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Construction of a Novel Shuttle Vector for Tetragenococcus species based on a Cryptic Plasmid from Tetragenococcus halophilus

  • Min Jae Kim;Tae Jin Kim;Yun Ji Kang;Ji Yeon Yoo;Jeong Hwan Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.211-218
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    • 2023
  • A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Emr), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

Resolution of Molecular Species of the Triacylglycerol Containing Petroselinic Acid $(cis-C_{18:1{\omega}12)$ by Silver Ion-HPLC

  • Joh, Yong-Goe;Kim, Seong-Jin
    • Journal of the Korean Applied Science and Technology
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    • v.22 no.4
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    • pp.339-348
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    • 2005
  • On the analysis of triacylglycerol (TG) from the kernels of Acanthopanax sessiliflorus by reversed phase-HPLC, it was separated into three main fractions of PN 44, 46 and 48, according to partition number (PN). On the contrary, it could be clearly classified into seven fractions of SMM, MMM, SMD, MMD, SDD, MDD and MDT by silver ion-HPLC by the number of double bond in the acyl chains of TG species. But resolution of so-called critical pairs of TG molecular species such as molecular pairs of $P_eLL$ $[C_{18:1{\omega}12}/(C_{18:2{\omega}6)2}]$ and OLL $[C_{18:1{\omega}9}/(C_{18:2{\omega}6)2}]$ and OOL $[(C_{18:1{\omega}9)2}/C_{18:2{\omega}6]$, and $P_eP_eL$ $[(C_{18:2{\omega}12)2}/C_{18:1{\omega}6]$ was not achieved $(P_e;$ petroselinic acid, L; linoleic acid, O; oleic acid). On the other hand, TG extracted from Aralia continentalis kernels were also fractionated into seven groups of SSM, SMM, MMM, SMD, MMD, SDD and MDD (S; saturated acid, M; monoenoic acid, D; dienoic acid) by silver ion-HPLC, although it's were classified into three groups of PN 44, 46 and 48 by reversed phase-HPLC. The fractions of SMM, MMM, MMD and MDD were divided into two subfractions, respectively; the fractions of SMM, MMM, MMD and MDD were resolved into the subfraction of $PP_e/P_e$ and POO (critical pairs from each other), that of $P_e/P_e/P_e$ and OOO, that of $P_e/P_e/L$ and OOL, and that of $P_e/L/L$ and OLL.

G1 Arrest of U937 Human Monocytic Leukemia Cells by Sodium Butyrate, an HDAC Inhibitor, Via Induction of Cdk Inhibitors and Down-regulation of pRB Phosphorylation (Cdk inhibitors의 발현 증가 및 pRB 인산화 저해에 의한 HDAC inhibitor인 sodium butyrate에 의한 인체백혈병세포의 G1 arrest유발)

  • Choi, Yung-Hyun
    • Journal of Life Science
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    • v.19 no.7
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    • pp.871-877
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    • 2009
  • We investigated the effects of sodium butyrate, a histone deacetylase inhibitor, on the cell cycle progression in human monocytic leukemia U937 cells. Exposure of U937 cells to sodium butyrate resulted in growth inhibition, G1 arrest of the cell cycle and induction of apoptosis in a dose-dependent manner as measured by MTT assay and flow cytometry analysis. The increase in G1 arrest was associated with the down-regulation in cyclin D1, E, A, cyclin-dependent kinase (Cdk) 4 and 6 expression, and up-regulation of Cdk inhibitors such as p21 and p27. Sodium butyrate treatment also inhibited the phosphorylation of retinoblastoma protein (pRB) and p130, however, the levels of transcription factors E2F-1 and E2F-4 were not markedly modulated. Furthermore, the down-regulation of phosphorylation of pRB and p130 by this compound was associated with enhanced binding of pRB and E2F-1, as well as p130 and E2F-4, respectively. Overall, the present results demonstrate a combined mechanism involving the inhibition of pRBjp130 phosphorylation and induction of Cdk inhibitors as targets for sodium butyrate that may explain some of its anti-cancer effects in U937 cells.

