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Mycelial Culture of Lentinus edodes Alleviates Rat Liver Toxicity Induced by Carbon Tetrachloride and Ethanol (표고버섯균사체의 사염화탄소 및 알콜로 처리된 흰쥐 간기능 보호 효과)

  • Ha, Yeong-L.;Kim, Young-S.;Ahn, Chae-R.;Kweon, Jung-M.;Park, Cherl-W.;Ha, Young-K.;Kim, Jeong-O.
    • Journal of Life Science
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    • v.20 no.1
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    • pp.133-141
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    • 2010
  • The protective effect of a mixed powder from solid-cultured and liquid-cultured Lentinus edodes mycelia (2:1, w/w) (designate LED) on the carbon tetrachloride ($CCl_4$)- and ethanol-induced hepatotoxicity of male Sprague-Dawley (SD) rat was investigated. In the $CCl_4$-induced rat hepatotoxicity experiment, rats of 4 groups (6 rats/group) were administere with Normal (0.2 ml distilled water), Control (0.2 ml distilled water), LED (LED 200 mg/kg BW + 0.2 ml distilled water), and Silymarin (200 mg/kg BW + 0.2 ml distilled water), p.o., daily for 2 weeks. Afterwards, all groups except for the Normal group were subjected to abdominal injection with $CCl_4$ ($CCl_4$ : corn oil, 1:1 v/v; 0.5 ml/kg BW). For the ethanol- induced rat hepatotoxicity experiment, rats were divided into 5 groups (5 rats/group): Normal; Pair-fed control (PFC); Control (ethanol); LED (ethanol + LED 200 mg/kg BW); and Silymarin (ethanol + silymarin 200 mg/kg BW). Rats of the Normal and PFC groups were fed a basal liquid diet, and rats of the Control, LED, and Silymarin groups were fed a liquid ethanol diet containing LED or Silymarin. Eight weeks later, blood and liver samples were collected to analyze biomarkers. In $CCl_4$-induced SD rats, LED elevated hepatic superoxide dismutase (SOD), catalase, and glutathione peroxidase (GSH peroxidase) activities and thiobarbituric reactive substances (TBARS) were reduced, resulting in the reduction of glutamate-oxalate transaminase (GOT), glutamate-pyruvate transaminase (GPT) and lactic dehydrogenase (LDH) activities in plasma. Similar results of these enzymes and biochemical markers in both liver tissues and plasma were seen in ethanol-induced hepatotoxicity of SD rats. In addition, elevated alcohol dehydrogenase (ADH) activity and reduced expression of cytochrome p450 mixed monooxygenase enzyme (CYP2E1) were seen in liver tissues from ethanol-treated rats by LED treatment. These effects of LED were similar to those of Silymarin. In in vitro experiments, LED showed antioxidant activity in a 2,2-diphenyl-1-picrylhydrazyl (DPPH) system and mouse liver mitochondria system induced by NADPH/$Fe^{2+}$ and cumine hydroperoxide (CuOOH). These results indicate that LED protected SD rat hepatotoxicity, induced by $CCl_4$ and ethanol, through its antioxidative activity and might be useful as a material for protection from hepatoxicity in humans.

Expression and Purification of Recombinant Human Interferon-gamma Produced by Escherichia coli (대장균이 생산한 재조합 인체 감마인터페론의 발현과 정제)

  • Park, Jung-Ryeol;Kim, Sung-Woo;Kim, Jae-Bum;Jung, Woo-Hyuk;Han, Myung-Wan;Jo, Young-Bae;Jung, Joon-Ki
    • KSBB Journal
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    • v.21 no.3
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    • pp.204-211
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    • 2006
  • For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

Effect of Production In Vitro Embryo using Boar Frozen Semen (돼지 동결 정액을 이용한 체외 수정란 생산 효율)

