• Journal of Veterinary Clinics
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• v.30 no.6
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• pp.442-448
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• 2013

The aim of the present study was to develop glycerol-free TRIS extender using glucose for dog sperm cryopreservation. We determined the appropriate concentration of glucose in glycerol-free TRIS and the exposure time in glycerol-free TRIS containing 0.3 M glucose at $4^{\circ}C$. Ejaculates of six dog sperm were cooled in glycerol-free TRIS through $4^{\circ}C$ for 100 min, cooled at $4^{\circ}C$ in TRIS with different glucose concentrations 0 M, 0.04 M, 0.1 M, 0.2 M and 0.3 M, respectively for 30 min followed by cryopreservation. After thawing at $37^{\circ}C$ for 25 sec, membrane and acrosome integrities of dog sperm were evaluated. In addition, the effect of exposure time (10, 30, 50 and 70 min) of sperm to glycerol-free TRIS containing 0.3 M glucose at $4^{\circ}C$ on progressive motility, viability, and DNA integrity following sperm cryopreservation was studied. Membrane integrity and acrosome integrity were assessed by 6-carboxyfluoresceindiacetate (6-CFDA)/propidium iodide (PI) fluorescent staining and Pisum sativum agglutinin conjugated to fluorescein isothiocyanate, respectively. DNA integrity was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling, using flow cytometry. Sperm frozen in glycerol-free TRIS supplemented with 0.2 M or 0.3 M glucose have an intact plasma membrane (CFDA+/PI-) after cryopreservation than sperm frozen in the extenders with lower glucose concentrations (p<0.05). Acrosome integrity was significantly higher in the 0.3 M group than less than 0.1 M groups (p<0.05). The sperm DNA fragmentation index did not differ according to exposure time, although progressive motility was significantly higher in the 50 min exposure group than the other groups (p<0.05). These results indicate that cryopreservation of dog sperm is feasible and yields more motile sperm following freezing and thawing in glycerol-free TRIS containing 0.3 M glucose with the exposure time for 50 min at $4^{\circ}C$.

• Korean Journal of Food Science and Technology
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• v.17 no.6
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• pp.500-505
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• 1985

A method to store concentrated milk in the liquid state at $-15^{\circ}C$ was developed, and quality changes during storage of milk were evaluated. Combined cryoprotectants (CCP) suitable for storing concentrated milk in the liquid state at $-15^{\circ}C$ were consisted of 17.74% sucrose, 8.87% glucose, 8.87% fructose, 2% glycerol, 0.25% sodium hexametaphosphate, 0.25% NaCl and 0.02% ascorbic acid. The amount of CCP to be added to concentrated milk to depress freezing point to $-15^{\circ}C$ was 38% by weight. Gelation due to protein denaturation was the most serious quality change during storage, which adversely affected appearance and utilization of the stored product. Gelation was observed after 3 weeks storage in the control, but it was not in milk with CCP throughout 18 weeks storage. Amount of protein precipitated increased in the control during storage, whereas there was no protein precipitated in milk with CCP. Surface color and peroxide value of the control and treatment did not change significantly during storage, and there were no marked differences between the control and treatment. These results indicated that quality of concentrated milk could be preserved, without gelation, by storing milk with CCP in the -liquid state at the frozen storage temperature. Besides, energy required for freezing preservation of milk could be significantly reduced by elimination of phase changes for freezing and thawing, and the stored product could be continuously processed for the final products without long waiting time for thawing.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.275-289
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• 2002

