• Clinical and Experimental Pediatrics
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• v.50 no.10
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• pp.1011-1017
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• 2007

Purpose : We tried to assess the optimal conditions to improve low transduction efficiency and their effect on target cells. Methods : Cultured NIH 3T3 cells were incubated with retroviral vectors bearing an enhanced green fluorescent protein (eGFP) gene. We varied the ratio of viral vectors to target cells (1:1-1:8) and the number of transfections (${\times}1$, ${\times}2$), and compared transduction efficiencies. Also, the effects of polybrene on transduction efficiency and viability of target cells were assessed. Transduction of the eGFP gene was evaluated by observing NIH 3T3 cells under a fluorescence microscope and efficiencies were measured by the percentage of eGFP positive cells using FACscan. Results : As the ratio of retroviral vectors to target cells increased, transduction efficiency was greatly improved, from 7% (1:1) to 38% (1:4). However, transduction efficiency did not increase any more when the ratio increased from 1:4 to 1:8. Cells transfected twice showed higher transduction efficiencies than cells transfected once, at a ratio of 1:8. The eGFP gene transduced to NIH 3T3 cells sustained its expression during repeated passages. However, after the third passage (day 9), the percentage of eGFP positive cells began to decline. The degree of this decline in eGFP expression was lower in cells transfected twice than in cells transfected once (P<0.05). The addition of polybrene did not have any toxic effect on NIH 3T3 cells and greatly increased transduction efficiency (P=0.007). In addition to vector component, transduction efficiency was very sensitive to culture confluence. Cells cultured and transfected in 24-well plate showed higher transduction efficiency, although cells cultured in 6- well plate proliferated more (P=0.024). Conclusion : Our data could be used as a basis for retrovirus-based gene therapy. Further study will follow using human cells as target cells.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.17-25
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• 2013

The thermotolerant methylotrophic yeast Hansenula polymorpha is an attractive model organism for various fundamental studies, such as the genetic control of enzymes involved in methanol metabolism, peroxisome biogenesis, nitrate assimilation, and resistance to heavy metals and oxidative stresses. In addition, H. polymorpha has been highlighted as a promising recombinant protein expression host, especially due to the availability of strong and tightly regulatable promoters. In this study, we investigated the possibility of employing human serum albumin (HSA) as the fusion tag for the secretory expression of heterologous proteins in H. polymorpha. A set of four expression cassettes, which contained the methanol oxidase (MOX) promoter, translational HSA fusion tag, and the terminator of MOX, were constructed. The expression cassettes were also designed to contain sequences for accessory elements including His8-tag, $2{\times}(Gly_4Ser_1)$ linkers, tobacco etch virus protease recognition sites (Tev), multi-cloning sites, and strep-tags. To determine the effects of the size of the HSA fusion tag on the secretory expression of the target protein, each cassette contained the HSA gene fragment truncated at a specific position based on its domain structure. By using the Green fluorescence protein gene as the reporter, the properties of each expression cassette were compared in various conditions. Our results suggest that the translational HSA fusion tag is an efficient tool for the secretory expression of recombinant proteins in H. polymorpha.

• Journal of Animal Science and Technology
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• v.48 no.3
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• pp.453-466
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• 2006

To comply with stricter regulations provoked by increasing odor nuisance, it is imperative to practice effective odor control for sustainable livestock production. This study was conducted to assess odor and odorous compounds emitted from liquid animal manure with different treatment methods such as Fresh Manure(without treatment, FM), Anaerobic Digestion(AD) and Thermophilic Aerobic Digestion(TAD) and their application to soil. Air samples were collected at the headspace of liquid manure, upland and paddy soil, and analyzed for odor intensity and offensiveness using an olfactometry; odor concentration index using odor analyser; nitrogen-containing compound such as ammonia(NH3) using fluorescence method; and sulfur containing compounds such as hydrogen sulfide(H2S), methyl mercaptan(MeSH), dimethyl sulfide(DMS) and dimethyl disulfide(DMDS) using gas chromatography-pulsed flame photometric detector, respectively. Odor intensity, offensiveness and concentration index from TAD liquid manure was statistically lower than those from FM and AD(p<0.01). Mean concentrations of H2S, MeSH, DMS, DMDS and NH3 were 65.93ppb, 18.55ppb, 5.26ppb, 0.33ppb and 10.57ppm for liquid manure with AD; and 5.15ppb, 0.97ppb, 0.80ppb, 0.56ppb and 1.34ppm for liquid manure with TAD, respectively. More than 60% of malodorous compounds related to nitrogen and sulfur were removed by heterotrophic microorganisms during TAD treatment. When liquid manure was applied onto upland and paddy soil, NH3 removal efficiencies ranged from 51 to 94% and 22 to 91% for AD and TAD liquid manure, respectively. The above results show that liquid manure with TAD is superior to AD and FM with respect to the odor reduction and odor problem caused by land applied liquid manure is directly related to the degree of odor generated by the manure treatment method.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.79-89
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• 2001

