• Title/Summary/Keyword: mFasL

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Overexpression of Recombinant Arylsulfatase Cloned from Pseudoalteromonas carrageenovora

  • Kim Jong-Oh;Kim Seok-Ryel;Lim Jae-Myung;Nam Soo-Wan;Kim Hyeung-Rak
    • Fisheries and Aquatic Sciences
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    • v.8 no.3
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    • pp.118-121
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    • 2005
  • Arylsulfatase cloned from a marine bacterium, Pseudoalteromonas carrageenovora, was over-expressed in Escherichia coli. Most of the recombinant arylsulfatase was found in the cell lysate with induction up to $10{\mu}M$ IPTG. However, enzyme activity was observed both in the culture supernatant and cell lysate by induction with IPTG concentration of $50-5,000{\mu}M$. Most of the recombinant enzyme was localized in the periplasmic space with $10{\mu}M$ IPTG induction, while half of the enzyme was distributed in the periplasmic space with $50{\mu}M$ IPTG induction. Cell growth and arylsulfatase activity did not change with the induction time, and the level of recombinant arylsulfatase expression was maintained at 4-5 U/mL after 6 to 14 hr of culture.

Antioxidant and Cholinesterase Inhibitory Activities of Aqueous Extract from Rainbow Trout Oncorhynchus mykiss

  • Baek, Jae-Min;Yoon, Na-Young;Kim, Yeon-Kye;Lee, Doo-Seog;Yoon, Ho-Dong;Park, Jeung-Sook
    • Fisheries and Aquatic Sciences
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    • v.14 no.2
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    • pp.89-92
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    • 2011
  • We investigated the antioxidant and cholinesterase inhibitory activities of the aqueous extract of rainbow trout Oncorhynchus mykiss. The antioxidant activity of O. mykiss aqueous extract was determined by in vitro peroxynitrite scavenging activity and reducing power assays. The aqueous extract of O. mykiss showed potent peroxynitrite radical scavenging activity ($IC_{50}=0.12{\pm}0.001\;mg/mL$) and reducing power (absorbance=$0.47{\pm}0.001$) at the concentration of 1 mg/mL. The in vitro cholinesterase inhibitory activity of O. mykiss aqueous extract was examined using spectrophotometric analyses of acetyl- and butyrylcholinesterase. The aqueous extract of O. mykiss showed acetylcholinesterase inhibitory activity ($IC_{50}=1.61{\pm}0.13\;mg/mL$), but did not exhibit inhibitory activity against butyrylcholinesterase. These results suggest that O. mykiss possesses antioxidant and acetylcholinesterase inhibitory activities and provide scientific evidence for the health benefits of O. mykiss aqueous extract.

Serial Particle Size Fractionation and Water Quality in a Recirculating Aquaculture System for Eel

  • Lee, Jin-Hwan
    • Fisheries and Aquatic Sciences
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    • v.13 no.2
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    • pp.133-139
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    • 2010
  • The effects of suspended solids size on culture water quality were determined in a commercial recirculating aquaculture system (RAS) for Japanese eel, Anguilla japonica. The particulate phase of the culture water was serially divided into six size fractions using 300, 200, 100, 75, 45, and 26 ${\mu}m$ pore size stainless sieves. The total, dissolved, and particulate nitrogen and phosphorus, and suspended solids for each fraction were determined. The concentration ranges in the fractions were: total nitrogen, 164-148 mg $L^{-1}$; total phosphorus, 20.4-15.5 mg $L^{-1}$; and total suspended solids, 8.1-6.1 mg $L^{-1}$. The concentration of total nitrogen and total phosphorus decreased significantly (P<0.05) with a 26 ${\mu}m$ and 200 ${\mu}m$ filter pore size, respectively. Nutrients from dissolved organic substances were much higher than from particulates. Analysis of particle size fractionation and its effects on water quality is useful to estimate removal efficiencies of a commercial effluent screening device for solid management and development of solid removal systems.

