• Title/Summary/Keyword: Sf cell

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Preparation of Sulfobetaine Chitosan, Silk Blended Films, and Their Properties (설포베타인 키토산의 실크 블렌드 필름의 제조 및 그들의 성질)

  • Koo, Ja-Sung;Cha, Jae-Ryung;Oh, Se-Heang;Gong, Myoung-Seon
    • Polymer(Korea)
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    • v.38 no.1
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    • pp.54-61
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    • 2014
  • Water-soluble sulfobetaine chitosan (SCs) was prepared for a blending film with Bombyx mori silk fibroin (SF) by reacting chitosan with 1,3-propanesultone. A series of SF/SCs blended films were successfully prepared by mixing aqueous solutions of B. mori SF and SCs. The SF/SCs blended films were examined through spectroscopic and thermal analysis to determine the morphological changes of SF in the SCs. The effects of the SF/SCs blend ratios on physical and mechanical properties were investigated to discover the feasibility of using these films as biomedical materials such as artificial skin and wound dressing. X-ray analysis showed good compatibility between the two biopolymers. The in vitro degradation behavior of the SF/SCs blended films was systematically investigated for up to 8 weeks in phosphate buffered saline solution at $37^{\circ}C$ and showed a mass loss of 46.4% after 8 weeks. All films showed no cytotoxicity by MC3T3-E1 assay. After 3 days of culture, the relative cell number on all the SF/SCs films was slightly lower than that of an optimized tissue culture plastic.

Microarray Analysis of Gene Expression Profile by Treatment of Schizandrae fructus Extract in Inflammation-induced Human Epithelial A549 Cells (염증이 유발된 인간기관지상피세포에서 오미자가 Microarray를 이용한 유전자 발현 분석에 미치는 영향)

  • Jung, Jin-Yong;Jung, Sung-Ki;Jung, Hee-Jae;Rhee, Hyung-Koo
    • The Journal of Internal Korean Medicine
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    • v.29 no.3
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    • pp.543-553
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    • 2008
  • Objective: The goal of this study was to determine the anti-asthma mechanism of SF on TNF-${\alpha}$ induced activation on A549 (human type II-like epithelial) cells. Using oligonucleotide microarray, we sought to establish the molecular mechanism of the protective effects of SF on A549 cells. Material & Methods : Cells were cultured in three different conditions: 1) negative control group was cultured in normal condition of DMEM, 2) positive control group was activated with TNF-${\alpha}$, IL-4. and IL-1${\beta}$, and 3) SF treated group was previously treated with 0.1${\mu}g/ml$ SF after TNF-${\alpha}$, IL-4. and IL-1 activation. Cells of positive control and SF treated groups were cultured for 30 min, 1hr, 3hr and 6hr. Results : The comparative analysis of the gene expression profile revealed that proinflammatory cytokines such as IL1F8, IL1F9, IL1R1. IL1RN, IL1RAPL1, IL8, TNFRSF4, TNFSF10c, TNFSF13, TRAF5, and TRAF7 and inflammation-related genes including MMP2, MMP11, MMP14, MMP15, MMP16, MMP19, MMP25, and MMP27 were down regulated with SF treatment. Cell adhesion molecule genes such as ITGB1, ITGBL1, selectin P ligand, selectin E, ICAM2, ICAM3, VCAM1, PECAM, FCER1G and MMP28 genes were also down-regulated in SF treated A549 cells. Conclusion : These results suggest that the anti-asthmatic effects of SF could be mediated by regulating specific genes related with cell adhesion, proinflammatory cytokine and inflammation-related genes in A549 cells.

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Cloning of tlrD, 23S rRNA Monomethyltransferase Gene, Overexpression in Eschepichia coli and Its Activity (235 rRNA Monomethyltransferase인 tlrD의 클로닝, 이의 대장균에서 대량생산과 활성 검색)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.166-172
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    • 2007
  • ERM proteins transfer the methyl group to $A_{2058}$ in 23S rRNA, which reduces the affinity of MLS (macrolide-lincosamide-streptogramin B) antibiotics to 23S rRNA, thereby confer the antibiotic resistance on micro-organisms ranging from antibiotic producers to pathogens and are classified into monomethyltransferase and dimethyltransferase. To investigate the differences between mono- and dimethyltransferase, tirD, a representative monomethylase gene was cloned in Escherichia coli from Streptomyces fradiae which contains ermSF, dimethylase gene as well to overexpress the TlrD for the first time. T7 promoter driven expression system successfully overexpress tlrD as a insoluble aggregate at $37^{\circ}C$ accumulating to around 55% of the total cell protein but unlike ErmSF, culturing at temperature as low as $18^{\circ}C$ did not make insoluble aggregate of protein into soluble protein. Coexpression of Thioredoxin and GroESL, chaperone was not helpful in turning into soluble protein either as in case of ErmSF. These results might suggest that differences between mono- and dimethylase could be investigated on the basis of the characteristics of protein structure. However, a very small amount of soluble protein which could not be detected by SDS-PAGE conferred antibiotic resistance on E. coli as in ErmSF which was expected from the activity exerted by monmethylase in a cell.