  • Cho, Sang-Rae;Kim, Hyun-Jong;Choe, Chang-Yong;Son, Dong-Soo;Choi, Sun-Ho;Son, Jun-Kyu;Kim, Sung-Jae;Kim, Jae-Bum;Han, Man-Hye;Jin, Hyun-Ju
    • Journal of Embryo Transfer
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    • v.24 no.3
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    • pp.199-205
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    • 2009
  • This study was carried out to investigate the effective genetic resources preservation system using the frozen boar semen. The porcine oocytes were matured for 44 hours in NCSU-23 medium with or without 10% Porcine Follicle Fluid (PFF), 0.5 ${\mu}g/ml$ porcine FSH, 0.5 ${\mu}g/ml$ equine LH, 1.0 ${\mu}g/ml$ 17 $\beta$-estradiol ($E_2$) and 10 ng/ml Epidermal Growth Factor (EGF) under mineral oil at $38.5^{\circ}C$ in humidified atmosphere of 5% $CO_2$ in air. After 44 h of culture, the oocytes were inseminated with frozen-thawed semen and fresh semen prepared with mTBM medium for 6 h. Later, set of 50 presumptive zygotes were transferred into 4-well dish (500 ${\mu}l$) of IVC medium. for embryos freezing, slow-freezing and vitrification methods were used as a cryopreservation. Differences among treatments were analyzed using General Linear Model Procedure by SAS Package (version 6.12) differences were considered significant when p<0.05. Following IVF and IVC, the rates of cleavage and blastocysts formation were significantly higher (p<0.05) in hormone supplemented group than that of hormone-free group (25.7 vs, 12.1). The development rates to cleavage and blastocysts were significantly higher in PZM-5 group than NCSU-23 group (60.3%, 46.6% vs 27.4%, 11.1%). Further improvement was achieved when PZM-5 was supplemented with FBS. Cleavage rates was significantly higher in fresh semen source group than frozen semen (66.7% vs 43.7%). However in blastocysts rates was similar two groups. Post-thaw survival rates of embryos were 1.2% and 2.2% in slow-frezing and vitrification groups, respectively. The results of our study suggest that it is still possible to improve the culture conditions and boar semen cryopreservation for enhance reproductive technology and animal genetic resources conservation.

A Novel Tannase from the Xerophilic Fungus Aspergillus niger GH1

  • Marco, Mata-Gomez;Rodriguez, Luis V.;Ramos, Erika L.;Renovato, Jacqueline;Cruz-Hernandez, Mario A.;Rodriguez, Raul;Contreras, Juan;Aguilar, Cristobal N.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.987-996
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    • 2009
  • Aspergillus niger GH1 previously isolated and identified by our group as a wild tannase producer was grown under solid-state (SSC) and submerged culture (SmC) conditions to select the enzyme production system. For tannase purification, extracellular tannase was produced under SSC using polyurethane foam as the inert support. Tannase was purified to apparent homogeneity by ultrafiltration, anion-exchange chromatography, and gel filtration that led to a purified enzyme with a specific activity of 238.14 IU/mg protein with a final yield of 0.3% and a purification fold of 46. Three bands were found on the SDS-PAG with molecular masses of 50, 75, and 100 kDa. PI of 3.5 and 7.1% N-glycosylation were noted. Temperature and pH optima were 600e and 6.0 [methyl 3,4,5-trihydroxybenzoate (MTB) as substrate], respectively. Tannase was found with a $K_M$ value of $0.41{\times}10^{-4}M$ and the value of $V_{max}$ was $11.03{\mu}$moL/min at $60^{\circ}C$ for MTB. Effects of several metal salts, solvents, surfactants, and typical enzyme inhibitors on tannase activity were evaluated to establish the novelty of the enzyme. Finally, the tannase from A. niger GH1 was significantly inhibited by PMSF (phenylmethylsulfonyl fluoride), and therefore, it is possible to consider the presence of a serine or cysteine residue in the catalytic site.

