• Title/Summary/Keyword: Single-Cell Analysis

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Proton Conducting Crosslinked Membranes by Polymer Blending of Triblock Copolymer and Poly(vinyl alcohol)

  • Lee, Do-Kyoung;Park, Jung-Tae;Choi, Jin-Kyu;Roh, Dong-Kyu;Lee, Jung-Hyun;Shul, Yong-Gun;Kim, Jong-Hak
    • Macromolecular Research
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    • v.16 no.6
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    • pp.549-554
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    • 2008
  • Proton conducting crosslinked membranes were prepared using polymer blends of polystyrene-b-poly(hydroxyethyl acrylate)-b-poly(styrene sulfonic acid) (PS-b-PHEA-b-PSSA) and poly(vinyl alcohol) (PVA). PS-b-PHEA-b-PSSA triblock copolymer at 28:21:51 wt% was synthesized sequentially using atom transfer radical polymerization (ATRP). FT-IR spectroscopy showed that after thermal ($120^{\circ}C$, 2 h) and chemical (sulfosuccinic acid, SA) treatments of the membranes, the middle PHEA block of the triblock copolymer was crosslinked with PVA through an esterification reaction between the -OH group of the membrane and the -COOH group of SA. The ion exchange capacity (IEC) decreased from 1.56 to 0.61 meq/g with increasing amount of PVA. Therefore, the proton conductivity at room temperature decreased from 0.044 to 0.018 S/cm. However, the introduction of PVA resulted in a decrease in water uptake from 87.0 to 44.3%, providing good mechanical properties applicable to the membrane electrode assembly (MEA) of fuel cells. Transmission electron microscopy (TEM) showed that the membrane was microphase-separated with a nanometer range with good connectivity of the $SO_3H$ ionic aggregates. The power density of a single $H_2/O_2$ fuel cell system using the membrane with 50 wt% PVA was $230\;mW/cm^2$ at $70^{\circ}C$ with a relative humidity of 100%. Thermogravimetric analysis (TGA) also showed a decrease in the thermal stability of the membranes with increasing PVA concentration.

Characterization of RAD4 Homologous Gene from Coprinus cinereus (균류 Coprinus cinereus에서 DNA 회복에 관여하는 RAD4 유사유전자의 분리와 특성)

  • Choi, In-Soon
    • Journal of Life Science
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    • v.13 no.4
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    • pp.522-528
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    • 2003
  • The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the RAD4 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. In order to investigation whether the increase of transcripts by DNA damaging agent, transcripts levels were examined after treating the cells. The level of transcript did not increase by untraviolet light (UV). This result indicated that the RAD4 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the RAD4 homologous gene is essential for cell viability.

Development of LED Module for Tooth Care with Effect of Promoting Scar Treatment and Analysis of Optical Properties (흉터 치료 촉진 효과를 갖는 치아 케어용 LED 모듈 개발과 광학적특성 분석)

  • Yoo, Kyun-Man;Son, Jeong-Hueon;Jo, Hyun-Min;Kang, So-Hi;Kang, Seong-Soo;Park, Sung-Jun
    • Journal of the Korean Society of Industry Convergence
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    • v.23 no.4_2
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    • pp.701-708
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    • 2020
  • In this paper, we analyzed the optical properties of the LED module for dental care, which has the effect of promoting cell regeneration and scar treatment. The LED module is a U-shaped module suitable for the shape of teeth. It is manufactured with an LED module (632 nm) and an LED module (632 nm + 870 nm), analyzes the results of optical properties, sets the irradiation distance, irradiation time, and is effective in healing skin wounds. Evaluation was conducted. It was tested in 6 test groups according to the light irradiation conditions, and light was irradiated to the scar site every other day for 7 days, 1 day and 3 days. As a result, it was confirmed that the effect of scar treatment was the highest when the combined wavelength of 632 nm + 870 nm was irradiated in pulse mode than when the single wavelength was irradiated and the composite wavelength was continuously irradiated. In group C group irradiated with PW Mode: pulse mode (period 36 ms, pulse width 35 ms) using a composite wavelength with LED module (632 nm + 870 nm) than group A without light irradiation, the length of scar reduction was 19 %, the area of the scar was further reduced by 10%, and it was confirmed that it is effective in treating scars in the wound area.