Mortality of Fishes and Shellfishes to Harmful Algal Blooms

  • Lee Sam Geun;Kim Hak Gyoon;Cho Eun Seob;Lee Chang Kyu
    • Fisheries and Aquatic Sciences
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    • v.6 no.3
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    • pp.160-163
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    • 2003
  • Mortality of several species of fish and shellfish exposed to Harmful Algal Blooms (HABs) caused by Cochlodinium polykrikoides, Heterosigam akashiwo, Alexandrium tamarense, Eutreptiella gymnastica, Heterocapsa triquetra and Prorocentrum micans was studied. When fish were exposed to a cell density of 8,000 cells $mL^{-1}$ in C. polykrikoides, $35\%$ of flatfish and darkbanded rockfish died within 48 hrs. However, jacopever rockfish had mortality of higher than $85\%$. Rock bream, filefish and red sea bream showed $100\%$ mortality within 10 hrs with an exposure cell density of 8,000 cells $mL^{-1}$. The rest of HABs except for C. polykrikoides showed that there was no fish and shellfish death throughout the 48 hrs even in the maximum cell density of 100,000 cells $mL^{-1}$ These results imply that C. polykrikoides can have a serious impact on fish mortality and it is regarded as an ichthyotoxic dinoflagellate. The fish death may be attributed to anoxia caused by a combination of the production of reactive oxygen species (ROS) and polysaccharide from C. polykrikoides during blooms.

Rockfish (Sebastes schlegeli) Vitellogenin: Purification, Characterization and Development of Sandwich ELISA System

  • Jung Jee Hyun;Kim Dae Jung;Han Chang Hee
    • Fisheries and Aquatic Sciences
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    • v.7 no.3
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    • pp.99-108
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    • 2004
  • Vitellogenin (VTG) was purified from serum of $estradiol-l7{\beta}-treated rockfish$(Sebastes schlegeli) by precipitation with $EDTA-Mg^{2+}$ and ammonium sulfate and two step chromatography (anion exchange chromatography and gel permeation chromatography) was performed on FPLC system. Rockfish VTG (rfVTG) was characterized and its properties were determined. The monomers have apparent, molecular mass of about 188 kDa as indicated by SDS-PAGE. Amino acid composition analysis of rfVTG was similar to VTG from other oviparous teleosts. Cysteine and lysine were present at relatively high level. Leucine was present at relatively lower level than in other species. The N-terminal amino acid sequence was evaluated to identify rfVTG. Western blot analysis using an antibody against the purified VTG showed that the antibody reacted with both plasma of $estradiol-l7{\beta}-treated rockfish$ treated male and purified VTG, whereas there was no reaction with male serum of the control. An ELISA was developed using monoclonal and polyclonal antibodies against rfVTG. The assay range was 3.2 ng/mL and 1,000 ng/mL and the value of the intra and inter assay variations were within $9.7{\%}$ and $11.2{\%}$, respectively. Recovery rate was $96.8{\%}$. The sandwich ELISA could be useful for the detection of VTG and could be good for screening of estrogenic compounds.

Red pepper seed water extract inhibits preadipocyte differentiation and induces mature adipocyte apoptosis in 3T3-L1 cells

