• Journal of Embryo Transfer
• /
• v.18 no.3
• /
• pp.195-201
• /
• 2003

This study was carried out to establish most suitable freezing condition, to evaluate the different condition of freezing and thawing rates on the viability and motility of frozen canine spermatozoa. The ejaculated semen was added to obtained 200∼400 ${\times}$ 10$^{6}$ /$m\ell$ with extender I and was cooled to 4$^{\circ}C$ over 30, 60 and 120 minutes. And then, semen was diluted with extender II containing 4, 6 and 8％(v/v) glycerol for 60 min, respectively and packaged in 0.5$m\ell$ straw, equilibrated far 30, 60 and 120 min at 4$^{\circ}C$ and cryopreserved in liquid nitrogen vapor at different distance(3, 5, 7 and 9 cm, respectively), plunged into nitrogen tank. Samples were thawed by placing straws into 27, 37, 47, 57$^{\circ}C$ water bath for 120, 20 and 12 sec, respectively. The results were as follows; 1. The survival and motility rate immediately post-thawing was significantly higher in samples frozen in 4％ glycerol than 6 or 8％ glycerol(P< 0.05). 2. According to equilibration time at 4$^{\circ}C$, the survival and motility rate immediately post-thawing was significantly higher in samples frozen after 60 min equilibration than 30 or 120 min equilibration(P<0.05). 3. Freezing in distance of 5 cm from liquid nitrogen yield better survival and motility rate than the others(3, 7 or 9 cm)(P<0.05). 4. The effect of thawing rate on sperm survival were higher when the thawing was done at 37$^{\circ}C$ for 120 sec(P<0.05).

• Journal of Veterinary Clinics
• /
• v.11 no.2
• /
• pp.545-552
• /
• 1994

This experiment was carried out to investigate the renditions to maintain good post-thaw motility and viability of canine spermatozoa when the semen was frozen using methanol. The semen from two male dogs which had been proven to be fertile in the previous one year was treated with different compositions of semen diluent and was frozen at different freezing temperatures, When canine semen was frozen at-2$0^{\circ}C$, -6$0^{\circ}C$, or -8$0^{\circ}C$, the spermatozoa frozen and stored at -2$0^{\circ}C$ showed very low post-thaw motility and viability from day 2 to 7 and showed no viability since day 15 after freezing. The spermatozoa frozen and stored at -6$0^{\circ}C$ or -8$0^{\circ}C$ showed higher post-thaw motility and viability on day 2, 1, 15 and 30 after freezing than that frozen and stored at-2$0^{\circ}C$(p<0.01), with no difference between two groups. Among different composition groups of the semen diluents of control(tris + egg yolk + glycerol), egg yolk-free, 히ycerol-free, and tris-free, Prior to freezing, the egg yolk-free diluent showed significantly love. motility and viability than the other diluents(p<0.05). On each thawing day (from day 2 to 15 after freezing), control group showed considerably higher motility and viability than the other groups(p<0.01). The canine spermatozoa frozen and stored at -6$0^{\circ}C$ and -8$0^{\circ}C$ showed gradual decrease of motility from day 2 to 30 after freezing and the spermatozoa of these two groups thawed on day 30 showed considerably love. motility than those thawed on day 2 after freezing, respectively(p<0.01). These results indicate that the freezing temperature of either -6$0^{\circ}C$ or -8$0^{\circ}C$ can be applicable to the freezing method using methanol and also all of the components of the semen diluent including cryoprotectant, buffer and cold-shock buffer are very important to maintain motility and viability of canine spermatozoa in the freezing and thawing procedure.

• Korean Journal of Soil Science and Fertilizer
• /
• v.28 no.2
• /
• pp.135-144
• /
• 1995

