• Asian-Australasian Journal of Animal Sciences
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• v.23 no.3
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• pp.318-325
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• 2010

Two experiments were designed to evaluate the effects of the amino acids glycine and cysteine on cryopreservation of ram spermatozoa. After primary evaluation of collected ejaculates, the semen samples were pooled and diluted 1:4 before cooling (experiment 1) and freezing (experiment 2) with Tris-Citrate-Fructose-Yolk (TCFY) extender supplemented with different concentrations of glycine and cysteine (5, 10, 15 and 20 mM). As the control, semen was diluted and frozen in the extender without amino acids. Motility, viability and membrane integrity were assessed as the parameters for semen quality in the first experiment. In the second experiment, motility, progressive motility, viability, membranes and acrosome integrity were evaluated after the freezing-thawing process. The results of the first experiment indicated that the addition of 10 and 15 mM cysteine compared to the control (basic) extender significantly increased (p<0.01) the motility, viability and membrane integrity of spermatozoa after cooling. However, further increasing these amino acids up to 20 mM had a significant negative effect (p<0.05). Our results showed no significant differences (p>0.05) between 5 mM glycine compared to 5 mM cysteine and between 20 mM glycine and 20 mM cysteine. The results of experiment 2 showed that the amino acids significantly improved post-thaw motility, progressive motility, viability, membranes and acrosome integrity of ram spermatozoa. These positive effects were observed at concentrations between 5 to 15 mM of glycine and cysteine, with the best results at 15 mM. Further increasing of amino acid concentrations significantly decreased the post-thaw characteristics of spermatozoa, but the results showed that cysteine was better than glycine and control extenders. The data indicated that addition of glycine or cysteine to the freezing extender can be recommended for cryopreservation of ram spermatozoa. However, further studies are still needed to determine the effect of such addition on fertility in farm animals.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.6-6
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• 2001

Oocyte freezing has become a prevalent source for related reproductive technologies. This study was carried out to evaluate viability of post-thawed bovine oocyte injected DTT-treated sperm following by two different activation stimuli (Group 1, 5 M ionomycin, 5 min ＋ CR1aa, 3 h . 1.9 mM dimetylaminopurine (DMP), 3 h; group 2, ionomycin ＋ 10 $\mu\textrm{g}$/$m\ell$ cycloheximide(CHX), 5h). The techniques of ultra-rapid freezing used in this study were essentially similar to those of described by Vajta et al (Theriogenology 1999; 52:939-948), Denuded oocytes at 22 h of culture were exposed to cryoprotectant (3.2 M Ethylene glycol, 2.36 M DMSO, 0.6 M sucrose), and followed by freezing in electron microscopic grid. After thawing the oocytes were transferred back into the drop of maturation medium and cultured for additional 2 h before being subjected to ICSI. All eggs were then cultured in CRlaa medium, and transferred into M199+10% FCS on day 4. The culture was maintained until day 9. In Experiment 1, frozen-ICSI eggs were compared on development into blastocyst to those of unfrozen and IVF control. Those eggs were activated with the method of group 2. A higher proportion of unfrozen-ICSI and IVF eggs developed into cleavage and blastocysts than of frozen-ICSI eggs (65% and 13%; 71% and 23% vs. 39% and 8%; P<0.05). In Experiment 2, development and ploidy of embryos made from group 1 were compared to those from group 2. Between groups there did not differ on the rates of development, however, chromosomal abnormality in group 1 was significantly higher than in group 2 (49% vs. 30%; P<0.05). The present result suggests that frozen bovine oocytes can be used for ICSI.

• Journal of Veterinary Clinics
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• v.35 no.2
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• pp.39-45
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• 2018

We evaluated the effects of green tea extract (GTE) supplementation at different dilution steps on boar sperm freezing and in vitro fertilization. Sperm intracellular hydrogen peroxide ($H_2O_2$), motility, viability, acrosome integrity and morphology were determined. In addition, sperm IVF parameters (penetration and monospermy) and glutathione (GSH) levels of presumptive zygotes (PZs) were evaluated. Semen was diluted in lactose egg yolk (LEY) and cooled at $5^{\circ}C$ for 3 h (first dilution step) and then diluted in LEY with 9% glycerol and maintained at $5^{\circ}C$ for 30 min (second dilution step). Four experimental groups were compared: first and second dilution steps without GTE (control), first dilution step with GTE (Step 1), second dilution step with GTE (Step 2) and first and second dilution step with GTE (Step 1+2). The spermatozoa were frozen in nitrogen vapor. Higher sperm motility, viability and acrosome integrity after thawing were observed in Step 1, Step 2 and Step 1+2 groups compared with the control (P < 0.05). Lower $H_2O_2$ level was observed in Step 1+2 compared with control and Step 1 (P < 0.05). For IVF, matured oocytes were co-cultured with spermatozoa frozen according to the experimental groups. GSH levels of PZs were significantly higher in Step 2 and Step 1+2 than in control and Step 1 (P < 0.05) without a significant difference in IVF parameters. In conclusion, supplementation with GTE in both first and second dilution steps during the freezing process resulted in better boar sperm cryopreservation and might be beneficial for further embryo development.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.127-135
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• 1999