  • Kim, Hwa-Jin;You, Mi-Kyoung;Lee, Young-Hyun;Kim, Hyun-Jung;Adhikari, Deepak;Kim, Hyeon-A
    • Nutrition Research and Practice
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    • v.12 no.6
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    • pp.494-502
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    • 2018
  • BACKGROUND/OBJECTIVES: Reducing the number of adipocytes by inducing apoptosis of mature adipocytes as well as suppressing differentiation of preadipocytes plays an important role in preventing obesity. This study examines the anti-adipogenic and pro-apoptotic effect of red pepper seed water extract (RPS) prepared at $4^{\circ}C$ (RPS4) in 3T3-L1 cells. MATERIALS/METHODS: Effect of RPS4 or its fractions on lipid accumulation was determined in 3T3-L1 cells using oil red O (ORO) staining. The expressions of AMP-activated protein kinase (AMPK) and adipogenic associated proteins [peroxisome proliferator-activated receptor-${\gamma}$ ($PPAR-{\gamma}$), CCAAT/enhancer-binding proteins ${\alpha}$ (C/EBP ${\alpha}$), sterol regulatory element binding protein-1c (SREBP-1c), fatty acid synthase (FAS), and acetyl-CoA carboxylase (ACC)] were measured in 3T3-L1 cells treated with RPS4. Apoptosis and the expression of Akt and Bcl-2 family proteins [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated death promoter (Bad), Bcl-2 like protein 4 (Bax), Bal-2 homologous antagonist/killer (Bak)] were measured in mature 3T3-L1 cells treated with RPS4. RESULTS: Treatment of RPS4 ($0-75{\mu}g/mL$) or its fractions ($0-50{\mu}g/mL$) for 24 h did not have an apparent cytotoxicity on pre and mature 3T3-L1 cells. RPS4 significantly suppressed differentiation and cellular lipid accumulation by increasing the phosphorylation of AMPK and reducing the expression of $PPAR-{\gamma}$, C/EBP ${\alpha}$, SREBP-1c, FAS, and ACC. In addition, all fractions except ethyl acetate fraction significantly suppressed cellular lipid accumulation. RPS4 induced the apoptosis of mature adipocytes by hypophosphorylating Akt, increasing the expression of the pro-apoptotic proteins, Bak, Bax, and Bad, and reducing the expression of the anti-apoptotic proteins, Bcl-2 and p-Bad. CONCLUSIONS: These finding suggest that RPS4 can reduce the numbers as well as the size of adipocytes and might useful for preventing and treating obesity.

Anti-obesity Effect of Aster Yomena Ethanol Extract in High Fat Diet-induced Obese Mice (고지방 식이로 유도된 비만 생쥐에서 쑥부쟁이 에탄올 추출물의 항비만 효과)

  • Lee, Ho Jae;Kim, Hyun Sik;Seo, Sang Wan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.31 no.6
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    • pp.348-355
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    • 2017
  • Aster yomena (AY) have been used as a traditional medicine to treat cough, bronchial asthma, and insect bites in Korea. In this study, we evaluated the inhibition of adipogenesis in 3T3-L1 cells and in high-fat diet (HFD)-induced obese mice by AY ethanol extract. Lipid accumulation measurement indicates that AY markedly inhibited adipogenesis in a dose-dependent manner. qRT-PCR results demonstrated that the mRNA expression of adipogenic transcription factors such as peroxisome proliferator-activated receptor-${\gamma}$ ($PPAR-{\gamma}$) in 3T3-L1 cells were significantly down-regulated by AY treatment. And inhibited the expression of FAS, a protein responsible for lipid synthesis, transport and storage. Oral administration of AY (100, 250, and 500 mg/kg, P.O/daily for 4 weeks) was conducted in high-fat diet induced obese mice and C57BL/6 mice. AY was orally administered for 4 weeks to extract liver and epididymal fat, and hematoxylin and eosin staining(H&E staining) was observed. Observation showed that the fat concentration of liver tissue tended to decrease dose-dependently and decreased significantly at 500 mg/kg concentration. The AY-administered group of HFD-induced mice had a lower body weight gain, along with decreased triglycerides and total cholesterol compared with the control mice, however, the HDL-cholesterol/total cholesterol ratio was increased. These results indicate that AY exhibits anti-obesity effects in obese mice by decreasing in serum lipid levels and lipogenesis related gene.

