a Soret maximum at 393 nm, which shifted to 424 nm with several clean isobestic points upon binding of OsAOS1 to imidazole. The spectral shift induced by imidazole correlated with inhibition of OsAOS1 activity, implying that imidazole may coordinate to ferric heme iron, triggering a heme-iron transition from high spin state to low spin state. The implications and significance of a putative type II ligand-induced spin state transition in OsAOS1 are discussed.

Quercetin induces dual specificity phosphatase 5 via serum response factor

  • Kanokkan Boonruang;Ilju Kim;Chaeyoung Kwag;Junsun Ryu;Seung Joon Baek
    • BMB Reports
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    • v.56 no.9
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    • pp.508-513
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    • 2023
  • The phytochemical quercetin has gained attention for its anti-inflammatory and anti-tumorigenic properties in various types of cancer. Tumorigenesis involves the aberrant regulation of kinase/phosphatase, highlighting the importance of maintaining homeostasis. Dual Specificity Phosphatase (DUSP) plays a crucial role in controlling the phosphorylation of ERK. The current study aimed to clone the DUSP5 promoter, and investigate its transcriptional activity in the presence of quercetin. The results revealed that quercetin-induced DUSP5 expression is associated with the serum response factor (SRF) binding site located in the DUSP5 promoter. The deletion of this site abolished the luciferase activity induced by quercetin, indicating its vital role in quercetin-induced DUSP5 expression. SRF protein is a transcription factor that potentially contributes to quercetin-induced DUSP5 expression at the transcriptional level. Additionally, quercetin enhanced SRF binding activity without changing its expression. These findings provide evidence of how quercetin affects anti-cancer activity in colorectal tumorigenesis by inducing SRF transcription factor activity, thereby increasing DUSP5 expression at the transcriptional level. This study highlights the importance of investigating the molecular mechanisms underlying the anti-cancer properties of quercetin, and suggests its potential use in cancer therapy.

Development of Dual-task Based Cognitive Function Screening Test for the Elderly (이중과제를 활용한 노인인지기능 선별검사 개발)

  • Kwak, Ho-Soung;Yoo, Eun-Young;Park, Hae Yean;Lee, Seo-Hyun;Han, Dae-Sung;Park, Ji-Hyuk
    • Therapeutic Science for Rehabilitation
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    • v.9 no.4
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    • pp.69-81
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    • 2020
  • Objective : The purpose of this study was to develop a Yonsei dual task cognitive screening test (Y-DuCog) for the elderly. Methods : The reliability and validity test of Y-DuCog (Yonsei Dual Task Cognitive Screening Test) was developed by 229 elderly people aged over 60 years from community organizations at Gyeonggi-do and Chungcheong-do from May 2019 to August 2019. In addition, the criteria for classifying elderly with cognitive impairment were presented. Results : The correlation analysis between MMSE-K, MoCA-K and Y-DuCog were a correlation between the DTE and CRR of Y-DuCog. As a result of internal consistency, Cronbach's-α values of DTE and CRR showed .848 (p<.01) and .916 (p<.01), respectively. The test-retest reliability was high. The screening point showed 88.7% sensitivity and 83.5% specificity at 31.76 seconds in total DTE, and 84.5% sensitivity and 76.6% specificity at 0.38 in total dual-task CRR. Conclusion : This study verified the reliability and validity of Y-DuCog. It was found that the level of education was not a barrier to the undertaking of this test. Furthermore, the test could be performed easily and quickly. It is also expected to be used to evaluate the effectiveness of cognitive function assessment and intervention methods in the elderly.

Dual-Isotope SPECT Imaging with Thallium-201 and Technetium-99m MIBI in Detecting Coronary Artery Disease (관상동맥질환 진단에서 Thallium-201과 Technetium-99m MIBI를 이용한 Dual-Isotope SPECT 영상)

