• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.409-419
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• 2001

This study was to examine whether the in vitro friability, motility and intact acrosome of frozen-thawed bovine and human sperm can be improved by adding Pentoxifylline (PF) or Fertilization Promoting Peptide (FPP). Human semen was frozen ultra-rapidly using Test yolk-buffer (TYB) freezing medium. Additive (PF, FPP) effects in frozen-thawed bovine and human sperm were analyzed by microscopic count for sperm motility and coomassie brilliant blue staining method f3r sperm acrosome intact. The in vitro motility of frozen-thawed bovine sperm with 5 mM PF treatment group (50.0%) was significantly higher than that of control (34.0%) (P＜0.05). In the frozen-thawed bovine sperm was examined, the intact acrosome rate of 50 nM FPP treatment (49.0%) was significantly higher than those of control (30.0%) and 25 nM FPP (38.0%) treatment groups (P＜0.01). In human semen, when in vitro motility of sperm with PF addition prior to freezing was examined, the result of 5 mM treatment group (51.0%) was significantly higher than those of control and 2.5 mM treatment group (39.0, 40.0%) (P＜0.01). In addition, 50 nM (75.5%) FPP adding in all treatment procedures for human semen freezing (before freezing, freezing and after thawing) was significant effect on maintenance of the sperm intact acrosome percentage (control: 45.0; 25 nM: 53.0; 100 nM: 68.0%) (P＜0.01). Also, the intact acrosome rate of human sperm with FPP (65.0%) was significantly higher than that with PF (43.0%) (P＜0.05), although sperm motility was slightly higher in PF treatment group. These results suggest that improved sperm motility and intact acrosome of frozen thawed bovine and human sperm can be obtained by addition of PF or FPP, and that the enhanced in vitro viability, motility and intact acrosome can be obtained by addition of FPP in all semen freezing procedures.

• The Journal of Engineering Geology
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• v.24 no.1
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• pp.111-122
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• 2014

For accurate laboratory evaluations of soil deposits, it is essential that the samples are undisturbed. An artificial ground-freezing system is the one of the most effective methods for obtaining undisturbed samples from sand deposits. The objective of this study is to estimate the shear strengths and the characteristics of elastic waves of frozen-thawed and unfrozen specimens through the undrained triaxial compression test. For the experiments, Jumunjin standard sands are used to prepare frozen and unfrozen specimens with similar relative densities (60% and 80%). The water pluviation method is used to simulate the fully saturated condition under the groundwater table. When thawing the frozen specimens, the temperature is measured every minute. After the specimens are completely thawed, undrained triaxial compression tests are conducted using the same procedures as for the unfrozen specimens. During the triaxial tests (saturation, consolidation, and shear phase), compressional and shear waves are measured. The results show that the freeze-thaw process has minor effects on the peak deviatoric stress and shear strength values, and that the process does not affect the internal friction angle. The compressional wave velocity increases with increasing B-value to 1800 m/s in the saturation phase, but tends to remain constant in the process of consolidation and shearing. The shear wave velocity decreases with increasing B-value in the process of saturation, but changes velocity in accordance with the change in effective stress in the processes of consolidation and shearing. The compressional wave velocity has similar values regardless of the freeze-thaw process, but values of shear wave velocity are slighly lower in frozen-thawed specimens than in unfrozen specimens. This study is a preliminary experiment for estimating the shear strength and characteristics of elastic wave velocity in undisturbed frozen specimens that have been obtained using the artificial ground-freezing method.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.2
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• pp.153-162
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• 1992

To solve the logistical problems of the sperm penetration assay (SPA) to provide just a sufficient number of hamster ova exactly when they are needed, a new method to cryopreserve the ova has been devised (1-step dehydration and 2-step thawing). After freezing & thawing of zona-intact (ZI) and zona-free (ZF) hamster ova according to this new method, the frozen-thawed ova were compared with fresh, control ova (FO) in terms of the degree of sperm penetration in SPA using semen samples from fertile donors, subfertile, and infertile male. Each sperm sample was capacitated for 42 hours inTEST-Yolk Buffer before insemination in SPA. In fertile doner, both the penetration rate and penetration index were lower in SPA using frozen ova (ZI; 92.4%, 6.2, ZF; 63.7%, 3.9) than those of SPA using fresh ova (99.3%, 8.4). There was a significant correlation between the penetration index of SPA using FO and ZI (p<0.001), and between those of SPA using FO and ZF and ova (p<0.001). In subfertile patient, both the penetration rate and penetration index were lowered in frozen ova (ZI; 62.3%, 1.3, ZF; 21.8%, 0.4) than those of fresh ova (74.8%, 1.8). There were significant correlation between the penetration rate and penetration index in ZI ova (p<0.05 and p<0.001, respectively). In infertile patient, both the penetration rate and penetration index were ZI; 3.1%, 0.0, ZF;0.0%, 0.0, respectively. There were significant correlation between the penetration rate and penetration index in ZI ova (p<0.05).

