• Journal of Embryo Transfer
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• v.29 no.3
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• pp.241-248
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• 2014

This study was carried out to evaluate the effects of embryonic stage, cryoprotectant, and freezing-thawing method on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Two to eight cell embryos were obtained from oviducts of mated $F_1$ hybrid female mice superovulated by PMSG and hCG. Two-step EG, DMSO and 4-step EG, DMSO were used as cryoprotectant and dehydration and rehydration method of embryos, and slow-cooling or rapid-cooling method was used as frozen program. The survival rates of embryos were measured after thawing and rehydration, and the developmental rates of embryos were compared and observed during culturing embryos for 24, 48, 72, 96 hrs. As for the survival and development rates of embryos according to embryonic stage, the survival rate of 2 cell stage in EG and DMSO was significantly higher than 4~8 cell (65.4% versus 61.2%, 81.1% versus 72.5%) (p<0.01, p<0.01), but the development rates of 4~8 cell embryos in EG and DMSO were significantly higher than 2 cell embryos for whole culture period (p<0.01) and the development rates of 4~8 cell embryos in EG were significantly higher than 2 cell embryos in DMSO (p<0.01). As for the survival and development rates of embryos according to cryoprotectant, the survival rate of 2 cell embryo in DMSO was significantly higher than that in EG(77.0% versus 64.4%) (p<0.01), whereas the development rate of embryos was not differ till 24 hrs. The development rate from morular to hatching blastocyst, however, was sinificantly higher in EG than in DMSO during 48 hr (p<0.01). The survival rate of 4~8 cell embryo was 62.5% in EG and 73.3% in DMSO. The development rates of embryo in EG were significantly higher for whole culture periods (p<0.01, 0.05). In respect to the effect of freezing and thawing program on the survival and development rates of embryos, method of slow cooling and rapid thawing was more effective than that of rapid cooling and rapid thawing. The survival rate of embryo in 2 cell stage was higher than in 4~8 cell stage, and EG appears more effective cryoprotectant than DMSO because EG showed better development rates of embryos in 2 and 4~8 cell stage. Moreover, slow cooling and rapid thawing method was considered as the best cryopreservation program.

• Journal of Pharmaceutical Investigation
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• v.37 no.2
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• pp.79-84
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• 2007

This study was aimed to establish analytical method of Bi to develop a guideline of the bioequivalence test of tripotassium dicitrato bismuthate (TDB). For this purpose, a simple, specific and sensitive inductively coupled plasma-mass spectrometry (ICP/MS) method were developed and validated in human plasma. Various concentrations of bismuth standard solution (0-25ng/mL) were prepared with distilled water and human blank plasma. To 10mL of the volumetric flasks, 2mL of blank plasma was added with 8ml of distilled water. Bi standard solution was added to prepare the calibration samples and injected into ICP-MS. The plasma samples obtained from volunteers given 3 tablets of bismuth (total 900mg as TDB) were analyzed as described above. As a result, the coefficients of variation were <20% in quantitation limit (0.2 ng/mL) and <15% at the rest of concentrations. The stability test by repeated freezing-thawing cycles showed that the samples were stable only for 24hr. The stability tested for samples with a short-term period of storage at room temperature and pre-treatment prior to the analysis showed very stable over 24hr. In 8 healthy Korean subjects received Denol tablets at the dose of 900mg bismuth, AUC, $C_{max},\;T_{max}$ and half-life $(t_{1/2})$ were determined to be $198.33{\pm}173.78 ng{\cdot}hr/mL,\;64.48{\pm}27.06 ng/mL,\;0.52{\pm}0.21 hr,\;and\;5.15{\pm}2.67 hr$, respectively, from the plasma bismuth concentration-time curves. In conclusion, the method was suitable for the determination of bismuth in human plasma samples and could be applied to bioequivalence test of bismuth tablet.