  • Lee, Gyu-Gwang;Cho, Ihn-Ho;Lee, Hyoung-Woo;Park, Jong-Sun;Won, Kyu-Chang;Sin, Dong-Gu;Kim, Young-Jo;Shim, Bong-Sup
    • Journal of Yeungnam Medical Science
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    • v.16 no.1
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    • pp.101-107
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    • 1999
  • We evaluated the results of sequential SPECT dual-isotope imaging with Tl-201 and Tc-99m MIBI in 24 patients. all of whom also had coronary angiography within the past one month. Coronary angiography showed that 12 patients had no CAD, 4 patients had one-vessel CAD, 7 patients had two-vessel CAD and 1 patient had three-vessel CAD. Serial studies of resting Tl-201 and dipyridamole stress Tc-99m MIBI were completed within 2 hours. When more than 50% of coronary artery narrowing was considered significant, the overall sensitivity and specificity of CAD detection were 91.7%. The sensitivity of CAD detection in patients with one-vessel and multi-vessel diseases was 75% and 100%. respectively. Therefore, sequential dual-isotope SPECT demonstrated high sensitivity and specificity for CAD detection. In conclusion, sequential dual-isotope imaging is feasible and can be completed in a short time and may therefore enhance laboratory throughput and patient convenience.

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Diagnostic testing for Duchenne/Becker Muscular dystrophy using Dual Priming Oligonucleotide (DPO) system (Dual Priming Oligonucleotide (DPO) system을 이용한 듀시엔/베커형 근이영양증 진단법)

  • Kim, Joo-Hyun;Kim, Gu-Hwan;Lee, Jin-Joo;Lee, Dae-Hoon;Kim, Jong-Kee;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.5 no.1
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    • pp.15-20
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    • 2008
  • Purpose : Large exon deletions in the DMD gene are found in about 60% of DMD/BMD patients. Multiplex PCR has been employed to detect the deletion mutation, which frequently generates noise PCR products due to the presence of multiple primers in a single reaction as well as the stringency of PCR conditions. This often leads to a false-negative or false-positive result. To address this problematic issue, we introduced the dual primer oligonucleotide (DPO) system. DPO contains two separate priming regions joined by a polydeoxyinosine linker that results in high PCR specificity even under suboptimal PCR conditions. Methods : We tested 50 healthy male controls, 50 patients with deletion mutation as deletion-positive patient controls, and 20 patients with no deletions as deletion-negative patient controls using DPO-multiplex PCR. Both the presence and extent of deletion were verified by simplex PCR spanning the promoter region (PM) and 18 exons including exons 3, 4, 6, 8, 12, 13, 17, 19, 43-48, 50-52, and 60 in all 120 controls. Results : DPO-multiplex PCR showed 100% sensitivity and specificity for the detection a deletion. However, it showed 97.1% sensitivity and 100% specificity for determining the extent of deletions. Conclusion : The DPO-multiplex PCR method is a useful molecular test to detect large deletions of DMD for the diagnosis of patients with DMD/BMD because it is easy to perform, fast, and cost-effective and has excellent sensitivity and specificity.

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Evaluate the diagnostic accuracy in the assessment of coronary artery stenoses using MDCT (MDCT를 이용한 관상동맥협착 진단의 정확성 평가)

  • Yang, Won-Seok;Sin, Sung-Gyu;Park, Jae-Hong
    • Journal of the Korean Society of Radiology
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    • v.6 no.4
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    • pp.275-279
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    • 2012
  • Methods and results : The suspected patient who have results of CTA and CAG examinations to evaluate coronary stenose to undergo each 16MD CT(n=93) and dual source CT(n=100). As a results of statistic, the highest rank of sensitivity, specificity, PPV, NPV and accuracy in coronary artery with using 16MDCT was displayed in LAD(73.5%), RCA(74.5%), LAD(66.7%), LCX(75%), LCX(67.7%). The mean diagnostic accuracy of dual source CT was more 17% than 16MDCT. Dual source CT was recorded 84% mean of accuracy. In addition to, segments of coronary artery did not show significant differences in all of them. However, distal segment become more and more accurate than proximal site.

Development of Two Quantitative Real-Time PCR Diagnostic Kits for HPV Isolates from Korea

  • Jeeva, Subbiah;Kim, Nam-Il;Jang, In-Kwon;Choi, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1350-1358
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    • 2012
  • Viral pathogens, alongside other pathogens, have major effects on crustacean aquaculture. Hepatopancreatic parvovirus (HPV) is an emerging virus in the shrimp industry and has been detected in shrimp farms worldwide. The HPV genome has greater diversity than other shrimp viruses owing to its wide host range and geographical distribution. Therefore, developing diagnostic tools is essential to detect even small copy numbers from the target region of native HPV isolates. We have developed two easy to use quantitative real-time PCR kits, called Green Star and Dual Star, which contain all of the necessary components for real-time PCR, including HPV primers, using the primers obtained from the sequences of HPV isolates from Korea, and analyzed their specificity, efficiency, and reproducibility. These two kits could detect from 1 to $1{\times}10^9$ copies of cloned HPV DNA. The minimum detection limits obtained from HPV-infected shrimp were $7.74{\times}10^1$ and $9.06{\times}10^1$ copies in the Green Star and Dual Star assay kits, respectively. These kits can be used for rapid, sensitive, and efficient screening for HPV isolates from Korea before the introduction of postlarval stages into culture ponds, thereby decreasing the incidence of early development of the disease.