• Journal of the Korea Concrete Institute
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• v.19 no.6
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• pp.701-706
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• 2007

This study evaluated the durability of high-performance shotcrete mixed in the proper proportions using alkali-free and cement mineral accelerators as a permanent support that maintains its strength for the long term. Durability tests were performed the chloride permeability, repeated freezing and thawing, accelerated carbonation, and the effects of salt environments. Test results showed that all the shotcrete mixes included silica fume had low permeability. In addition, after 300 freeze/thaw cycles, the shotcrete mix had excellent freeze/thaw resistance more than the 85% relative dynamic modulus of elasticity. The accelerated carbonation test results were no effect of accelerator type but, the depth of carbonation was greater in the shotcrete mix containing silica fume. No damage was seen in a salt environments. Therefore, the high performance shotcrete mix proportions used in this study showed excellent durability.

• Journal of Embryo Transfer
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• v.23 no.4
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• pp.239-243
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• 2008

This study was conducted to investigate the effects of type of the sugar supplemented to the extender on the vigor, viability and intact acrosomal rates of frozen-thawed dog spermatozoa. The ejaculated semen was diluted with TRIS-citric acid extender containing 200mM TRIS, 73mM citric acid, 6% (v/v) glycerol, 20% (v/v) egg yolk, 1% (v/v) antibiotics (streptomycin/penicillin), 44 mM sugar, which was either glucose, fructose or glucose-fructose combination, and distilled water to make the final volume of 100ml. Extended semen samples were cooled at $4^{\circ}C$ for an hour, packaged in 0.25ml straws, equilibrated for 10 minutes in liquid nitrogen vapor, and frozen in liquid nitrogen. Samples were thawed by placing straws into $37^{\circ}C$ water for 120 seconds. After thawing, vigor, viability and intact acrosomal rates of frozen-thawed semen were compared according to type of sugar. No significant differences were observed between glucose and fructose groups. In addition, combination of the 2 sugars also did not show any significant differences in the vigor, viability and intact acrosomal rates. In conclusion, glucose and fructose were equally efficient as sugar supplements for freezing extender.

• Journal of Pharmaceutical Investigation
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• v.33 no.4
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• pp.293-298
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• 2003

This study was attempted to develop the physicochemical ad morphological stability test methods for the cream and lotion formulations among the cosmetic foundations and to provide the guidance for the stability methods with respect to basic emulsions and creams. With these developed stability test methods, we can evaluate the expired date or life time of the available basic cosmetics, especially basic lotions ad creams. Also, the stability test methods established in this study can be used as a guideline to test physical and morphological stability of cosmetics in the future. Thus, we selected two types of basic cosmetics such as lotions and creams made by four different cosmetic companies ad applied them to the stability test methods depending on the temperature changes such as temperature cycling and freezing-thawing cycling test. After the temperature changes, the conductivity, turbidity, particle size, creaming ratio and pH changes of the creams and lotions were evaluated and morphological changes such as crystal formation, odor, color and feeling of the creams and lotions were also tested. As the results of the stability tests, all the tested creams and lotions except for one lotion were stable. Therefore, it may be concluded that these short-term accelerated stability tests as physical stability test depending on the temperature change study were suitable for the stability testing methods for the basic cosmetics and may be useful for the establishment of the guideline for the stability test of cosmetics.

• Journal of Microbiology and Biotechnology
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• v.15 no.4
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• pp.831-837
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• 2005

Low-temperature adaptation and cryoprotection were studied in Leuconostoc mesenteroides SYl, a strain isolated from Kimchi. L. mesenteroides SY1 cells grown in exponential growth phase at $30^{\circ}C$ were exposed to $15^{\circ}C,\;10^{\circ}C$, and $5^{\circ}C$ for 2, 4, and 6 h, respectively, and then frozen at $- 70^{\circ}C$ for 24 h. Survival ratio was measured after the cells were thawed. The freezing-thawing cycles were repeated four times. Preadapted cells survived better than non-adapted control cells, and the highest survival ratio ($96\%$) was observed for cells preadapted for 2 h at $5^{\circ}C$, whereas control cells showed only $22\%$. The 2D gel showed that two proteins (spots A and B) were induced in cells preadapted at lower temperatures. Spots A and B have the same molecular weight (7 kDa), but the pI was 4.6 for spot A and 4.3 for spot B. The first 29 and 15 amino acid sequences from spots A and B were determined, and they were identical, except for one amino acid. A csp gene was cloned, and nucleotide sequencing confirmed that the gene encoded spot A cold shock protein.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.17-17
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• 2003

