• Title/Summary/Keyword: Ta-C

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Effects of Mutagenesis for Glycosylation Sites of Recombinant Human EPO During Production from Cultured CHO Cell

  • Lee, Hyun-Gi;Seong, Hwan-Hoo;Im, Seok-Ki;Chung, Hee-Kyoung;Lee, Poongyeon;Lee, Yeun-Kun;Min, Kwan-Sik;Chang, Won-Kyoung;Lee, Hoon-Taek
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.97-97
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    • 2002
  • Human eryhropoietin (EPO) is acidic glycoprotein hormone that plays key role in hematopoiesis by facilitating differentiation of erythrocyte and formation of hemoglobin (Hb) and is used for the treatment of anemia. Human EPO is consist of 166 amino acids which is modified by three N-glycosylations (24, 38, 83) and single O-glycosylation (126). N-glycosylation is reported to be related to the cellular secretion and activity of EPO. In this study, we examined effects of mutagenesis in glycosylation site of recombinat hEPO for the cellular secretion during production from cultured CHO cell. We produced rhEpo which was cloned by PCR from human liver cDNA (TaKaRa) in cultured CHO cell. Using supernatant of the culture, ELISA assay and western analysis were performed. To estimate biological activity, 20IU of rhuEpo was subcutaneously injected into four ICR mice. After 8 days, HCT level was increased average 13 per cent, RBC was increased ca. 2${\times}$10$\^$6//${\mu}\ell$. In disease model Rat (anemia c-kit, WSRC-WS/WS), HCT was increased ca. 12%, RBC was increased ca. 1.6${\times}$10$\^$6//${\mu}\ell$. These results suggests that rhEpo we produced has biological activity. To remove glycosylation site by substituting 24, 38, 83, and 126th asparagine (or serine) with glutamic acid, overlapping -extension site-directed mutagenesis was performed. To add novel glycosylation sites, 69, 105th leucine was mutated to asparagine. Mutant EPO construct was transfected into CHO cell. Supernatant of the cell culture was analyzed using ELISA assay with monoclonal anti-EPO antibody (Medac, Germany). Since, several reports for mutagenesis of glycosylation sites showed case-by-case results, we examined both transient expression and stable expression. Addition of novel glycosylation sites resulted no secretion while deletion mutants had little effect except some double deletion mutants (24/83 and 38/83) and triple mutant. We suggest that not single but combination of glycosyl group affect secretion of EPO.

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Effects of Vitamin C on Oxidative Stress Due to Anesthesia and Surgical Trauma in Dogs (개 마취와 수술 창상에 따른 산화스트레스에 대한 비타민 C의 효과)

  • Choi, Kyeong-Ha;Lee, Jae-Yeon;Jeong, Seong-Mok;Joo, Gregory K.;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.28 no.5
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    • pp.473-478
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    • 2011
  • The study evaluated the antioxidant effects of ascorbic acid (AA) against oxidative stress during laparotomy in dogs under general anesthesia using isoflurane. Twelve dogs were randomly assigned to either the ascorbic acid group (AAG) or the sham group (SG). AAG received intravenous dosage of 100 mg/animal of standard AA 10 minutes before anesthesia. Plasma levels of cortisol, glucose, total oxidant status (TOS), total antioxidant status (TAS), and oxidative stress index (OSI) were measured. Cortisol level increased significantly with time in both groups (p < 0.05). The change of glucose levels was not significantly different between both groups. TOS and OSI increased significantly with time in the SG (p < 0.05), whereas it did not significantly change with time in the AAG. The TOS and OSI of the AAG were significantly lower than in the SG (p < 0.05). However, TAS did not significantly change between both groups. The results suggest that the decrease of TOS in the AAG might be related to the conversion of antioxidants to oxidants. The decrease of OSI indicates that the decrease of reactive oxygen species (i.e., oxidative stress) produced at the site of injury is able to decrease surgical side effects of abnormal blood circulation, organ failure, and inflammation. Therefore, AA can be used to protect a surgical patient from oxidative stress in canine laparotomy.

