• Title/Summary/Keyword: Cell retention

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Enhanced and Targeted Expression of Fungal Phytase in Saccharomyces cerevisiae

  • LIM, YOUNG-YI;EUN-HA PARK;JI-HYE KIM;SEUNG-MOON PARK;HYO-SANG JANG;YOUN-JE PARK;SEWANG YOON;MOON-SIK YANG;DAE-HYUK KIM
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.915-921
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    • 2001
  • Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. In order to express a high level of fungal phytase in Saccharomyces cerevisiae, various expression vectors were constructed with different combinations of promoters, translation enhancers, signal peptides, and terminator. Three different promoters fused to the phytase gene (phyA) from Aspergillus niger were tested: a galactokinase (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, and yeast hybrid ADH2-GPD promoter consisting of alcohol dehydrogenase II (ADH2) and a GPD promoter. The signal peptides of phytase, glucose oxidase (GO), and rice amylase 1A(RAmy1A) were included. Plus, the translation enhancers of the ${\Omega}$ sequence and UTR70 from the tobacco mosaic virus (TMV) and spinach, respectively, were also tested. Among the recombinant vectors, pGphyA06 containing the GPD promoter, the ${\Omega}$ sequence, RAmy1A, and GAL7 terminator expressed the highest phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase was also performed by inserting an endoplasmic reticulum (ER) retention signal, KDEL sequence, into the C-terminus of the phytase within the vector pHphyA-6. It appeared that the KDEL sequence directed most of the early expression of phytase into the intracellular compartment yet more than $60\%$ of the total phytase activity was still retained within the cell even after the prolonged (>3 days) incubation of the transformant. However, the intracellular enzyme activity of the transformant without a KDEL sequence was as high as that of the extracellular one, thereby strongly suggesting that the secretion of phytase in S. cerevisiae appeared to be the rate-limiting step for the expression of a large amount of extracellular recombinant phytase, when compared with other yeasts.

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Different Sources and Levels of Copper Supplementation on Performance and Nutrient Utilization of Castrated Black Bengal (Capra hircus) Kids Diet

  • Mondal, M.K.;Biswas, P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.7
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    • pp.1067-1075
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    • 2007
  • Twenty eight 3-4 month old castrated Black Bengal kids (Capra hircus) were used to determine the effects of source and level of dietary copper (Cu) concentration on their performance and nutrient utilization. Cu was supplemented (0, 10, 20 and 30 mg/kg diet DM) as copper sulfate ($CuSO_4$, $5H_2O$) or copper proteinate (Cu-P). Kids were fed a basal diet containing maize (19.5%), soybean (17.0%), deoiled rice bran (56.5%), molasses (4.0%), di-calcium phosphate and salt (1.0% each) and mineral and vitamin mixture (0.5% each) supplements at 3.5% of body weight to meet NRC (1981) requirements for protein, energy, macro minerals and micro minerals, excluding Cu. The basal diet contained 5.7 mg Cu/kg, 122.5 mg Fe/kg, 110 mg Zn/kg, 0.26 mg Mo/kg and 0.32% S. $CuSO_4$ or Cu-P was added to the basal diet at the rate of 10, 20 and 30 mg/kg. Kids were housed in a well ventilated shed with facilities for individual feeding in aluminum plated metabolic cages. Blood samples were collected from the jugular vein on d 0, 30, 60 and 90 to determine hemoglobin (Hb), packed cell volume (PCV), total erythrocyte count (TEC), total leukocyte count (TLC) and serum enzymes (alkaline phosphatase, alanine transferase and aspertate transferase). A metabolism trial of 6 days duration was conducted after 90 days of experimental feeding. Statistical analysis revealed that source and level of Cu supplementation improved live weight gain (p<0.04) and average daily gain (p<0.01). No significant contribution of source and level of Cu to alter serum serum enzymes was evident. Goats fed Cu-P tended to have higher Hb, PCV and TEC than with $CuSO_4$ supplementation. Cu-P increased digestibility of ether extract (EE, p<0.02) and crude fiber (p<0.05) and showed an increasing trend (p<0.09) for digested crude protein (CP) and crude fiber (CF). Supplemental dose of Cu linearly improved (p<0.02) digestibilities of dry matter (DM), organic matter (OM), EE and nitrogen free extract (NFE). Though the absorption of nitrogen (N) was not affected (p>0.10) by both source and dose of Cu, N retention was affected (p<0.04) and there was a significant $Source{\times}Dose$ interaction (p<0.05). Final body weight (BW) was not influenced (p>0.10) by the source of Cu but increasing dose of Cu increased (p<0.04) the BW of kids. TDN intake (g/kg $W^{0.75}$) was higher (p<0.05) with the increased dose of Cu and there was a significant $Source{\times}Dose$ interaction. It was concluded that supplementation of Cu from different sources and varying dose level in a concentrate based diet may improve performance, nutrient utilization and plane of nutrition in castrated Black Bengal kids. The effects on performance and nutrient utilization are more pronounced with Cu-P than $CuSO_4$ supplementation. Higher dose of Cu showed better result than lower dose.

