• Journal of Veterinary Clinics
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• v.21 no.4
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• pp.363-368
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• 2004

To investigate the effects of seeding during freezing procedure on post-thaw viability, motility, and acrosome integrity of boar spermatozoa, semen from 5 Yorkshire boars were collected for this experiment. Raw semen were diluted with Merck I, subsequently added with cooling diluent containing lactose and egg yolk and with freezing diluent containing glycerol. The diluted semen were frozen on the rack in the styrofoam box filled with liquid nitrogen at the distance of 5 cm or I cm above LN2 level. Seeding was performed to only a group of straws frozen at 5 cm away on the surface of LN2. The frozen semen were thawed in $50^{\circ}C$C water and the viability and local motility were analyzed by sperm analysis imaging system. A part of thawed semen was taken for the examination of morphology of apical ridge of the acrosome to compare with the effect of seeding between the seeding-treated and non treated groups. I. Post-thaw viability was considerably higher in seeding-treated sperm than non-seeding group (p<0.01), however, no difference of local motility was obtained among the groups. 2. At three hours after thawing, viability was also higher in seeding-treated group than non-treated group (p<0.05), along with no difference of motility among the groups. 3. Higher normal acrosome integrity was obtained in the seeding-treated sperm than non-treated groups (p<0.01). 4. Between non-seeded groups, higher normal acrosome integrity was obtained in the sperm group frozen at 5cm upper on the surface of LN2 than that frozen at 1cm away (p<0.01). These results indicated that seeding treatment during freezing boar spermatozoa was beneficial to post-thaw viability and normal acrosome integrity.

• Korean Journal of Animal Reproduction
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• v.24 no.3
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• pp.263-268
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• 2000

The development of single blastomeres isolated from 2-, 4- and 8-cell mouse embryos and the ability of such blastomeres to survive slow freezing were studied. Of 223, 60 and 188 single blastomeres isolated from 2-, 4- and 8-cell mouse embryos, respectively, 111 blastomeres (49.8%) from 2-cell embryos, 12 blastomeres (20.0%) from 4-cell embryos and blastomeres (16.5) from 8-cell embryos developed into blastocysts after culture for 96 hrs. The recovery rate was 54.2% (65/120), 46.4% (13/28) and 24.3% (17/70) of blastomeres derived from 2-, 4- and 8-cell embryos following freezing and thawing and the survival of frozen-thawed blastomeres was 27.1% (16/59), 36.4% (4/11) and 17.6% (3/17), and respectively. The apparently six normal fetuses were obtained from frozen-thawed blastomere from 2-cell embryos after transferring into the recipients. These results indicate that mouse btastomeres isolated from preimplatation stage embryos can survive storage in liquid nitrogen following slow freezing.

• Korean Journal of Animal Reproduction
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• v.23 no.2
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• pp.165-174
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• 1999

Boar semen can be frozen successfully. However, there is a large variability in the extent of damage boar semen samples experiences during cryopreservation. This experiment was undertaken to find out factors that affect a post-thaw viability of boar spermatozoa. For this purpose, cryodiluents(BF5, LEY, Soejima and M-Soejima), cryoprotectants(glycerol. ethylene glycol, and propylene glycol), pre-freezing method(dryice-pellet, dryice-straw and L$N_2$vapour-st-raw) and total time required for freezing(2. 5, and 7 h) were compared as a factors. To investigate quality of semen during freezing process, motility(%), normal apical ridges(%, NAR), and proportion of living sperm(%) by flow cytometic analysis were assessed after collection, cooled, pre-frozen and post-thawing. Post-thaw motility of semen diluted with M-Soejima was 52.0%, respectively. When heparin, caffeine or heparin＋caffeine was added to 2nd cryodiluent of M-Soejima during freezing process, the highest motility after thawing was shown at the addition of caffeine (2mM), with 61.7$\pm$2.9% of motility. M-Soejima with heparin or caffeine was significantly higher than that of controI(p＜0.05). The result using glycerol(Gly), ethylene glycol(EG), propylene glycol(PG), and their mixture (Gly＋EG and Gly＋PG) as cryoprotectants, the highest motility was shown at the mixture treatment with Gly plus PG. However, the highest proportion of live spermatozoa was shown at Gly＋EG, there was no significantly difference among treatments(p＞0.05). When semen was pre-frozen with three manners(dryice-pellet, dryice-straw, and L$N_2$ vapor-straw), motility(%) of post-thaw spermatozoa was the highest in the L$N_2$ vapor-straw pre-freezing method of M-Soejima cryodiluent with 57.5% of motility, For a simple, economical and timesaving approach to freezing boar semen, total time required for freezing were 2, 5, and 7 hours, post-thaw motility were 43.8, 45.0 and 38.8%, NAR were 19.5, 22.7 and 28.5%, and viability were 20.8, 19.9 and 22.1%, respectively. This data suggests that boar semen diluted with M-Soejima cryodiluent contained caffeine, using mixture of glycerol and propylene glycol or ethylene glycol as cryoprotectants, frozen with 2 hours, can be taken better motility, NAR, and proportion of live spermatozoa.