Improvement of Shelf-life and Quality in Fresh-Cut Tomato Slices:

  • Hong Ji Heun
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2004.10a
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    • pp.67-72
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    • 2004
  • Quality of fresh-cut tomato slices was compared during cold storage under various modified atmosphere packaging conditions. Chilling injury of slices in containers sealed with Film A was higher than with Film B; these films had oxygen transmission rates of 87.4 and 60.0 ml $h^{-1}\;m^{-2}\;atm^{-1}$ at $5^{\circ}C\;and\;99\%$ RH, respectively. While slices in containers with an initial atmospheric composition of air, $4\%\;CO_2+1\;or\;20\%\;O_2,\;8\%\;CO_2+1\;or\;20\%\;O_2,\;or\;12\%\;CO_2+20\%\;O_2$ showed fungal growth, slices in containers with $12\%\;CO_2+1\%\;O_2$ did not. Low ethylene in containers enhanced chilling injury. Modified atmosphere packaging provided good quality tomato slices with a shelf-life of 2 weeks or more at $5^{\circ}C$. Experiments were conducted to compare changes in quality of slices of red tomato (Lycopersicon esculentum Mill. 'Sunbeam') fruit from plants grown using black polyethylene or hairy vetch mulches under various foliar disease management systems including: no fungicide applications (NF), a disease forecasting model (Tom-Cast), and weekly fungicide applications (WF), during storage at $5^{\circ}C$ under a modified atmosphere. Slices were analyzed for firmness, soluble solids content (SSC), titratable acidity (TA), pH, electrolyte leakage, fungi, yeasts, and chilling injury. With both NF and Tom-Cast fungicide treatments, slices from tomato fruit grown with hairy vetch (Vicia villosa Roth) mulch were firmer than those from tomato fruit grown with black polyethylene mulch after 12 days storage. Ethylene production of slices from fruit grown using hairy vetch mulch under Tom-Cast was about 1.5- and 5-fold higher than that of slices from WF and NF fungicide treatments after 12 days, respectively. The percentage of water-soaked areas (chilling injury) for slices from tomato fruit grown using black polyethylene mulch under NF was over 7-fold that of slices from tomato fruit grown using hairy vetch under Tom-Cast. When stored at $20^{\circ}C$, slices from light-red tomato fruit grown with black polyethylene or hairy vetch mulches both showed a rapid increase in electrolyte leakage beginning 6 hours after slicing. However, slices from tomato fruit grown using the hairy vetch mulch tended to have lower electrolyte leakage than those grown with black polyethylene mulch. These results suggest that tomato fruit from plants grown using hairy vetch mulch may be more suitable for fresh-cut slices than those grown using black polyethylene mulch. Also, use of the disease forecasting model Tom-Cast, which can result in lower fungicide application than is currently used commercially, resulted in high quality fruit for fresh-cut processing. Experiments were conducted to determine if ethylene influences chilling injury, as measured by percentage of slices exhibiting water-soaked areas in fresh-cut tomato slices of 'Mountain Pride' and 'Sunbeam' tomato (Lycopersicon esculentum Mill.). Ethylene concentration in containers without ventilation significantly increased during storage at $5^{\circ}C$, whereas little or no accumulation of ethylene occurred in containers with one or six perforations. Chilling injury was greatest for slices in containers with six perforations, compared to slices in containers with one perforation, and was over 13-fold greater than that of slices in control containers with no perforations. An experiment was also performed to investigate the effectiveness of including an ethylene absorbent pad in containers on subsequent ethylene accumulation and chilling injury. While ethylene in the no-pad controls increased continually during storage of both 'Mountain Pride' and 'Sunbeam' tomatoes at $5^{\circ}C$ under modified atmosphere conditions, no increase in accumulation of ethylene was observed in containers containing ethylene absorbent pads throughout storage. The ethylene absorbent pad treatment resulted in a significantly higher percentage of chilling injury compared with the no-pad control. In studies aimed at inhibiting ethylene production using AVG during storage of slices, the concentration of ethylene in control containers (no AVG) remained at elevated levels throughout storage, compared to containers with slices treated with AVG. Chilling injury in slices treated with AVG was 5-fold greater than that of controls. Further, we tested the effect of ethylene pretreatment of slices on subsequent slice shelf-life and quality. In slices treated with ethylene (0, 0.1, 1, or $10\;{mu}L\;L^{-1}$) immediately after slicing, ethylene production in non-treated controls was greater than that of all other ethylene pre-treatments. However, pretreatment of slices 3 days after slicing resulted in a different pattern of ethylene production during storage. Ihe rate of ethylene production by slices treated with 1 L $L^{-1}$ ethylene 3 days after slicing was greater during storage than any of the other ethylene treatments. With slices pre-treated with ethylene, both immediately and 3 days after slicing, the rate of ethylene production tended to show an negative correlation with chilling injury. Chemical name used: 1-aminoethoxyvinylglycine (AVG).

