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Domain Characterization of Cyclosporin Regio-Specific Hydroxylases in Rare Actinomycetes

  • Woo, Min-Woo;Lee, Bo-Ram;Nah, Hee-Ju;Choi, Si-Sun;Li, Shengying;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1634-1639
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    • 2015
  • Cytochrome P450 hydroxylase (CYP) in actinomycetes plays an important role in the biosynthesis and bioconversion of various secondary metabolites. Two unique CYPs named CYP-sb21 and CYP-pa1, which were identified from Sebekia benihana and Pseudonocardia autotrophica, respectively, were proven to transfer a hydroxyl group at the 4th or 9th N-methyl leucine position of immunosuppressive agent cyclosporin A (CsA). Interestingly, these two homologous CYPs showed different CsA regio-selectivities. CYP-sb21 exhibited preferential hydroxylation activity at the 4th position over the 9th position, whereas CYP-pa1 showed the opposite preference. To narrow down the CYP domain critical for CsA regio-selectivity, each CYP was divided into four domains, and each domain was swapped with its counterpart from the other CYP. A total of 18 hybrid CYPs were then individually tested for CsA regio-selectivity. Although most of the hybrid CYPs failed to exhibit a significant change in regio-selectivity in the context of CsA hydroxylation, hybrid CYP-pa1 swapped with the second domain of CYP-sb21 showed a higher preference for the 9th position. Moreover, hybrid CYPsb21 containing seven amino acids from the 2nd domain of CYP-pa1 showed higher preference for the 4th position. These results imply that the 2nd domain of CsA-specific CYP plays a critical role in CsA regio-selectivity, thereby setting the stage for biotechnological application of CsA regio-selective hydroxylation.

Transportation Time is Significantly Decreased in Acute Ischemic Stroke Patients Under Drip-and-Ship Paradigm for Thrombolysis

  • Kim, Jeong-Yeon;Cha, Jae-Kwan;Kim, Dae-Hyun;Nah, Hyun-Wook;Jeong, Jin-Heon
    • Journal of Neurocritical Care
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    • v.11 no.2
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    • pp.86-92
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    • 2018
  • Background: A delay of transfer for patients with acute stroke needing emergent revascularization is a huge hurdle for efficacy of revascularization. The objective of this study was to investigate changes of transportation time calculated by image to door (ITD) time (from checking brain images at first contact hospital to arriving at our emergency center) before and after 2015. Methods: This study was performed in a retrospective manner from 2013 into 2017. Acute ischemic stroke (AIS) patients having intravenous thrombolysis and/or mechanical thrombectomy during the observation period were enrolled. Among them, those who had revascularization under 'Drip-and-Ship' or 'Ship-and-Drip' paradigm were selected. Results: During the observation period, 225 patients were treated under 'Drip-and-Ship' or 'Ship-and-Drip' paradigm. Twenty-three were excluded due the lack of detailed data. Among 202 patients, 73 and 129 were treated under Drip-and-Ship and Ship-and-Drip paradigms, respectively. In 2013, 35 patients from 18 hospitals (median distance, 25 km) were transferred to our regional stroke center and their median ITD time was 116 minutes. It was gradually decreased after 2015. In 2017, ITD time was significantly (P<0.01) shortened to 85 minutes without significant changes in transfer distance. The median onset to puncture time was also significantly (P=0.03) decreased from 365 minutes in 2013 to 270 minutes in 2017. Conclusion: Our results implicate that many hospitals in our stroke region might have recognized the importance of rapid transportation for AIS after 2015.

Streptomyces BAC Cloning of a Large-Sized Biosynthetic Gene Cluster of NPP B1, a Potential SARS-CoV-2 RdRp Inhibitor