Effects of soy-based formula on infants' growth and blood laboratory values spanning 3 years after birth (대두 영유아식이 생후 3세까지 신체 계측치 및 혈액검사에 미치는 영향)

  • Yoon, Ji Eun;Kim, Mi-Jung;Han, Heon-Seok
    • Clinical and Experimental Pediatrics
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    • v.52 no.1
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    • pp.28-35
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    • 2009
  • Purpose : We compared body growth, blood cell counts, and chemistry among infants receiving soy-based formula (SF), breast milk (BM), and casein-based formula (CF). Methods : Full-term neonates delivered at our hospital from June 2001 to August 2003 were recruited and divided into 3 feeding groups (BM=20, SF=19, CF=12 ) according to the parents wishes. Breast feeding or artificial formulae were given during the initial 3 months of age; thereafter, weaning foods were added freely. Height, weight, head circumference, skin-fold thickness, and mid-arm circumference were measured at birth and 1, 2, 4, 5, 12, and 36 months of age. Cell counts and blood chemistry were analyzed at 5, 12, and 36 months of age. Result : At 5 months of age, body weight was the lowest in the SF group; the height was similar among all groups. Thereafter, there were no differences in height or weight among the groups until 36 months of age. Hemoglobin was the lowest in the BM group at 5 and 12 months of age (P< 0.05 ). At 5 months of age, serum cholesterol, BUN, phosphates, and K+ were significantly lower in the SF group; thereafter, all chemical parameters were similar until 36 months of age. Conclusion : Infants fed with SF showed normal growth during the first 3 years of life as compared to infants fed with BM and CF. Low values of serum phosphates and K+ at 5 months of age in the SF group, despite the high mineral content, suggest that further investigation is needed for effective mineral absorption.

Pituitary Adenylate Cyclase-activating Polypeptide (PACAP) Treatment during Pre-maturation Increases the Maturation of Porcine Oocytes Derived from Small Follicles

  • Park, Kyu-Mi;So, Kyoung-Ha;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.33 no.1
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    • pp.1-11
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    • 2018
  • Cellular cyclic adenosine-3' 5'-monophosphate (cAMP) modulator is known as meiotic inhibitor and can delays spontaneous maturation in IVM experiment. Among many cAMP modulators, the role of Pituitary adenylate cyclase activating polypeptide (PACAP) on IVM isn't known. The purpose of this study is to improve the maturation of oocytes derived from follicles ${\leq}3mm$ in diameter through PACAP as meiotic inhibitor during pre-in vitro maturation (pre-IVM). First, we checked PACAP and its receptors in cumulus cells and, to establish the optimal phase and concentration of PACAP for pre-IVM, we conducted chromatin configuration assessments. As a result, the rate of GV (Germinal Vesicle) according to duration of pre-IVM was significantly decreased 12 h and 18 h after IVM (87.1 and 84.1%, respectively) compared to 0 h (99.4%). When COC was cultured for 18 h, the GV rate in the $1{\mu}M$ of PACAP treatment group (82.1%) was significantly higher than any other PACAP treatment groups (60.5, 64.1, 74.4 and 69.9 %, respectively). So, we divided into four groups as follows; MF (the conventional IVM group, obtained from follicle from 3 to 6 mm in diameter), SF (the conventional IVM group, obtained from follicle ${\leq}3mm$ in diameter), Pre-SF(-)PACAP (IVM group including 18 h pre-IVM without $1{\mu}M$ of PACAP, obtained from follicle ${\leq}3mm$ in diameter) and Pre-SF(+)PACAP (IVM group including 18 h pre-IVM with $1{\mu}M$ of PACAP, obtained from follicle ${\leq}3mm$ in diameter). To examine the effect of PACAP during pre-IVM, we investigated analysis of nuclear maturation, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels. In cumulus cells, PACAP receptors, ADCYAP1R1 and VIPR1 were detected but were not detected in oocytes. After IVM, the Pre-SF(+)PACAP had the highest Metaphase II rate (91.7%) among all groups (P<0.05). The GSH levels in the MF and Pre-SF(+)PACAP were significantly higher than in the other groups (P<0.05) and ROS levels was no significant difference among all groups. In conclusion, these results indicated that even though the oocytes were derived from SF, pre-IVM application of PACAP improved meiotic and cytoplasmic maturation by regulating intracellular oxidative stress.