Pear pomace water extract inhibits adipogenesis and induces apoptosis in 3T3-L1 adipocytes

  • Rhyu, Jin;Kim, Min Sook;You, Mi-Kyoung;Bang, Mi-Ae;Kim, Hyeon-A
    • Nutrition Research and Practice
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    • v.8 no.1
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    • pp.33-39
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    • 2014
  • Obesity occurs when a person's calorie intake exceeds the amount of energy burns, which may lead to pathologic growth of adipocytes and the accumulation of fat in the tissues. In this study, the effect and mechanism of pear pomace extracts on 3T3-L1 adipocyte differentiation and apoptosis of mature adipocytes were investigated. The effects of pear pomace extract on cell viability and the anti-adipogenic and proapoptotic effects were investigated via MTT assay, Oil red O staining, western blot analysis and apoptosis assay. 3T3-L1 preadipocytes were stimulated with DMEM containing 10% FBS, 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), $5{\mu}g/ml$ insulin and $1{\mu}M$ dexamethasone for differentiation to adipocytes. 3T3-L1 cells were cultured with PBS or water extract of pear pomace. Water extract of pear pomace effectively inhibited lipid accumulations and expressions of PPAR-${\gamma}$ and $C/EBP{\alpha}$ in 3T3-L1 cells. It also increased expression of p-AMPK and decreased the expression of SREBP-1c and FAS in 3T3-L1 cells. The induction of apoptosis was observed in 3T3-L1 cells treated with pear pomace. These results indicate that pear pomace water extract inhibits adipogenesis and induces apoptosis of adipocytes and thus can be used as a potential therapeutic substance as part of prevention or treatment strategy for obesity.

Growth Enhancement of the Microalga Tetraselmis suecica by an Extract of the Green Alga Monostroma nitidum

  • Cho Ji Young;Choi Jae Suk;Kang Se Eun;Ahn Sang Jung;Kim Joong Kyun;Hong Yong Ki
    • Fisheries and Aquatic Sciences
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    • v.8 no.1
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    • pp.1-5
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    • 2005
  • Cell growth of the marine microalga Tetraselmis suecica was regulated by the addition of seaweed extracts in its culture medium. Of 26 species of seaweed tested, water-soluble extracts from Monostroma nitidum and Pachymeniopsis elliptica and methanol-soluble extract from Sargassum confilsum enhanced the growth of T suecica cells. The water extract of M. nitidum was the most effective, producing up to a 2-fold increase in cell density with the addition of 1 mg/mL of extract to the culture medium. Cell size, gross biochemical composition, fatty acids, and digestion efficiency all differed marginally between cultures of T suecica grown with and without the M. nitidum extract.

Experimental and theoretical justification of passive heat removal system for irradiated fuel assemblies of the nuclear research reactor in a spent fuel pool

  • Ta Van Thuong;O.L. Tashlykov;S.M. Glukhov;D.E. Shumkov;Yu.V. Volchikhina
    • Nuclear Engineering and Technology
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    • v.55 no.6
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    • pp.2088-2095
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    • 2023
  • The safety of nuclear installations is largely determined by the tightness of fuel elements cladding. As the Fukushima nuclear accident showed, the main task in case of loss of power supply is to ensure reliable removal of residual heat release from spent fuel pool (SFP) with irradiated fuel assemblies (IFAs). The paper presents the results of calculated-experimental studies and thermal-hydraulic modeling of temperature storage modes of IFAs in SFP. Experimental studies of SFP's temperature regime and calculated evaluation of residual heat removal due to the thermal conductivity of building structures surrounding the SFP were performed. To ensure the safe operation of research reactors, it's necessary to know the IFA's residual heat power (RHP) in the reactor and SFP, which is determined depending on the operating time of fuel assemblies (FAs) and the IFAs calculated holding time. The FAs operating time depends on the reactor energy output. The IFAs calculated holding time is determined by the fuel burnup, U-235 mass in the fuel, and reactor utilization factor. The IFAs fuel burnup was calculated using the MCU-PTR program. Also presented are the RHP's calculation results using some of the empirical dependencies. The concept of a passive heat removal system (PHRS) based on thermosyphon's operating principle was proposed.