  • Park, Ji-Hee;Park, Heung-Soon;Nah, Hee-Ju;Kang, Seung-Hoon;Choi, Si-Sun;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.7
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    • pp.911-917
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    • 2022
  • As valuable antibiotics, microbial natural products have been in use for decades in various fields. Among them are polyene compounds including nystatin, amphotericin, and nystatin-like Pseudonocardia polyenes (NPPs). Polyene macrolides are known to possess various biological effects, such as antifungal and antiviral activities. NPP A1, which is produced by Pseudonocardia autotrophica, contains a unique disaccharide moiety in the tetraene macrolide backbone. NPP B1, with a heptane structure and improved antifungal activity, was then developed via genetic manipulation of the NPP A1 biosynthetic gene cluster (BGC). Here, we generated a Streptomyces artificial chromosomal DNA library to isolate a large-sized NPP B1 BGC. The NPP B1 BGC was successfully isolated from P. autotrophica chromosome through the construction and screening of a bacterial artificial chromosome (BAC) library, even though the isolated 140-kb BAC clone (named pNPPB1s) lacked approximately 8 kb of the right-end portion of the NPP B1 BGC. The additional introduction of the pNPPB1s as well as co-expression of the 32-kb portion including the missing 8 kb led to a 7.3-fold increase in the production level of NPP B1 in P. autotrophica. The qRT-PCR confirmed that the transcription level of NPP B1 BGC was significantly increased in the P. autotrophica strain containing two copies of the NPP B1 BGCs. Interestingly, the NPP B1 exhibited a previously unidentified SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) inhibition activity in vitro. These results suggest that the Streptomyces BAC cloning of a large-sized, natural product BGC is a valuable approach for titer improvement and biological activity screening of natural products in actinomycetes.

Effects of Ginsenosides on $GABA_A$ Receptor Channels Expressed in Xenopus Oocytes

  • Choi, Se-Eun;Choi, Seok;Lee, Jun-Ho;Paul J.Whiting;Lee, Sang-Mok;Nah, Seung-Yeol
    • Archives of Pharmacal Research
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    • v.26 no.1
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    • pp.28-33
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    • 2003
  • Ginsenosides, major active ingredients of Panax ginseng, are known to regulate excitatory ligand-gated ion channel activity such as nicotinic acetylcholine and NMDA receptor channel activity. However, it is not known whether ginsenosides affect inhibitory ligand-gated ion channel activity. We investigated the effect of ginsenosides on human recombinant $GABA_A$ receptor (${\alpha}_1{\beta}_1{\gamma}_{2s}$) channel activity expressed in Xenopus oocytes using a two-electrode voltage-clamp technique. Among the eight individual ginsenosides examined, namely, $Rb_1$, $Rb_2$, Rc, Rd, Re, Rf, $Rg_1$ and $Rg_2$, we found that Rc most potently enhanced the GABA-induced inward peak current ($I_{GABA}$). Ginsenoside Rc alone induced an inward membrane current in certain batches of oocytes expressing the $GABA_A$ receptor. The effect of ginsenoside Rc on $I_{GABA}$ was both dose-dependent and reversible. The half-stimulatory concentration ($EC_{50}$) of ginsenoside Rc was 53.2$\pm$12.3 $\mu$M. Both bicuculline, a $GABA_A$ receptor antagonist, and picrotoxin, a $GABA_A$ channel blocker, blocked the stimulatory effect of ginsenoside Rc on $I_{GABA}$. Niflumic acid (NFA) and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), both $CI^{-1}$ channel blockers, attenuated the effect of ginsenoside Rc on I$I_{GABA}$. This study suggests that ginsenosides regulated $GABA_A$ receptor expressed in Xenopus oocytes and implies that this regulation might be one of the pharmacological actions of Panax ginseng.

Evaluation of slip coefficient of slip critical joints with high strength bolts

  • Nah, Hwan-Seon;Lee, Hyeon-Ju;Kim, Kang-Seok;Kim, Woo-Bum
    • Structural Engineering and Mechanics
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    • v.32 no.4
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    • pp.477-488
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    • 2009
  • A slip critical joint has various values to adopt the proper slip coefficient in various conditions of faying surfaces in the following codes: AISC, AIJ and Eurocode 3. However, the Korean Building Code still regulates the unique slip coefficient, 0.45, regardless of the diverse faying conditions. In this study, the slip resistance test, including five kinds of surface treatments were conducted to obtain the proper slip coefficients available to steel plate KS SM490A. The faying surfaces were comprised of a clean mill, rust, red lead paint, zinc primer, and shot blast treatment. The candidates for high strength bolts were torque-shear bolts, torque-shear bolts with zinc coating, and ASTM A490 bolts. Based on the test results, the specimens with a shot blasted surface and rusted surface exhibited $k_s$, 0.61, and 0.5, respectively. It is recommended that the specimens with zinc primer exhibit $k_s{\geq}0.40$. The clean mill treated surface had prominently lower values, 0.27. For red lead painted treatment, the thickness of the coating affects the determinant of slip coefficient, so it is necessary to establish a minimum $k_s$ of 0.2, with a coating thickness of 65 ${\mu}m$. During 1,000 hours of relaxation, the uncoated surfaces exhibited the loss of clamping force behind 3%, while the coated surfaces within a certain limited thickness exhibited the loss of clamping within a range of 4.71% and 8.37%.