Incorporation of Winter Rapeseed (Brassica napus) as Green Manure on Mineralization and Uptake of Nitrogen to Succeeding Corn (Zea mays L.) (유채의 녹비 환원에 의한 질소무기화 및 옥수수의 질소 흡수)

  • Choi, Bong-Su;Hong, Ki-Chan;Sung, Jwa-Kyung;Nam, Jae-Jak;Lim, Jung-Eun;Lee, Hyeon-Yong;Yang, Jae-E.;Ok, Yong-Sik
    • Korean Journal of Organic Agriculture
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    • v.17 no.3
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    • pp.381-391
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    • 2009
  • Crop production can be secured by the cycle of green manure crops as an alternative of the chemical fertilizer. Recently, rapeseed (Brassica napus L.) has been cultivated in the south part of Korea for the production of biodiesel. In this research, we focused on recycling rapeseed residue, which is produced after harvesting the rapeseed for biodiesel, as a potential source of nitrogen to the succeeding crop. Pot experiment was conducted to evaluate the effects of winter rapeseed as green manure on mineralization and uptake of nitrogen to the succeeding corn (Zea mays L.). Result showed that total nitrogen and C/N ratio of rapeseed at the harvesting stage was 0.54% and 63, respectively. The incorporation of rapeseed without decomposition period slightly inhibited nitrogen uptake to the succeeding corn compared to those with 30 days decomposition period. The pH and EC values of soils increased by increasing the period of decomposition of rapeseed from 5.2 to 6.4 and from 0.05 dS/m to 0.21 dS/m, respectively. Significant amounts of $NH_4^+$ and $NO_3^-$ are released by incorporation of rapeseed. The succeeding corn took up 86% and 88% of inorganic nitrogen released from the rapeseed with and without decomposition period, respectively. The overall results suggested that the utilization of rapeseed residue as green manure can be an alternative source of nitrogen in corn-rapeseed double cropping system.

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Study on Meat Tenderizer -Part 1. Tenderizing ability of commercial meat tenderizer- (Meat Tenderizer 제조에 관한 연구 -(제1보) 시판(市販) meat tenderizer의 연육효과-)

  • Yang, Ryung;Kim, Kun-Wha;Lee, Jung-Hee;Youn, Jung-Eae;Yu, Ju-Hyun
    • Korean Journal of Food Science and Technology
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    • v.7 no.4
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    • pp.221-228
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    • 1975
  • Meat tenderness is one of the most important factors in meat products because it plays a major role in the palatability of meat. To get information on the role of commercial meat tenderizer, the tenderizing ability of commercial meat tenderizer was measured with various substrates. The results obtained are as follows. 1. Content of crude protein in a commercial meat tenderizer was 4.9%. 2. Optimum temperature for proteolytic activity of meat tenderizer was $60{\sim}70^{\circ}C$. 3. Maximal activity of proteinase was obtained at pH $6{\sim}7$. 4. Proteolytic enzyme was activated by KCN, NaCN, EDTA. Thus, it was concluded that protease system of commercial meat tenderizer composed of plant origin proteinases. 5. Proteinase activity was completely inhibited by 10mM of N-Ethylmaleimide. 6. Commercial meat tenderizer showed stronger proteolytic activity on casein than on the water soluble fraction of meat protein, whereas it hydrolyzed the myofibrillar protein less efficiently.

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Evaluation of Near-infrared Fluorescence-conjugated Peptides for Visualization of Human Epidermal Receptor 2-overexpressed Gastric Cancer

