• Title/Summary/Keyword: Cell density

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Production of Antifreeze Protein from Antarctic Bacterium Flavobacterium frigoris PS1 by using Fed-batch Culture of Recombinant Pichia pastoris (재조합 Pichia pastoris의 유가식 배양을 통한 남극세균 Flavobacterium frigoris PS1 유래 결빙방지단백질의 생산)

  • Kim, Eun Jae;Do, Hackwon;Lee, Jun Hyuck;Lee, Sung Gu;Kim, Hak Jun;Han, Se Jong
    • KSBB Journal
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    • v.29 no.4
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    • pp.303-306
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    • 2014
  • Antifreeze proteins (AFP) inhibit ice growth to permit the survival of polar organisms in the cold environments. The recombinant AFP from an Antarctic bacterium, Flavobacterium frigoris PS1, FfIBP (Flavobacterium frigoris ice-binding protein), was produced using Pichia pastoris expression system. The optimum fermentation temperature ($30^{\circ}C$) and pH (5) for FfIBP production were determined using a fed-batch culture system. The maximal cell density and purified FfIBP were 112 g/L and 70 mg/L, respectively. The thermal hysteresis (TH) activity (0.85) of FfIBP obtained using a glycerol-methanol fed-batch culture system was 2-fold higher than that of the LeIBP (Leucosporidium ice-binding protein). This work allows for large-scale production of FfIBP, which could be extended to further application studies using recombinant AFPs.

Large Increase in Leuconostoc citreum KM20 Dextransucrase Activity Achieved by Changing the Strain/Inducer Combination in an E. coli Expression System

  • Ko, Jin-A;Jeong, Hyung-Jae;Ryu, Young-Bae;Park, Su-Jin;Wee, Young-Jung;Kim, Do-Man;Kim, Young-Min;Lee, Woo-Song
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.510-515
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    • 2012
  • A recombinant putative dextransucrase (DexT) was produced from Leuconostoc citreum KM20 as a 160 kDa protein, but its productivity was very low (264 U/l). For optimization, we examined enzyme activity in 7 Escherichia coli strains with inducer molecules such as lactose or IPTG. E. coli BL21-CodonPlus(DE3)-RIL exhibited the highest enzyme activity with lactose. Finally, DexT activity was remarkably increased by 12-fold under the optimized culture conditions of a cell density to start induction ($OD_{600}$) of 0.95, a lactose concentration of 7.5 mM, and an induction temperature of $17^{\circ}C$. These results may effectively apply to the heterologous expression of other large DexT genes.

Comparative Effects of $PKB-{\alpha}$ and $PKC-{\zeta}$ on the Phosphorylation of GLUT4-Containing Vesicles in Rat Adipocytes

  • Hah, Jong-Sik
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.6
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    • pp.487-496
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    • 2000
  • Insulin stimulates glucose transport in muscle and fat cells by promoting the translocation of glucose transporter (GLUT4) to the cell surface. Phosphatidylinositide 3-kinase (PI3-kinase) has been implicated in this process. However, the involvement of protein kinase B (PKB)/Akt and $PKC-{\zeta}$, those are known as the downstream target of PI3-kinase in regulation of GLUT4 translocation, is not known yet. An interesting possibility is that these protein kinases phosphorylate GLUT4 directly in this process. In the present study, $PKB-{\alpha}$ and $PKC-{\zeta}$ were added exogenously to GLUT4-containing vesicles purified from low density microsome (LDM) of the rat adipocytes by immunoadsorption and immunoprecipitation for direct phosphorylation of GLUT4. Interestingly GLUT4 was phosphorylated by $PKC-{\zeta}$ and its phosphorylation was increased in insulin stimulated state but GLUT4 was not phosphorylated by $PKB-{\alpha}.$ However, the GST-fusion proteins, GLUT4 C-terminal cytoplasmic domain (GLUT4C) and the entire major GLUT4 cytoplasmic domain corresponding to N-terminus, central loop and C-terminus in tandem (GLUT4NLC) were phosphorylated by both $PKB-{\alpha}$ and $PKC-{\zeta}.$ The immunoblots of $PKC-{\zeta}$ and $PKB-{\alpha}$ antibodies with GLUT4-containing vesicles preparation showed that $PKC-{\zeta}$ was co-localized with the vesicles but not $PKB-{\alpha}.$ From the above results, it is clear that $PKC-{\zeta}$ interacts with GLUT4-containing vesicles and it phosphorylates GLUT4 protein directly but $PKB-{\alpha}$ does not interact with GLUT4, suggesting that insulin-elicited signals that pass through PI3-kinase subsequently diverge into two independent pathways, an Akt pathway and a $PKC-{\zeta}$ pathway, and that later pathway contributes, at least in part, insulin stimulation of GLUT4 translocation in adipocytes via a direct GLUT4 phosphorylation.