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Spatial Distribution of Transparent Exopolymer Particles(TEP) and Their Relation to Carbon Species in the Euphotic Layer of the Northern East Sea (동해 북부해역 유광층에서 TEP 분포와 이산화탄소 인자와의 상호관련성)

  • Jeon, Hyun-Duck;Rho, Tae-Keun;Lee, Tong-Sup
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.17 no.2
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    • pp.33-44
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    • 2012
  • Transparent exopolymer particles (TEP) are formed by aggregation of polysaccharide products excreted by phytoplankton and have sticky character like gel. They play important role in the production of marine snow in water column. To study the distribution pattern of TEP concentration and its role in carbon cycle in the surface ocean, we measured pH, Total alkalinity (TA), and chlorophyll-a in addition to physical characteristics of seawater within the surface water column. TEP concentrations ranged from nearly undetectable values to $338{\mu}g\;Xeq\;l^{-1}$. They were considerably lower than previously reported values from costal sites, but showed similar values observed in other oceanic region during phytoplankton bloom periods. The spatial distribution of TEP concentrations were similar to those of chlorophyll-a, which indicate that the production of TEP were closely related to phytoplankton. Calculated total dissolved inorganic carbon ($TCO_2$) from the pH and TA was normalized to 35 psu of salinity ($NTCO_2$) and showed negative linear relationship with temperature. Biological drawdown of $NTCO_2$ ($NTCO_{2bio}$) was estimated from the difference between theoretical $NTCO_2$ values and observed $NTCO_2$. In the warm region located south of $40^{\circ}N$ along the $132.5^{\circ}N$ meridional lines, $NTCO_{2bio}$ showed negative value and TEP concentrations were high. This suggested that negative $NTCO_{2bio}$ may be attributed to the biological processes. At the stations located between 44 and $46^{\circ}N$, TEP concentrations showed high concentration at the chlorophyll-a maximum layer within the water column while they showed low concentration in the surface layer. Carbon content of TEP constituted about 40% of $NTCO_{2bio}$ at the chlorophylla maximum layer. In this study, we could not observe any positive and negative relationship between TEP concentration and $NTCO_2$ or pH. It is obvious that we should consider the importance of TEP in the biological carbon cycling processes within surface layer.

