• The Korean Journal of Mycology
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• v.38 no.1
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• pp.48-53
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• 2010

Agrobacterium-mediated transformation was conducted in order to generate DNA insertional mutants of Flammulina velutipes. Agrobacterium tumefaciens AGL-1 harboring pBGgHg was transformed into gill tissues of Flammulina velutipes strain KACC42777. The transformants resistant on hygromycine ($30\;{\mu}g/ml$) were confirmed by PCR. The targeted insertional sites were amplified by inverse PCR and sequenced. To find the phenotype variation of all generated transformants, bottle cultivation which followed by the standard cultivation protocol were conducted. Color variation was observed on the cultivated fruiting bodies. Furthermore, the transformant pool will be used as a good genetic resources for studying gene function.

• Toxicological Research
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• v.24 no.1
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• pp.37-44
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• 2008

The in vitro cell transformation assays (CTA) were performed using BALB/3T3 murine fibroblasts and HaCaT human keratinocytes in order to evaluate concordance between both in vitro CTAs and carcinogenicity with compounds differing in their genotoxic and carcinogenic potential. Six test articles were evaluated, two each from three classes of compounds: genotoxic carcinogens (2-amino-5-nitrophenol and 4-nitroquinoline-N-oxide), genotoxic noncarcinogens (8-hydroxyquinoline and benzyl alcohol), and nongenotoxic carcinogens (methyl carbamate and N-nitrosodiphenylamine). Any foci of size $\geq$2 mm regardless of invasiveness and piling was scored as positive in CTA with BALB/3T3. As expected, four carcinogens regardless of their genotoxicity had positive outcomes in two-stage CTA using BALB/3T3 cells. However, of the two genotoxic noncarcinogens, benzyl alcohol was positive CTA finding. We concluded that, of the 6 chemicals tested, the sensitivity for BALB/3T3 system was reasonably high, being 100%. The respective specificity for BALB/3T3 assay was 50%. We also investigated the correlation between results of BALB/3T3 assay and results from HaCaT assay in order to develop a reliable human cell transformation assay. However, evaluation of staining at later time points beyond the confluency stage did not yield further assessable data because most of HaCaT cells were detached after $2{\sim}3$ days of confluency. Thus, after test article treatment, HaCaT cells were split before massive cell death began. In this modified protocol for this HaCaT system, growing attached colonies were counted instead of transformed foci 3 weeks since last subculture. Compared to BALB/3T3 assay, HaCaT assay showed moderate low sensitivity and high specificity. Despite these differences in specificity and sensitivity, both cell systems did exhibit same good concordance between in vitro CTA and rodent carcinogenicity findings (overall 83% concordant results). At present the major weakness of these in vitro CTA is lack of validation for regulatory acceptance and use. Thus, more controlled studies will be needed in order to be better able to assess and quantitatively estimate in vitro CTA data.

• Journal of Plant Biotechnology
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• v.38 no.4
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• pp.251-257
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• 2011

Rice is the most important crop as a model plant for functional genomics of monocotyledons. Rice is usually transformed using Agrobacterium tumefaciens. However, the transformation efficiency using previous method is still low. In this study, we established a new method by modifying the general Agrobacterium protocol especially in the inoculation and co-cultivation step. We directly inoculated Agrobacterium containing a CIPK15 gene under the control of CaMV 35S promoter and NOS terminator in the pCAM1300 vector into the pre-soaked seeds in N6D media for 24 hours. After 7 days of culture at $25^{\circ}C$, calli were formed on seeds cultured on the co-cultivation medium containing an antioxidant compound (1 mM dithiothreitol) and of Agrobacterium growth-inhibiting agent (3 mg/L silver nitrate). We obtained 35 and 22 transgenic plants in rice cultivars, Gopumbyeo and Ilpumbyeo, with increase of transformation efficiency by 30.4% and 22.6%, respectively compared to the general transformation method. The new method in this study would lead to reduction of substantial labor and time to generate transgenic plants.

• Journal of Plant Biotechnology
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• v.40 no.1
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• pp.37-42
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• 2013

Rice is one of the most important cereal crops as a model plant for functional genomics of monocotyledons and usually transformed using Agrobacterium tumefaciens. However, the transformation's process using previous method is still time consuming and uneconomical, low efficiency. In this study, we established a new method by modifying the general Agrobacterium protocol especially in the infection and co-cultivation, Agrobacterium elimination, infected calli's selection steps using liquid media. We directly inoculated Agrobacterium containing a ZjLsL gene under the control of constitutive promoter into the 1- to 3-week-old rice calli derived from mature seeds. After 3 days of co-cultivation, the infected calli were transferred onto liquid media of Agrobacterium elimination and calli's selection for 3 days. The calli were transferred to calli's growth solid media for 14 days and then the calli transferred to shoot induction and root induction media. Putative transformants were initially selected on the medium containing phosphinothricin, and the PAT protein verified by PAT strip test. This method in this study would lead to reduction of substantial labor and time to generate transgenic plants.