The Crystal Structure of a Sulfur Sorption Complex of the Dehydrated Partially $Co^{2+}$-Exchanged Zeolite A

  • 염영훈;송성환;김양
    • Bulletin of the Korean Chemical Society
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    • v.16 no.9
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    • pp.823-826
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    • 1995
  • The crystal structure of a sulfur sorption complex of the dehydrated partially Co2+ exchanged zeolite A (a=12.058(2) Å) has been determined by single-crystal X-ray techniques. The crystal structure was solved and refined in cubic space group Pm3m at 21(1) ℃. Ion Exchange with aqueous 0.05 M Co(NO3)2 was done by the static method. The crystal of Na4Co4-A was dehydrated at 380 ℃ and 2 × 10-6 Torr for 2 days, followed by exposure to about 100 Torr of sulfur at 330 ℃ for 72 h. Full matrix least-squares refinement converged to R1=0.084 and Rw=0.074 with 102 reflections for which I > 3σ(I). Crystallographic analysis shows that 2.8 Co2+ ions and 4 Na+ ions per unit cell occupy 6-ring sites on the threefold axes. 1.2 Co2+ ions occupy the 8-ring sites on fourfold axes. 2.8 Co2+ ions at Co(1) are recessed 0.66 Å into the large cavity and 4 Na+ ion at Na(1) are recessed 0.77 Å into the sodalite cavity from the (111) plane of O(3)'s. Approximately 16 sulfur atoms were sorbed per unit cell. Two S8 rings, each in a butterfly form, are found in the large cavity. The bond length between S and its adjacent S is 2.27(3) Å. The distance between 6-ring Co2+ ion and its adjacent sulfur is 2.53 (2) Å and that between 8-ring Co2+ ions and its adjacent sulfur is 2.72(9) Å. The angles of S-S'-S and S'-S-S'/ in octasulfur rings are 119.0(2)°and 113.0(2)°, respectively.

Rab25 Deficiency Perturbs Epidermal Differentiation and Skin Barrier Function in Mice

  • Jeong, Haengdueng;Lim, Kyung-Min;Goldenring, James R.;Nam, Ki Taek
    • Biomolecules & Therapeutics
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    • v.27 no.6
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    • pp.553-561
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    • 2019
  • Rab25, a member of the Rab11 small GTPase family, is central to achieving cellular polarity in epithelial tissues. Rab25 is highly expressed in epithelial cells of various tissues including breast, vagina, cervix, the gastrointestinal tract, and skin. Rab25 plays key roles in tumorigenesis, mainly by regulating epithelial differentiation and proliferation. However, its role in skin physiology is relatively unknown. In this study, we demonstrated that Rab25 knock-out (KO) mice show a skin barrier dysfunction with high trans-epidermal water loss and low cutaneous hydration. To examine this observation, we investigated the histology and epidermal differentiation markers of the skin in Rab25 KO mice. Rab25 KO increased cell proliferation at the basal layer of epidermis, whereas the supra-basal layer remained unaffected. Ceramide, which is a critical lipid component for skin barrier function, was not altered by Rab25 KO in its distribution or amount, as determined by immunohistochemistry. Notably, levels of epidermal differentiation markers, including loricrin, involucrin, and keratins (5, 14, 1, and 10) increased prominently in Rab25 KO mice. In line with this, depletion of Rab25 with single hairpin RNA increased the expression of differentiation markers in a human keratinocyte cell line, HaCaT. Transcriptomic analysis of the skin revealed increased expression of genes associated with skin development, epidermal development, and keratinocyte differentiation in Rab25 KO mice. Collectively, these results suggested that Rab25 is involved in the regulation of epidermal differentiation and proliferation.