• Proceedings of the Korean Vacuum Society Conference
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• 2012.02a
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• pp.571-571
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• 2012

We discovered the formation of C60 aggregates in solution by means of photoluminescence spectroscopic study on C60 in solutions. From the in-depth investigation of temperature dependence of the luminescence of C60 in toluene, benzene and CS2 solutions, we reported that the C60 aggregates are formed during cooling at the freezing temperature of these solvents. Furthermore, the C60 aggregates can be changed to stable structures by irradiating with UV pulse-laser (Nd:YAG laser, 355nm). As a consequence, we could obtain nano-scale photo-polymerized C60 clusters, which appear as round-shaped nano- scale particles in high resolution transmission electron-microscopy (HRTEM) images. However, the yield of the nano-scale C60 clusters obtained by this method is too small. So we designed and developed a system to obtain C60 cluster of macroscopic quantity by using ultrasonic nebulizer. In this system, C60 solution was vaporized to several micro-sized droplets in vacuum, resulting in the formation of C60 aggregates by evaporating solvent (toluene). The system was invented to produce nano-scale carbon clusters by the irradiation of UV light upon C60 aggregates in vacuum. We have characterized the products, C60 cluster, obtained from the system by using UV absorption spectra and HPLC spectra. Although the products have a possibility of inclusion various forms of C60 cluster, results support that the product formed from the system by using vaporizer method establishes a new method to obtain C60 cluster in macroscopic quantity. In the presentation, the details of the system and the results of characterization are reported.

• Geotechnical Engineering
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• v.9 no.2
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• pp.15-26
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• 1993

Frost heave on foundation(or ice jacking) is defined that foundation is uplifted by heav- ing force exerted around foundation from freezing of soils. This phenominon is often occurred in the light -weight structure e.g, small building, electro - telegraph pole, street light, pipe line, budge and reference point of survey. Frost heave is the most important factor in design of foundation of structure and the key issue in understanding mechanism of foundation in permafrost area. In this study is reviewed the state of the art on the analysis method of frost heave in USSR and is suggested the design method of benchmark pile. On the basis of above results, this study suggests a design chart able to do esign simply the benchmark pile in Yakutsk region.

• Macromolecular Research
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• v.12 no.4
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• pp.367-373
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• 2004

We have developed a rigorous heat treatment method to improve the biocompatibility of chitosan as a tissue-engineered scaffold. The chitosan scaffold was prepared by the controlled freezing and lyophilizing method using dilute acetic acid and then it was heat-treated at 110$^{\circ}C$ in vacuo for 1-3 days. To explore changes in the physicochemical properties of the heat-treated scaffold, we analyzed the degree of deacetylation by colloid titration with poly(vinyl potassium sulfate) and the structural changes were analyzed by scanning electron microscopy, Fourier transform infrared (FT-IR) spectroscopy, wide-angle X-ray diffractometry (WAXD), and lysozyme susceptibility. The degree of deacetylation of chitosan scaffolds decreased significantly from 85 to 30% as the heat treatment time increased. FT-IR spectroscopic and WAXD data indicated the formation of amide bonds between the amino groups of chitosan and acetic acids carbonyl group, and of interchain hydrogen bonding between the carbonyl groups in the C-6 residues of chitosan and the N-acetyl groups. Our rigorous heat treatment method causes the scaffold to become more susceptible to lysozyme treatment. We performed further examinations of the changes in the biocompatibility of the chitosan scaffold after rigorous heat treatment by measuring the initial cell binding capacity and cell growth rate. Human dermal fibroblasts (HDFs) adhere and spread more effectively to the heat-treated chitosan than to the untreated sample. When the cell growth of the HDFs on the film or the scaffold was analyzed by an MTT assay, we found that rigorous heat treatment stimulated cell growth by 1.5∼1.95-fold relative to that of the untreated chitosan. We conclude that the rigorous dry heat treatment process increases the biocompatibility of the chitosan scaffold by decreasing the degree of deacetylation and by increasing cell attachment and growth.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.179-186
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• 2003

Human embryonic stem (hES) cell lines have been derived from human blastocysts and are expected to have far-reaching applications in regenerative medicine. The objective of this study is to improve freezing method with less cryo-injuries and best survival rates in hES cells by comparing various vitrification conditions. For the vitrifications, ES cells are exposed to the 4 different cryoprotectants, ethylene glycol (EG), 1,2-propanediol (PROH), EG with dime-thylsulfoxide (DMSO) and EG with PROH. We compared to types of vehicles, such as open pulled straw (OPS) or electron microscopic cooper grids (EM grids). Thawed hES cells were dipped into sequentially holding media with 0.2 M sucrose for 1 min, 0.1 M sucrose for 5 min and holding media for 5 min twice and plated onto a fresh feeder layer. Survival rates of vitrified hES cells were assessed by counting of undifferentiated colonies. It shows high survival rates of hES cells frozen with EG and DMSO (60.8％), or EG and PROH(65.8％) on EM grids better than those of OPS, compared to those frozen with EG alone (2.4％) or PROH alone (0％) alone. The hES cells vitrified with EM grid showed relatively constant colony forming efficiency and survival rates, compared to those of unverified hES cells. The vitrified hES cells retained the normal morphology, alkaline phosphates activity, and the expression of SSEA-3 and 4. Through RT-PCR analysis showed Oct-4 gene expression was down-regulated and embryonic germ layer markers were up-regulated in the vitrified hES cells during spontaneous differentiation. These results show that vitrification method by using EM grid supplemented with EG and PROH in hES cells may be most efficient at present to minimize cyto-toxicity and cellular damage derived by ice crystal formation and furthermore may be employed for clinical application.