• Journal of Pharmaceutical Investigation
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• v.40 no.2
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• pp.125-131
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• 2010

A bioequivalence study of LANIDIEM$^{(R)}$ tablet 4 mg (Samil. Co., Ltd.) to Vaxar$^{(R)}$ tablet 4 mg (GlaxoSmithKline Co., Ltd.) was conducted according to the guidelines of Korea Food and Drug Administration (KFDA). Forty healthy male Korean volunteers were enrolled in the study and thirty six volunteers completed the study according to the protocol. Thirty six volunteers received each medicine at the lacidipine dose of 4 mg in a $2{\times}2$ crossover study. There was one week wash-out period between the doses. Plasma concentrations of lacidipine were monitored by a high performance liquid chromatography - tandem mass spectrometry (LC-MS/MS) for over a period of 24 hours after drug administration. $AUC_t$ (the area under the plasma concentration-time curve from time zero to 24 hr) was calculated by the linear trapezoidal rule method. $C_{max}$ (maximum plasma drug concentration) and $T_{max}$ (time to reach $C_{max}$) were compiled from the plasma concentration-time data. Analysis of variance was carried out using logarithmically transformed $AUC_t$ and $C_{max}$. No significant sequence effect was found for all of the bioavailability parameters indicating that the crossover design was properly performed. The 90% confidence intervals of the $AUC_t$ ratio and the $C_{max}$ ratio for LANIDIEM$^{(R)}$/Vaxar$^{(R)}$ were log 0.8102~log 1.0417 and log 0.8493~log 1.1439, respectively. These values were within the acceptable bioequivalence intervals of log 0.80~log 1.25. Thus, our study demonstrated the bioequivalence of LANIDIEM$^{(R)}$ tablet 4 mg and Vaxar$^{(R)}$ tablet 4 mg with respect to the rate and extent of absorption.

• Clean Technology
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• v.28 no.4
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• pp.293-300
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• 2022

The remarkable mechanical, electrical, and thermal properties of graphene have recently sparked tremendous interest in various research fields. One of the most promising methods to produce large quantities of graphene dispersion is liquid-phase exfoliation (LPE) which utilizes ultrasonic waves or shear stresses to exfoliate bulk graphite into graphene flakes that are a few layers thick. Graphene dispersion produced via LPE can be transformed into graphene ink to further boost graphene's applications, but producing high-quality graphene more economically remains a challenge. To overcome this shortcoming, an advanced LPE process should be developed that uses relatively cheap natural graphite as a graphene source. In this study, a flow-LPE process was used to exfoliate natural graphite to produce graphene that was three times cheaper and seven times larger than synthetic graphite. The optimal exfoliation conditions in the flow-LPE process were determined in order to produce high-quality graphene flakes. In addition, the structural and electrical properties of the flakes were characterized. The electrical properties of the exfoliated graphene were investigated by carrying out an ink formulation process to prepare graphene ink suitable for inkjet printing, and fabricating a printed graphene pattern. By utilizing natural graphite, this study offers a potential protocol for graphene production, ink formulation, and printed graphene devices in a more industrial-comparable manner.

• The Korean Journal of Nuclear Medicine Technology
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• v.14 no.1
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• pp.46-53
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• 2010

Purpose: The static image of nuclear medicine study should be acquired without a motion, however, it is difficult to acquire static image without movement for the serious patients, advanced aged patients. These movements cause decreases in reliability for quantitative and qualitative analysis, therefore re-examination was inevitable in the some cases. Consequently, in order to improve the problem of motion artifacts, the authors substituted the dynamic acquisition technique for the static acquisition, using motion correction. Materials and Methods: A capillary tube and IEC body phantom were used. First, the static image was acquired for 60 seconds while the dynamic images were acquired with a protocol, 2 sec/frame${\times}$30 frames, under the same parameter and the frames were summed up into one image afterwards. Also, minimal motion and excessive motion were applied during the another dynamic acquisition and the coordinate correction was applied towards X and Y axis on the frames where the motion artifact occurred. But the severe blurred images were deleted. Finally, the resolution and counts were compared between the static image and the summed dynamic images which before and after applying motion correction, and the signal of frequency was analysed after frequency spatial domain was transformed into 2D FFT. Supplementary examination, the blind test was performed by the nuclear medicine department staff. Results: First, the resolution in the static image and summed dynamic image without motion were 8.32 mm, 8.37 mm on X-axis and 8.30 mm, 8.42 mm on Y-axis, respectively. The counts were 484 kcounts, 485 kcounts each, so there was nearly no difference. Secondly, the resolution in the image with minimal motion applying motion correction was 8.66 mm on X-axis, 8.85 mm on Y-axis and had 469 kcounts while the image without motion correction was 21.81 mm, 24.02 mm and 469 kcounts in order. So, this shows the image with minimal motion applying motion correction has similar resolution with the static image. Lastly, the resolution in the images with excessive motion applying motion correction were 9.09 mm on X-axis, 8.83 mm on Y-axis and had 469 kcounts while the image without motion correction was 47.35 mm, 40.46 mm and 255 kcounts in order. Although there was difference in counts because of deletion of blurred frames, we could get similar resolution. And when the image was transformed into frequency, the high frequency was decreased by the movement. However, the frequency was improved again after motion correction. In the blind test, there was no difference between the image applying motion correction and the static image without motion. Conclusion: There was no significant difference between the static image and the summed dynamic image. This technique can be applied to patients who may have difficulty remaining still during the imaging process, so that the quality of image can be improved as well as the reliance for analysis of quantity. Moreover, the re-examination rate will be considerably decreased. However, there is a limit of motion correction, more time will be required to successfully image the patients applying motion correction. Also, the decrease of total counts due to deletion of the severe blurred images should be calculated and the proper number of frames should be acquired.