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Water Quality and Phytoplankton Standing Crops in the Daechung Reservoir (대청호의 수질과 식물플랑크톤 현존량)

  • 이진환;오희목;맹주선
    • Korean Journal of Environmental Biology
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    • v.18 no.3
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    • pp.355-365
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    • 2000
  • In order to study the relationship between water quality and phytoplankton standing crops, the authors investigated the physicochemical factors, phytoplankton standing crops and chlorophyll-a at two stations in the Daechung Reservoir during the period from May to November 1998. Nutrients (nitrite, nitrate, ammonia and phosphate) have gradually increased more than those of the previous reports. Concentrations of the total nitrogen, phosphorus, chlorophyll -a and transparency exceeded the standard value of eutrophication. It could be assumed that the eutrophication of the Daechung reservoir was caused by various pollutants. Chlorophyll-a concentration was under the influence of phytoplankton standing crops, but very tiny cell-sized blue-green algae were less important in regards to the chlorophyll-a. Except at station 1 from July to August, phytoplankton standing crops bloomed during the time of the survey. Spring phytoplankton blooms caused by nitrate and phosphate in the surface layer. High concentrations of the suspended solids in July and August were caused by an inflow of them due to heavy rains. Density of heterotrophic bacteria was high in August owing to the high water temperatures and the organic nutrients which were from the nearby reservoir.

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Clinical Study of 61 cases Benign Lung Tumor (폐양성 종양 61례의 외과적 고찰)

  • 박종원
    • Journal of Chest Surgery
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    • v.22 no.2
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    • pp.272-289
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    • 1989
  • Benign lung tumors compose a heterogeneous group of solid growths that present variations in clinical features, depending on whether the origin is within the bronchus on lung parenchyma or from visceral pleura. Benign tumors of the lung are relatively uncommon, and series are to be found in the literature and the classification of benign tumors of the lung continues to be controversial because of disagreement concerning the origin and prognosis of many common lesions. We adopt Liebows original classification but excluded bronchial adenoma which no longer considered as benign tumor and added pulmonary A-V fistula and congenital cystic adenomatoid malformation. We analyzed 61 cases of benign tumors which were composed of 16 original Korean cases and 45 cases which were reported on journal of Thoracic & Cardiovascular Surgery. The results were. [1] Incidence; Of 61 cases, chondromatous hamartoma was 2 cases [41 %], congenital cystic adenomatoid malformation 10 cases [16.4 %], pulmonary A-V fistula 5 cases [8.1 %], sclerosing hemangioma 4 cases [6.5 %], teratoma, plasma cell granuloma & mesothelioma were 3 cases [4.9%], Castlemans disease 2 cases [3.3%], and mucous gland adenoma, paraganglioma, and leiomyoma 1 case [1.6 %]. [2] Age & Sex distribution; Male 30 cases and female 31 cases. Mean age was 31.4 years old. [3] Main symptom; was coughing, 32.8%, and no symptom, 24.6%. [4] Sixty eight percentage of chest film showed mass density, and 4 cases showed calcification, 2 cases had lobulation. [5] Size of mass was large and multiple mass was 2 cases. Endobronchial tumors were 9 cases, 14.9 %. [6] Three cases of endobronchial tumor were preoperatively diagnosed by bronchoscopy and 2 cases of pulmonary A-V fistula were diagnosed by pulmonary arteriography. [7] Seven cases, 11.5%, had associating diseases such as bronchogenic cyst, thymic cyst, Schwannoma, situs inversus, bronchiectasis and bronchogenic carcinoma. [8] Minor resection such as excision 8i: wedge resection were 15 cases, 26.2 %, and 6 cases, 75.4 %, of lobectomy were performed including 5 cases of pneumonectomy 5 cases had. [9] Postoperative complications; One case, 1.6 %, expired due to respiratory insufficiency. Two cases had re-operation due to bleeding and hemoptysis.