• Journal of Embryo Transfer
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• v.24 no.4
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• pp.253-257
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• 2009

To enhance the embryo preservation technology and better application of embryo transfer technique to the field (dairy science or animal reproduction. etc.), we examined the viabilities of bovine embryos produced in vitro and in vivo after cryopreservation according to their developmental stage and thawing temperature. Bovine embryos from in vivo/vitro fertilization (Hanwoo) were examined at day 7, 8, and 9. Survival rates and total cell numbers of in vivo fertilized embryos were as follows: morulae 68.8% and $67\;{\pm}\;6.0$; blastocysts 80.5% and $120\;{\pm}\;10$; expanded blastocysts 77.4% and $138\;{\pm}\;9.7$, respectively. Rates of embryo development for blastocysts and expanded blastocysts after thawing were significantly higher than that of morula stage embryos (p<0.05). While survival rates of in vitro fertilized embryos according to developmental stage showed no significant difference among groups (morula 67.9%; blastocyst 74.3%; and expanded blastocyst 79.4%), total cell numbers were significantly lower than those of other groups (morula $64\;{\pm}\;5.9$; blastocyst $116\;{\pm}\;8.7$; and expanded blastocyst $135\;{\pm}\;9.1$) For the viability according to thawing temperature, survival rate was higher in $37^{\circ}C$.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.1
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• pp.9-17
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• 2004

Objective: The aim of this study were to compare the effects of EG and PROH on cryopreservation of mouse and human embryos, and to find the optimal protocol for embryo freezing. Methods: Human embryos derived from fertilized eggs showing 3 pronuclei (PN) and mouse embryos were divided into two groups respectively: dehydrated with 1.5 M EG + 0.2 M sucrose or 1.5 M PROH + 0.2 M sucrose using the slow freezing method. Moreover mouse embryos were controlled the exposure time of cryoprotectant during dehydration or rehydration steps. Results: The survival rates of human embryos were 79.2% (84/106) in EG group and 77.9% (88/113) in PROH group. In mouse embryos, the survival and development rates up to blastocyst were 70.6% (245/347), 44.1% (123/279) in EG group and 62.1% (198/319), 45.1% (123/279) in PROH group, respectively. However, in EG group, partially damaged embryos after thawing were decreased compared to PROH group. In combination group, when the exposure time during dehydration and rehydration were reduced, the survival and embryonic developments were increased slightly, but not significant. Conclusion: Cryopreservation of mouse and human embryos at cleavage stage by using EG or PROH exhibited no statistical difference in the survival rate and/or developmental rate to blastocyst. However, the use of EG for cryopreservation of embryos might reduce the exposure time of the cryoprotectant because of a high permeation of EG and result in lessen its toxic effects.

• Journal of the Korea institute for structural maintenance and inspection
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• v.9 no.2
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• pp.173-180
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• 2005

Metakaolin is a cementitious material for producing high-strength concrete. This material is now used as substitute for silica-fume. In this paper, we did the mechanical and durability test such as compressive/tensile/flexural strength test, chloride ion diffusion, chemical attack and repeated freezing and thawing, carbonation test. In the mechanical tests, 10~15% for binder is optimum substitute rate. And, in the chloride ion diffusion test, according to the increase of substitute of metakaolin & silica-fume for binder, the diffusion coefficient was more reduced. In the chemical attack test, by the filler effect of fine powder such metakaolin and silica-fume, the resistance is more excellent than normal concrete. In the other durability test, the concrete using metakaolin also compared with those of silica-fume substitute concrete. Through these tests, we recognized that metakaolin can be used as a substitute for silica-fume.

