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Crystal Structure of LysB4, an Endolysin from Bacillus cereus-Targeting Bacteriophage B4

  • Hong, Seokho;Son, Bokyung;Ryu, Sangryeol;Ha, Nam-Chul
    • Molecules and Cells
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    • v.42 no.1
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    • pp.79-86
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    • 2019
  • Endolysins are bacteriophage-derived enzymes that hydrolyze the peptidoglycan of host bacteria. Endolysins are considered to be promising tools for the control of pathogenic bacteria. LysB4 is an endolysin produced by Bacillus cereus-infecting bacteriophage B4, and consists of an N-terminal enzymatic active domain (EAD) and a C-terminal cell wall binding domain (CBD). LysB4 was discovered for the first time as an L-alanoyl-D-glutamate endopeptidase with the ability to breakdown the peptidoglycan among B. cereus-infecting phages. To understand the activity of LysB4 at the molecular level, this study determined the X-ray crystal structure of the LysB4 EAD, using the full-length LysB4 endolysin. The LysB4 EAD has an active site that is typical of LAS-type enzymes, where $Zn^{2+}$ is tetrahedrally coordinated by three amino acid residues and one water molecule. Mutational studies identified essential residues that are involved in lytic activity. Based on the structural and biochemical information about LysB4, we suggest a ligand-docking model and a putative endopeptidase mechanism for the LysB4 EAD. These suggestions add insight into the molecular mechanism of the endolysin LysB4 in B. cereus-infecting phages.

Local Silencing of Connective Tissue Growth Factor by siRNA/Peptide Improves Dermal Collagen Arrangements

  • Cho Lee, Ae-Ri;Woo, Inhae
    • Tissue Engineering and Regenerative Medicine
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    • v.15 no.6
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    • pp.711-719
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    • 2018
  • BACKGROUND: Collagen organization within tissues has a critical role in wound regeneration. Collagen fibril diameter, arrangements and maturity between connective tissue growth factor (CTGF) small interfering RNA (siRNA) and mismatch scrambled siRNA-treated wound were compared to evaluate the efficacy of CTGF siRNA as a future implement for scar preventive medicine. METHODS: Nanocomplexes of CTGF small interfering RNA (CTGF siRNA) with cell penetrating peptides (KALA and $MPG^{{\Delta}NLS}$) were formulated and their effects on CTGF downregulation, collagen fibril diameter and arrangement were investigated. Various ratios of CTGF siRNA and peptide complexes were prepared and down-regulation were evaluated by immunoblot analysis. Control and CTGF siRNA modified cells-populated collagen lattices were prepared and rates of contraction measured. Collagen organization in rabbit ear 8 mm biopsy punch wound at 1 day to 8 wks post injury time were investigated by transmission electron microscopy and histology was investigated with Olympus System and TS-Auto software. CONCLUSION: CTGF expression was down-regulated to 40% of control by CTGF siRNA/KALA (1:24) complexes (p<0.01) and collagen lattice contraction was inhibited. However, down-regulated of CTGF by CTGF $siRNA/MPG^{{\Delta}NLS}$ complexes was not statistically significant. CTGF KALA-treated wound appeared with well formed-basket weave pattern of collagen fibrils with mean diameter of $128{\pm}22nm$ (n = 821). Mismatch siRNA/KALA-treated wound showed a high frequency of parallel small diameter fibrils (mean $90{\pm}20nm$, n = 563). CONCLUSION: Controlling over-expression of CTGF by peptide-mediated siRNA delivery could improve the collagen orientation and tissue remodeling in full thickness rabbit ear wound.

Structure of a Copper(Ⅱ) Hexaazamacrotricyclic Complex : (1,3,6,9,11,14-Hexaazatricyclo[12.2.1.16,9]octadecane)-copper(Ⅱ) Perchlorate

