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Cloning and Characterization of Pseudomonas mucidolens Exoinulinase

  • Kwon, Young-Man;Kim, Hwa-Young;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.238-243
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    • 2000
  • An exoinulinase (${\beta}-D-fructofuranosidase$) gene was cloned by chromosome walking along the upstream region of the endoinulinase gene of Pseudomonas mucidolens isolated from soil. the exoinulinase gene consisted of an ORF of 0,506 bp encoding a polypeptide of 501 amino acids with a deduced molecular weight of 55,000. The exoinulinase produced by the recombinant Escherichia coli $DH5{\alpha}$ strain was also purified to homogeneity as determined by SDS-PAGE and a zymogram. The molecular weight of the purified exoinulinase according to both SDS-PAGE and gel filtration matched the deduced molecular weight of the protein described above, thereby indicating that the native form of the exoinulinase was a monomer. The purified enzyme hydrolyzed activity value of 2.0. Furthermore, no inulo-oligomers were liberated from the inulin substrate in the enzymatic reaction mixtures incubated for 90 min at $55^{\circ}C$. Taken together, these results indicate that the purified ${\beta}-D-fructofuranosidase$ was an exoinulinase. The pH and temperature optima of the exoinulinase were pH 6.0 and $55^{\circ}C$, respectively. the enzymehad no apparent requirement for a cofactor, and its activity was completely inactivated by $Ag^{+},{\;}Hg^{2+},{\;}and{\;}Zn^{2+}$. Kinetic experiments gave $K_m,{\;}V_{max},{\;}and{\;}K_{cat}$ values for inulin of 11.5 mM, 18 nM/s, and $72{\;}s^{-1}$, respectively. the exoinulinase was fairly stable in broad pH conditions (pH 5-9), and at pH 6.0 it showed a residual activity of about 70% after 4 h incubation at $55^{\circ}C$.

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Identification and Characterization of the Acid Phosphatase HppA in Helicobacter pylori

  • Ki, Mi-Ran;Yun, Soon-Kyu;Choi, Kyung-Min;Hwang, Se-Young
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.483-493
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    • 2011
  • An acid phosphatase (HppA) activated by $NH_4Cl$ was purified 192- and 34-fold from the periplasmic and membrane fractions of Helicobacter pylori, respectively. SDS-polyacrylamide gel electrophoresis revealed that HppA from the latter appears to be several kilodaltons larger in molecular mass than from the former by about 24 kDa. Under acidic conditions (pH${\leq}$4.5), the enzyme activity was entirely dependent on the presence of certain mono- and/or divalent metal cations (e.g., $K^+$,$ NH_4{^+}$, and/or $Ni^{2+}$). In particular, $Ni^{2+}$ appeared to lower the enzyme's $K_m$ for the substrates, without changing $V_{max}$. The purified enzyme showed differential specificity against nucleotide substrates with pH; for example, the enzyme hydrolyzed adenosine nucleotides more rapidly at pH 5.5 than at pH 6.0, and vice versa for CTP or TTP. Analyses of the enzyme's N-terminal sequence and of an $HppA^-$ H. pylori mutant revealed that the purified enzyme is identical to rHppA, a cloned H. pylori class C acid phosphatase, and shown to be the sole bacterial 5'-nucleotidase uniquely activated by $NH_4Cl$. In contrast to wild type, $HppA^-$ H. pylori cells grew more slowly. Strikingly, they imported $Mg^{2+}$ at a markedly lowered rate, but assimilated urea rapidly, with a subsequent increase in extracellular pH. Moreover, mutant cells were much more sensitive to extracellular potassium ions, as well as to metronidazole, omeprazole, or thiophenol, with considerably lowered MIC values, than wild-type cells. From these data, we suggest that the role of the acid phosphatase HppA in H. pylori may extend beyond 5'-nucleotidase function to include cation-flux as well as pH regulation on the cell envelope.