Ginsentology III;Identifications of Ginsenoside Interaction Sites for Ion Channel Regulation

  • Choi, Sun-Hye;Shin, Tae-Joon;Lee, Byung-Hwan;Lee, Jun-Ho;Hwang, Sung-Hee;Pyo, Mi-Kyung;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.32 no.2
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    • pp.99-106
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    • 2008
  • A ligand - whether an endogenous hormone, neurotransmitter, exogenous toxin or synthetic drug - binds to plasma membrane proteins (e.g., ion channels, receptors or other functional proteins) to exert its physiological or pharmacological effects. Ligands can also have functional groups, showing stereospecificity for interaction sites on their counterpart plasma membrane proteins. Previous reports have shown that the ginsenoside Rg$_3$, a bioactive ginsenoside, meets these criteria in that: 1) an aliphatic side chain of $Rg_3$ plays a role as a functional group, 2) Rg$_3$ regulates voltage- and ligand-gated ion channels in a stereospecific manner with respect to carbon-20, and 3) $Rg_3$ regulates subsets of ligand-gated and voltage-gated ion channels through specific interactions with identified amino acid residues inside the channel pore, in the outer pore entryway, or in toxin binding sites. Rg$_3$, therefore, could be a candidate for a novel ginseng-derived glycosidic ligand regulating ion channels and receptors. This review will examine how Rg$_3$ regulates voltage-gated and ligand-gated ion channels through interactions with its target proteins in the plasma membrane. Hopefully, this review will advance understanding of ginseng pharmacology at the cellular and molecular levels.

Development of a Smartphone Application for Promoting Bone Health in Korean Young Adult Women: A Pilot Study (성인초기 여성의 골건강 증진을 위한 스마트폰 앱 개발 및 적용: 예비연구)

  • Park, Young-Joo;Lee, Sook-Ja;Shin, Nah-Mee;Shin, Hyunjeong;Yoon, Sungjin;Jeon, Songi;Cho, Inhae
    • Korean Journal of Adult Nursing
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    • v.27 no.4
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    • pp.459-471
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    • 2015
  • Purpose: The purpose of this study was to develop a smartphone application and evaluate it's effectiveness and usability for promoting bone health in Korean young adult women. Methods: A smartphone application was developed covering exercise, nutrition, and health habits related to bone health. A pilot testing was conducted using a one-group pre/posttest design. A total of 22 female college students with low bone mass participated in the 8-week use of a pilot testing. Results: The "Strong bone Fit body (SbFb)" application was developed using Android platform. In the pilot testing, bone mineral density, serum CTX, and 25 (OH)Vit.D were significantly decreased. There is no significant change in serum osteocalcin and calcium. Serum phosphorus concentration, nutrient intakes (fiber, VitB6, and pantotenic acid), and all bone composition profiles were significantly increased. Bone-health knowledge was significantly increased but no change in health belief and self-efficacy. Conclusion: The results of this pilot testing suggest the effectiveness of the SbFb application to enhance and promote self-managing abilities for bone health. It can provide continuous feedback and high-quality bone health information. However, some contents and working methods of the application need to be modified and its effectiveness on bone health requires further investigations.