Study on Introduction to Predicting Indicator of Cyanobacteria Dominance in Algae Bloom Warning System of Hangang Basin (한강유역 조류경보제에 남조류 우점 예측인자 도입에 관한 연구)

  • Kim, Tae Kyun;Choi, Jae Ho;Lee, Kyung Ju;Kim, Young Bae;Yu, Sung Jong
    • Journal of Korean Society of Environmental Engineers
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    • v.36 no.5
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    • pp.378-385
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    • 2014
  • The chlorophyll-a concentration in algae bloom warning system of Hangang basin did not predict the cyanobacteria dominance. In this study, suggest the predicting indicator of cyanobacteria dominance through analyzing the environmental factors affecting on the cell count of cyanobacteria. Firstly, the dominance of algae was analyzed with seasonal variation during Jan. 2012~Sep. 2013. The diatom dominated phytoplankton communities during the period of January~April. In the May~June, the green algae dominated. And, the dominance of algae was changed to cyanobacteria in the July~August. Also, the environmental factors affecting to cyanobacteria blooms ; nutrients (TN, TP), temperature, precipitation, dam-discharge were evaluated during the study period. Rather than temperature factor, relatively low dam discharge causes cyanobacteria to grow rapidly and create a blooms. The low dam-discharge may increase the water retention time. Finally, it is proved that a low ratio of TN to TP (<29:1) can favour the development of cyanobacteria blooms. Thus, the predicting indicator (TN:TP) have need to apply to the alarm bloom warning system of Hangang basin.

Peritoneal Dialysis in Dogs: 20 cases (2006-2008) (개에서 복막투석 적용: 20 증례 (2006-2008))

  • Nam, So-Jeong;Choi, Ran;Oh, Won-Seok;Hyun, Chang-Baig
    • Journal of Veterinary Clinics
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    • v.26 no.1
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    • pp.23-28
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    • 2009
  • Peritoneal dialysis (PD) is a clinical technique that therapeutically removes toxic solutes from body fluids and normalizes endogenous solutes whose aberrant concentrations disrupt normal physiology. This study retrospectively evaluated clinical outcomes and complications of PD in 20 dogs with renal failure. Blood works (total count of red blood cells (RBC), packed cell volume (PCV), the serum biochemical, and electrolyte values related to renal insufficiency) and complications associated with peritoneal dialysis, and clinical outcomes were recorded before and after PD. Additionally, creatinine reduction ratio (CRR) and urea nitrogen reduction ratio (URR) were calculated for evaluating the efficacy of PD. PD resulted in a significant (p < 0.05) reduction in blood urea nitrogen (BUN) concentration in 19 dogs, while a significant (p < 0.05) reduction in creatinine concentration in 17 dogs. The complications of PD were hypoalbuminemia (12/20, 60%), anemia (10/20, 50%), subcutaneous dialysate leakage (9/20, 45%), bacterial peritonitis (6/20, 30%), dialysate retention (5/20, 25%) and limb edema (4/20, 20%). This study demonstrated that PD was effective in reducing the magnitude of azotemia in dogs with renal failure especially in acute phase, although the complication rate was high but manageable.

Electrochemical Performance on the H3BO3 Treated Soft Carbon modified from PFO as Anode Material (음극소재로 PFO에서 개질된 붕산처리 소프트 카본의 전기화학적 성능)

  • Lee, Ho Yong;Lee, Jong Dae
    • Korean Chemical Engineering Research
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    • v.54 no.6
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    • pp.746-752
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    • 2016
  • In this study, soft carbon was prepared by carbonization of carbon precursor (pitch) obtained from PFO (pyrolysis fuel oil) heat treatment. Three carbon precursors prepared by the thermal reaction were 3903 (at $390^{\circ}C$ for 3 h), 4001 (at $400^{\circ}C$ for 1 h) and 4002 (at $400^{\circ}C$ for 2 h). After the prepared soft carbon was ground to a particle size of $25{\sim}35^{\circ}C$, the soft carbon was synthesised by the chemical treatment with boric acid ($H_3BO_3$). The prepared soft carbon were analysed by XRD, FE-SEM and XPS. Also, the electrochemical performances of soft carbon were investigated by constant current charge/discharge test, cyclic voltammetry and impedance tests in the electrolyte of $LiPF_6$ dissolved inorganic solvents (EC:DMC=1:1 vol%+VC 3 wt%). The coin cell using soft carbon of $25{\sim}35^{\circ}C$ with 3903 soft carbon ($H_3BO_3$/Pitch=3:100 in weight) has better initial capacity and efficiency (330 mAh/g, 82%) than those of other coin cells. Also, it was found that the retention rate capability of 2C/0.1C was 90% after 30 cycles.