Understanding on nutrient solute movement during the course of freezing and thawing was attempted through laboratory and field obsevations. Small sectioned tubes with 5cm inner diameter, 0.2cm thick and 1cm long were connected to 30cm long soil columns for laboratory study. The columns were filled with soil, and treated with 20mmol/kg $KNO_3$ for upper 5cm. The upper end was set in the freezing section, and the lower end was set in the refrigerating section of a refrigerator. Temperature was controlled at $-7({\pm}1)^{\circ}C$ and $1.5({\pm}1)^{\circ}C$, respectively. After top 5cm soil was frozen, the columns were sectioned, and analyzed for $NO_3^-$, $NH_4^+$ and $K^+$. For field study, the 20cm inner diameter and lm long soil columns were installed in Chuncheon and Daegwanryung, where the altitude was 74m and 840m, respectively. The soils used were silt loam and clay loam. The top 20cm soils were treated with 50mmol/kg as $KNO_3$. The soil columns were taken during winter freezing and after thawing. By laboratiry study, upward movement of $NO_3^-$ and $K^+$ during the course of freezing was confirmed. The upward movement of $K^+$ was, however, one fifth to one tenth of $NO_3^-$. The upward movement of inorganic nitrogen as well as laboratory during the course of freezing, but large amount of nitrogen was lost from the profile after thawing in early spring. Leached nitrogen from the upper 20cm to lower part was 17 to 24 percents. The maximum depth of leaching during the experiment was 50cm for all soils. The net loss of inorganic nitrogen from the whole profile ranged 8.7 to 39.5 percents. The net loss was greater in Daegwanryung where temperature was lower and snowfall was larger than Chuncheon, and the loss was greater from the silt loam soil than clay loam soil of which percolation rate was small. The results implied that reasons for nitrogen loss during the winter might include surface washing by snow melt as well as leaching and denitrification.

• Journal of the Korea institute for structural maintenance and inspection
• /
• v.17 no.5
• /
• pp.31-39
• /
• 2013

In this study, performance of the post-installed anchor system was evaluated with reduced strength of concrete and anchor. One of the post-installed anchors was selected to performance evaluation. Concrete strength was reduced by freeze-thawing test, and the post-installed anchor strength was reduced by corrosion test. The post-installed Anchor was installed in concrete of freezing and thawing and original concrete, and corroded anchor was installed in original concrete only. Anchor diameter and installation depth of the anchor were the variable for each specimen. Performance of post-installed anchor system of each specimen was evaluated by pullout test. Anchor diameter and installation depth of anchor, it may affect the performance of the post-installed anchor system from the experimental test results. Fracture mode of each post-installed anchor system had occurred differently depending on the durability of concrete and anchor. The anchor pullout strength from the experimental test results was used in order to compare with the results of CCD (Concrete Capacity Design) method, and CCD equation was modified. Modified equation was able to predict the anchor pullout strength of post-installed anchor system in Original concrete and freezing and thawing of concrete.

• Proceedings of the Korea Concrete Institute Conference
• /
• 2008.04a
• /
• pp.161-164
• /
• 2008

In recent years, carbon fiber-reinforced polymer (CFRP) has been widely used for repairing and/or strengthening structural elements in concrete. Not enough test data, however, are available to predict the long-term performance of the repaired and improved structures exposed to weathering. The objective of this research is to study the effect of freeze-thaw cycling on the behavior of reinforced concrete (RC) beams strengthened in shear with carbon fiber sheet. Six small-scale RC beams (100mm${\times]$100mm${\times]$400mm) were strengthened with CFRP in shear, subjected to up to 400 cycles freeze-thawing from -17${\sim}4^{\circ}C$, and tested to failure in four-point bending. Test result, there was no significant damage to carbon fiber sheet strengthened concrete beams had been suffered 30 cycles of freeze-thawing, and more over 60 cycles of freezing-thawing brought about a reduction in resistance of only 25% of the initial level.

• Journal of Microbiology
• /
• v.41 no.3
• /
• pp.262-265
• /
• 2003

Although enumerating bacterial cells is a fundamental step in understanding microbial ecosystems in marine environments, substantial decrease in bacterial counts with increasing sample storage time hampers the accurate estimation of bacterial biomass. We compared the variations in bacterial cell numbers caused by freezing and thawing of sample bottles or slides. Bacterial counts of seawater samples frozen only once in a sampling bottle yielded approximately 95％ of the original numbers after 90 days, whereas 80％ of the original count was obtained for samples prepared on slides. Only 67％ and 58％ of the original counts were recovered in samples repeatedly frozen and thawed in bottles or on slides, respectively. The results indicated that freezing a seawater sample in a bottle increased the consistency of the epifluorescence microscopic enumeration of bacterial cells.