• Journal of the Korean Recycled Construction Resources Institute
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• v.7 no.2
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• pp.138-144
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• 2019

To analyze the present state of buildings suspended in Gangwon and estimate the compressive strength by visual inspection and Schmidt hammer method in order to analyze durability etc.. In this study, we analyzed the problems that existed in the location where construction is suspended and efficient management method. Expected construction restoration of construction will be restarted, and important parts of the construction site should be given the best protection measures so that the quality can be maintained thoroughly. The construction of the suspended construction is exposed to the freezing and thawing damage over time. Therefore, it is necessary to take measures such as maintenance, and take protective measures by establishing a plan to improve the durability of buildings that are under construction.

• Journal of Advanced Navigation Technology
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• v.27 no.6
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• pp.865-870
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• 2023

Recently, accidents caused by black ice, a road freezing phenomenon caused by natural power, are increasing. Black ice is difficult to identify directly with the human eye and is more likely to misunderstand it as standing water, so there is a high accident rate caused by car sliding. To solve this problem, this paper presents a method of detecting black ice centered on LiDAR sensors. With a small, inexpensive, and high-accuracy light detection and ranging (LiDAR) sensor, the temperature and inclination angle are set differently to detect black ice and asphalt by setting different reflection angles of asphalt and black ice differently in temperatures and inclinations. The LIDARO carried out in the study points out that additional research and improvement are needed to increase accuracy, and through this, more reliable black ice detection methods can be suggested. This method suggests a method of detecting black ice through early system design research by preventing accidents caused by black ice in advance.

• International Journal of Computer Science & Network Security
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• v.24 no.3
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• pp.125-134
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• 2024

The novel coronavirus 2019 is called COVID-19 has outspread swiftly worldwide. An early diagnosis is more important to control its quick spread. Medical imaging mechanics, chest calculated tomography or chest X-ray, are playing a vital character in the identification and testing of COVID-19 in this present epidemic. Chest X-ray is cost effective method for Covid-19 detection however the manual process of x-ray analysis is time consuming given that the number of infected individuals keep growing rapidly. For this reason, it is very important to develop an automated COVID-19 detection process to control this pandemic. In this study, we address the task of automatic detection of Covid-19 by using a popular deep learning model namely the VGG19 model. We used 1300 healthy and 1300 confirmed COVID-19 chest X-ray images in this experiment. We performed three experiments by freezing different blocks and layers of VGG19 and finally, we used a machine learning classifier SVM for detecting COVID-19. In every experiment, we used a five-fold cross-validation method to train and validated the model and finally achieved 98.1% overall classification accuracy. Experimental results show that our proposed method using the deep learning-based VGG19 model can be used as a tool to aid radiologists and play a crucial role in the timely diagnosis of Covid-19.

• Journal of Microbiology and Biotechnology
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• v.6 no.3
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• pp.209-212
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• 1996

A two-step cooling technique has been developed for cryopreservation of suspension cultured Scutellaria baicalensis cells. Efficient regrowth of cryopreserved cells was obtained in cryoprotected cells with a mixture of 1.5 M glycerol and 0.4 M sucrose in Schenk and Hildebrandt medium without pretreatment in high osmotic medium. Optimum freezing conditions were found to be a cooling rate of $0.5^{\circ}C$ min from $4^{\circ}C$ to $-40^{\circ}C$, and then retaining samples at $-40^{\circ}C$ for 30 min prior to plunging into liquid nitrogen. A regrowth rate of approximately 95$%$ was obtained after three month storage in liquid nitrogen. Callus cultures established from the cryopreserved cells were found to produce the same patterns of flavonoid accumulation and retain their baicalin producing activity.