Electrochemical Characteristics of PFO pitch Anode prepared by Chemical Activation for Lithium Ion Battery (리튬이온전지용 화학적 활성화로 제조된 석유계 피치 음극소재의 전기화학적 특성)

  • Hwang, Jin Ung;Lee, Jong Dae
    • Korean Chemical Engineering Research
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    • v.55 no.3
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    • pp.307-312
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    • 2017
  • In this study, the electrochemical performance of surface modified carbon using the PFO (pyrolyzed fuel oil) was investigated by chemical activation with KOH and $K_2CO_3$. PFO was heat treated at $390{\sim}400^{\circ}C$ for 1~3h to prepared the pitch. Three carbon precursors (pitch) prepared by the thermal reaction were 3903 (at $390^{\circ}C$ for 3h), 4001(at $400^{\circ}C$ for 1h) and 4002 (at $400^{\circ}C$ for 2h). Also, the effect of chemical activation catalysts and mixing time on the development of porosity during carbonization was investigated. The prepared carbon was analyzed by BET and FE-SEM. It was shown that chemical activation with KOH could be successfully used to develop carbon with specific surface area ($3.12m^2/g$) and mean pore size (22 nm). The electrochemical characteristics of modified carbon as the anode were investigated by constant current charge/discharge, cyclic voltammetry and electrochemical impedance tests. The coin cell using pitch (4002) modified by KOH has better initial capacity (318 mAh/g) than that of other pitch coin cells. Also, this prepared carbon anode appeared a high initial efficiency of 80% and the retention rate capability of 2C/0.1 C was 92%. It is found that modified carbon anode showed improved cycling and rate capacity performance.

Bombyx mori Protein Disulfide Isomerase (bPDI) Protects Sf9 Cells from Endoplasmic Reticulum (ER) Stress (소포체 스트레스에 대한 Protein Disulfide Isomerase의 세포보호효과)

  • Goo, Tae-Won;Yun, Eun-Young;Kim, Sung-Wan;Choi, Kwang-Ho;Kang, Seok-Woo;Kwon, Ki-Sang;Kwon, O-Yu
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1129-1134
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    • 2007
  • In the previous our study, a cDNA that encodes protein disulfide isomerase from Bombyx mori (bPDI)was isolated and characterized. bPDI has an open reading frame of 494 amino acids contained two PDI-typical thioredoxin active site of WCGHCK and ER (endoplasmic reticulum) retention signal of the KDEL motif at its C-terminal. Recent studies have demonstrated that misfolded proteins are accumulated in many diseases including Alzheimer’s, goiter, emphysema, and prion infections. bPDI was over-expressed or knock-downed in Sf9 cells to study the relationship between bPDI expression and protections against protein misfolding. bPDI gene was cloned in insect expression vector pIZT/V5-His for over-expression and bPDI double-stranded RNA (dsRNA) was generated for knock-down. Over-expression of bPDI significantly improved survival rate, but bPDI dsRNA transfection significantly reduced survival rate after 48 hours exposure. In mock-transfected or wild-type cells had no significant effect. The results support the view that bPDI is one of the important intracellular components for cell protect mechanism, especially, against ER stress such as protein misfolding.

Study on Bread-making Quality with Barley Sourdough in Composite Bread (보리 Sourdough의 제빵성 연구)

  • Ryu, Chung-Hee;Kim, Sun-Young
    • Korean journal of food and cookery science
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    • v.21 no.5
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    • pp.733-741
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    • 2005
  • Waxy barley flour was fermented by two kinds of starter cultures; L. plantarum and L. brevis, alone or in combination and the effect was evaluated on waxy barley and wheat composite bread quality. In all three barley sourdoughs, fermentation decreased the pH, total sugar and reducing sugar, and increased lactic acid bacteria cell numbers. However yeasts (S. cerevisiae) were reduced. There was significant difference in physicochemical characteristics between the reference(composite barley dough containing improvement agent) and the barley sourdoughs (p <0.05). Barley sourdough fermented by L. plantarum showed more desirable farinogram properties of peak time, stability and elasticity than that of the reference. The rheofermentometer data for L. brevis produced the most $CO_2$ release curve, whereas L. plantarum held maximum $CO_2$ retention differed significantly from that of the breads made with barley sourdoughs fermented with the respectives starter cultures (p < 0.05). Barley sourdough bread fermented with L. plantarum resulted in better bread quality than the reference bread. The positive effect of fermentation with L. plantarum on bread quality was evident when comparing the well developed protein-starch matrix structure of the bread baked with barley sourdough with the reference bread.