  • Cheon Manseog;Suh Paik Myunghyun;Shin Whanchul
    • Bulletin of the Korean Chemical Society
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    • v.13 no.4
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    • pp.363-367
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    • 1992
  • The crystal structure of (1,3,6,9,11,14-hexaazatricycol[12.2.1.$1^{6,9}$]octadecane)copper(Ⅱ) perchlorate, Cu($C_{12}H_{26}N_6$)$(ClO_4)_2$, has been determined by the X-ray diffraction methods. The crystal data are as follows: Mr=516.9, triclinic, ${\alpha}=8.572\;(2)$, b=8.499 (3), c=15.204 (3) ${\AA}$, ${\alpha}=80.42\;(5),\;{\beta}=73.57\;(3),\;{\gamma}=69.82\;(4)^{\circ},\;V=994.2\;{\AA}^3,\;D_C=1.726\;gcm^{-3}$, space group $P{\tilde{1}},\;Z=2,\;{\mu}=21.27\;cm^{-1}&, F(000)=534 and T=297 K. The structure was solved by direct methods and refined by full-matrix least-squares methods to and R value of 0.081 for 1608 observed reflections measured with graphite-mono-chromated Mo Ka radiation on a diffractometer. There are two independent complexes in the unit cell. The two copper ions lie at the special positions (1/2, 0, 0) and (0, 1/2, 1/2)and each complex possesses crystallographic center of symmetry. Each Cu ion is coordinated to four nitrogen donors if the hexaazamacrotricyclic ligand and weakly interacts with two oxygen atoms of the perchlorate ions to form a tetragonally distorted octahedral coordination geometry. The Cu_N (sec), Cu_N(tert) and Cu_O coordination distances are 1.985 (14), 2.055 (14) and 2.757 (13) ${\AA}$ for the complex A and 1.996 (10), 2.040 (11) and 2.660 (13) ${\AA}$ for the complex B, respectively. The macrocycles in the two independent cations assume a similar conformation with the average r.m.s. deviation of 0.073 ${\AA}$. Two 1,3-diazacyclopentane ring moieties of the hexaazamacrotricyclic ligand are placed oppositely and almost perpendicularly to the square coordination plane of the ruffled 14-membered macrocycle. The secondary N atoms are hydrogen-bonded to the perchlorate O atoms with distances of 3.017 (23) and 3.025 (19) ${\AA}$ for the complexes A and B, respectively.

Prevalence of Accident Occurrence Among Scientific Laboratory Workers of the Public University in Lebanon and the Impact of Safety Measures

  • Nasrallah, Inaam M.;El Kak, Assem K.;Ismaiil, Lina A.;Nasr, Rihab R.;Bawab, Wafa T.
    • Safety and Health at Work
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    • v.13 no.2
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    • pp.155-162
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    • 2022
  • Background: Workers are exposed to several risks in academic laboratories due to the presence of potentially hazardous substances. The main objective of this study was to assess the prevalence of accident occurrence and associated risk factors among laboratory workers at the scientific laboratories of the public university in Lebanon and the impact of safety measures training and availability. Methods: In this observational study, a survey was conducted for one year in scientific laboratories at faculties of the public university. Results: Among the participants (N = 220), 45.0% have had accidents; the main cause was exposure to chemicals (73.7%) and more specifically by inhalation (45.4%). Females (85.9%) were more exposed to accidents than males. Laboratory workers with a master's degree, a full-time schedule, and more than ten years of experience were significantly more exposed to accidents (p < 0.05). A significant association was found between accident occurrence and training on management of hazardous products (p = 0.044), risks related to workplace (p = 0.030), eyewash and emergency shower (p < 0.001), first aid (p = 0.012), and facial protection availability (p = 0.019). In spite of the lack of safety culture and efficient training on laboratory safety, participants have shown a very good perception regarding safety measures to be applied in case of work accidents. Conclusion: Based on our findings, the prevalence of accident occurrence is elevated among lab workers at the public university. The impact of regular training on laboratory safety preventive measures is of great importance to ensure the efficiency of occupational health and safety in scientific laboratories.

Performances of Li-Ion Batteries Using LiNi1-x-yCoxMnyO2 as Cathode Active Materials in Frequency Regulation Application for Power Systems

  • Choi, Jin Hyeok;Kwon, Soon-Jong;Lim, Jungho;Lim, Ji-Hun;Lee, Sung-Eun;Park, Kwangyong
    • KEPCO Journal on Electric Power and Energy
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    • v.6 no.4
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    • pp.461-466
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    • 2020
  • There are many application fields of electrical energy storage such as load shifting, integration with renewables, frequency or voltage supports, and so on. Especially, the frequency regulation is needed to stabilize the electric power system, and there have to be more than 1 GW as power reserve in Korea. Ni-rich layered oxide cathode materials have been investigated as a cathode material for Li-ion batteries because of their higher discharge capacity and lower cost than lithium cobalt oxide. Nonetheless, most of them have been investigated using small coin cells, and therefore, there is a limit to understand the deterioration mode of Ni-rich layered oxides in commercial high energy Li-ion batteries. In this paper, the pouch-type 20 Ah-scale Li-ion full cells are fabricated using Ni-rich layered oxides as a cathode and graphite as an anode. Above all, two test conditions for the application of frequency regulation were established in order to examine the performances of cells. Then, the electrochemical performances of two types of Ni-rich layered oxides are compared, and the long-term performance and degradation mode of the cell using cathode material with high nickel contents among them were investigated in the frequency regulation conditions.