Stress distribution in premolars restored with inlays or onlays: 3D finite element analysis

  • Yang, Hongso;Park, Chan;Shin, Jin-Ho;Yun, Kwi-Dug;Lim, Hyun-Pil;Park, Sang-Won;Chung, Hyunju
    • The Journal of Advanced Prosthodontics
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    • v.10 no.3
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    • pp.184-190
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    • 2018
  • PURPOSE. To analyze stress distribution in premolars restored with inlays or onlays using various materials. MATERIALS AND METHODS. Three-dimensional maxillary premolar models of abutments were designed to include the following: 1) inlay with O cavity (O group), 2) inlay with MO cavity (MO group), 3) inlay with MOD cavity (MOD group), and 4) onlay (ONLAY group). A restoration of each inlay or onlay cavity was simulated using gold alloy, e.max ceramic, or composite resin for restoration. To simulate masticatory forces, a total of 140 N static axial force was applied onto the tooth at the occlusal contact areas. A finite element analysis was performed to predict the magnitude and pattern of stresses generated by occlusal loading. RESULTS. Maximum von Mises stress values generated in the abutment teeth of the ONLAY group were ranged from 26.1 to 26.8 MPa, which were significantly lower than those of inlay groups (O group: 260.3-260.7 MPa; MO group: 252.1-262.4 MPa; MOD group: 281.4-298.8 MPa). Maximum von Mises stresses generated with ceramic, gold, and composite restorations were 280.1, 269.9, and 286.6 MPa, respectively, in the MOD group. They were 252.2, 248.0, 255.1 MPa, respectively, in the ONLAY group. CONCLUSION. The onlay design (ONLAY group) protected tooth structures more effectively than inlay designs (O, MO, and MOD groups). However, stress magnitudes in restorations with various dental materials exhibited no significant difference among groups (O, MO, MOD, ONLAY).

Load-bearing capacity of various CAD/CAM monolithic molar crowns under recommended occlusal thickness and reduced occlusal thickness conditions

  • Choi, Sulki;Yoon, Hyung-In;Park, Eun-Jin
    • The Journal of Advanced Prosthodontics
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    • v.9 no.6
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    • pp.423-431
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    • 2017
  • PURPOSE. The goal of this study was to evaluate the fracture resistances of various monolithic crowns fabricated by computer-aided design and computer-aided manufacturing (CAD/CAM) with different thickness. MATERIALS AND METHODS. Test dies were fabricated as mandibular molar forms with occlusal reductions using CAD/CAM. With different occlusal thickness (1.0 or 1.5 mm), a polymer-infiltrated ceramic network (Enamic, EN), and zirconia-reinforced lithium silicate (Suprinity, SU and Celtra-Duo, CD) were used to fabricate molar crowns. Lithium disilicate (e.max CAD, EM) crowns (occlusal: 1.5 mm) were fabricated as control. Seventy crowns (n=10 per group) were bonded to abutments and stored in water for 24 hours. A universal testing machine was used to apply load to crown until fracture. The fractured specimens were examined with a scanning electron microscopy. RESULTS. The type of ceramics and the occlusal thickness showed a significant interaction. With a recommended thickness (1.5 mm), the SU revealed the mean load similar to the EM, higher compared with those of the EN and CD. The fracture loads in a reduced thickness (1.0 mm) were similar among the SU, CD, and EN. The mean fracture load of the SU and CD enhanced significantly when the occlusal thickness increased, whereas that of the EN did not. CONCLUSION. The fracture loads of monolithic crowns were differently influenced by the changes in occlusal thickness, depending on the type of ceramics. Within the limitations of this study, all the tested crowns withstood the physiological masticatory loads both at the recommended and reduced occlusal thickness.

A Study on the Convective Heat Transfer in a Regenerative Ice Energy System by a Bundle of the Heat-pipes. (히이트파이프 다발을 이용한 냉축열시스템에서의 대류열전달에 관한 연구)

  • 권형정;김경석;김경근
    • Journal of Advanced Marine Engineering and Technology
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    • v.14 no.4
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    • pp.57-66
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    • 1990
  • In the design of an electric power plant, the capacity to meet the peak load demand is one of the important factors to be considered. This peak load usually occurs when the most of the cooling air conditioning systems are being operated during daytime in summer season, which inevitably entails the construction of an additional electric power plant. This study is aimed to carry out a basic experiment for the development of a cooling air conditioning system using the ice energy by the surplus electric power during the night-time. The experimental apparatus consists of four major parts; (1) the heating section consisting of the air duct and I.D. fan, (2) the cold section with the ice chamber, (3) the bundle of heat pipes made in a form of the staggered arrangement with ${C_y}/{d_o}$=2.0 and ${C_x}/{d_o}$=1.73, (4) the refrigerator system to cool down the ice chamber. This study involves an intensive experiment concerning the convective heat transfer of the air flow surrounding the bundle of heat pipes. This major experimental parameters are the amount of working fluid, the velocity of air and the working temperature. The major findings of the present study are as follows; (1) The optimum amount of the working fluid necessary for the horizontal heat pipes is much more than that for the vertical type. (2) The convective heat transfer coefficients of the air are coincided with the empirical equations of Grimson and ${\breve{Z}ukauskas}$. (3) The equation of the mean heat transfer coefficient obtained in the present study is ${N_um}=0.32 {Re_max^{0.63}}$.