The objective of this study was to develop an in vitro assessment of sperm fertilizing capacity of bulls and investigate the factors influencing sperm function and characteristics of frozen-thawed bovine spermatozoa. in vitro fertilization (IVF), the evaluation of motility and normal morphology, HOST (hypoosmotic swelling test), Ca-ionophore induced acrosome reaction, luminol and lucigenin-dependent chemiluminescence for the measurement of reactive oxygen species (ROS), the measurement of malondialdehyde formation for the analysis of lipid peroxidation (LPO), and the evaluation of DNA fragmentation using the method of 747-mediated nick end labelling (TUNEL) by flow cytometry were performed in frozen-thawed bovine spermatozoa. Correlations between the rates of fertilization, blastocyst formation after IVF and the values of respective assays were investigated. 1. IVF rate and blastocyst formation rate averaged 64.4% and 34.3% for spermatozoa from high -fertility bull group and averaged 18.5% and 6.2% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. Sperm motility and percentage acrosome reaction averaged 79.0% and 66.2% for spermatozoa from high-fertility bull group and averaged 40.7% and 22.9% for spermatozoa from low-fertility bull group, respectivitely. There were not different between two bull groups. 2. Luminol depenent chemiluminescence, LPO and DNA fragementation averaged 6.4, 2.0 nmol and 2.6% from spermatozoa from high-fertility bull group and averaged 6.5, 3.1 nmol and 7.4% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. There was no significant difference in lucigenin dependent chemiluminescence between two bull groups. 3. Fertilization rate was positively correlated with motility and the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between fertilization rate and the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. 4. Blastocyst formation rate was positively correlated with the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between blastocyst formation rate and motility, the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. In conclusion, these data suggest that ROS significantly impact semen quality. The assays of this study may provide a basis fur improving in vitro assessment of sperm fertilizing capacity.

• Pediatric Infection and Vaccine
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• v.14 no.1
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• pp.91-96
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• 2007

Purpose : This study was performed to evaluate a new rapid diagnostic kit (BIOLINE $RSV^{TM}$; Standard Diagnostics Inc., Yongin, Korea), a lateral-flow immunoassay, in the detection of respiratory syncytial virus (RSV) from the nasopharyngeal aspirates (NPA) of children with lower respiratory tract infections (LRTIs) in comparison with other diagnostic methods. Methods : Three hundred and nineteen NPAs were selected from a large pool of NPAs that had been obtained from children with LRTIs. All specimens had already been tested for RSV by culture and immunofluorescent (IF) test, and had been kept frozen. Tests with BIOLINE $RSV^{TM}$ were performed at least twice. All who conducted the experiments or interpreted the test results were blinded to the results of both culture and IF tests. Results : One hundred seven (97.3%) of 110 specimens that were positive for RSV by both culture and IF test, 29 (87.9%) of 33 that were positive by IF test only, 20 (76.9%) of 26 that were positive by culture only, and 140 (93.3%) of 150 that were negative by both methods were negative for RSV by BIOLINE $RSV^{TM}$. By combining the above results, the following 5 diagnostic values of BIOLINE $RSV^{TM}$ were determined in comparison with viral culture or IF test; sensitivity, 92.3% (156/169, 95% confidence interval [CI], 87.1-97.5%); specificity, 93.3% (140/150, 95% CI, 88.4-98.2%); positive predictive value, 94.0% (156/166, 95% CI, 89.5-98.5%); negative predictive value, 91.5% (140/143, 95% CI, 86.0-97.0%); and agreement, 95.9% (306/319, 95% CI, 92.1-99.7%), respectively. Conclusion : This study revealed that BIOLINE $RSV^{TM}$ demonstrated good sensitivity and specificity for the detection of RSV antigen from NPAs of children with LRTIs. Because of simple methods and quick results, this test may be useful for the diagnosis of RSV infection during the epidemic periods.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1273-1273
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• 2001