Cryopreservation is able to store the surplus pre-embryos for freezing and furthermore thawing and transfer in a subsequent cycle. Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps, freezing media and embryonic stages on the rates of viability and development of cryopreserved mouse embryos. Female ICR mice ($6{\sim}8$ weeks old) were induced to superovulate by sequential intraperitoneal injection of 5 IU PMSG and 5 IU hCG 48h apart. Mouse embryos were collected according to its developmental stage after the injection of hCG. Embryos were cryopreserved not only by cryoprotectant step (1 step${\sim}$4 step) but also in a variety of media (HTF, IVF medium, D-PBS) and cell stage. The results were as follows: There is no clear advantage in these freezing media of rapid method, but 4 cell and 8 cell of slow method (2, 3, 4 step) have advantage in D-PBS. The development of embryos according to cell stage become greater in 8 cell stage. In the treatment steps of cryopreservation, the development of embryo to blastocyst was similar among rapid method, but the development of 4 cell and 8 cell embryos to blastocyst according to slow method was better than rapid method.

• Journal of Korean Society of Forest Science
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• v.26 no.1
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• pp.19-22
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• 1975

Freezing resistance of ten cultivars of Chestnut (Castanea crenata S. et Z.) collected from four different sites of Kyunggi Province, Korea on March 2, 1975, was measured to find out the differences among tissue parts, and those among cultivars. The freezing and thawing rates were controlled lower than $6^{\circ}C/hr$. which occurs in nature. The resistance to low temperature was in order from lowest to highest; winter bud, cambium, xylum ray parenchyma and bark cortex. The difference in cold hardiness among cultivars was not consistent among tissue parts of twig stem except in cultivar Dan-Taeck of which all tissue parts showed highest cold-hardiness. The importance of the study on the seasonal variation in cold hardiness of different tissue parts was discussed in terms of choosing the most cold resistant Chestnut culitivar in Korea.

• Applied Chemistry for Engineering
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• v.1 no.1
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• pp.23-29
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• 1990

The effect of a rapid freeze pretreatment of organic ion exchange resins on their grinding properties was studied. It was found that the structure of ion exchange resins was defected by freezing pressure formed in the process of rapid freezing. The defected resins didn't recover their own structure after thawing and those could be easy to be broken at room temperature by small force. Therefore, organic ion exchange resins could be ground readily at room temperature after rapid-freezing the fully swelled resins using by solid carbon dioxide, or liquid nitrogen. The rapid freeze pretreatment of cation exchange resins was very effective on grinding in particular. However, the effect of the pretreatment of anion exchange resins on grinding was less than that of cation exchange resins. In case of anion exchange resins, the ionic form of affected the grindability remarkably.

• Journal of Embryo Transfer
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• v.24 no.4
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• pp.275-279
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• 2009

The beneficial effect of glycerol as a cryoprotectant, especially for sperm cryopreservation, has been shown in many studies. However, glycerol is toxic to living cells, and boar sperm in particular show greater sensitivity to glycerol than sperm from other domestic animals. Amides have been studied as alternative cryoprotectants for freezing stallion sperm. Sperm frozen in methylformamide or dimethylformamide as cryoprotectants show similar motility when thawed compared with sperm frozen in glycerol. We evaluated the cryoprotective effects of dimethylformamide on boar sperm freezing. To test the effect of amides, the concentration of boar semen was adjusted to $10^9sperm/mL$, and seminal plasma was removed using Hulsen solution. After centrifugation, the pellet was diluted in modified-Modena B extender. Lactose-egg yolk (LEY) extender was used as the cooling extender. The freezing extender was madeed aaddition of the optimal amount of glycerol and amides to LEY-Glycerol-Orvus ES Paste extender, and this extender was used for the second dilution. Diluted sperm were frozen in liquid nitrogen using the 0.5 mL straw method. Sperm frozen in extender with glycerol as a cderol were compared with those frozen in extender including the different amides. Sperm were tested for motility, viability, the sperm chromatin structure assay, and normal apical ridge after thawing. The percent of motile sperm diluted in glycerol was as high as that in the stallion study (61%). Dimethylformamide showed positive effects on sperm quality and was better than glycerol. Methylformamide provided similar sperm quality as glycerol. Therefore, dimethylformamide is useful for reducing cryoinjury in boar sperm and is expected to be useful as an alternative cryoprotectant.

