• Asian-Australasian Journal of Animal Sciences
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• v.21 no.4
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• pp.494-498
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• 2008

MTT (3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) reduction assay is a method that validates the viability of an active cell. Dehydrogenase in mitochondria converts yellow colored insoluble tetrazolium salt to purple colored water-soluble formazan. Sperm also have mitochondria in the midpiece, therefore sperm viability could be evaluated by MTT reduction assay. Several studies have already demonstrated the capability of application of the MTT reduction assay to sperm of several species in Hepes-BSA buffer. Because most liquid semen was diluted in extender like BTS (Beltsville Thawing Solution), Modena or Androhep when it is used or transferred, semen needed another dilution in Hepes-BSA buffer to assess sperm viability. In this study, we evaluated boar sperm viability especially in BTS extended semen and compared the efficiency of this test with eosin-nigrosin staining. We used the fresh BTS extended semen from a local A.I center. Semen sample was diluted to $3.0{\times}10^7$ sperms/ml in BTS. The rates of formazan production were measured in 96-well microtiter plates immediately and 1h after incubation at $17^{\circ}C$ using a spectrophotometer at wave length 560 nm. Simultaneously, split samples of the same semen were tested, using eosin-nigrosin staining to compare the efficiency of the MTT assay of sperm viability in BTS. The correlation between the results of these tests was calculated using Student-t test and ANOVA. The results revealed a strong correlation between the results of MTT reduction rate and the results that were simultaneously determined by eosin-nigrosin staining at 1 h. In conclusion, the MTT reduction test was an effective and simple method to validate sperm viability and it could be used as a simple tool to evaluate sperm viability in the local A.I center and laboratory.

• Journal of Animal Reproduction and Biotechnology
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• v.34 no.3
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• pp.247-252
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• 2019

We observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. According to the results from each freezing extender, the sperm membrane integrity (HOST: Hypoosmotic Swelling Test) analysis in TCGGD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Glycerol 3%, Dimethylsulpoxide 3.5 M) is 59.8 ± 0.7, TCGSD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Sucrose 0.1 M, Dimethylsulpoxide 3.5 M) is 59.3 ± 0.5 were significantly higher (p < 0.05) among the experimental groups. And MMPs analysis result, we observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. The expression of active MMP-2 was the highest in sperms frozen in TCGSD and TCGD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Dimethylsulpoxide 3.5 M), Meanwhile, sperms from the TCGGD and TCGED (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Ethylene glycol 3%, Dimethylsulpoxide 3.5 M) group showed lower level of active MMP-2 expression. Together, these results indicate that adding glycerol or sucrose to the sperm freezing buffer would not only suppress MMPs expression but also minimize DNA fragmentation, providing a mean to improve the success rate in the in vitro manipulation of rabbit sperms. Therefore, these results suggest that TCGGD or TCGSD extender method for freezing-thawing of rabbit sperm increased the viability after thawing.

• Polymer(Korea)
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• v.34 no.5
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• pp.442-449
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• 2010

Polyurethane(PU) prepolymer was synthesized from ethylene oxide/propylene oxide(EO/PO) random polyether polyol, toluene diisocyanate and chain extender such as ethylene glycol and 1,4-butanediol. PU foams having various compositions were fabricated from PU prepolymers with different hard segment contents(%) and mixed foaming solution of different compositions. PU foam from chain extender-introduced PU prepolymer and mixed foaming solution containing glycerin showed better mechanical property than other groups. Various PU foams were tested on their mechanical property, moisture vapor transmission rate, absorption speed, absorptivity, morphology and cell culture test. According to the test, the PU foam fabricated from chain extender-introduced PU prepolymer and mixed foaming solution containing optimum composition of F-68, glycerin and CMC was found to have the best property for wound dressing materials. From in vivo animal study, it was confirmed that above PU foam showed rapid wound recovery.

• Polymer(Korea)
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• v.34 no.1
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• pp.8-13
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• 2010

The physical properties of polymeric foams depend on the density of foams, physical properties of base polymers, the content of open cells, and cell structures including the size and its distribution, the shape of cell, and the thickness of skin layer. The foam density is affected by the chemistry of raw materials, the concentration of crosslinking agent and the blowing agent as well as the operating parameters during production process. In this study, the basic formulations of foams are composed of polyester polyol, MDI, amine catalyst, tin catalyst, silicone surfactant, and water. Cross-linking density of polyurethane was increased by using chain extenders. Also, the mechanical properties of polyurethane foam were improved by using the inorganic fillers (silica 1,2 and talc 1,2) having different $SiO_2$ contents and particle sizes. We investigated the properties of modulus, tensile strength, compressive strength and hardness of foams obtained by changing kind of inorganic filler and chain extender, and observed the distribution of inorganic filler as well as variation of cell size within the foams by electron microscopy.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.1
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• pp.16-18
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• 2002