A 12b 100 MS/s Three-Step Hybrid Pipeline ADC Based on Time-Interleaved SAR ADCs

  • Park, Jun-Sang;An, Tai-Ji;Cho, Suk-Hee;Kim, Yong-Min;Ahn, Gil-Cho;Roh, Ji-Hyun;Lee, Mun-Kyo;Nah, Sun-Phil;Lee, Seung-Hoon
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.14 no.2
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    • pp.189-197
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    • 2014
  • This work proposes a 12b 100 MS/s $0.11{\mu}m$ CMOS three-step hybrid pipeline ADC for high-speed communication and mobile display systems requiring high resolution, low power, and small size. The first stage based on time-interleaved dual-channel SAR ADCs properly handles the Nyquist-rate input without a dedicated SHA. An input sampling clock for each SAR ADC is synchronized to a reference clock to minimize a sampling-time mismatch between the channels. Only one residue amplifier is employed and shared in the proposed ADC for the first-stage SAR ADCs as well as the MDAC of back-end pipeline stages. The shared amplifier, in particular, reduces performance degradation caused by offset and gain mismatches between two channels of the SAR ADCs. Two separate reference voltages relieve a reference disturbance due to the different operating frequencies of the front-end SAR ADCs and the back-end pipeline stages. The prototype ADC in a $0.11{\mu}m$ CMOS shows the measured DNL and INL within 0.38 LSB and 1.21 LSB, respectively. The ADC occupies an active die area of $1.34mm^2$ and consumes 25.3 mW with a maximum SNDR and SFDR of 60.2 dB and 69.5 dB, respectively, at 1.1 V and 100 MS/s.

Lymphoid Interstitial Pneumonia Associated with Primary Sjogren's Syndrome - A Case Report (일차성 Sjogren씨 증후군에서 발생한 림프구성 간질성 폐렴 1예: 증례보고와 국내문헌고찰)

  • Mo, Sang-Il;Lee, Hyeok-Gyu;Cho, A-Ra;Chung, Hye-Kyoung;Lee, Ho-Sung;Choi, Jae-Sung;Seo, Ki-Hyun;Nah, Seong-Su;Kim, Yong-Hoon;Na, Ju-Ock
    • Tuberculosis and Respiratory Diseases
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    • v.69 no.5
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    • pp.375-380
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    • 2010
  • Lymphoid interstitial pneumonia (LIP) is a rare benign lymphoproliferative interstitial lung disease. LIP has been associated with autoimmune disorders, HIV, viral infections, and so on. Once underlying systemic diseases have been excluded, a diagnosis of idiopathic LIP can be made. Although 6 cases of pathologically confirmed LIP have occurred in Korea, thus far none has been associated with primary Sjogren's syndrome. A 44-year-old man was admitted to hospital due to a dry cough and dypsnea on exertion that had been ongoing for 2 months. A chest radiography showed multiple and variable-sized cystic lesions, on both lungs and both interstitial infiltration and consolidation in both lower lung fields. Tests for autoantibody showed positive results of anti-nuclear antibody and anti-Ro/La antibody. The patient underwent a video assisted thoracoscopic surgery biopsy and pathologically confirmed LIP. We report the first known case of LIP-associated with primary Sjogren's syndrome in Korea.

TOWARD A NEXT GENERATION SOLAR CORONAGRAPH: DIFFRACTED LIGHT SIMULATION AND TEST RESULTS FOR A CONE OCCULTER WITH TAPERED SURFACE

  • Yang, Heesu;Bong, Su-Chan;Cho, Kyung-Suk;Choi, Seonghwan;Park, Jongyeob;Kim, Jihun;Baek, Ji-Hye;Nah, Jakyoung;Sun, Mingzhe;Gong, Qian
    • Journal of The Korean Astronomical Society
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    • v.51 no.2
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    • pp.27-36
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    • 2018
  • In a solar coronagraph, the most important component is an occulter to block the direct light from the disk of the sun. Because the intensity of the solar outer corona is $10^{-6}$ to $10^{-10}$ times of that of the solar disk ($I_{\odot}$), it is necessary to minimize scattering at the optical elements and diffraction at the occulter. Using a Fourier optic simulation and a stray light test, we investigated the performance of a compact coronagraph that uses an external truncated-cone occulter without an internal occulter and Lyot stop. In the simulation, the diffracted light was minimized to the order of $7.6{\times}10^{-10}I_{\odot}$ when the cone angle ${\theta}_c$ was about $0.39^{\circ}$. The performance of the cone occulter was then tested by experiment. The level of the diffracted light reached the order of $6{\times}10^{-9}I_{\odot}$ at ${\theta}_c=0.40^{\circ}$. This is sufficient to observe the outer corona without additional optical elements such as a Lyot stop or inner occulter. We also found the manufacturing tolerance of the cone angle to be $0.05^{\circ}$, the lateral alignment tolerance was $45{\mu}m$, and the angular alignment tolerance was $0.043^{\circ}$. Our results suggest that the physical size of coronagraphs can be shortened significantly by using a cone occulter.