Investigation of the Insulin-like Growth Factor System in Breast Muscle during Embryonic and Postnatal Development in Langshan and Arbor Acres Chickens Subjected to Different Feeding Regimens

  • Lu, F.Z.;Chen, J.;Wang, X.X.;Liu, Honglin
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.4
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    • pp.471-482
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    • 2009
  • Nutrient availability may control muscle growth directly and indirectly through its influence on regulatory factors. We analyzed the effects of nutrient availability on the breast muscle insulin-like growth factor system. Real time RT-PCR was used to quantify the level of transcription in breast muscle from Langshan (LS) layer and Arbor Acres (AA) broiler chickens subjected to different feeding regimens during embryonic and postnatal development. The AA chickens were fed AA diet (AA, control group) while the LS chickens were either fed LS diet (LL) or AA diet (LA). According to our results, insulin-like growth factor (IGF)-II (embryonic day 16 (E16) - postnatal day 42 (P42)), IGF-I receptor (IGF-IR, E18-P42), and IGF binding protein (IGFBP)-2 (E18-P42), -5 (E16-P14), -7 (E12-P0), and -3 (E12-P0) were positively correlated with IGF-I, while IGFBP-3 (P0-P28) was negatively correlated with IGF-I. In comparison, IGF-IR (E18-P42), IGFBP-2 (E18-P42), IGFBP-5 (E14-P0), and IGFBP-3 (E16-P0) were positively correlated with IGF-II, while IGF-IR (E10-E16) and IGFBP-3 (P0-P28) were negatively correlated with IGF-II. Moreover, IGFBP-2 (E16-P42), -7 (E10-E16), and -3 (E10-E16) were positively correlated with IGF-IR, while IGFBP-3 (P0-P28) was negatively correlated with IGF-IR. Finally, IGFBP-7 (E12-P0) was positively correlated with IGFBP-3, while IGFBP-2 (P0-P28) and -7 (P0-P42) were negatively correlated with IGFBP-3. Overall, the AA chickens exhibited higher levels of IGF-I, IGF-IR, and IGFBP-2 mRNA expression than the LL chickens, while the opposite was true for IGFBP-7. No strain differences in IGF-I, IGF-IR, and IGFBP-7 mRNA expression were detected between LA and AA chickens; however, a strain difference was observed for IGFBP-2. LA chickens exhibited higher levels of IGFBP-2 than LL chickens, while the opposite was true for IGFBP-7. Our data show the first evidence that certain genes may be correlated during specific developmental periods and that strain differences in the expression of those genes in LS and AA chickens are due to differential responses to the same diet.

Alteration of Lipid Metabolism by Ginseng Supplements With Different Levels of Vitamin E in High Cholesterol-Fed Rats

  • Do, Kyung-Min;Park, Yong-Bok;Bok, Song-Hae;Lee, Mi-Kyung;Jeong, Tae-Sook;Choi, Myung-Sook
    • Preventive Nutrition and Food Science
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    • v.6 no.1
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    • pp.66-72
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    • 2001
  • Ginseng may have antioxidant and pharmacologic effects similar to those of vitamin E. The interactive effect of ginseng and vitamin E was studied with respect to cholesterol metabolism and the antioxidant status. A ginseng supplement (0.1%, wt/wt) with comparable levels of vitamin E was provided with a high-cholesterol (1%, wt/wt) diet to rats for 5 weeks. The amount of vitamin E included in the ginseng-free and ginseng diets was either a low (low-E) or a normal (normal-E) level. The ginseng supplements significantly (p<0.05) altered the concentrations of plasma triglycerides in both the low-vitamin E group and normal-vitamin E group compared to the each ginseng-free group. The hepatic triglyceride and cholesterol content were not significantly (p>0.05) different between groups regardless of the vitamin E level in the diet. The hepatic HMG-CoA reductase activity was significantly (p<0.05) lowered by the ginseng supplement in both the low-vitamin E and the normal-vitamin E groups compared to the ginseng-free group. The HMG-CoA reductase activity was also significantly (p<0.05) lowered with in increase of the dietary vitamin E in the ginseng-free group. The excretion of fecal neutral sterol was significantly (p<0.05) lower in the normal-E ginseng group than th low-E ginseng-free group. Neither dietary ginseng nor vitamin E significantly changed the hepatic antioxidant enzymes activity. This data indicates that ginseng supplements lower the concentration of plasma triglyceride and hepatic HMG-CoA reductase activity regardless of eh dietary vitamin E level. This information may contribute to understanding the interactive effect of ginseng and vitamin E on cholesterol biosynthesis in high cholesterol-fed rats.