Signals of MLCK and ROCK Pathways Triggered via Lymphotoxin β Receptor are Involved in Stress Fiber Change of Fibroblastic Reticular Cells (FRC에서 Lymphotoxin β receptor의 자극은 MLCK와 ROCK의 이중 신호전달 경로를 통해 stress fiber 변화에 관여)

  • Kim, Dae Sik;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.29 no.2
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    • pp.256-264
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    • 2019
  • Lymphotoxin ${\beta}$ receptor ($LT{\beta}R$), a member of the tumor necrosis factor receptor family, plays an important role in lymphoid tissue's architecture and organogenesis. In contrast, MLCK and ROCK play critical roles in the regulation of stress fiber (SF) formation in cells. To determine whether $LT{\beta}R$ stimulation in fibroblastic reticular cells (FRCs) is involved in these signaling pathways, myosin light chain kinase inhibitor-7 (ML-7) was used to inhibit them. ML7-treated FRCs completely blocked SFs and showed retraction and shrinkage processes comparable to those observed in agonistic anti-$LT{\beta}R$ antibody-treated cells. The inhibition of ROCK activity with Y27632-induced changes in actin cytoskeleton organization and cell morphology in FRCs. Actin bundles rearranged into SFs, and phospho-myosin light chain (p-MLC) co-localized in FRCs. We checked the level of Rho-guanosine diphosphate (RhoGDP)/guanosine triphosphate (GTP) exchange activity using FRC lysate. When $LT{\beta}R$ was stimulated with agonistic anti-$LT{\beta}R$ antibodies, Rho-GDP/GTP exchange activity was markedly reduced. Regarding $LT{\beta}R$ signaling with a focus on MLCK inhibition, we showed that the phosphorylation of MLCs was reduced by $LT{\beta}R$ stimulation in FRCs. Cytoskeleton components, such as tubulin, b-actin, and phospho-ezrin proteins acting as membrane-cytoskeleton linkers, were produced in de-phosphorylation, and they reduced expression in agonistic anti-$LT{\beta}R$ antibody-treated FRCs. Collectively, the results suggested that MLCK and ROCK were simultaneously responsible for SF regulation triggered by $LT{\beta}R$ signaling in FRCs.

Tryptic Digestion and Cytochalasin B Binding Assay of the Human HepG2-Type Glucose Transporter Expressed in Spodoptera frugiperda Clone 21-AE Cells

  • Lee Chong-Kee
    • Biomedical Science Letters
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    • v.11 no.1
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    • pp.57-61
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    • 2005
  • The number of sites at which a protein can be readily cleaved by a proteolytic enzyme is greatly influenced by its three-dimensional structure. For native, properly-folded proteins both the rate of cleavage and number of sites at which cleavage takes place are usually much less than for the denatured protein. In order to compare the tertiary structure of recombinant HepG2 type glucose transporter with that of its native counterpart in the erythrocyte, the pattern of tryptic cleavage of the protein expressed in insect cell membranes was therefore examined. After 30 minutes digestion, a fragment of approximate Mr 19,000-21,000 was generated. In addition to this, there were two less intensely stained fragments of apparent Mr 28,000 and 17,000. The pattern of labelling was similar up to 2 hours of digestion. However, the fragments of Mr 19,000-21,000 and Mr 17,000 were no longer detectable after 4 hours digestion. The observation of a very similar pattern of fragments yielded by tryptic digestion of the HepG2 type transporter expressed in insect cells suggests that the recombinant protein exhibits a tertiary structure similar if not identical to that of its human counterpart. Also, the endogenous sugar transporter(s) present in Sf21 cells did not bind cytochalasin B, the potent transporter inhibitor. Therefore, the baculovirus/Spodoptera frugiperda (Sf) cell expression system could be very useful for production of large amounts of human glucose transporters, heterologously.