Errors in Net Ecosystem Exchanges of CO2, Water Vapor, and Heat Caused by Storage Fluxes Calculated by Single-level Scalar Measurements Over a Rice Paddy (단일 높이에서 관측된 저장 플럭스를 사용할 때 발생하는 논의 이산화탄소, 수증기, 현열의 순생태계교환량 오차)

  • Moon, Minkyu;Kang, Minseok;Thakuri, Bindu Malla;Lee, Jung-Hoon
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.17 no.3
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    • pp.227-235
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    • 2015
  • Using eddy covariance method, net ecosystem exchange (NEE) of $CO_2$ ($F_{CO_2}$), $H_2O$ (LE), and sensible heat (H) can be approximated as the sum of eddy flux ($F_c$) and storage flux term ($F_s$). Depending on strength and distribution of sink/source of scalars and magnitude of vertical turbulence mixing, the rates of changes in scalars are different with height. In order to calculate $F_s$ accurately, the differences should be considered using scalar profile measurement. However, most of flux sites for agricultural lands in Asia do not operate profile system and estimate $F_s$ using single-level scalars from eddy covariance system under the assumption that the rates of changes in scalars are constant regardless of the height. In this study, we measured $F_c$ and $F_s$ of $CO_2$, $H_2O$, and air temperature ($T_a$) using eddy covariance and profile system (i.e., the multi-level measurement system in scalars from eddy covariance measurement height to the land surface) at the Chengmicheon farmland site in Korea (CFK) in order to quantify the differences between $F_s$ calculated by single-level measurements ($F_s_{-single}$ i.e., $F_s$ from scalars measured by profile system only at eddy covariance system measurement height) and $F_s$ calculated by profile measurements and verify the errors of NEE caused by $F_s_{-single}$. The rate of change in $CO_2$, $H_2O$, and Ta were varied with height depending on the magnitudes and distribution of sink and source and the stability in the atmospheric boundary layer. Thus, $F_s_{-single}$ underestimated or overestimated $F_s$ (especially 21% underestimation in $F_s$ of $CO_2$ around sunrise and sunset (0430-0800 h and 1630-2000 h)). For $F_{CO_2}$, the errors in $F_s_{-single}$ generated 3% and 2% underestimation of $F_{CO_2}$ during nighttime (2030-0400 h) and around sunrise and sunset, respectively. In the process of nighttime correction and partitioning of $F_{CO_2}$, these differences would cause an underestimation in carbon balance at the rice paddy. In contrast, there were little differences at the errors in LE and H caused by the error in $F_s_{-single}$, irrespective of time.

Pathophysiological Regulation of Vascular Smooth Muscle Cells by Prostaglandin F2α-dependent Activation of Phospholipase C-β3 (Prostaglandin F2α 의존적 phospholipase C-β3 활성화에 의한 혈관평활근세포의 병태생리 조절 연구)

  • Kang, Ki Ung;Oh, Jun Young;Lee, Yun Ha;Lee, Hye Sun;Jin, Seo Yeon;Bae, Sun Sik
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1516-1522
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    • 2018
  • Atherosclerosis is an obstructive vessel disease mainly caused by chronic arterial inflammation to which the proliferation and migration of vascular smooth muscle cells (VSMCs) is the main pathological response. In the present study, the primary responsible inflammatory cytokine and its signaling pathway was investigated. The proliferation and migration of VSMCs was significantly enhanced by the prostaglandin $F_{2{\alpha}}$ ($PGF_{2{\alpha}}$), while neither was affected by tumor necrosis factor ${\alpha}$. Prostacyclin $I_2$ was seen to enhance the proliferation of VSMCs while simultaneously suppressing their migration. Both prostaglandin $D_2$ and prostaglandin $E_2$ significantly enhanced the migration of VSMCs, however, proliferation was not affected by either of them. The proliferation and migration of VSMCs stimulated by $PGF_{2{\alpha}}$ progressed in a dose-dependent manner; the $EC_{50}$ value of both proliferation and migration was $0.1{\mu}M$. VSMCs highly expressed the phospholipase isoform $C-{\beta}3$ ($PLC-{\beta}3$) while others such as $PLC-{\beta}1$, $PLC-{\beta}2$, and $PLC-{\beta}4$ were not expressed. Inhibition of the PLCs by U73122 completely blocked the $PGF_{2{\alpha}}$-induced migration of VSMCs, and, in addition, silencing $PLC-{\beta}3$ significantly diminished the $PGF_{2{\alpha}}$-induced proliferation and migration of VSMCs. Given these results, we suggest that $PGF_{2{\alpha}}$ plays a crucial role in the proliferation and migration of VSMCs, and activation of $PLC-{\beta}3$ could be involved in their $PGF_{2{\alpha}}$-dependent migration.