  • Jeong, Kyoungyun;Kong, Seong-Ho;Bae, Seong-Woo;Park, Cho Rong;Berlth, Felix;Shin, Jae Hwan;Lee, Yun-Sang;Youn, Hyewon;Koo, Eunhee;Suh, Yun-Suhk;Park, Do Joong;Lee, Hyuk-Joon;Yang, Han-Kwang
    • Journal of Gastric Cancer
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    • v.21 no.2
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    • pp.191-202
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    • 2021
  • Purpose: A near-infrared (NIR) fluorescence imaging is a promising tool for cancer-specific image guided surgery. Human epidermal receptor 2 (HER2) is one of the candidate markers for gastric cancer. In this study, we aimed to synthesize HER2-specific NIR fluorescence probes and evaluate their applicability in cancer-specific image-guided surgeries using an animal model. Materials and Methods: An NIR dye emitting light at 800 nm (IRDye800CW; Li-COR) was conjugated to trastuzumab and an HER2-specific affibody using a click mechanism. HER2 affinity was assessed using surface plasmon resonance. Gastric cancer cell lines (NCI-N87 and SNU-601) were subcutaneously implanted into female BALB/c nu (6-8 weeks old) mice. After intravenous injection of the probes, biodistribution and fluorescence signal intensity were measured using Lumina II (Perkin Elmer) and a laparoscopic NIR camera (InTheSmart). Results: Trastuzumab-IRDye800CW exhibited high affinity for HER2 (KD=2.093(3) pM). Fluorescence signals in the liver and spleen were the highest at 24 hours post injection, while the signal in HER2-positive tumor cells increased until 72 hours, as assessed using the Lumina II system. The signal corresponding to the tumor was visually identified and clearly differentiated from the liver after 72 hours using a laparoscopic NIR camera. Affibody-IRDye800CW also exhibited high affinity for HER2 (KD=4.71 nM); however, the signal was not identified in the tumor, probably owing to rapid renal clearance. Conclusions: Trastuzumab-IRDye800CW may be used as a potential NIR probe that can be injected 2-3 days before surgery to obtain high HER2-specific signal and contrast. Affibody-based NIR probes may require modifications to enhance mobilization to the tumor site.

Numerical Estimation of Wind Loads on FLNG by Computational Fluid Dynamics (전산유체역학을 이용한 FLNG의 풍하중 추정에 관한 연구)

  • Sang-Eui, Lee
    • Journal of Navigation and Port Research
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    • v.46 no.6
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    • pp.491-500
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    • 2022
  • It has been noted that an accurate estimation of wind loads on offshore structures such as an FLNG (Liquefied Natural Gas Floating P roduction Storage Offloading Units, LNG FPSOs) with a large topside plays an important role in the safety design of hull and mooring system. Therefore, the present study aims to develop a computational model for estimating the wind load acting on an FLNG. In particular, it is the sequel to the previous research by the author. The numerical computation model in the present study was modified based on the previous research. Numerical analysis for estimating wind loads was performed in two conditions for an interval of wind direction (α), 15° over the range of 0° to 360°. One condition is uniform wind speed and the other is the NPD model reflecting the wind speed profile. At first, the effect of sand-grain roughness on the speed profile of the NPD model was studied. Based on the developed NPD model, mesh convergence tests were carried out for 3 wind headings, i.e. head, quartering, and beam. Finally, wind loads on 6-degrees of freedom were numerically estimated and compared by two boundary conditions, uniform speed, and the NPD model. In the present study, a commercial RANS-based viscous solver, STAR-CCM+ (ver. 17.02) was adopted. In summary, wind loads in surge and yaw from the wind speed profile boundary condition were increased by 20.35% and 34.27% at most. Particularly, the interval mean of sway (45° < α <135°, 225° < α < 315°) and roll (60° < α < 135°, 225° < α < 270°) increased by 15.60% and 10.89% against the uniform wind speed (10m/s) boundary condition.

Artificial Neural Network with Firefly Algorithm-Based Collaborative Spectrum Sensing in Cognitive Radio Networks