Aeroelastic testing of a self-supported transmission tower under laboratory simulated tornado-like vortices

  • Ezami, Nima;El Damatty, Ashraf;Hamada, Ahmed;Hangan, Horia
    • Wind and Structures
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    • v.34 no.2
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    • pp.199-213
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    • 2022
  • The current study investigates the dynamic effects in the tornado-structure response of an aeroelastic self-supported lattice transmission tower model tested under laboratory simulated tornado-like vortices. The aeroelastic model is designed for a geometric scale of 1:65 and tested under scaled down tornadoes in the Wind Engineering, Energy and Environment (WindEEE) Research Institute. The simulated tornadoes have a similar length scale of 1:65 compared to the full-scale. An extensive experimental parametric study is conducted by offsetting the stationary tornado center with respect to the aeroelastic model. Such aeroelastic testing of a transmission tower under laboratory tornadoes is not reported in the literature. A multiaxial load cell is mounted underneath the base plate to measure the base shear forces and overturning moments applied to the model in three perpendicular directions. A three-axis accelerometer is mounted at the level of the second cross-arm to measure response accelerations to evaluate the natural frequencies through a free-vibration test. Radial, tangential, and axial velocity components of the tornado wind field are measured using cobra probes. Sensitivity analyses are conducted to assess the variation of the structural dynamic response associated with the location of the tornado relative to the lattice transmission tower. Three different layouts representing the change in the orientation of the tower model relative to the components of the tornado-induced loads are considered. The structural responses of the aeroelastic model in terms of base shear forces, overturning moments, and lateral accelerations are measured. The results are utilized to understand the dynamic response of self-supported transmission towers to the tornado-induced loads.

Cloning and Characterization of Porcine Uroplakin II Gene

  • D. N. Kwon;H. K. Shin;C. K. Hwang;D. W. Ok;Kim, J. H.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.19-19
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    • 2001
  • Mammalian urothelium undergoes unique membrane specialization by making the asymmetric unit membrane (AUM) that is covered with the apical cell surface during terminal differentiation. The AUM contains several major integral membrane proteins including uroplakin Ia, Ib, II and III. The genes for uroplakins have been cloned from humans and mice, but not from porcine. In this study, we report the cloning of the UPII genomic DNA, which codes for the full length open reading frame for the uroplakin II protein. The deduced amino acid sequence encodes of a hydrophobic NH$_2$-terminal peptide, a prosequence, and a mature protein. The prosequence contains three potential N-glycosylation sites and a RGRR cleavage site that may be involved in uroplakin II processing and maturation. Northern and immunohistochemistry analyses showed that the porcine UPII gene is only expressed in urothelium and that the protein was specifically localized in urothelial superficial cells. A 2kb of upstream in the promoter sequence contains multiple transcription factor binding sites, including GC-box, SPI, AP2, and GATA-box sites, but not for TATA or CAAT-box sequences. Comparison of the porcine UPII promoter sequence with that of the murine by MEME system presented two conserved motifs, suggesting a cis-acting regulatory role for the conserved sequences. Sequence homology between two species in motif A and B was 79% and 80% respectively, although their relative locations were different. During the gestation, mouse bladder at estrus stages and day 10 after parturition showed higher UPII expression, while showed lower expression at peri-implantation stage. Taken together, our results showed that the porcine UPII gene was expressed highly and specifically in the bladder urothelium and that steroid hormones for implantation changed the expression of UPII in the bladder, although the biological significance of UPII remains to be not determined.