During the process of freezing, spermatozoa suffer cold shock which increases their susceptibility to lipid peroxidation which plays an important role in ageing of spermatozoa, shortening their life span and affecting the preservation of semen. An experiment was therefore conducted to study the effect of addition of natural antioxidants into semen diluents on the preservability of buffalo semen. Split semen samples were extended in milk egg yolk diluents fortified with vitamin E (MYE), vitamin C (MYC) and control group (MYO); Tris-egg yolk diluents fortified with vitamin E (TYE), vitamin C (TYC) and control group (TYO) and evaluated for their preservabilities at 4-7$^{\circ}C$ and $37^{\circ}C$. Overall least squares mean of percent motility observed after 0, 24, 48, 72 and 96 h of preservation at 4-7$^{\circ}C$ were 66.70, 54.00, 36.80, 21.90 and 12.50, respectively while the estimates for semen extended in MYE, MYC, MYO, TYE, TYC and TYO were 44.80, 42.70, 38.70, 36.00, 35.20 and 33.00 percent, respectively. The results showed that motility was significantly (p<0.01) affected by extender (extender-antioxidant combination) and preservation interval. Overall least squares mean percent motility observed after 0, 4, 8, 12 and 24 h of preservation at $37^{\circ}C$ were 68.50, 58.90, 45.00, 38.10 and 18.10 percent, respectively, while the estimates for semen extended in MYE, MYC, MYO, TYE, TYC and TYO were 48.20, 49.30, 46.80, 45.30, 42.30 and 42.50 percent, respectively. Extender and storage interval were found to be significantly (p<0.01) affecting spermatozoa motility on room temperature preservation. The results indicated that the incorporation of antioxidants, especially vitamin E, had beneficial effect on preservability of buffalo semen.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.12
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• pp.1716-1721
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• 2006

The present study was conducted to observe the effect of osmolality of glycerolated TEST-yolk glycerol extenders on post-thawing sperm kinematics of ram spermatozoa of the native Malpura breed maintained in a semi-arid tropical environment. Good quality semen obtained from adult rams was pooled, split and diluted to 1,000 million spermatozoa per ml in complete TEST-yolk-glycerol extenders of 900, 1,200, 1,500 and 1,800 mOsm/kg osmolality. Diluted semen samples were loaded in 0.25 ml straws and cooled down to $-125^{\circ}C$ freezing temperature at the rate of $-25^{\circ}C$ per minute under controlled conditions before plunging into liquid nitrogen for storage. The thawing of straws was performed at $50^{\circ}C$ in a water bath for 10 seconds and sperm kinematics of the frozen-thawed spermatozoa were assessed by a computer-assisted sperm analysis technique. Osmolality of diluent had no significant effect on post-thawing % motility, % rapid, % medium and % slow moving frozen-thawed spermatozoa but significantly (p< 0.05) affected the % linearity and % straightness. The post-thawing % motility and % rapid motile spermatozoa were highest in samples extended in diluent of 1,500 mOsm/kg osmolality and lowest in 900 mOsm/kg. The curvilinear velocity of spermatozoa was significantly (p<0.05) higher for samples extended in 1,800 mOsm/kg, compared to those in 900 and 1,200 mOsm/kg, but the effect was not significantly different to those extended in diluent of 1,500 mOsm/kg osmolality. The study indicated that ram spermatozoa could tolerate a wide osmolality range for dilution in the complete TEST-yolk-glycerol extender for their cryosurvival. The highest recovery of motile spermatozoa following thawing was achieved in samples extended in the TEST-yolk-glycerol diluent of 1,500 mOsm/kg osmolality.

• Journal of the Korean Wood Science and Technology
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• v.12 no.2
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• pp.35-43
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• 1984

This study was carried out to investigate the block shear strength of the 2ply laminated wood composed of Pinus thunbergii with ureaformaldehyde resin as adhesives according to pressure (6, 9, 12, 15kg/$cm^2$), pressing time(5, 10, 20, 30 hrs.), amount of spread(54, 108, 217, 324g/$cm^2$), hardener(10, 20, 30, 40% of 10% $NH_4CL$ on resin) and extender(0, 5, 10, 15% wheat flour on resin), and the bending strength and bending young's modulus of laminated beam according to the number of ply. The results were summarized as follows; 1. According to pressing pressure with amount of spread 217g/$cm^2$ both dry and wet shear strength of laminated wood showed the highest in 15kg/$cm^2$, and hot-cold soaking treatment showed the highest in 9kg/$cm^2$, while all shear strength of dry, wet and hot-cold soaked laminated wood have been reduced with the increasing of pressing time. 2. According to amount of spread, adhesion strength with the dry, wet and hot-cold soaking treatments revealed the highest in 217g/$cm^2$ and have been reduced under or over 217g/$cm^2$ of spread. 3. According to addition of hardener and extender, all shear strength of laminated block with the dry, wet and hot-cold soaking treatments have been reduced in increasing of addition amount of hardener and extender. The bending strength of beam according to the number of ply showed the highest in 2ply laminated wood and horizontal loading beam to glue line had the higher in strength than the vertical loading.