• Proceedings of the Korean Institute of Building Construction Conference
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• 2022.11a
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• pp.175-176
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• 2022

By studying the characteristics of matrix insulated through heat generated through oxidation of iron powder, the basic research results on the possibility of buffering and applicability of Cold weather concrete as a curing method are presented. In order to prevent freezing due to a sharp decrease in temperature in the initial stage of curing, iron powder (Fe), powder activated carbon, which is a small amount of porous carbonaceous adsorbent, and salt (NaCl) as an oxidizing agent are replaced with iron powder admixture. As the curing temperature increases, the strength tends to increase, and when replacing the admixture at the same curing temperature, the strength slightly decreases. This is determined as a result of generating iron oxide through an oxidation reaction of iron powder, activated carbon, and NaCl generating a large amount of pores in the matrix. In addition, the internal temperature tends to increase as the mixing substitution rate increases, and it is judged that the oxidation heat of the iron powder mixture affects the increase of the internal temperature during curing. The higher the replacement rate of the iron powder mixture, the slightly lower the strength, but it is determined that freezing and melting that may occur in the early stage of curing can be prevented due to an increase in the initial internal temperature.

• Food Science of Animal Resources
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• v.33 no.6
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• pp.763-771
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• 2013

This study was carried out to investigate the effects of various temperatures of electro-magnetic resonance and air blast freezing methods on the physicochemical quality of meat. Beef (loin and round), pork (belly and ham) and chicken (breast and leg) were purchased at a commercial market, and the meat was frozen using three methods: air blast freezing ($-20^{\circ}C$ and $-45^{\circ}C$) and electro-magnetic resonance quick freezing. Changes in the physicochemical properties of meat were analyzed by drip loss, cooking loss, water holding capacity (WHC) and proximate compositions. In comparison, regardless of the animal species and cuts of meat, electro-magnetic resonance quick freezing (2 h) resulted in a completely frozen product in a much shorter time than $-20^{\circ}C$ and $-45^{\circ}C$ air blast freezing (24 h and 8 h, respectively). Drip loss of loin which had underwent electro-magnetic resonance quick freezing were significantly (p<0.05) lower than those of the other two treatments, but cooking loss and water holding capacity were the highest at 43.7% and 60.7%, respectively (p<0.05). Characteristics such as crude protein, crude fat and moisture compositions showed significant differences, depending on the cuts and freezing methods (p<0.05). The fat composition of electro-magnetic resonance quick frozen loin and round were significantly low (p<0.05). However, moisture content was the highest compared to other freezing methods, as 67.1% and 71.9%, respectively (p<0.05). Electro-magnetic resonance quick freezing was an appropriate way to reduce the deterioration of meat quality due to freezing, and the drip loss was least for the part with low moisture, low protein, and high fat.

• Food Science of Animal Resources
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• v.27 no.2
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• pp.190-196
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• 2007

This study was carried out to investigate the effects of pressure assisted freezing(PAF) on physicochemical properties of pork meat. Pork meat was frozen under pressure up to 200 MPa at $-60^{\circ}C$, and compared with fresh control. Phase transition temperature decreased with increasing pressure level, while pressure level had no effect on supercooling extent. Increasing pressure level increased pH of meat significantly(p<0.05). Thawing losses of all treatments were significantly higher(p<0.05) than control with the exception of PAF at 200 MPa. Water holding capacity(WHC) was increased significantly(p<0.05) with increasing pressure level up to 100 MPa. Cooking loss tended to decrease with increasing pressure level. In color, CIE $L^*-\;and\;b^*-value$ increased with increasing pressure level, while CIE $a^*-value$ decreased significantly(p<0.05). Increasing pressure level up to 150 MPa increased shear force significantly(p<0.05), however, no significant difference between 150 and 200 MPa in shear force was found(p>0.05). Therefore, the results indicated that excessive pressure level in PAF caused several losses in meat qualities, while PAF at mild pressure level improved meat qualities compared to atmosphere freezing.