The efficiency of the luminal fermentation process influences overall efficiency of luminal production, animal health and reproduction. Ruminant production systems have a significant impact on the global environment, as well. Animal wastes contribute to pollution of the environment as ammonia volatilized to the air and nitrate leached to ground water. Microbial protein synthesis in the rumen satisfies a large proportion of the protein requirements of animals. Quantifying the microbial synthesis is possible by using markers for lumen bacteria and protozoa such as nucleic acids, purine bases, some specific amino acids, or by isotopic $^{15}N,^{32}P,\;and\;^{35}S$ labelled feeds. All those methods require cannulated animals, they are time-consuming and some methods are very expensive as well. Many attempts have been made to find an alternative method for indirect measurement of microbial synthesis in intact animals. The present investigations aimed to assess possibilities of NIRS for prediction of purine nitrogen excretion and ruminal microbial nitrogen synthesis by NIR spectra of urine. Urine samples were collected from 12 growing sheep,6 of them male, and 6- female. The sheep were included in feeding experiment. The ration consisted of sorghum silage and protein supplements -70:30 on dry matter basis. The protein supplements were chosen to differ in protein degradability. The urine samples were collected daily in a vessel containing $60m{\ell}$ 10% sulphuric acid to reduce pH below 3 and diluted with tap water to 4 liters. Samples were stored in plastic bottles and frozen at $-20^{\circ}C$ until chemical and NIRS analysis. The urine samples were analyzed for purine derivates - allantoin, uric acid, xantine and hypoxantine content. Microbial nitrogen synthesis in the lumen was calculated according to Chen and Gomes, 1995. Transmittance urine spectra with sample thickness 1mm were obtained by NIR System 6500 spectrophotometer in the spectral range 1100-2500nm. The calibration was performed using ISI software and PLS regression, respectively. The following statistical results of NIRS calibration for prediction of purine derivatives and microbial protein synthesis were obtained.(Table Omitted). The result of estimation of purine nitrogen excretion and microbial protein synthesis by NIR spectra of urine showed accuracy, adequate for rapid evaluation of microbial protein synthesis for a large number of animals and different diets. The results indicate that the advantages of the NIRS technology can be extended into animal physiological studies. The fast and low cost NIRS analyses could be used with no significant loss of accuracy when microbial protein synthesis in the lumen and the microbial protein flow in the duodenum are to be assessed by NIRS.

• Journal of Korean Tunnelling and Underground Space Association
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• v.20 no.2
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• pp.255-268
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• 2018

Subsea tunnel can be highly vulnerable to seawater intrusion due to unexpected high-water pressure during construction. An artificial ground freezing (AGF) will be a promising alternative to conventional reinforcement or water-tightening technology under high-water pressure conditions. In this study, the freezing energy and required time was calculated by the theoretical model of the heat flow to estimate the total amount of refrigerant required for the artificial ground freezing. A lab-scale freezing chamber was devised to investigate changes in the thermal and mechanical properties of sandy soil corresponding to the variation of the salinity and water pressure. The freezing time was measured with different conditions during the chamber freezing tests. Its validity was evaluated by comparing the results between the freezing chamber experiment and the numerical analysis. In particular, the freezing time showed no significant difference between the theoretical model and the numerical analysis. The amount of refrigerant for artificial ground freezing was estimated from the numerical analysis and the freezing efficiency obtained from the chamber test. In addition, the energy ratio for maintaining frozen status was calculated by the proposed formula. It is believed that the energy ratio for freezing will depend on the depth of rock cover in the subsea tunnels and the water temperature on the sea floor.

• Current Optics and Photonics
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• v.1 no.2
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• pp.107-112
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• 2017

