• Nuclear Medicine and Molecular Imaging
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• v.43 no.5
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• pp.468-477
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• 2009

Purpose: Hydrodynamic-based procedure is a simple and effective gene delivery method to lead a high gene expression in liver tissue. Non-invasive imaging reporter gene system has been used widely with herpes simplex virus type 1 thymidine kinase (HSV1-tk) and its various substrates. In the present study, we investigated to image the expression of HSV1-tk gene with 5-(2-iodovinyD-2'-deoxyuridine (IVDU) in mouse liver by the hydrodynamicbased procedure. Materials and Methods: HSV1-tk or enhanced green fluorescence protein (EGFP) encoded plasmid DNA was transferred into the mouse liver by hydrodynaminc injection. At 24 h post-injection, RT-PCR, biodistribution, fluorescence imaging, nuclear imaging and digital wholebody autoradiography (DWBA) were performed to confirm transferred gene expression. Results: In RT-PCR assay using mRNA from the mouse liver, specific bands of HSV1-tk and EGFP gene were observed in HSV1-tk and EGFP expressing plasmid injected mouse, respectively. Higher uptake of radiolabeled IVDU was exhibited in liver of HSV1-tk gene transferred mouse by biodistribution study. In fluorescence imaging, the liver showed specific fluorescence signal in EGFP gene transferred mouse. Gamma-camera image and DWBA results showed that radiolabeled IVDU was accumulated in the liver of HSV1-tk gene transferred mouse. Conclusion: In this study, hydrodynamic-based procedure was effective in liver-specific gene delivery and it could be quantified with molecular imaging methods. Therefore, co-expression of HSV1-tk reporter gene and target gene by hydrodynamic-based procedure is expected to be a useful method for the evaluation of the target gene expression level with radiolabeled IVDU.

• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.161-167
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• 2007

In this study, we constructed and tested retrovirus vectors designed to express the human thrombopoietin gene under the control of the tetracycline-inducible promoters. To increase the hTPO gene expression at him-on state, WPRE sequence was also introduced into retrovirus vector at downstream region of either the hTPO gene or the sequence encoding reverse tetracycline-controlled transactivator (rtTA). Primary culture cells (PFF, porcine fetal fibroblast; CEF, chicken embryonic fibroblast) infected with the recombinant retrovirus were cultured in the medium supplemented with or without doxycycline for 48hr, and induction efficiency was measured by comparing the hTPO gene expression level using RT-PCR, western blot and ELISA. Higher hPTO expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hTPO (in CEF) or rtTA(in PFF) gene. This resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

• Korean Journal of Environmental Agriculture
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• v.10 no.1
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• pp.1-10
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• 1991

The object of this stduy was to investigate the pollution of Pb in paddy field soil with different distance from roadside and to find out the relationship between the ratio of chemical fractions of total Pb and soil characteristics. Lead from automobiles is exhausted as particulates composed primarily of halide compounds (PbBrCl, $PbBr_2$, $PbCl_2$). The samples of soil were collected directly from the paddy fields with different distance from the roadside of highway and expressway which are located in Kyungpook province. A sequential extraction procedure was used to fractionate Pb in paddy field soil into the disignated forms of water soluable, exchangeable, organically bounded, carbonate, sulfide, and residual Pb. Results obtained are summerized as follows. 1. The content of Pb in paddy field soil was the highest in Chungdo, 30.0 ppm, the lowest in Koryung, 14.8 ppm, and the total average content was 21.9 ppm. The effect of traffic volume was not clear, but a slight difference according to the order of opened year of roads was showed. 2. The effect of distance from roadside was not clear. The content of Pb in paddy field soil with different distance from roadside was 22.2 ppm within l0m, 22.1 ppm in 10∼30m, 22.2 ppm in 30∼50m. and 21.3 ppm beyond 50m. 3. The distribution of Pb fractions in soil showed a wide difference depending on soil properties. The average content of exchangeable. organically bounded, carbonate, sulfide, and residual Pb was 8.6%, 33.6%, 29.8%, 21.5%, and 6.7%, of total Pb in the soil, respectively. 4. The content of organically bounded Pb in soil showed highly positive correlation with organic matter and CEC, while the content of exchangeable Pb was highly negative correlation. 5. With higher soil organic matter and CEC, organically bounded Pb fraction tend to be higher but exchangeable Pb fraction tend to be lower. Other forms of Pb showed no difference with soil organic matter contend and CEC. The distribution of Pb fraction related to CEC showed similiar tendency with that of organic matter content.

