• Journal of Embryo Transfer
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• v.23 no.3
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• pp.197-201
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• 2008

In order to provide information of genetic variants for Haptoglobin (Hp) gene, which may be related to weight traits in pig, a total of 235 animals from National Institute of Animal Science (NIAS) were screened with 3 primers. The primer sequences were selected using the porcine cDNA sequences based on NM_214000, and the exon boundaries were estimated. Genetic variants were observed using direct sequencing analysis, and there were 9 SNPs detected at nucleotide positions 503 (A/G), 509 (A/G), 709 (C/T), 734 (C/A), 742 (G/A), 769 (A/G), 840 (C/T), 876 (C/T) and 882 (C/A). All the SNPs were located in coding regions, and mutations caused amino acid changes at nucleotide positions 503, 509, 734, 742 and 769. Allele frequencies of SNPs were estimated for all segments. The SNPs at nucleotide position 509 (p<0.0001) and 734 (p<0.05) were significantly associated with average daily gain, but no significance was observed with other SNPs. From the results, the identified SNPs may be a useful candidate marker for the porcine weight gain traits.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.7
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• pp.917-922
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• 2008

A disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif (ADAMTS1) plays a critical role in follicular rupture and represents a major advance in the proteolytic events that control ovulation. In this study, a 9,026-bp DNA sequence containing the full coding region, all 8 introns and part of the 5'and 3' untranslated region of the porcine ADAMTS1 gene was obtained. Analysis of the ADAMTS1 gene using the porcine radiation hybrid panel indicated that pig ADAMTS1 is closely linkage with microsatellite marker S0215, located on SSC13q49. The open reading frame of its cDNA covered 2,844 bp and encoded 947 amino acids. The coding region of porcine ADAMTS1 as determined by sequence alignments shared 85% and 81% identity with human and mouse cDNAs, respectively. The deduced protein contained 947 amino acids showing 85% sequence similarity both to the human and mouse proteins, respectively. Comparative sequencing of three pig breeds revealed one single nucleotide polymorphism (SNP) within exon 7 of which a G-C substitution at position 6006 changes a codon for arginine into a codon for proline. The substitution was situated within a PvuII recognition site and developed as a PCR-RFLP marker for further use in population variation investigations and association analysis with litter size. Allele frequencies of this SNP were investigated in seven pig breeds/lines. An association analysis in a new Qingping female line suggested that different ADAMTS1 genotypes have significant differences in litter size (p<0.01).

• Reproductive and Developmental Biology
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• v.28 no.2
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• pp.77-82
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• 2004

The purpose of this study was performed to establish the in vitro culture system of porcine in vitro maturation and in vitro fertilization(IVM/IVF) embryo. These studies was to determine the effects of antioxidants(aesculetin, taurine and melatonin) in porcine IVM/IVF embryos. In routine porcine IVM/IVF procedure, oocytes were cultured for 40∼44h incubation in NCSU 23 mediumand matured oocytes were inseminated with frozen semen. Then 2 to 8 cell embryos were removed cumulus cell and were allotted randomly to NCSU 23 containing different concentration of antioxidants in 5% $O_2$ and 5% $CO_2$ at 38.5$^{\circ}C$. Cell numbers of blastocyst were also counted using double fluorescence stain method. Aesculetin were added to NCSU 23 medium at concentration of 1 ug, 5 ug, and 10 ug, when treated with 10 ug(35.7%) of aesucletin at the rate of embryos of the morula plus blatocsyts were higher than those of any other groups (30.2%, 29.5% and 29.2%)(P<0.05). The developmental rates beyond morula stage of porcine embryos in NCSU 23 medium supplemented with taurine 0, 2.5 and 5.0 mM were 26.1%, 26.9% and 31.7%, respectively. The addition of 5.0 mM taurine was higher the developmental rate beyond morula stage than in any other groups. In NCSU 23 medium treated with melatonin 0, 1, 5 and 10 nM, the developmental rate of morula plus blastocysts were 33.3%, 39.1%, 33.3% and 27.9%, respectively. The developmental rate of morula and blascytocys treated with 1nM melatonin was higher than in any other groups(P<0.05). Cell numbers of blastocyst in NCSU 23 treated with melatonin 0, 1, 5 and 10nM were 41.0, 42.6, 39.6 and 33.0, respectively. These results indicate that aesculetin, taurine and melation can increase the developmental rate beyond the morulae and blastocysts in porcine embryos.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.9
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• pp.1257-1261
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• 2006