Steroid hormones control the expression of many cellular regulators, and a role thor estrogen in mouse oocytes has been well documented. The preovulatory $E_2$increment is generally accepted as the endocrine process regulating induction of in vivo oocyte maturation To address whether the activity of the T-type $Ca^{2+}$ channel is altered by 17 beta-estradiol ( $E_2$), we examined the actions of $E_2$on the calcium channel of mouse oocytes and early embryos. Oocrtes were collected from the oviduct of mice treated with pregnant mare's serum gonadotropin (PMSG) and human choronic gonadotropin (hCG). Whole cell voltage clamp technique and confocal microscopy were used to examine that $E_2$increase intracellular $Ca^{2+}$ concentration ([C $a^{2+}$]$_{i}$ ) via voltage dependent $Ca^{2+}$ channel (VDC) and estrogen receptor (FSR), and $E_2$concentration by the use of radioimmunoassay (RIA) were examined in mouse. The results obtained were as follows: The peak of $Ca^{2+}$ current induced by $E_2$increased 122% to 1.50$\pm$0.03 nA from 1.23$\pm$0.21 nA (n=15) in the presence of 5 mM extracellular $Ca^{2+}$ concentration ([C $a^{2+}$]$_{o}$ ). The increased $Ca^{2+}$ current was temporally associated with $Ca^{2+}$ transients. The intracellular $Ca^{2+}$ level increased 207%~30 s following the addition of 1${\mu}{\textrm}{m}$ $E_2$(relative fluorescence intensity: 836.4$\pm$131.2 for control, n=10, 1736.4$\pm$192.0 in the presence of $E_2$, n=10). $E_2$increased amplitude of $Ca^{2+}$ current and [C $a^{2+}$]$_{i}$ . $E_2$-induced $Ca^{2+}$ current and $E_2$concentration in blood were showed difference on the stage of embryo. These results suggest that $E_2$modulate $Ca^{2+}$ channel to increase $Ca^{2+}$ influx.$Ca^{2+}$ influx.

• Korean Journal of Veterinary Research
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• v.22 no.2
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• pp.175-185
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• 1982

Since bovine lymphosarcoma causes considerable economic loss to the dairy industry, seroepidemiological survey on bovine leucosis virus (BLV) was carried out for the dairy herds throughout the country to observe the epidemiological situation of the disease by using immunodiffusion test. Attempts were simultaneously made to detect bovine leucosis virus in the lymphocytes from BLV antibody-positive cattle by means of fluorescent antibody techniques, syncytium assay and electron microscopy. In immunodiffusion test for BLV antibody in 2003 heads of dairy cattle selected randomly from 164 herds, the prevalence of positive reactors by regions were 37.8% in Central, 27.2% in Honam (Southwest), 28.0% in Youngnam (Southeast) and 25.2% in Youngdong (East coast)and averaging 29.7%. By provinces, Chungcheong appeared the highest prevalence of BLV antibody carriers (41.8%), while Jeonbug revealed the lowest incidence rate (24.4%). When the results of serological studies were analyzed by age groups and the sizes of herds, the number of reactors increased gradually with the advance in the age of cattle and the herd size. The highest rate of BLV carriers was found in the ages between 6 and 8 years, and in the size of herds with 20 to 50 heads. One hundred and seventeen breeding bulls from the central regions were tested for BLV antibody. Four out of 70 bulls (5.7%) of Korean cattle and 14 out of 39 bulls (35.9%) of Holstein were reactive for BLV antigens. Of 164 dairy herds examined, 17 herds (10.4%) have no BLV antibody-positive cattle, while 42 herds (25.6%) were included in the range of 20 to 40% of the positive rate and 10 herds (6.1%) in the range of over 80% of the rate. When the lymphocytes from the BLV antibody carrying cattle were cultured in the presence of phytohemagglutinin and stained with FITC-conjugated sheep anti-BLV serum, 8 out of 11 cases (72.7%) of BLV positive cattle revealed specific fluorescence for BLV in the lymphocytes. In syncytium assay of the peripheral lymphocytes of the cattle, 5 out of 7 (71.4%) lymphocytes from BLV antibody carriers induced syncytia in the indicators of bovine embryonic splenic cells. The cultured lymphocytes were examined with an electron microscope to detect the BLV particles. Two out of 6 specimens (33.3%) from the reactors showed the typical type C virus with the size of 90 to 110 nm around microvilli and in intracytoplasmic vacuoles.