The milk of transgenic animals may provide an attractive vehicle for large-scale production of hEPO. Since glycosylation is cell type specific, recombinant human EPO (rhEPO) produced in different host cells contain different patterns of oligosaccharides, which could affect the biological functions. However, there have been no reports on the characteristics of rhEPO derived from milk of transgenic animals. To address this objective, several transgenic mice by using pWAPhEPO and/or pBC1hEPO expression vector were produced. However, 2 lines of pWAPhEPO founder female mouse died during late gestational day (day 18) before offspring could be obtained. They showed a severe splenomegaly, Unlike those of pWAPhEPO, mammary gland epithelial cells from biopsies of lactating pBC1hEPO transgenic mice had marked immunoreactivity to EPO and any activity was not detected in other tissues. The expression level of rhEPO is about 0.7% of mammary gland cellular total soluble proteins and an amount of 300~500 mg/L rhEPO is secreted into milk. Furthermore, the pBC1hEPO transgenic mice transmitted this character to their progeny in mendelian manner. In order to determine the extent of glycosylation variation, N-linked oligosaccharide structures present in the milk-derived rhEPO were characterized. Most of milk-derived rhEPO is fully glycosylated. the biological activity of milk-derived rhEPO was comparable to that of purified CHO-derived rhEPO, and milk-derived rhEPO showed relatively stable after freezing and thawing. Taken together, the results illustrate the potential of transgenic animals in the large-scale production of biopharmaceuticals.

• Journal of Embryo Transfer
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• v.31 no.4
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• pp.355-365
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• 2016

The cryopreservation of sperm has become the subject of research for successful artificial insemination technologies. Antifreeze proteins (AFPs), one of the factors necessary for effective cryopreservation, are derived from certain Antarctic organisms. These proteins decrease the freezing point of water within these organisms to below the temperature of the surrounding seawater to protect the organism from cold shock. Accordingly, a recent study found that AFPs can increase the motility and viability of spermatozoa during cryopreservation. To evaluate this relationship, we performed cryopreservation of boar sperm with AFPs produced in the Arctic yeast Leucosporidium sp. AFP expression system at four concentrations (0, 0.01, 0.1, and $1{\mu}g/ml$) and evaluated motility using computer assisted sperm analysis. DNA damage to boar spermatozoa was measured by the comet assay, and sperm membrane integrity and acrosome integrity were evaluated by flow cytometry. The results showed that motility was positively affected by the addition of AFP at each concentration except $1{\mu}g/ml$ (p<0.001). Although cryopreservation with AFP decreased the viability of the boar sperm using, the tail DNA analyses showed that there was no significant difference between the control and the addition of 0.1 or $0.01{\mu}g/ml$ AFP. In addition, the percentage of live sperm with intact acrosomes showed the least significant difference between the control and $0.1{\mu}g/ml$ AFP (p<0.05), but increased with $1{\mu}g/ml$ AFP (p<0.001). Our results indicate that the addition of AFP during boar sperm cryopreservation can improve viability and acrosome integrity after thawing.

• Magazine of the Korean Society of Agricultural Engineers
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• v.24 no.2
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• pp.67-75
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• 1982

A study on the effect of water reducing agent on the various characteristics of concrete has been conducted. The experimental results of the study are summarized as follows. 1. Slump test for the concrete added water reducing setretarding agent in proper quantity have been conducted. According to the test results, the decreasing rate of slump value become bigger than plain concrete with increase of the unit weight of cement and elapse of time 2. In case the proper quantity content of maximum compressive strength in Fig. 5 of water reducing set retarding agent is added, unit weight of water is decreased about 15% or so as compared with plain concrete. with the increase of water reducing set accelerating agent content unit weight of water is decreased much more, And other hand, amount of air entraining shows the increasing tendency with the increase of water reducing agent content. 3. The adding rate of water reducing agent which produce maximum strength shows that WR-CH and WR-SA which is water reducing set-starding agent is 0.2% and WR-CO is 0.5% and that WS-PO which is water reducing set accelerating agent is 0.5 4. compressive strength jof the concrete made of sulfate resistant cement shows less than the strength of normal portland cement at initial strength but the strength of both cement shows almost same at curing age of 28 days. 5. when proper quantity of water reducing set retarding agent is used, boned strength is increased about 15% at curing age of 28days. 6. According to the result of durability test, dynamic young's mudulus of elasticity at plain concrete is decreased about 50% as compared with initial step at 300 cycle of freezing and thawing after curing age of days. on the contarary the concrete used water reducing agent is decreased less than 7%.