• Journal of Food Hygiene and Safety
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• v.32 no.4
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• pp.290-297
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• 2017

The purpose of this study was developed for the determination of tridemorph in agricultural commodities samples. Tridemorph residues in samples were extracted with acetonitrile, partitioned with saline water, and then purified using and aminopropyl ($NH_2$) SPE catridge. The purified samples were quantified and confirmed via liquid chromatograph-tandem mass spectrometer (LC-MS/MS) in positive ion mode using multiple reaction monitoring (MRM). Matrix-matched calibration curves were linear over the calibration ranges (0.005~2.5 ng) into a blank extract with $r^2$ > 0.999. The limits of detection and quantification were 0.001 and 0.005 mg/kg, respectively. The average recovery ranged between 75.9% and 103.7% at different concentration levels (LOQ, 10 LOQ, 50 LOQ, n = 5) with relative standard deviations (RSDs) less than 9.0%. An interlaboratory study was conducted to validate the method by Korea Advanced Food Research Institute. The average recovery ranged between 87.0% and 109.2% at different concentration levels (LOQ, $10{\times}LOQ$, $50{\times}LOQ$, n = 5) with relative standard deviations (RSDs) less than 8.0%. All values were consistent with the criteria ranges requested in the Codex guidelines (CAC/GL40, 2003) and Food Safety Evaluation Department guidelines (2016). The results prove that the developed analytical methods is accurate, effective and sensitive for tridemorph determination.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.415-422
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• 2005

library of the mutants was prepared by transposon mutagenesis of the Mycobacterium bovis BCG. We screened this library for the resistance to an anti-tuberculosis antibiotic, PA-824. Most of the mutants resistant to the PA-824 were not able to synthesize the coenzyme $F_{420}$ which is normally produced by the wild type M. bovis BCG strains. HPLC analysis of the cellular extract showed that one of those mutants which lost the ability to synthesize $F_{420}$ still produced F0. The insertion site of the transposon in this mutant was determined by an inverse PCR and the transposon was found to be inserted in the Rv2435c open reading frame (ORF). Rv2435c ORF is predicted to encode an 80.3 kDa protein. Rv2435c protein appears to be bound to the cytoplasmic membrane, its N-terminal present in the periplasm and C-terminal in the cytoplasm. The C-terminal portion of this protein is highly homologous with the adenylyl cyclases of both prokaryotes and eukaryotes. There are 15 ORFs which have homology with the class III AC proteins in the genome of the M. tuberculosis and M. bovis. Two of those, Rv1625c and Rv2435c, are highly homologous with the mammalian ACs. We cloned the cytoplasmic domain of the Rv2435c ORF and expressed it with six histidine residues attached on its C-terminal in Escherichia coli to find out if this protein is a genuine AC. Production of that protein in E. coli was proved by purifying the histidine-tagged protein by using the Ni-NTA resin. This protein, however, failed to complement the cya mutation in E. coli, indicating that this protein lacks the AC activity. All of the further attempts to convert this protein to a functional AC by a mutagenesis with UV or hydroxylamine, or construction of several different fusion proteins with Rv1625c failed. It is, therefore, possible that Rv2435c protein might affect the conversion of F0 to $F_{420}$ not by synthesizing cAMP but by some other way.