Isoforms of Glucose 6-Phosphate Dehydrogenase in Deinococcus radiophilus

  • Sung, Ji-Youn;Lee, Young-Nam
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.318-325
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    • 2007
  • Glucose 6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) in Deinococcus radiophilus, an extraordinarily UV-resistant bacterium, was investigated to gain insight into its resistance as it was shown to be involved in a scavenging system of superoxide $(O_2^{-1})$ and peroxide $(O_2^{-2})$ generated by UV and oxidative stresses. D. radiophilus possesses two G6PDH isoforms: G6PDH-1 and G6PDH-2, both showing dual coenzyme specificity for NAD and NADP. Both enzymes were detected throughout the growth phase; however, the substantial increase in G6PDH-1 observed at stationary phase or as the results of external oxidative stress indicates that this enzyme is inducible under stressful environmental conditions. The G6PDH-1 and G6PDH-2 were purified 122- and 44-fold (using NADP as cofactor), respectively. The purified G6PDH-1 and G6PDH-2 had the specific activity of 2,890 and 1,033 U/mg protein (using NADP as cofactor) and 3,078 and 1,076 U/mg protein (using NAD as cofactor), respectively. The isoforms also evidenced distinct structures; G6PDH-1 was a tetramer of 35 kDa subunits, whereas G6PDH-2 was a dimer of 60kDa subunits. The pIs of G6PDH-1 and G6PDH-2 were 6.4 and 5.7, respectively. Both G6PDH-1 and G6PDH-2 were inhibited by both ATP and oleic acid, but G6PDH-1 was found to be more susceptible to oleic acid than G6PDH-2. The profound inhibition of both enzymes by ${\beta}-naphthoquinone-4-sulfonic$ acid suggests the involvement of lysine at their active sites. $Cu^{2+}$ was a potent inhibitor to G6PDH-2, but a lesser degree to G6PDH-1. Both G6PDH-1 and G6PDH-2 showed an optimum activity at pH 8.0 and $30^{\circ}C$.

Evaluation of p16INK4a/Ki-67 Dual Immunostaining in Liquid-based Cytology for Diagnosis of Uterine Cervical Dysplasia and Cancer (자궁경부 이형성증과 암의 진단을 위한 액상세포 검체에서 p16INK4a/Ki-67 이중면역염색의 평가)

  • Sung, Mi Hee;Lee, Hoon Taek;Shin, Min Shik;Oh, Seo Young;Kim, Wook Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.3
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    • pp.132-139
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    • 2015
  • Recently, $p16^{INK4a}$/Ki-67 dual immunostaining has been introduced as a new biomarker protocol for early detection of uterine cervical dysplasia and cancer in liquid-based cytology (LBC). We performed the $p16^{INK4a}$/Ki-67 dual immunostaining using a CINtec$^{(R)}$ PLUS kit in a total of 109 LBC cases of cervicovaginal smear and compared its results with those from LBC, HPV hybrid capture II (HC II) test and histological diagnosis. Expression of $p16^{INK4a}$ and Ki-67 was significantly associated with cases of LSIL or higher in cytological diagnosis and cases of cervical intraepithelial neoplasia (CIN) 1 or higher in histological diagnosis (p<0.001 and p<0.001, respectively). Among forty-six cases of atypical squamous cells of undetermined significance (ASCUS) in LBC, $p16^{INK4a}$ and Ki-67 was expressed in 31 (67.4%), which were positively associated with cases of CIN I lesion or higher in histology. The sensitivity of $p16^{INK4a}$/Ki-67 dual immunostaining for finding lesions of CIN 1 or higher was 89.0%, which was higher than LBC. The specificity was 73.5%, which was higher than that of the HC II test. Based on these results, the $p16^{INK4a}$/Ki-67 dual immunostaining method can be a useful diagnostic marker for improving the sensitivity of LBC and the specificity of HC II test.