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Fatigue Design of Spot Welded Lap Joint Considered Residual Stress (잔류응력을 고려한 점용접이음재의 피로설계)

  • Son, Il-Seon;Bae, Dong-Ho;Hong, Jeong-Gyun;Lee, Beom-No
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.24 no.3 s.174
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    • pp.743-751
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    • 2000
  • Because welding residual stress is formidable result in electric resistance spot welding process, and it detrimentally affect to fatigue crack initiation and growth at nugget edge of spot welded la p joints, it should be considered in fatigue analysis. Thus, accurate prediction of residual stress is very important. In this study, nonlinear finite element analysis on welding residual stress generated in process of the spot welding was conducted, and their results were compared with experimental data measured by X-ray diffraction method. By using their results, the maximum principal stress considered welding residual stress at nugget edge of the spot welded lap joint subjected to tension-shear load was calculated by superposition method. And, the $\Delta$P- $N_f$ relations obtained through fatigue, tests on the IB-type spot welded lap joints was systematically rearranged with the maximum principal stress considered welding residual stress. From the results, it was found th2at fatigue strength of the IB-type spot welded lap joints could be systematically and more reasonably rearranged by the maximum principal stress($\sigma$1max-res considered welding residual stress at nugget edge of the spot welding point.

Efficient Power Reduction Technique of LiDAR Sensor for Controlling Detection Accuracy Based on Vehicle Speed (차량 속도 기반 정확도 제어를 통한 차량용 LiDAR 센서의 효율적 전력 절감 기법)

  • Lee, Sanghoon;Lee, Dongkyu;Choi, Pyung;Park, Daejin
    • IEMEK Journal of Embedded Systems and Applications
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    • v.15 no.5
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    • pp.215-225
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    • 2020
  • Light detection and ranging (LiDAR) sensors detect the distance of the surrounding environment and objects. Conventional LiDAR sensors require a certain amount of a power because they detect objects by transmitting lasers at a regular interval depending on a constant resolution. The constant power consumption from operating multiple LiDAR sensors is detrimental to autonomous and electric vehicles using battery power. In this paper, we propose two algorithms that improve the inefficient power consumption during the constant operation of LiDAR sensors. LiDAR sensors with algorithms efficiently reduce the power consumption in two ways: (a) controlling the resolution to vary the laser transmission period (TP) of a laser diode (LD) depending on the vehicle's speed and (b) reducing the static power consumption using a sleep mode depending on the surrounding environment. A proposed LiDAR sensor with a resolution control algorithm reduces the power consumption of the LD by 6.92% to 32.43% depending on the vehicle's speed, compared to the maximum number of laser transmissions (Nx·max). The sleep mode with a surrounding environment-sensing algorithm reduces the power consumption by 61.09%. The proposed LiDAR sensor has a risk factor for 4-cycles that does not detect objects in the sleep mode, but we consider it to be negligible because it immediately switches to an active mode when a change in surrounding conditions occurs. The proposed LiDAR sensor was tested on a commercial processor chip with the algorithm controlling the resolution according to the vehicle's speed and the surrounding environment.

Red-Orange Emissive Cyclometalated Neutral Iridium(III) Complexes and Hydridoiridium(III) Complex Based on 2-Phenylquinoxaline : Structure, Photophysics and Reactivity of Acetylacetone Towards Cyclometalated Iridium Dimer