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A study on the change of the views on E.M.T. profession of the E.M.T. college students (일부 전문대 응급구조학생의 응급구조전문직관의 변화에 대한 연구)

  • Kim, Hak-Soo;Choi, Eun-Sook;Kim, Jin-Hue;Shin, Dong-Min;Lee, In-Soo
    • The Korean Journal of Emergency Medical Services
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    • v.1 no.1
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    • pp.28-41
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    • 1997
  • The purpose of this study was to identify the change of perceptions related to E.M.T. profession of E.M.T. student and to provide the basic data for development of curriculum. The subjects of the survey were 216 students from 3 junior college by questionaire from November 25 to November 29, 1996 in Kwang-Ju, Cheong-Ju, and Kong-Ju. The results were analyzed using the SAS and can be summerized as follows : 1. The view on profession was a statistically significant difference inspecial knowledge & skill training (t=-1.686, P<.001), high income (t=-0.753, P<.05) 2. The view on competence as a E.M.T. was a statistically significant differncein sufficient professional knowledge on the E.M.T. (t=-1.144, P<.001), own's health (t=-0.808, P<.001), sincerity and responsibility (t=-1.429, P<.01), satisfaction and effort (t=-1.335, P<.01), boundary of affairs (t=-1.356, P<.01) 3. The view on E.M.T. profession was a statistically significant difference inprotect of pt. life & to relieve the pain (t=-2.388, P<.001) 4. The view on necessary knowledge of E.M.T. was a statistically significant difference in patient assessment (t=-2.168, P<.001), ACLS (t=1.678, P<.001), hemorrhage & shock (t=-3.252, P<.001), trauma (t=-4.284, P<.001), cardiopulmonary disturbance (t=-2.897, P<.001), acute abdomen (t=-4.284, P<.001), neonatal disease (t=-4.032, P<.001), OBGY disease(t=-4.151, P<.001), emergency delivery (t=-2.825, P<.001), infectious disease (t=-2.930, P<.001), environmental emergency (t=-3.123, P<.05), recording(t=2.22, P<.05) 5. The view on ideology of E.M.T. was a statistically significant difference in belief & faith (t=-2.344, P<.001), knowing of own (t=-2.142, P<.01), humanity (t=-2.581, P<.05), knowing of patient (t=-2.079, P<.05).

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A Study on the Development of Game Platform in Web 3.0: Focused on P2E, C2E Models (웹 3.0에서의 메타버스 플랫폼 발전 방향에 대한 연구: P2E, C2E 모델을 중심으로)

  • Hyo Won Moon;Seon Bin Lim;Hee Dong Yang
    • Journal of Information Technology Services
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    • v.22 no.1
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    • pp.75-93
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    • 2023
  • The game industry has grown not only by combining them with various technologies also introducing new types of business models such as P2P, F2P, and P2W. Furthermore, games which implemented X2E model with blockchain technology are recently in the spotlight of the public attention. As domestic game companies have also prospect the blockchain games feasible, they are seeking ways to expand their global market share by strengthening the X2E model. Hence, by carrying this new business model out, it is expected to diversify their global revenue stream, which was previously confined to Asia region. This study analyzed the case of companies that have implemented the P2E and C2E models in order to suggest the direction of development for the game platform in Web 3.0 era. The cases of P2E game platform, which constitute of Axie Infinity and Mir 4, encompass the compensation structure, the stabilization mechanism of the in-game token economy, and future strategies regarding blockchain gaming. Likewise, the platform structure, business model, and future growth potential was discussed in terms of C2E scheme, focusing on the ZEPETO and Roblox cases. Based on the above case analysis, this study attempted to provide information on the current limitations and development directions of the P2E and C2E platforms. The current limitations in legal and industrial aspects should be addressed to facilitate the blooming of blockchain and P2E game industry. In addition, the necessity of not only social support also improvement on the technology and social stigma of full-time creators is ought to be emphasized in an effort to encourage the development of C2E platforms.