• Korean Journal of Environmental Agriculture
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• v.34 no.4
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• pp.318-327
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• 2015

BACKGROUND: Trinexapac-ethyl is a plant growth regulator (PGR) that inhibits the biosynthesis of plant growth hormone (gibberellin). It is used for the prevention of lodging, increasing yields of cereals, and reducing mowing of turf. The experiment was conducted to establish a determination method for trinexapac-ethyl and its metabolites trinexapac in agricultural products using LC-MS/MS.METHODS AND RESULTS: Trinexapac-ethyl and trinexapac were extracted from agricultural products with methanol/ distilled water and the extract was partitioned with dichloromethane and then detected by LC-MS/MS. Limit of detection(LOD) was 0.003 mg/kg and limit of quantification(LOQ) was 0.01 mg/kg, respectively. Matrix matched calibration curves were linear over the calibration ranges (0.01-1.0 mg/L) for all the analytes into blank extract withr²> 0.997. For validation purposes, recovery studies were carried out at three different concentration levels (LOQ, 10LOQ, 50LOQ,n=5). Recoveries of trinexapacethyl and trinexapac were within the range of 73.6-106.9%, 72.7-99.2%, respectively. The relative standard deviations (RSDs) were less than 9.0%. All values were consistent with the criteria ranges requested in the CODEX guideline(CAC/GL 40, 2003).CONCLUSION: The proposed analytical method was accurate, effective and sensitive for trinexapac-ethyl and trinexapac determination and it can be used to as an official method in Korea.

• Korean journal of food and cookery science
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• v.17 no.4
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• pp.344-352
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• 2001

The purpose or this study was to investigate the effect or H $_2$O fraction or Dioscorea japonica Thunb(DJT) and selenium(Se) on the lipid peroxidation in streptozotocin (STZ)-induced diabetic rats. Male Sprague-Dawley rats weighing 180∼220g were divided into 5 groups: One normal rat group and 4 diabetic rat groups(the STZ-Control group, the DJT group, the DJT-Se group and the Se group). Diabetes was induced in the male rats by injection of STZ into tail vein at a dose of 45 mg/kg. The H$_2$O-fraction of DJT(500 mg/kg) was administered orally for 14 days. The supplementation was achieved with the AIN-76 recommendation diet by adding 2 mg/kg diet of selenium as Na$_2$SeO$_3$ which was prepared freshly everyday. The levels of glycogen in liver and muscle and protein in kidney, liver and muscle were measured. The liver concentrations of cholesterol and triglyceride were analyzed. Also, the malondialdehyde(MDA) levels in kidney, liver and lung were determined. The glycogen levels of liver and muscle in diabetic groups were not significantly different from the normal group. The protein concentrations of kidney, liver and muscle were not significantly different either, but the level of muscle protein was higher than STZ-Control group. The levels of liver cholesterol were significantly different between normal and STZ-Control groups and decreased in all diabetic experimental groups fed on H$_2$O-fraction of DJT and Se supplementation compared with the STZ-Control group. The levels of liver triglyceride were higher in the DJT-Se group than the STZ-Control group. The concentrations of MDA in lung decreased greatly by the administration of Se among all and the concentration of liver MDA was significantly reduced and that of DJT-Se group was the lowest. In conclusion, the results indicated that the administration of H$_2$O-fraction of DJT with selenium supplementation has a synergistic antioxidative effect by influencing on lipid metabolites and peroxidation especially in liver.

• Journal of Life Science
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• v.27 no.2
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• pp.155-163
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• 2017

Mori Folium, the leaf of Morus alba, is a traditional medicinal herb that shows various pharmacological activities such as antiinflammatory, antidiabetic, antimelanogenesis, antioxidant, antibacterial, antiallergic, and immunomodulatory activities. However, the mechanisms of their inhibitory effects on adipocyte differentiation and adipogenesis remain poorly understood. In the present study, we investigated the inhibition of adipocyte differentiation and adipogenesis by ethanol extracts of Mori Folium (EEMF) in 3T3-L1 preadipocytes. Treatment with EEMF suppressed the terminal differentiation of 3T3-L1 preadipocytes in a dose-dependent manner, as confirmed by a decrease in the lipid droplet number and lipid content through Oil Red O staining. EEMF significantly reduced the accumulation of cellular triglyceride, which is associated with a significant inhibition of pro-adipogenic transcription factors, including sterol regulatory element-binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor-${\gamma}$ ($PPAR{\gamma}$), and CCAAT/enhancer-binding proteins ${\alpha}$ ($C/EBP{\alpha}$) and ${\beta}$ ($C/EBP{\beta}$). In addition, EEMF potentially downregulated the expression of adipocyte-specific genes, including adipocyte fatty acid binding protein (aP2) and leptin. Furthermore, EEMF treatment effectively increased the phosphorylation of the AMP-activated protein kinase (AMPK) and acetyl CoA carboxylase (ACC); however, treatment with a potent inhibitor of AMPK, compound C, significantly restored the EEMF-induced inhibition of pro-adipogenic transcription factors and adipocyte-specific genes. These results together indicate that EEMF has preeminent effects on the inhibition of adipogenesis through the AMPK signaling pathway, and further studies will be needed to identify the active compounds in Mori Folium.