Nutrient Intake, Its Utilization, Rumen Fermentation Pattern and Blood Bio-Chemical Constituents of Sheep Fed Urea Treated Mustard (Brassica campestris) Straw

  • Misra, A.K.;Karim, S.A.;Verma, D.L.;Mishra, A.S.;Tripathi, M.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.12
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    • pp.1674-1680
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    • 2000
  • A study was conducted to compare the feeding value of urea treated and untreated mustard straw (MS) for sheep. Treated MS was prepared by adding urea-N at 1.84% and followed by packing in a pit silo for 21 days. Two groups of six empty Avikaline ewes were fed untreated (UTMS) and treated (TMS) mustard straw along with 200 g concentrate per head daily for 90 days. Untreated MS had 0.41% N and the urea treatment increased its N value to 1.58 %. The cell wall constituents were decreased in the TMS except for cellulose which remained unaffected. Dry matter intake of TMS was consistently higher than that of UTMS. Digestibility of DM, OM and fibre fractions of MS improved by the urea treatment. Ewes in both groups were in positive N balance while % N retention was lower in UTMS (26.30%) than in TMS (52.14%). The TMS fed group on average consumed 30.2 g DM, 2.9 g digestible crude protein and $0.2MJ\;DE\;per\;kg\;BW\;day^{-1}$ and maintained their weight whereas, the UTMS fed ewes lost weight. The VFA concentration in rumen liquor was higher in TMS than in UTMS. Total-N, ammonia-N and TCA-precipitable-N were also higher in TMS fed ewes. Blood glucose concentrations in the two groups were similar at initiation of the study. However the glucose concentration of UTMS fed group was significantly (p<0.01) lower than those fed UTMS at the termination of the study. Urea-N concentration was also higher in TMS fed group after 90 days of feeding period. It is concluded that urea treatment of MS improved N value of MS from 0.41% to 1.58% along with sizable improvement in nutritive value and in conjunction with 200 g concentrate, TMS can serve as maintenance ration for sheep. ($ME_{lakt}/ME_{m}=1.46$).

Structural and Electrochemical Properties of Li2Mn0.5Fe0.5SiO4/C Cathode Nanocomposite

  • Chung, Young-Min;Yu, Seung-Ho;Song, Min-Seob;Kim, Sung-Soo;Cho, Won-Il
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4205-4209
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    • 2011
  • The $Li_2Mn_{0.5}Fe_{0.5}SiO_4$ silicate was prepared by blending of $Li_2MnSiO_4$ and $Li_2FeSiO_4$ precursors with same molar ratio. The one of the silicates of $Li_2FeSiO_4$ is known as high capacitive up to ~330 mAh/g due to 2 mole electron exchange, and the other of $Li_2FeSiO_4$ has identical structure with $Li_2MnSiO_4$ and shows stable cycle with less capacity of ~170 mAh/g. The major drawback of silicate family is low electronic conductivity (3 orders of magnitude lower than $LiFePO_4$). To overcome this disadvantage, carbon composite of the silicate compound was prepared by sucrose mixing with silicate precursors and heat-treated in reducing atmosphere. The crystal structure and physical morphology of $Li_2Mn_{0.5}Fe_{0.5}SiO_4$ was investigated by X-ray diffraction, scanning electron microscopy, and high resolution transmission electron microscopy. The $Li_2Mn_{0.5}Fe_{0.5}SiO_4$/C nanocomposite has a maximum discharge capacity of 200 mAh/g, and 63% of its discharge capacity is retained after the tenth cycles. We have realized that more than 1 mole of electrons are exchanged in $Li_2Mn_{0.5}Fe_{0.5}SiO_4$. We have observed that $Li_2Mn_{0.5}Fe_{0.5}SiO_4$ is unstable structure upon first delithiation with structural collapse. High temperature cell performance result shows high capacity of discharge capacity (244 mAh/g) but it had poor capacity retention (50%) due to the accelerated structural degradation and related reaction.