  • Velmurugan., S;P. Ezhumalai;E.A. Mary Anita
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.17 no.7
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    • pp.1951-1975
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    • 2023
  • Recent advances in Cognitive Radio Networks (CRN) have elevated them to the status of a critical instrument for overcoming spectrum limits and achieving severe future wireless communication requirements. Collaborative spectrum sensing is presented for efficient channel selection because spectrum sensing is an essential part of CRNs. This study presents an innovative cooperative spectrum sensing (CSS) model that is built on the Firefly Algorithm (FA), as well as machine learning artificial neural networks (ANN). This system makes use of user grouping strategies to improve detection performance dramatically while lowering collaboration costs. Cooperative sensing wasn't used until after cognitive radio users had been correctly identified using energy data samples and an ANN model. Cooperative sensing strategies produce a user base that is either secure, requires less effort, or is faultless. The suggested method's purpose is to choose the best transmission channel. Clustering is utilized by the suggested ANN-FA model to reduce spectrum sensing inaccuracy. The transmission channel that has the highest weight is chosen by employing the method that has been provided for computing channel weight. The proposed ANN-FA model computes channel weight based on three sets of input parameters: PU utilization, CR count, and channel capacity. Using an improved evolutionary algorithm, the key principles of the ANN-FA scheme are optimized to boost the overall efficiency of the CRN channel selection technique. This study proposes the Artificial Neural Network with Firefly Algorithm (ANN-FA) for cognitive radio networks to overcome the obstacles. This proposed work focuses primarily on sensing the optimal secondary user channel and reducing the spectrum handoff delay in wireless networks. Several benchmark functions are utilized We analyze the efficacy of this innovative strategy by evaluating its performance. The performance of ANN-FA is 22.72 percent more robust and effective than that of the other metaheuristic algorithm, according to experimental findings. The proposed ANN-FA model is simulated using the NS2 simulator, The results are evaluated in terms of average interference ratio, spectrum opportunity utilization, three metrics are measured: packet delivery ratio (PDR), end-to-end delay, and end-to-average throughput for a variety of different CRs found in the network.

Downregulatory Effect of Extracts from Mistletoe (Viscum album) and Pueraria Root (Pueraria radix) on Cellular NF-κB Activation and heir Antioxidant Activity (겨우살이(Viscum album)와 칡뿌리(Pueraria radix) 추출물의 NF-κB활성 억제 및 항산화 효과)

  • Song, Hee-Sun;Park, Yeon-Hee;Kim, Seung-Kyoon;Moon, Won-Kuk;Kim, Dong-Woo;Moon, Ki-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.10
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    • pp.1594-1600
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    • 2004
  • Effects of mistletoe (Viscum album) extract and pueraria (Pueraria radix) extract on cellular NF-$textsc{k}$B activity were evaluated in human malignant keratinocytes (SCC-13) to elucidate the possible correlation of NF-$textsc{k}$B with antioxidant activity. The antioxidant activities of these natural extracts were examined in four different evaluation methods, i.e., lipid peroxidation value (POV) evaluation test, I,l-diphenyl-2-picrylhydrazyl radical (DPPH), nitric oxide (NO) scavenging test, and reducing power assay. Pueraria extract (0.5 mg) and mistletoe extract (5 mg) downregulated the cellular NF-$textsc{k}$B activation up to 35% and 10% compared to the control, respectively, although their effects were lower than the known NF-$textsc{k}$B downregulator, vitamin C (8.8 mg, 53%) in a cell-based NF-$textsc{k}$B activity assay system. In the POV test, relative antioxidant activities of mistletoe extract (86%) and pueraria extract (75%) were significantly higher than the known antioxidant, vitamin C (48%) at the same concentration (10 mg) and the degree of activity increased in a dose-dependent manner. Pueraria extract showed more potential radical scavenging activities than those of mistletoe extract evaluated in both DPPH and NO test. Especially, the NO radical scavenging activity of pueraria extract ($SC_{50}$/, 88 $\mu$g) was comparable to that of vitamin C ($SC_{50}$/, 77 $\mu$g). Even pueraria extract possessed a much less reducing power compared to vitamin C, it also revealed higher reducing power than that of mistletoe extract. These results indicate that mistletoe extract and pueraria extract may serve as an useful natural antioxidant agents, and led to suggest the hypothesis that compounds having an antioxidant activity, i.e., radical scavenging activity or reducing power may be correlated with the downregulation of NF-$textsc{k}$B activation in human keratinocytes.