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Nanoscale Characterization of a Heterostructure Interface Properties for High-Energy All-Solid-State Electrolytes (고에너지 전고체 전해질을 위한 나노스케일 이종구조 계면 특성)

  • Sung Won Hwang
    • Journal of the Semiconductor & Display Technology
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    • v.22 no.1
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    • pp.28-32
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    • 2023
  • Recently, the use of stable lithium nanostructures as substrates and electrodes for secondary batteries can be a fundamental alternative to the development of next-generation system semiconductor devices. However, lithium structures pose safety concerns by severely limiting battery life due to the growth of Li dendrites during rapid charge/discharge cycles. Also, enabling long cyclability of high-voltage oxide cathodes is a persistent challenge for all-solid-state batteries, largely because of their poor interfacial stabilities against oxide solid electrolytes. For the development of next-generation system semiconductor devices, solid electrolyte nanostructures, which are used in high-density micro-energy storage devices and avoid the instability of liquid electrolytes, can be promising alternatives for next-generation batteries. Nevertheless, poor lithium ion conductivity and structural defects at room temperature have been pointed out as limitations. In this study, a low-dimensional Graphene Oxide (GO) structure was applied to demonstrate stable operation characteristics based on Li+ ion conductivity and excellent electrochemical performance. The low-dimensional structure of GO-based solid electrolytes can provide an important strategy for stable scalable solid-state power system semiconductor applications at room temperature. The device using uncoated bare NCA delivers a low capacity of 89 mA h g-1, while the cell using GO-coated NCA delivers a high capacity of 158 mA h g−1 and a low polarization. A full Li GO-based device was fabricated to demonstrate the practicality of the modified Li structure using the Li-GO heterointerface. This study promises that the lowdimensional structure of Li-GO can be an effective approach for the stabilization of solid-state power system semiconductor architectures.

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Hyaluronic acid and proteoglycan link protein 1 suppresses platelet-derived growth factor-BB-induced proliferation, migration, and phenotypic switching of vascular smooth muscle cells

  • Dan Zhou;Hae Chan Ha;Goowon Yang;Ji Min Jang;Bo Kyung Park;Bo Kyung Park;In Chul Shin;Dae Kyong Kim
    • BMB Reports
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    • v.56 no.8
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    • pp.445-450
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    • 2023
  • The development of atherosclerotic cardiovascular disease is associated with the phenotypic switching of vascular smooth muscle cells (SMCs) from a contractile to a synthetic state, leading to cell migration and proliferation. Platelet-derived growth factor-BB (PDGF-BB) modulates this de-differentiation by initiating a number of biological processes. In this study, we show that gene expression of hyaluronic acid (HA) and proteoglycan link protein 1 (HAPLN1) was upregulated during differentiation of human aortic SMCs (HASMCs) into a contractile state, but downregulated upon during PDGF-BB-induced dedifferentiation. This is the first study showing that the treatment of HASMCs with full-length recombinant human HAPLN1 (rhHAPLN1) significantly reversed PDGF-BB-induced decrease in the protein levels of contractile markers (SM22α, α-SMA, calponin, and SM-MHC), and inhibited the proliferation and migration of HASMCs induced by PDGF-BB. Furthermore, our results show that rhHAPLN1 significantly inhibited the phosphorylation of FAK, AKT, STAT3, p38 MAPK and Raf mediated by the binding of PDGF-BB to PDGFRβ. Together, these results indicated that rhHAPLN1 can suppress the PDGF-BB-stimulated phenotypic switching and subsequent de-differentiation of HASMCs, highlighting its potential as a novel therapeutic target for atherosclerosis and other vascular diseases.

Analysis of Monitoring Results and Back Analysis for Rigid Diaphragm Wall Supported by Ground Anchor (지반앵커로 지지된 강성 지하연속벽체의 상세계측 결과분석 및 역해석 평가)

  • Lee, Jong-Sung;Hwang, Eui-Suk;Cho, Sung-Hwan;Lee, Jun-Hwan
    • Journal of the Korean Geotechnical Society
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    • v.25 no.5
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    • pp.39-46
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    • 2009
  • In this study, behavior of a rigid continuous wall, earth pressure distribution with construction stage, and axial force of earth anchors were evaluated based on field monitoring data and numerical analysis results. For this purpose, a construction site excavated using the diaphragm wall was selected and full instrumentation system was introduced. From monitoring results, it was found that the values of horizontal displacement of the wall measured from the inclinometers, which were installed within the diaphragm wall were similar to analytical value. The earth pressure increased with excavation progress due to jacking force of the ground anchors installed in previous excavation stages. When the excavation depth reached 60% of the final depth, observed earth pressure distribution was similar to that estimated from Peck's apparent earth pressure distribution. When the excavation depth was around 90% of the final depth, values of observed earth pressure showed middle values between those of Peck's and Tschebotarioffs apparent earth pressures. It was also observed that, when excavation depth is deep, values of the earth pressures from the rigid wall were similar to those estimated from conventional earth pressure distribution shape proposed for flexible walls.