An adaptive optics system can be simulated or analyzed to predict its closed-loop performance. However, this type of prediction based on various assumptions can occasionally produce outcomes which are far from actual experience. Thus, every adaptive optics system is desired to be tested in a closed loop on an optical test bench before its application to a telescope. In the close-loop test bench, we need an atmospheric simulator that simulates atmospheric disturbances, mostly in phase, in terms of spatial and temporal behavior. We report the development of an atmospheric turbulence simulator consisting of two point sources, a commercially available deformable mirror with a $12{\times}12$ actuator array, and two random phase plates. The simulator generates an atmospherically distorted single or binary star with varying stellar magnitudes and angular separations. We conduct a simulation of a binary star by optically combining two point sources mounted on independent precision stages. The light intensity of each source (an LED with a pin hole) is adjustable to the corresponding stellar magnitude, while its angular separation is precisely adjusted by moving the corresponding stage. First, the atmospheric phase disturbance at a single instance, i.e., a phase screen, is generated via a computer simulation based on the thin-layer Kolmogorov atmospheric model and its temporal evolution is predicted based on the frozen flow hypothesis. The deformable mirror is then continuously best-fitted to the time-sequenced phase screens based on the least square method. Similarly, we also implement another simulation by rotating two random phase plates which were manufactured to have atmospheric-disturbance-like residual aberrations. This later method is limited in its ability to simulate atmospheric disturbances, but it is easy and inexpensive to implement. With these two methods, individually or in unison, we can simulate typical atmospheric disturbances observed at the Bohyun Observatory in South Korea, which corresponds to an area from 7 to 15 cm with regard to the Fried parameter at a telescope pupil plane of 500 nm.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.3
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• pp.213-217
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• 2001

In order to develop a new separation technology, supercritical fluid extraction process was used to produce high purity pigments and fatty acids from seaweed (Undaria pinnatifida). Supercritical carbon dioxide was used as a solvent and ethanol as an entrainer. The sample was treated by a frozen drier and experiments were conducted with a semi-batch flow system at various operating conditions (pressure range, $10.3\~17.2$ MPa; temperature range, $30\~45^{\circ}C$: particle size, $500\~1,000{\mu}m$ extraction time, 60 min). Characteristics of the recovered pigment (chlorophyll a) and fatty acids were determined by UV-spectrophotometry and gas chromatography, respectively. The highest extraction efficiency for fatty acids and pigments was achieved at 12.4 MPa, $35^{\circ}C$, $500{\mu}m$of seaweed size.

• Journal of Embryo Transfer
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• v.30 no.3
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• pp.121-128
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• 2015

The objective of this study was to evaluate the efficiency of sperm cryosurvival in boar sperm separated by Percoll containing antioxidant enzymes. The boar semen was collected into a pre-warmed ($37^{\circ}C$) thermos bottle by gloved-hand method and was separated by 65% Percoll with superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) before freezing. The frozen sperm was thawed at $38.5^{\circ}C$ for 45 sec in water-bath for sperm characteristic analysis. The sperm were estimated with SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction, Rhodamine123/PI double staining for mitochondrial integrity and were analyzed using flow cytometry. In results, sperm viability, acrosome reaction and mitochondrial integrity were improved in separated sperm groups compared with unseparated sperm by Percoll (UP) group. Especially, viability was significantly higher in sperm separated by Percoll containing 400 IU CAT group compared with other groups (P<0.05). And acrosome reaction was decreased in sperm separated by Percoll with 300 IU SOD, 400 IU CAT and 0.5 mM GSH groups compared with other groups, however, there were no significantly difference mitochondrial integrity among sperm separated by Percoll with antioxidant enzymes. In conclusion, we suggest that use of Percoll containing antioxidant enzymes for sperm separation will be beneficial for sperm cryopreservation in pigs.

• Journal of Service Research and Studies
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• v.6 no.3
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• pp.29-40
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• 2016

Since the late 20th century, there has been much effort to improve the market value of media contents which are commercialized in a digital format, by fusing digital data of video, audio, numerals, characters with IT technology together. Then by what criteria and methodologies could the market value for the drama "Sons of the Sun" or the animated film 'Frozen', often referred to in the meida, be estimated? In the circumstances there has been little or no research on the valuation framework of media contents and the status of their valuation system development to date, we propose a practical valuation models for various purposes such as contents trading, review of investment adequacy, etc., by formalizing and presenting a contents valuation framework for the four types of media of movies, online games, and broadcasting commercials, and animations. Therefore, we develope computational methods of cash flows which includes production cost by media content types, provide reference databases associated with key variables of valuation (economic life cycle, discount rates, contents contribution and royalty rates), and finally propose the valuation framework of media contents based on both income approach and relief-from-royalty method which has been applied to valuation of intangible assets so far.