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Regeneration of PCB Etchants and Copper Recovery in a Batch-type Electrolytic Cell (회분식 전해조에서 PCB 식각폐수의 재생 및 구리의 회수)

  • Nam, Sang Cheol;Nam, Chong Woo;Tak, Yongsug;Oh, Seung Mo
    • Applied Chemistry for Engineering
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    • v.8 no.2
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    • pp.161-171
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    • 1997
  • Anodic regeneration of PCB enchant and cathodic deposition of copper using electrochemical method has been studied. Cu(I)/Cu(II) concentration ratio as a function of Cu(I) oxidation at the anode was measured from the potential difference between platinum and Ag/AgCl/4M KCl electrodes. Chlorine gas evolution was minimized by maintaining Cu(I) concentration above a specific concentration and using non-porous graphite electrode. Dendritic copper deposition was observed at the cathode and the optimum conditions for Cu deposition was identified as the current density of $360mA/cm^2$, and copper concentration of 12 g/l. Titanium was the most effective cathode material which showed a higher current efficiency and copper recovery. The current efficiency decreased with increasing temperature, but the highest power efficiency was achieved at $50^{\circ}C$.

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Effect of Herbicide Paraquat on Electron Donor and Acceptor (제초제 Paraquat의 전자수용 및 방출에 대한 영향)

  • Kim Mi-Lim;Choi Kyung-Ho
    • Journal of Life Science
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    • v.15 no.2 s.69
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    • pp.311-315
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    • 2005
  • When paraquat was added to the bacterial membrane or mitochondrial suspension, the mixture turned dark blue, but the color was disappeared by aeration. The same phenomenon was seen when electrons were supplied to the paraquat. Blue color appeared from near the cathode, and then spreaded to whole transit system. Coloration was accelerated by addition of alkali, but the color was reduced by addition of acid or oxygen. Paraquat exhibited absorption at ultraviolet region by electron transfer at the concentrations as low as 1.0 mM which did not exert difficulty in showing color reaction. Paraquat caused the increase of the optical density at 340 nm by electron transit, and an aspect of that had a strong resemblance to NADH. The acute toxic action of paraquat seemes to depend on inhibition of energy metabolism cased by paraquat action of electron donor and acceptor.

Preparation of Seasoned and Semi-Dried Right-Eyed Flounder and Quality of Its Product during Storage (참가자미 조미 반건조제품의 제조와 저장 중 품질안정성)

  • 양승택
    • Journal of Life Science
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    • v.9 no.1
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    • pp.44-49
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    • 1999
  • A study was conducted to investigate the possibility of processing seasoned and semi-dried right-eyed flounder (Pseudopleuronects herzensteini) and to examine its quality during storage at $\pm$$3^{\circ}C$ by analyzing the factors such as moisture content, pH, volatile basic nitrogen (VBN), viable cell count and peroxide value (POV). For processing of the seasoned and semi-dried product, optimum conditions of drying temperature and drying time were $20^{\circ}C$ and 4 hrs, respectively, under 50$\pm$5$\%$ of relative humidity and 2 m/sec of air blast speed. The shelf-lives of the products by vacuum-packaging method in nylon/polyethylene/linear low density polyethylene (0.015/0.045/0.040 mm) laminated film bag were 35 and 16 days during storage at $-3^{\circ}C$ and $3^{\circ}C$, respectively.