• Journal of Pharmaceutical Investigation
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• v.31 no.4
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• pp.289-295
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• 2001

Carvedilol is an antihypertensive and antianginal compound that combines nonselective beta-adrenoceptor blocking and vasodilation properties and is devoid of intrinsic sympathomimetic activity. The purpose of the present study was to evaluate the bioequivalence of two carvedilol tablets, $Dilatrend^{TM}$ (Chong Kun Dang Pharmaceutical Co., Ltd.) and $Carvelol^{TM}$ (Dae Won Pharmaceutical Co., Ltd.), according to the prior and revised guidelines of Korea Food and Drug Administration (KFDA). The carvedilol release from the two carvedilol tablets in vitro was tested using KP VII Apparatus II method with various different kinds of dissolution media (pH 1.2, 4.0, 6.8 buffer solution, water and blend of PSB80 into water). Eighteen normal male volunteers, $24.22{\pm}1.86$ years in age and $64.81{\pm}4.56\;kg$ in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After one tablet containing 25 mg of carvedilol was orally administered, blood was taken at predetermined time intervals and the concentrations of carvedilol in serum were determined using HPLC method with fluorescence detector. The dissolution profiles of two carvedilol tablets were very similar at all dissolution media. Besides, the pharmacokinetic parameters such as $AUC_t$, $C_{max}$ and $T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using non-transformed and logarithmically transformed $AUC_t$ and $C_{max}$. The results showed that the differences in $AUC_t$, $C_{max}$ and $T_{max}$ between two tablets based on the $Dilatrend^{TM}$ were 2.23%, -2.00% and 0.00%, respectively. Minimum detectable differences $({\Delta})$ at ${\alpha}=0.05$ and $1-{\beta}=0.8$ were less than 20% (e.g., 13.55% and 17.61% for $AUC_t$ and $C_{max}$, respectively). The powers $(l-{\beta})$ at ${\alpha}=0.05$, ${\Delta}=0.2$ for $AUC_t$ and $C_{max}$ were 98.08% and 88.81%, respectively. The 90% confidence intervals were within ${\pm}20%$ (e.g., $-5.69{\sim}10.16$ and $-12.30{\sim}8.30$ for $AUC_t$ and $C_{max}$, respectively). There were no sequence effect between two tablets in logarithmically transformed $AUC_t$ and $C_{max}$. The 90% confidence intervals using logarithmically transformed were within the acceptance range of log(0.8) to log(1.25) (e.g., $0.95{\sim}1.11$ and $0.89{\sim}1.09$ for $AUC_t$ and $C_{max}$, respectively). Two parameters met the criteria of prior and revised KFDA guideline for bioequivalence, indicating that $Carvelol^{TM}$ tablet is bioequivalent to $Dilatrend^{TM}$ tablet.

• Journal of Conservation Science
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• v.26 no.2
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• pp.171-182
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• 2010

The slag excavated from Gyesil-ri in Gongju, Yeonje-ri in Cheongwon and Beopcheonsaji (temple) site in Wonju are analyzed by X-ray Fluorescence Analyzer, metallurgical microscope, SEM-EDS etc., for chemical composition and microstructure to figure out the raw material and the iron manufacturing technique. First of all, as a result of principal component analysis, the total Fe-content of slag from Gyesil-ri is 39 to 44% and the modified rate is 15 to 21%, which is common in ancient iron slag. Yeonje-ri site is found the ancient iron-smelting furnace. The total Fe-content of slag from Yeonje-ri is 41 to 43% and modified rate is 18~30%, which is also the general value in the ancient slag. However only slag is excavated in the residential area at Beopcheonsaji site and there is no iron making relic. In addition, the result of principal component analysis contains that the total Fe-content of Beopcheonsaji site is 52 to 57%, and modified rate is 8 to 14%. It shows that the total Fe-content of Beopcheonsaji site is higher than relic from Gyesil-ri and Yeonje-ri and the modified rate is lower than other sites. This results mean that recollecting rate of Fe in Beopcheonsaji site is lower than other sites. Also, as a result of minor elements analysis, the slag from Gyesil-ri has the higher level of Ti, V and Zr than other sites and the microstructure are observed as magnetite and ulvospinel, so that the raw material of slag is iron sand. But the slag from Yeonje-ri and Beopcheonsaji site are identified to use iron ore. As a result of microstructure observation, fayalite, gray-columnar crystal, is found in the slag from Yeonje-ri and big wustite as main phase is observed in the slag from Beopcheonsaji site. This study show that the slag from Yeonje-ri is made of smelt ash produced during smelting works and the slag from Beopcheonsaji site is made of forging ash produced during forging work concerning the excavated location and the microstructure.