Physiological Effect of Yogurt with Powder Two Stage Fermented Dioscorea batatas Dence by Monascus sp. and Lactobacillus sp. (Monascus sp.와 Lactobacillus sp.를 이용한 2단 발효마 분말 첨가 요구르트의 생리활성 효과)

  • Jeon, Chun-Pyo;Lee, Jung-Bok;Choi, Chung-Sig;Kwon, Gi-Seok
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.151-157
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    • 2011
  • This study was to examine the quality characteristics and biological activities of the yogurt containing dual fermentation MFCY (Monascus-Fermented Chinese Yam) powder. Six different contents (0, 0.1, 0.3, 0.5, 1.0, and 2.0%, w/v) of MFCY powder were added to raw milk and 5% skim milk followed by fermentation with lactic acid bacteria at $37^{\circ}C$ for 12 h, and then the biological activities of the samples were investigated. Biological activities of the prepared yogurt were evaluated for acid production (pH, titratable acidity), number of viable cells, total polyphenol contents, DPPH radical scavenging activity, reducing power, angiotensin converting enzyme inhibitory effects and GABA contents. In this study, the results show that after fermentation in raw milk and 5% skim milk added with MFCY powder. The yogurt can produced pH, titratable acidity, number of viable cells, total polyphenol contents, DPPH radical scavenging activity and reducing power at 4.12-4.25 (pH), 0.94-0.97% (TA), $7.50{\times}10^9-1.14{\times}10^{10}$ CFU/ml, 161.4-329.9 (TP, mg/L), 54-94% (DPPH), and 1.13-1.53 (R.P), respectively. In addition, the showed ACE inhibitory activity and GABA contents was 67.1-87.7% and 304.6-685.4 (mg/L).

Assessment of VOCs Emission Characteristics from Building Materials such as Wall Paper, Paints, and Adhesives Using Small Chamber Method (소형챔버법을 이용한 건축자재 중 벽지, 페인트 및 접착제의 VOCs 방출특성 평가)

  • Lee Suk-Jo;Jang Seong-Ki;Cho Yong-Sung;Jung Kyung- Mi;Jeong Gi-Ho
    • Journal of Korean Society for Atmospheric Environment
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    • v.21 no.2
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    • pp.191-204
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    • 2005
  • Building and furnishing materials and consumer product are important sources of volatile organic compounds(VOCs) and other aldehydes in the indoor environment. Some available evidence indicates that VOCs can cause adverse health effects to the building occupants and contribute to some of the symptoms of what we call, 'Sick House Syndrome' in Korea. The aims of this study were to evaluate the efficiency of emission system and to investigate comparison of the emission characteristics of different building materials such as wall-papers, paints, and adhesives. The emission of VOCs from building materials were determined in the small chambers defining the temperature, relative humidity, and ventilation rate in this study. VOCs were sampled for 20 minutes using Tenax-TA tubes and analysed by GC-MS with thermal desorption. The stability of conditions for temperature and relative humidity in this small chamber system showed that the fluctuation of temperature was between 25.4$\pm$0.3$^{\circ}C$ and that of relative humidity was 50.2$\pm$0.6$\%$ under the airflow rate of 167 mL/min. The emission tests from building materials resulted in TVOC emission rates of 0.011 $\~$ 3.108 mg/m$^{2}$h after 7 days. The general wall-papers emitted toluene abundantly and the natural wall-papers mainly emitted n-butanol and a minor amount of alkanes compound such as n -tetradecane. The remainder consisted of toluene, m,p -xylene, and styrene. The paints mainly emitted toluene and the adhesives mainly emitted chloroform as well as toluene. As a result, this study is expected to suggest meaningful data for future studies in exposure control through selecting healthy building materials and for the establishment of guidelines for various building materials in Korea.