  • Sengottuvelan, Nallathambi;Yun, Seong-Jae;Kang, Sung-Kwon;Kim, Young-Inn
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4321-4326
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    • 2011
  • A new series of heteroleptic cyclometalated iridium(III) complexes has been synthesized and characterized by absorption, emission and cyclic voltammetry studies: $(pqx)_2Ir(acac)$ (1), $(dmpqx)_2Ir(acac)$ (2) and $(dfpqx)_2Ir(acac)$ (3) where pqx=2-phenylquinoxalinate, dmpqx=2-(2,4-dimethoxyphenyl)quinoxalinate, dfpqx=2-(2,4-difluorophenyl) quinoxalinate and acac=acetylacetonate anion. The reaction of excess acetylacetone with ${\mu}$-chloride-bridged dimeric iridium complex, $[(C\^N)_2Ir({\mu}-Cl)]_2$, gives a complex 1 and an unusual hydridoiridium(III) complex, $(pqx)IrH(acac)_2$ (4). The complex 1, 2 and 3 show their emissions in an orangered region (${\lambda}_{PL,max}$ = 583-616 nm), and the emission maxima can be tuned by the change of substituent at phenyl ring of 2-phenylquinoxaline ligand. The phosphorescent line shape indicates that the emissions originate predominantly from $^3MLCT$ states with little admixture of ligand-based $^3({\pi}-{\pi}^*)$ excited states. The structures of complex 3 and 4 are additionally characterized by a single crystal X-ray diffraction method. The complex 3 shows a distorted octahedral geometry around iridium(III) metal ion. A strong trans influence of the phenyl ring is examined. In complex 4, there are two discrete molecules which are mirror images each other at the ratio of 1:1 in an unit cell. We propose that the phosphorescent complex 1, 2 and 3 are possible candidates for the phosphors in OLEDs applications.

Improvement of Magnetic Properties of Nd-Fe-B Type Sintered Magnet (Nd-Fe-B계 소결자석의 자기적특성 향상 연구)

  • Kim, Yoon-B.;Jung, W.S.;Jeung, W.Y.
    • Journal of the Korean Magnetics Society
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    • v.12 no.2
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    • pp.57-63
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    • 2002
  • In order to increase the magnetic properties of a Nd-Fe-B sintered magnet, the general factors including particle size and its distribution, volume fraction of Nd$_2$Fe$_{14}$B phase, degree of alignment of Nd$_2$Fe$_{14}$B grain, oxygen content and grain size etc. should be optimized by controlling the composition of Nd-Fe-B alloy as well as the manufacturing process. In this study, fabrication of the Nd-Fe-B sintered magnet was carried out in a laboratory scale by controlling the composition of Nd-Fe-B alloy and the manufacturing process. The optimum milling condition was found by investigating the milling media, milling time and ball size. The addition of FeGa was effective to increase the coercivity of the Nd-Fe-B sintered magnet. A remanence of 14.4 kG, a coercivity of 9.4 kOe and a maximum energy product of 47 MGOe were obtained from the sintered magnet.

Identification and Classification of the Muscarinic Receptors in the Uterus (자궁 무스카린수용체의 확인 및 분류)

  • Lee, Shin-Woong;Lee, Jeung-Soo;Park, Young-Joo
    • YAKHAK HOEJI
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    • v.36 no.3
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    • pp.220-229
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    • 1992
  • The muscarinic acetylcholine receptors of the dog unpregant uterus were characterized using $[^3H]quinuclidinyl$ benzilate(QNB) as a radioligand and the binding of muscarinic receptor agonists and antagonists in the uterus was compared to that in the urinary bladder which contains almost exclusively the M2 receptors in order to determine the receptor subtypes in the uterus. $[^3H]QNB$ binding to uterus and bladder was rapid, saturable and reversible. Scatchard analysis of the saturation data gave linear plots and the Hill coefficients were close to unit, which indicated that each preparation contained a single population of specific binding sites for $[^3H]QNB$. The KD values(120 pM) for QNB were almost identical in both organs, whereas the $B_{max}$ value of 256 fmol/mg protein in the uterus was significantly different from that of 563 fmol/mg protein in the bladder. Muscarinic agonists and antagonists inhibited in a competitive manner the $[^3H]QNB$ binding to the same extent in both organs. The competition binding studies using antagonists(atropine and pirenzepine) exhibited a single binding site and this site had a low affinity for pirenzepine with the Ki value of about 330 nM. However, high and low affinity binding sites were observed with carbachol, methacholine and oxotremorine. These binding studies with agonists and antagonists did not show any differences in drug affinities between uterus and bladder. These results indicate that the muscarinic receptors in the uterus are M2 receptors which have a low affinity for pirenzepine.

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