• Journal of Embryo Transfer
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• v.23 no.2
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• pp.115-118
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• 2008

Glycerol is the cryoprotectant most frequently used to freeze semen in several of species. The objective of the present study was to compare the effect of three different glycerol concentrations (4, 6 or 8%, v/v) on frozen-thawed dog sperm survival rate. Ejaculates from 9 dogs collected by digital manipulation were pooled and assessed by macroscopic and microscopic criteria. Semen was divided into 3 aliquots, which were centrifuged and the sperm pellets rediluted with first Tris-glucose-citric acid extender. After 1 h cooling at $4^{\circ}C$, second extender containing 4, 6 or 8% glycerol was added, respectively. The semen was loaded into 0.25 ml straws and frozen and stored in liquid nitrogen and thawed. Sperm vigor, live:dead spermatozoa ratio using HOS test, and sperm morphology using $Spermac^{(R)}$ stain were evaluated. After thawing, there were no significant differences among groups in vigor, viability and morphology. In conclusion, the three glycerol concentrations (4, 6 or 8%) can be used successfully in cryopreservation of canine semen. Therefore the use of 4% glycerol in the extender has less toxic effect and reduces of freezing injuries.

• Journal of the Korean Wood Science and Technology
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• v.1 no.2
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• pp.3-10
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• 1973

This study was carried out to examine the characeristics of waste sanders(S)from. plywood and pre-finishing plywood surface sanding and double saw finishing, as a new extander in urea-formaldehyde resin(UFR) in plywood adhesive, and to focus, adhesive strength using the glue extended with milled sanders(MS) as extender, leveling the optimum amount of MS to be added, and examining the physical properties of glue extended MS & S. Also economical good feasibility of substitution for wheat flour(WF) with MS as a new extender is analyzed and presented in details. Selecting three standard samples of 80, 100 and 180 mesh, sorking them in distilled water at $20^{\circ}C$, 24 hours, redrying at $105^{\circ}C$ and rescreening the sample with standard screen, again, the 3 samples of 80, 100, and 180 mesh are passed 23 percent through 80 mesh sander standard sample 27 percent through on 100 mesh and only 10.9 percent through 180 mesh, respectively. The particle size of retained parts are greater in size of redried form. It seems undoubtly that particles to be extended in glue are got swollen and become greater in size and coarser in shape. The shape of fresh S particles are irregular thin needle with small scale, as shown in Figure 5. PFS are so finer than plywood S that only 9.8 percent of the S retained on 100 mest screen, 24. 30 percent on 100-160 mesh, and 65.9% on 160-180 mesh. But particle size of the fresh S is large enough to make the viscosity of glue direct extended with S too high to apply it glue spreader. The glue extended with milled sanders(MS). 3 hours milled PFS or 6 hours milled plywood S, having particle sizes shown in Tables 7 and 8, as ratio of Reain/MS/WF/water: 100/8/8/10, indicate good viscosity of 16 to 24 ps, as shown in Figure 5, for applying direct to glue spreader, have high tensile-shear strength (adhesive strength), 102.4 kp/$cm^2$, and 94 percent wood failure.

• Korean Journal of Veterinary Research
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• v.40 no.2
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• pp.403-414
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• 2000

To improve the semen production, the selenium(Se) and vitamin E(Vit. E) were administrated into Hanwoo young sire for intensification an antioxidant system and the taurine were supplemented into semen extender for improving the semen characteristics. The 16 heads ranging from twenty to thirty two months of age were randomly assigned to control group, Se-admistrated group(Se-group), Vit. E-administrated group(Vit. E-Group) and Se and Vit. E administrated group(Se and Vit. E-group). Se and Vit. E dministrated 3 times every 30 days by intramuscular injection. The administration of Se, Vit. E, and Se and Vit. E didn't affect on semen volume, sperm concentration, and total sperm number among all groups(p>0.05). Adiministration of Se improved sperm motility and viability. Motility of Se-group and control were 26.01% and 19.20%, respectively(p<0.05). Viability of Se-group(47.07%) was higher than control group(35.73%), Vit. E group(36.55%)(p<0.05). The administration of Se and Vit. E didn't affect sperm capacitation and acrosome reaction. The 100mM taurine supplement into semen extender increased the motility of frozen/thawed semen in the Vit. E-group(p<0.05) and had a beneficial effects on decreasing abnormality of frozen/thawed semen in all groups(p<0.05). These results indicate that Se administration improve sperm motility and viability, and the taurine supplement into semen extender decrease abnormality in Hanwoo young sire.