Clinical Patterns of Acute Poststreptococcal Glomerulonephritis: A Single Center's Experience (단일 기관 내의 급성 연쇄구균감염후 사구체신염 발생의 임상적 고찰)

  • Yu, Ri-Ta;Park, Se-Jin;Shin, Jae-Il;Kim, Kee-Hyuck
    • Childhood Kidney Diseases
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    • v.15 no.1
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    • pp.49-57
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    • 2011
  • Purpose: The aim of this study is to investigate the change in incidence and clinical characteristics of acute poststreptococcal glomerulonephritis (APSGN) through a single center's experience. Methods: We retrospectively analyzed the medical records of 53 children who were diagnosed with glomerular nephritis at National Health Insurance Corporation Ilsan Hospital between March 2000 and December 2009. Twenty-six of 53 patients falled to meet the dragnostic criteria of APSGN and were excluded. Chief complaints, physical examinations, urinalysis, urine culture, laboratory results, chest x-ray, and clinical manifestations were all retrieved from the remaining 27 patients. Results: The incidence of APSGN by age was highest in children between six to seven years of age, with similar distributions in both genders, and seasonal incidence was higher from November to February when compared to other months, which was similar to the previously reported studies. However, the annual number of APSGN patients did not show a significant decrease from 2000 to 2009. The incidences of edema and hypertension were 59.3% and 48.1% respectively, suggesting lower incidences than those from previous studies. Six patients (22.2%) experienced hematuria for more than 6 months. Hypertensive encephalopathy, one of APSGN critical complications occurred in one patient but resolved with conservative treatment. Conclusion: Our study shows that APSGN still occur as a common renal disease but the disease severity seems to decrease with milder clinical manifestations and less complications compared to past studies.

Characteristics of the Crystal Structure and Electrical Properties of Metal/Ferroelectric/Insulator/Semiconductor (Metal/Ferroelectric/Insulator/Semiconductor 구조의 결정 구조 및 전기적 특성에 관한 연구)

  • 신동석;최훈상;최인훈;이호녕;김용태
    • Journal of the Korean Vacuum Society
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    • v.7 no.3
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    • pp.195-200
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    • 1998
  • We have investigated the crystal structure and electrical properties of Pt/SBT/$CeO_2$/Si(MFIS) and Pt/SBT/Si(MFS) structures for the gate oxide of ferroelectric memory. XRD spectra and SEM showed that the SBT film of SBT/$CeO_2$/Si structure had larger grain than that of SBT/Si structure. Furthermore HRTEM showed that SBT/$CeO_2$/Si had 5 nm thick $SiO_2$layer and very smooth interface but SBT/Si had 6nm thick $SiO_2$layer and 7nm thick amorphous intermediate interface. Therefore, $CeO_2$film between SBT film and Si substrate is confirmed as a good candidate for a diffusion barrier. The remanent polarization decreased and coercive voltage increased in Pt/SBT/$CeO_2/Pt/SiO_2$/Si structure. This effect may increase memory window of MFIS structure directly related to the coercive voltage. From the capacitance-voltage characteristics, the memory of Pt/SBT(140 nm)/$CeO_2$(25 nm)/Si structure were in the range of 1~2 V at the applied voltage of 4~6 V. The memory window increased with the thickness of SBT film. These results may be due to voltage applied at SBT films. The leakage currents of Pt/SBT/$CeO_2$/Si and Pt/SBT/Si were $ 10^8A/\textrm{cm}^2$ and $ 10^6 A/\textrm{cm}^2$, respectively.

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