• Journal of Life Science
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• v.28 no.5
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• pp.605-614
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• 2018

Bee pollen has an outer wall which is resistant to both acidic and basic solutions and even the digestive enzymes in the gastrointestinal tract. Therefore, the oral bioavailability of bee pollen is only 10-15%. A previous study reported on wet-grinding technology which increased the extraction of active ingredients from bee pollen by 11 times. This study was designed to investigate the safety of wet-ground bee pollen. First, a single dose of wet-ground bee pollen was tested in both rats and beagle dogs at dosages of 5, 10, and 20 g/kg and 1.5, 3, and 6 g/kg, respectively. In rats, compound-colored stools were found in those administered 10 g/kg or more of wet-ground bee pollen. In beagle dogs, 6 g/kg of wet-ground bee pollen induced diarrhea in one male for four hours. However, no obvious clinical signs were found through the end of the experiment in rats and beagle dogs. In addition, no histological abnormality was found in all animals. The data indicates that a single dose of up to 20 g/kg of wet-ground bee pollen is safe. Next, the genetic toxicity of nano-sized bee pollen was tested. This study employed a bacterial reverse mutation test, a micronucleus assay, and a chromosomal aberration assay. In the micronucleus assay, there was no genetic toxicity up to the dosage of 2 g/kg. There was also no genetic toxicity in the bacterial reverse mutation test and chromosomal aberration assay. This data provides important information in developing nano-sized bee pollen into more advanced functional foods and herbal medicines.

• Restorative Dentistry and Endodontics
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• v.30 no.3
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• pp.178-183
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• 2005

This study was done to evaluate whether vital bleaching agents could influence on the translucency of the bovine enamel. The anterior bovine teeth that were extracted one day before and without any gross discoloration were obtained and then were preserved in physiologic saline. 6 mm cylindrical tooth specimens were fabricated with diamond puncher perpendicularly on labial surface of bovine tooth. After embedded in transparent acrylic resin with labial surface being exposed, they were cut to a thickness of 1.2 mm with low speed diamond saw (Isomat, Buehler Co., Lake Bluff, IL, USA). They were smoothly ground to 1 mm thickness of enamel with sandpaper. 24 specimens were randomly divided into 3 groups and control group respectively. Opalescence (10% carbamide peroxide, Ultradent, South Jordan, USA), Rembrandt (10% carbamide peroxide, DenMat, USA) and Opalescence F (15% carbamide peroxide with fluoride, Ultradent, USA) were applied on labial sides of the bovine enamel for 7 days (bleaching agents were reapplied every 24 hours) and the opposite surface was contacted to cotton that soaked in distilled water. The control group was soaked in distilled water. Three stimulus value X, Y and Z were evaluated with colorimeter (Color ' Color Differencerneter, Model TC-6FX, Tokyo Denshoku Co., Japan) on the labial surface of all specimen three times on white and black background plate before the bleaching agents were applied and on 3rd, 5th and 7th day after applied. The degree of translucency was normally assessed by measuring the inverse property, opacity (contrast ratio). 10% Opalescence, 15% Opalescence-F, and control group showed no significant variation in the translucency of bovine enamel, However Rembrandt decreased the translucency of it (p < 0.01).