Autophagy localization and cytoprotective role in cisplatin-induced acute kidney injury

  • Karunasagara, Shanika;Hong, Geum-Lan;Jung, Da-Young;Ryu, Si-Yun;Jung, Ju-Young
    • Korean Journal of Veterinary Research
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    • v.59 no.3
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    • pp.133-139
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    • 2019
  • Autophagy is a fundamental cellular process that maintains homeostasis and cell integrity, under stress conditions. Although the involvement of autophagy in various conditions has been elucidated, the role of autophagy in renal structure is not completely clarified. Our aim was to investigate the cytoprotective effect of autophagy against acute kidney injury (AKI) through cisplatin deteriorative pathway, which leads to AKI via renal cell degradation. For in vivo experiments, male Sprague Dawley rats were divided in to 2 groups (n = 6/group) as control, Cis-5D. Following a single intraperitoneal injection of cisplatin, rats were sacrificed after 5 days. Blood urea nitrogen (BUN), creatinine (Cr) and histological alterations were examined. Further, expression of key regulators of autophagy, light-clain 3 (LC3), p62, and Beclin1, was evaluated by immunohistochemistry (IHC). The rats exhibited severe renal dysfunction, indicated by elevated BUN, Cr. Hematoxylin and eosin staining revealed histological damages in cisplatin-treated rats. Furthermore, IHC analysis revealed increased expression of LC3, Beclin1 and decreased expression of p62. Furthermore, expression of aforementioned autophagy markers was restricted to proximal tubule. Taken together, our study demonstrated that cisplatin can cause nephrotoxicity and lead to AKI. This phenomenon accelerated autophagy in renal proximal tubules and guards against AKI.

Crystal Structure of Dehydrated Partially Ag$^+$-Exchanged Zeolite A treated with Cesium Vapor at 250${^{\circ}C}$

  • Kim, Duk-Soo;Song, Seong-Hwan;Kim, Yang
    • Bulletin of the Korean Chemical Society
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    • v.10 no.3
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    • pp.234-238
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    • 1989
  • The crystal structure of partially $Ag^+$-exchanged zeolite A, $Ag_{3.2}Na_{8.8}$-A, vacuum dehydrated at $360^{\circ}C$ and then exposed to 0.1 torr of cesium vapor for 12 hours at $250^{\circ}C$ has been determined by single-crystal X-ray diffraction techniques in the cubic space group Pm3m (a = 12.262(2)${\AA})\;at\;21(1)^{\circ}C$. The structure was refined to the final error indexes $R_1=0.068\;and\;R_2=0.072$ by using 338 reflections for which $I_o\;>\;3{\sigma}(I_o)$ and the composition of unit cell is $Ag_{3.2}Cs_{8.8}-A.\;3\;Cs^+$ ions lie on the centers of the 8-rings at sites of D4h symmetry. Two crystallographycally different 6-ring $Cs^+$ ions were found: 1.5 $Cs^+$ ions at Cs(2) are located inside of sodalite cavity and 4.3 $Cs^+$ ions at Cs(3) are located in the large cavity. The fractional occupancies observed at Cs(2) and Cs(3) indicate that the existence of at least three types of unit cells with regard to the 6-ring $Cs^+$ ions. For example, 50% of unit cells may have two $Cs^+$ ions at Cs(2) and 4 $Cs^+$ ions at Cs(3). 30% of unit cells may have one Cs+ ion at Cs(2) and 5 $Cs^+$ ions at Cs(3). The remaining 20% would have one $Cs^+$ ion at Cs(2) and 4 $Cs^+$ ions at Cs(3). On threefold axes of the unit cell two non-equivalent Ag atom positions are found in the large cavity, each containing 0.64 and 1.92 Ag atoms, respectively. A crystallographic analysis may be interpreted to indicate that 0.64 $(Ag_5)^+$ clusters are present in each large cavity. This cluster may be viewed as a tetrasilver molecule $(Ag_4)^0$(bond length, 2.84${\AA}$) stabilized by the coordination of one $Ag^+$ ion.