• Journal of Conservation Science
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• v.34 no.5
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• pp.333-343
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• 2018

This study was conducted to evaluate the conservation environment by monitoring temperature and humidity for two years and mapping the remaining pigments of mural paintings to diagnose the conservation state of mural paintings of Royal Tombs in Neungsan-ri. We evaluated the characteristics of condensation in the tomb. Compared with the results of a 2008 survey, we conducted state change of mural paintings in the tomb. The temperature in the main room, which has an annual average soil temperature distribution at 5 m depth in Korea, is maintained at $13{\sim}18^{\circ}C$. The temperature range of the main room was between less than $0.1^{\circ}C$ to $0.5^{\circ}C$, and the diurnal variation of temperature between summer (June to September) and winter (December to January) is the greatest. Condensation is more concentrated in the summer because the outdoor air was typically at higher temperatures than the main room inflows in the tomb. Mapping the remaining pigment composition and particle distribution of mural paintings showed that it was in the range of 36.72~39.53% of the wall area. The pigment range was confirmed to be the same as it was in 2008, through ultraviolet fluorescence reaction and infrared ray investigation. Therefore, the underground environment that receives dew condensation in the summer has been stable since 2008. However, continuous monitoring is needed because the deterioration of mural painting proceeds considerably after excavation and only a small percentage of the pigments survive.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.63 no.2
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• pp.140-148
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• 2018

As the global warming causing desertification increase, there is growing concern about damage of crops. It was to investigate how the treatment with hydrogen peroxide before leaf development affects the growth and yield of sorghum for minimizing a damage of crops to drought. The germination experiment was conducted at alternating temperature of $25^{\circ}C/20^{\circ}C$(12 hr/12 hr) under water stress condition of 0 ~ -0.20 MPa adjusted with PEG solution containing 0 and 10 mM $H_2O_2$. In order to know the effect of foliar application of hydrogen peroxide on the growth of sorghum, 10 mM hydrogen peroxide was treated to leaves at 3-leaf stage of sorghum growing in greenhouse conditions. Seed germination rate was increased by 20% in hydrogen peroxide treatment as compared to the Control. under water stress conditions (-0.15 ~ -0.20 MPa). The length of seedlings was also on the rise by the hydrogen peroxide treatment. In the greenhouse pot experiment, the morphological characteristics (plant height, stem diameter, leaf length, and leaf number) and physiological characteristics (chlorophyll content, chlorophyll fluorescence (Fv/Fm), stomatal conductance) were higher in the plants treated with hydrogen peroxide under the drought stress condition than those of plants of $H_2O$ treatment. Experiment conducted with the soil moisture gradient system showed that the foliar application of hydrogen peroxide increased photosynthetic ability of sorghum plant with respect to SPAD value and stomatal conductance and rooting capacity (root weight and root length) under drought condition. Generally, hydrogen peroxide treatment in sorghum increased the tolerance to drought stress and maintained better growth due to ameliorating oxidative stress.

• Journal of Food Hygiene and Safety
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• v.32 no.4
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• pp.329-335
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• 2017

Analysis method was presented for the simultaneous determination of nine bisphenol A related compounds such as bisphenol A (BPA), phenol, p-tert-butylphenol, bisphenol A diglycidyl ether (BADGE), $BADGE{\cdot}2H_2O$, $BADGE{\cdot}2HCl$, bisphenol F diglycidyl ether (BFDGE), $BFDGE{\cdot}2H_2O$ and $BFDGE{\cdot}2HCl$ migrated from inner coatings of metal food cans by high performance liquid chromatography (HPLC) with fluorescence detection. The method was validated by examining the linearity of calibration curve, the limit of detection (LOD), the limit of quantification (LOQ), recovery and uncertainty. The migration tests of nine BPA related compounds were carried out with four food simulants; deionized water (DW), 4% acetic acid, 50% ethanol and n-heptane. There was not any compound detected in DW, 4% acetic acid and 50% ethanol at $60^{\circ}C$ for 30 min and n-heptane at $25^{\circ}C$ for 60 min. BPA and phenol were migrated into 4% acetic acid and 50% ethanol at $95^{\circ}C$ for 30 min. The concentrations were ranged from 0 to $10.77{\mu}g/L$ of BPA and from 0 to $2.35{\mu}g/L$ of phenol. Canned foodstuffs mostly have long-term shelf life. We investigated migration of nine BPA related compounds according to the variation in storage periods (0~90 days) and temperatures (4, 25 and $60^{\circ}C$). All compounds were not founded during 90 days at $4^{\circ}C$ and $25^{\circ}C$, respectively. However BPA and $BADGE{\cdot}2H_2O$ were founded in DW and 4% acetic acid at $60^{\circ}C$. The migration levels of BPA and $BADGE{\cdot}2H_2O$ were close to the value of LOQ, respectively and did not change significantly as storage period. It was founded from results that the migration of BPA related compounds from metal food cans was controlled to a safe level.