• Journal of The Korean Society of Agricultural Engineers
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• v.57 no.4
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• pp.11-19
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• 2015

The important properties of EVA (ethylene vinyl acetate) redispersible polymer was waterproof, densification of internal pore space of concrete and ball bearing and micro filler. Also, the significant role of polypropylene(PP) fiber was crack control and blockade of movement for deterioration factors. The most studies for EVA were limited in the field of mortar and PP fiber reinforced concrete had been studied in the state of being restricted unit water content, rich mix and mixing much of the fiber without considering construction site. Therefore, the control mix design were applied in ready mixed concrete using 10 % fly ash of total cement weight used in batch plant. On the basis of control mix design, EVA contents ranging from 0 % to 10 % of total cement weight and PP fiber contents ranging from 0 % to 0.5 % of EVA concrete volume were used in the mix designs. The results showed the maximum compressive strength value was measured at EVA 5.0 % and PP fiber 0.1 %, the minimum water absorption ratio was at EVA 10 % and PP fiber 0 %, the durability factor for freezing and thawing resistance was at EVA 5.0 % and PP fiber 0.3 % and the minimum weight reduction ratio of resistance to sulfuric acid attack was at EVA 10 % and PP fiber 0.5 % after curing age 42days. Meanwhile, From these results, PP fiber reinforced EVA concrete would be very benefit, if each optimal mix types were used in hydraulic structures, underground utilities and agricultural structures.

• Geomechanics and Engineering
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• v.13 no.1
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• pp.25-41
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• 2017

In recent years, the amount of sludge ash (SA) has considerably increased due to rapid urbanization and population growth. In addition, its storage in landfills induces environmental pollution and health problems. Therefore, its disposal in an environmentally friendly way has become more important. The main goal of this study is to investigate the reusability of sludge ash as an additive with polypropylene fiber (PF) to stabilize marginal sand based on the compressive strength performances from UCS tests. For this purpose, a series of UCS tests was conducted. Throughout the experimental study, the used inclusion rates were 10, 15, 20 and 30% for sludge ash and 0, 0.5 and 1% for polypropylene fiber by total dry weight of the sand+sludge ash mixture and the prepared samples were cured for 7 and 14 days prior to the testing. Freezing and thawing resistance of the mixture including 10% sludge ash and 0, 0.5 and 1% polypropylene fiber was also examined. On the basis of UCS testing results, it is said that sludge ash inclusion remarkably enhances UCS performance of sand. Moreover, the addition of polypropylene fiber to the admixtures including sand and sludge ash significantly improves their stress-strain characteristics and post-peak strength loss as well as UCS. As a result of this paper, it is suggested that sludge ash be successfully reused with polypropylene fiber for stabilizing sand in soil stabilization applications. It is also believed that the findings of this study will contribute to some environmental concerns such as the disposal problem of sludge ash, recycling, sustainability, environmental pollution, etc. as well as the cost of an engineering project.

• Journal of the Korea Concrete Institute
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• v.18 no.2 s.92
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• pp.199-204
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• 2006

Recently, The sludge water of ready mixed concrete has been investigated because of environmental pollution and disposal cost. So, sludge water is partially reused as mixing water. However, if sludge water is reused too much that would influence the qualify of concrete. KS specification limits the amount of sludge content up to 3% of cement weight. In this study, the effect of ready mixed concrete sludge on the characteristics of concrete is compared to raise the reuse ratio of sludge of ready mixed concrete. According to this experiment results, as blending ratio of re-mi-con sludge increases, workability is decreases. However, the sludge of ready mixed concrete water have a positive effect on the strength development. The drying shrinkage and the resistance of freezing and thawing have a minor effect.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.98-98
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• 2003

Recently, human embryonic stem (hES) cells have become very important resources for ES cell basic research, cell replacement therapy, and other medical applications; thus, efficient cryopreservation methods for these cells are needed. This study examined whether a newly developed minimum volume cooling (MVC) vitrification method, which was tested through cryopreservation of sensitive bovine oocytes, can be used for freezing hES cells. Feeder-free cultured hES cell (MB03) colonies were mechanically dissected into several small clumps following enzymatic treatment. We compared the freezing efficiency of a slow-cooling method using a cryo-module (0.4-0.6C/min, 20-30 clumps/vial) and MVC vitrification using a modified 0.5-ml French mini-straw designated as a MVC straw (>$20,000{\circ}C$/min, 10 clumps/straw) After thawing, in vitro survival of hES cell clumps was higher for MVC-vitrified cells (80.8%, 97/120) than for slow-cooled cells (38.2%, 39/102). Further, the proliferation rate of surviving MVC-vitrified cells was similar to that of control hES cells from 2 weeks after thawing. In addition, vitrified-thawed hES cells demonstrated a normal karyotype, were positively immunostained for surface marker antibodies (AP, SSEA-4 and TRA-1-60) and the Oct-4 antibody, and could differentiate into all three embryonic germ layer cells in vitro. This result demonstrates that hES cell clumps can be successfully cryopreserved by a newly developed MVC vitrification method without loss of human cell characteristics.