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Cyclooxygenase-2 over-expression is associated with increased mast cells in CCl4-induced hepatic fibrosis

  • Jekal, Seung-Joo;Lee, Jae-Hyoung;Park, Seung-Teack
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.4
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    • pp.229-238
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    • 2012
  • Cyclooxygenase(COX-2) is an inducible enzyme that catalyzes the synthesis of prostaglandins (PGs) from arachidonic acid. Over-expression of COX-2 has been reported to be associated with progressive hepatic fibrosis in chronic hepatic C infection and rat liver fibrosis induced by carbon tetrachloride($CCl_4$). Recently, it is well known that mast cell products can stimulate the proliferation of hepatic stellate cells and key players in liver fibrosis. But little is known regarding their role in $CCl_4$-induced liver fibrosis in rat. Our aim was to investigate the relation between COX-2 expression and mast cells during liver fibrosis after $CCl_4$ treatment. Thirty Wistar rats were divided into five groups (non-treated 0, 2, 4, 6 and 8-week after $CCl_4$-treatment). Reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to assess the expression of ${\alpha}$-smooth muscle actin (${\alpha}$-SMA), collagen-1 and COX-2 in liver tissue from $CCl_4$-treated rats. The density of collagen and mast cells were determined using a computerized image analysis system in liver sections stained with picrosirius red and toluidine blue, respectively. The expression levels of ${\alpha}$-SMA, collagen-1 and COX-2 mRNA were significantly higher at 2 wk in $CCl_4$-treated groups than non-treated group. The number of mast cells in liver tissues increased gradually from 2 wk to 6 wk depending on the fibrosis severity but decreased abruptly at 8 wk. The significant increase of collagen-1 and ${\alpha}$-SMA mRNA expression in $CCl_4$-treated rats was continued until 6 wk while the COX-2 mRNA was significantly decreased at 8 wk. These results suggest that increased mast cells are closely associated with COX-2 over-expression during hepatic fibrogenesis of $CCl_4$-treated rats.

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Visual Analysis for Detection and Quantification of Pseudomonas cichorii Disease Severity in Tomato Plants

  • Rajendran, Dhinesh Kumar;Park, Eunsoo;Nagendran, Rajalingam;Hung, Nguyen Bao;Cho, Byoung-Kwan;Kim, Kyung-Hwan;Lee, Yong Hoon
    • The Plant Pathology Journal
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    • v.32 no.4
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    • pp.300-310
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    • 2016
  • Pathogen infection in plants induces complex responses ranging from gene expression to metabolic processes in infected plants. In spite of many studies on biotic stress-related changes in host plants, little is known about the metabolic and phenotypic responses of the host plants to Pseudomonas cichorii infection based on image-based analysis. To investigate alterations in tomato plants according to disease severity, we inoculated plants with different cell densities of P. cichorii using dipping and syringe infiltration methods. High-dose inocula (${\geq}10^6cfu/ml$) induced evident necrotic lesions within one day that corresponded to bacterial growth in the infected tissues. Among the chlorophyll fluorescence parameters analyzed, changes in quantum yield of PSII (${\Phi}PSII$) and non-photochemical quenching (NPQ) preceded the appearance of visible symptoms, but maximum quantum efficiency of PSII ($F_v/F_m$) was altered well after symptom development. Visible/near infrared and chlorophyll fluorescence hyperspectral images detected changes before symptom appearance at low-density inoculation. The results of this study indicate that the P. cichorii infection severity can be detected by chlorophyll fluorescence assay and hyperspectral images prior to the onset of visible symptoms, indicating the feasibility of early detection of diseases. However, to detect disease development by hyperspectral imaging, more detailed protocols and analyses are necessary. Taken together, change in chlorophyll fluorescence is a good parameter for early detection of P. cichorii infection in tomato plants. In addition, image-based visualization of infection severity before visual damage appearance will contribute to effective management of plant diseases.