• Korean Journal of Animal Reproduction
• /
• v.12 no.2
• /
• pp.77-83
• /
• 1988

This study was done with mouse embryo to assess effects of freezing media containing sucrose, freezing metods(1-F, 0.3$^{\circ}C$/min;2-F, 3-5$^{\circ}C$/min;3-F, 15$^{\circ}C$min;4-F, LN2 vapour) and cell freezers on the embryo survival determined using the FDA test. The results are summarized as follows. 1. The FDA score obtained with 1, 2, 3 and 4-F was 3.8, 3.6, 3.2 and 3.2, respectively. There was a significant difference(P<0.05) between 1-F, 3-F and 2-F, 4-F. 2. The score at the morular stage(3.8) higher(P<0.005) than the blastocyst stage of embryos(3.2). 3. No difference (P>0.05) was found between the score obtained with a automatic embryo freezer(4.0) and a liquid nitrogen container(3.7).

• Journal of Embryo Transfer
• /
• v.11 no.1
• /
• pp.35-40
• /
• 1996

This study was carried out to select the best cryoprotectant and to establish optimal concentration of the cryoprotectant in ultrarapid freezing of mouse 4-cell embryos and morulae, respectively. We investigated survival of ultrarapid frozen embryos according to various cryoprotectants such as glycerol, ethylene glycol, propylene glycol and dimethyl sulfoxide (DMSO). Suvival of the embryos frozen at different concentrations (3.0, 4.0 and 5.0 M) of indivisual cryoprotectant was also tested. Preimplantation developmental rate (96.3%, 83/86) of 4-cell mouse embryos treated with 4.0 M ethylene glycol after ultrarapid freezing and thawing was higher than those of other cryoprotectants (glycerol, propylene glycol and DMSO). In the ultrarapid freezing of mouse morulae, the highest developmental rate (98.8%, 89 /90) of the embryos to blastocysts was shown in the group of 5.0 M glycerol. Thus, these results demonstrate that 4.0 M ethylene glycol and 5.0 M glycerol are optimal for ultrarapid freezing of 4-cell mouse embryos and morulae, respectively.

• Journal of the Korean Society for Railway
• /
• v.8 no.1
• /
• pp.57-62
• /
• 2005

The frost heaving pressure can be a problem for weakening of the railway roadbed material. This study was initiated to investigate the soils frost heaving pressure and physical characteristics(Liquid limit, permeability, SEM analysis) resulting from freezing and freezing-thawing cycle process. Therefore, upon freezing a saturated soil in a closed-system from the top, a considerable pressure was developed. Weathered granite soils, sandy soil were used in the laboratory freezing test which sometimes subjected to thermal gradients under closed-systems. The frost heaving pressure arising within the soil samples and the temperature of the samples inside were monitored with elapsed time. The degree of saturation versus heaving pressure curve is also presented for weathered granite soil and the maximum pressure is closely related to this curve. Based on the laboratory test results, fine-grained soils with strong attractive forces between soil grains md water molecules, and additional water is attracted into the pores leading to further volume changes and ice segregation.

• Clinical and Experimental Reproductive Medicine
• /
• v.50 no.4
• /
• pp.213-222
• /
• 2023

Cryopreservation is an option for the preservation of pre- or post-pubertal female or male fertility. This technique not only is beneficial for human clinical applications, but also plays a crucial role in the breeding of livestock and endangered species. Unfortunately, frozen germ cells, including oocytes, sperm, embryos, and spermatogonial stem cells, are subject to cryoinjury. As a result, various cryoprotective agents and freezing techniques have been developed to mitigate this damage. Despite extensive research aimed at reducing apoptotic cell death during freezing, a low survival rate and impaired cell function are still observed after freeze-thawing. In recent decades, several cell death pathways other than apoptosis have been identified. However, the relationship between these pathways and cryoinjury is not yet fully understood, although necroptosis and autophagy appear to be linked to cryoinjury. Therefore, gaining a deeper understanding of the molecular mechanisms of cryoinjury could aid in the development of new strategies to enhance the effectiveness of the freezing of reproductive tissues. In this review, we focus on the pathways through which cryoinjury leads to cell death and propose novel approaches to enhance freezing efficacy based on signaling molecules.