Impact of Remanent Polarization and Coercive Field on Threshold Voltage and Drain-Induced Barrier Lowering in NCFET (negative capacitance FET) (NCFET (negative capacitance FET)에서 잔류분극과 항전계가 문턱전압과 드레인 유도장벽 감소에 미치는 영향)

  • Hakkee Jung
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.37 no.1
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    • pp.48-55
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    • 2024
  • The changes in threshold voltage and DIBL were investigated for changes in remanent polarization Pr and coercive field Ec, which determine the characteristics of the P-E hysteresis curve of ferroelectric in NCFET (negative capacitance FET). The threshold voltage and DIBL (drain-induced barrier lowering) were observed for a junctionless double gate MOSFET using a gate oxide structure of MFMIS (metal-ferroelectric-metal-insulator-semiconductor). To obtain the threshold voltage, series-type potential distribution and second derivative method were used. As a result, it can be seen that the threshold voltage increases when Pr decreases and Ec increases, and the threshold voltage is also maintained constant when the Pr/Ec is constant. However, as the drain voltage increases, the threshold voltage changes significantly according to Pr/Ec, so the DIBL greatly changes for Pr/Ec. In other words, when Pr/Ec=15 pF/cm, DIBL showed a negative value regardless of the channel length under the conditions of ferroelectric thickness of 10 nm and SiO2 thickness of 1 nm. The DIBL value was in the negative or positive range for the channel length when the Pr/Ec is 25 pF/cm or more under the same conditions, so the condition of DIBL=0 could be obtained. As such, the optimal condition to reduce short channel effects can be obtained since the threshold voltage and DIBL can be adjusted according to the device dimension of NCFET and the Pr and Ec of ferroelectric.

Molecular Analysis of Salmonella Enterotoxin Gene Expression

  • Lim, Sang-Yong;Seo, Ho-Seong;Yoon, Hyun-Jin;Choi, Sang-Ho;Heu, Sung-Gi;Ryu, Sang-Ryeol
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.598-606
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    • 2003
  • Salmonella encodes an enterotoxin (Stn) which possesses biological activity similar to the cholera toxin. Stn contributes significantly to the overall virulence of S. typhimurium in a murine model. The production of Stn is enhanced in a high-osmolarity medium and by contact with epithelial cells. In the present study, the in vitro and in vivo transcriptional regulations of the sin promoter revealed two promoters, P1 and P2. The P1 promoter identified by a primer extension analysis of stn mRNA exhibited a switching mechanism in vivo. Depending on the growth stage, transcription was initiated from different start sites termed $P1_S\;and\;P1_E$. $P1_S$, recognized by RNA polymerase containing ${\sigma}^S(E{\sigma}^S),\;and\;P1_E$, recognized by $E{\sigma}^70$, were activated during the stationary and exponential phases, respectively, while $P1_S\;and\;P1_E$ were both negatively regulated by CRPㆍcAMP and H-NS. Results revealed that $P1_S$ was the responsible promoter activated under a high osmolarity and low pH. The P2 promoter was identified 45 nucleotides downstream from $P1_E$ and negatively controlled by CRPㆍcAMP in vitro. No P2 activity was detected in vivo. The regulation of stn expression monitored using a Pstn::egfp fusion indicated that $E{\sigma}^S$ was required for the induction of stn and various factors were involved in stn regulation inside animal cells.

Effect of Packaging Material and Oxygen Absorbant on Quality Properties of Yukwa (포장재질 및 탈산소재가 유과의 품질특성에 미치는 영향)

  • Lee, Yong-Hwan;Kum, Jun-Seok;Ahn, Yong-Sik;Kim, Woo-Jung
    • Korean Journal of Food Science and Technology
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    • v.33 no.6
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    • pp.728-736
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    • 2001
  • Effects of packaging material and oxygen absorbant on physical and chemical properties of Yukwu were studied during storage to develop packaging conditions. The packaging materials used were PET/EVOH $(16\;{\mu}m)/PL$ : P1 and PET/EVOH $(24\;{\mu}m)/PL$ : P2 with or without oxygen absorbent (E1A : P1 and E2A : P2 for w/ $O_2$, absorbent, E1EA : P1 and E2EA : P2 for w/o $O_2$, absorbent). Color values for Yukwu indicated that L values of E1A, E1EA, E2A and E2EA were decreased during storage while those b values were increased. Hardness and chewiness of Yukwa were generally decreased, however those of E1A and E1EA were rather increased after 6 weeks of storage. Acid value of E2A had maintained less than 2.0 during 12 weeks of storage. E1A, E2A had the below of 20 in peroxide during 12 weeks. Aroma data by using electronic nose showed that there was no difference after 6 week storage time in different packaging materials. Sensory evaluation (Yukwa odor and rancid odor) showed very similar results with electronic nose one. E2A had the highest value of overall acceptability for sensory evaluation. Hardness and cheweness in physical measurement results had the highest correlation with hardness, crispness, overall-acceptability in sensory evaluation results.

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