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Characterization of Surface Textured Silicon Substrates by SF6/O2 Gas Mixture (SF6/O2 혼합가스에 의한 실리콘 웨이퍼의 표면 텍스쳐링 특성)

  • Kang, Min-Seok;Joo, Sung-Jae;Koo, Sang-Mo
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.25 no.5
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    • pp.345-348
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    • 2012
  • The optical losses associated with the reflectance of incident radiation are among the most important factors limiting the efficiency of a solar cell. Therefore, photovoltaic cells normally require special surface structures or materials, which can reduce reflectance. In this study, nano-scale textured structures with anti-reflection properties were successfully formed on silicon. The surface of sicon wafer was etched by the inductively coupled plasma process using the gaseous mixture of $SF_6+O_2$. We demonstrate that the reflection characteristic has significantly reduced by ~0% compared with the flat surface. As a result, the power efficiency $P_{max}$ of the nano-scale textured silicon solar cell were enhanced up to 20%, which can be ascribed primarily to the improved light trapping in the proposed nano-scale texturing.

Saucerneol F, a New Lignan Isolated from Saururus chinensis, Attenuates Degranulation via Phospholipase Cγ1 Inhibition and Eicosanoid Generation by Suppressing MAP Kinases in Mast Cells

  • Lu, Yue;Son, Jong-Keun;Chang, Hyeun Wook
    • Biomolecules & Therapeutics
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    • v.20 no.6
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    • pp.526-531
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    • 2012
  • During our on-going studies to identify bioactive compounds in medicinal herbs, we found that saucerneol F (SF), a naturally occurring sesquilignan isolated from Saururus chinensis (S. chinensis), showed in vitro anti-inflammatory activity. In this study, we examined the effects of SF on the generation of 5-lipoxygenase (5-LO) dependent leukotriene $C_4$ ($LTC_4$), cyclooxygenase-2 (COX-2) dependent prostaglandin $D_2$ ($PGD_2$), and on phospholipase $C{\gamma}1$ ($PLC{\gamma}1$)-mediated degranulation in SCF-induced mouse bone marrow-derived mast cells (BMMCs). SF inhibited eicosanoid ($PGD_2$ and $LTC_4$) generation and degranulation dose-dependently. To identify the molecular mechanisms underlying the inhibition of eicosanoid generation and degranulation by SF, we examined the effects of SF on the phosphorylation of $PLC{\gamma}1$, intracellular $Ca^{2+}$ influx, the translocation of cytosolic phospholipase $A_2$ ($cPLA_2$) and 5-LO, and on the phosphorylation of MAP kinases (MAPKs). SF was found to reduce intracellular $Ca^{2+}$ influx by inhibiting $PLC{\gamma}1$ phosphorylation and suppressing the nuclear translocations of $cPLA_2$ and 5-LO via the phosphorylations of MAPKs, including extracellular signal-regulated protein kinase-1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38. Taken together, these results suggest that SF may be useful for regulating mast cell-mediated inflammatory responses by inhibiting degranulation and eicosanoid generation.

Implementation of WLAN Baseband Processor Based on Space-Frequency OFDM Transmit Diversity Scheme (공간-주파수 OFDM 전송 다이버시티 기법 기반 무선 LAN 기저대역 프로세서의 구현)

  • Jung Yunho;Noh Seungpyo;Yoon Hongil;Kim Jaeseok
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.42 no.5 s.335
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    • pp.55-62
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    • 2005
  • In this paper, we propose an efficient symbol detection algorithm for space-frequency OFDM (SF-OFDM) transmit diversity scheme and present the implementation results of the SF-OFDM WLAN baseband processor with the proposed algorithm. When the number of sub-carriers in SF-OFDM scheme is small, the interference between adjacent sub-carriers may be generated. The proposed algorithm eliminates this interference in a parallel manner and obtains a considerable performance improvement over the conventional detection algorithm. The bit error rate (BER) performance of the proposed detection algorithm is evaluated by the simulation. In the case of 2 transmit and 2 receive antennas, at $BER=10^{-4}$ the proposed algorithm obtains about 3 dB gain over the conventional detection algorithm. The packet error rate (PER), link throughput, and coverage performance of the SF-OFDM WLAN with the proposed detection algorithm are also estimated. For the target throughput at $80\%$ of the peak data rate, the SF-OFDM WLAN achieves the average SNR gain of about 5.95 dB and the average coverage gain of 3.98 meter. The SF-OFDM WLAN baseband processor with the proposed algorithm was designed in a hardware description language and synthesized to gate-level circuits using 0.18um 1.8V CMOS standard cell library. With the division-free architecture, the total logic gate count for the processor is 945K. The real-time operation is verified and evaluated using a FPGA test system.