A Numerical Study of Cathode Block and Air Flow Rate Effect on PEMFC Performance (고분자전해질 연료전지의 환원극 블록과 공기 유량 영향에 대한 전산 해석 연구)

  • Jo, Seonghun;Kim, Junbom
    • Applied Chemistry for Engineering
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    • v.33 no.1
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    • pp.96-102
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    • 2022
  • Reactants of PEMFC are hydrogen and oxygen in gas phases and fuel cell overpotential could be reduced when reactants are smoothly transported. Numerous studies to modify cathode flow field design have been conducted because oxygen mass transfer in high current density region is dominant voltage loss factor. Among those cathode flow field designs, a block in flow field is used to forced supply reactant gas to porous gas diffusion layer. In this study, the block was installed on a simple fuel cell model. Using computational fluid dynamics (CFD), effects of forced convection due to blocks on a polarization curve and local current density contour were studied when different air flow rates were supplied. The high current density could be achieved even with low air supply rate due to forced convection to a gas diffusion layer and also with multiple blocks in series compared to a single block due to an increase of forced convection effect.

Production of Cloned Bovine Embryos Carrying with Human Thrombopoietin Gene

  • K.I. Wee;B.H. Son;Park, Y.H.;Park, J.S.;D.H. Ko;Lee, K.K.;Y.M. Han
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.60-60
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    • 2001
  • Human thrombopoietin (hTPO) is a cytokine that plays a central role in megakaryopoiesis by influencing on the development and maturation of megakaryocyte and platelet production. To induce hTPO production in the mammary gland, expression vector was constructed by combining the promoter of bovine beta-casein gene, cDNA of hTPO and neomycine resistance gene for transfection into fibroblasts. Bovine fibroblast cells derived from female ear skin were transfercted with the expression vector using Lipofectamine (Life Technology, NY). Transected cells resistant to G4l8 treatment (600 $\mu\textrm{g}$/$m\ell$) were recovered and colony formation was initiated at 13 days. The colonies with about 1 cm diameter were picked and analysed by PCR. Single transfected cells were individually transferred to enucleated oocytes. After electrofusion, the reconstructed embryos were exposed to calcium ionophore (5uM) for 5 min followed by treatment with 6-DMAP (2.5 mM) for 4h. The nuclear transfer embryos were cultured in CRlaa medium at 38.5C, 5% $CO_2$ for 7 days. Twenty three of 29 (79.3%) colonies were proved to be hTPO transfectants by PCR. The colonies were further passaged and used to produce transgenic embryos using nuclear transfer. Cleavage and developmental rates of reconstructed embryos to the blastocyst stage were 65.1% and 39.4%, respectively Of 22 blastocysts that developed from reconstructed embryos with the transfected cell, 20 embryos (90.9%) were positive for hTPO by using PCR analysis. The results suggest that somatic cell nuclear transfer is efficient for production of transgenic embryos.

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Promoter Analysis of the Cell Surface-abundant and Hypoviral-regulated Cryparin Gene from Cryphonectria parasitica

  • Kim, Myoung-Ju;Kwon, Bo-Ra;Park, Seung-Moon;Chung, Hea-Jong;Yang, Moon-Sik;Churchill, Alice C.L.;Van Alfen, Neal K.;Kim, Dae-Hyuk
    • Molecules and Cells
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    • v.26 no.5
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    • pp.496-502
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    • 2008
  • Cryparin, encoded as a single copy gene (Crp) of the chestnut blight fungus Cryphonectria parasitica, is the most abundant protein produced by this fungus. However, its accumulation is decreased remarkably in C. parastica strains containing the double-stranded (ds) RNA virus Cryphonectria hypovirus 1. To characterize the transcriptional regulatory element(s) for strong expression and viral regulation, promoter analysis was conducted. Serial deletion of the Crp promoter region resulted in a step-wise decrease in promoter activity, indicating a localized distribution of genetic elements in the cryparin promoter. Promoter analysis indicated two positive and a repressive cis-acting elements. Among them, the promoter region between nt -1,282 and -907 appeared to be necessary for hypoviral-mediated down-regulation. An electrophoretic mobility shift assay (EMSA) on the corresponding promoter region (-1,282/-907) indicated two regions at (-1,257/-1,158) and (-1,107/-1,008) with the characteristic AGGAGGA-N42-GAGAGGA and its inverted repeat TCCTCTC-N54-TCCTCCT, respectively, appeared to be specific binding sites for cellular factors.