Pleiomorphic adenoma gene-like1 (PLAGL1) encodes a zinc-finger (ZF) protein with seven ZFs of the C2H2-type which is a regulator of apoptosis and cell cycle arrest, and also regulates the secretion of insulin. In both human and mouse, PLAGL1 is a candidate gene for tumor suppressor and transient neonatal diabetes mellitus (TNDM). In this study, a 2,238 bp fragment covering the complete coding region was obtained and deposited to GenBank (accession number: DQ288899). The reverse transcriptase-polymerase chain reaction (RT-PCR) indicated that PLAGL1 was expressed almost equally in heart, liver, spleen, lung, kidney, stomach, small intestine, skeletal muscle, fat, uterus and ovary. Comparing the sequences of Large White and Meishan pigs, a C-T transition in exon 6 was found. The polymorphism could be detected by TaqI and was genotyped in five purebreds (Large White, Landrace, Meishan, Tongcheng and Bamei). Association analysis was performed between the polymorphism and carcass traits in 276 pigs of a "Large White${\times}$Meishan" F2 resource population. As a consequence, significant associations of the genotypes with shoulder backfat thickness (SFT) and internal fat rate (IFR) were observed. Pigs with TT genotype had low SFT and high IFR compared with TC or CC genotypes.

• Fisheries and Aquatic Sciences
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• v.12 no.1
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• pp.16-23
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• 2009

Lipids were extracted from marine rotifer, Brachionus rotundiformis in order to examine the functionality of lipid enzymatic modification. The fatty acids, palmitic, linoleic, oleic and stearic acids were the dominant forms accounting for approximately 35.8%, 21.5%, 15.9% and 7.7% of the total lipid content, respectively. Lipid fractions were categorized as neutral lipids (38.5%), glycolipids (45.9%) and phospholipids (17.6%), and after extraction from the rotifer were isolated by thin-layer chromatography (TLC) as free fatty acids (FFA), monoacylglycerol (MAG), diacylglycerol (DAG) and triacylglycerol (TAG). The production of polyunsaturated fatty acid (PUFA) concentrate from rotifer lipids was studied using lipase-catalyzed hydrolysis. In addition, rotifer lipids were modified by hydrolysis using lipases such as porcine pancreas, Candida rugosa and Rhizomucor miehei. The lipase from Rhizomucor miehei was effective in extracting linoleic acid (C 18:2), while the lipase from Candida rugosa was effective in palmitic acid (C16:0) extraction.

• Korean Journal of Agricultural Science
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• v.23 no.2
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• pp.206-211
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• 1996

This study was conducted to investigate the effects of medium and cumulus cell on in vitro fertilization of porcine follicular oocytes. The results obtained are as follows ; 1. The normal fertilization rates of in vitro matured follicular oocytes cultured in 00, mT ALP and TCM-HEPES medium were 14.0~24.3%, 30.8~32.7% and 21.4~23.9%, respectively. These data indicated that the optimal medium for fertilization of porcine oocytes in vitro was the mTALP medium 2. The normal fertilization rates of epididymal sperm were 24.3%(80), 30.8%(mTALP) and 23.9%(TCM-HEPES), and those of ejaculated sperm were 14.0%(B0), 32.7%(mTALP) and 21.4%(TCM-HEPES). 3. The sperm penetration rates of cumulus-enclosed and cumulus-free oocytes on in vitro fertilization were 54.0% and 72.0%. The normal fertilization rates of cumulus-enclosed and cumulus-free oocytes were 11.9% and 21.5%. The normal fertilization rate of cumulus-enclosed oocytes was significantly(P<0.05) higher than that of cumulus-free oocytes.

• BMB Reports
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• v.37 no.6
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• pp.684-690
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• 2004

Heparin lyase I was purified to homogeneity from Bacteroides stercoris HJ-15 isolated from human intestine, by a combination of DEAE-Sepharose, gel-filtration, hydroxyapatite, and CM-Sephadex C-50 column chromatography. This enzyme preferred heparin to heparan sulfate, but was inactive at cleaving acharan sulfate. The apparent molecular mass of heparin lyase I was estimated as 48,000 daltons by SDS-PAGE and its isoelectric point was determined as 9.0 by IEF. The purified enzyme required 500 mM NaCl in the reaction mixture for maximal activity and the optimal activity was obtained at pH 7.0 and $50^{\circ}C$. It was rather stable within the range of 25 to $50^{\circ}C$ but lost activity rapidly above $50^{\circ}C$. The enzyme was activated by $Co^{2+}$ or EDTA and stabilized by dithiothreitol. The kinetic constants, $K_m$ and $V_{max}$ for heparin were $1.3{\times}10^{-5}\;M$ and $8.8\;{\mu}mol/min{\cdot}mg$. The purified heparin lyase I was an eliminase that acted best on porcine intestinal heparin, and to a lesser extent on porcine intestinal mucosa heparan sulfate. It was inactive in the cleavage of N-desulfated heparin and acharan sulfate. In conclusion, heparin lyase I from Bacteroides stercoris was specific to heparin rather than heparan sulfate and its biochemical properties showed a substrate specificity similar to that of Flavobacterial heparin lyase I.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.11
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• pp.1651-1654
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• 2007

The mouse myxovirus resistance protein 1 (Mx1) is known to be sufficient to confer resistance to influenza viruses, and the gene encoding Mx1 is, therefore, an interesting candidate gene for disease resistance in farm animals. The porcine Mx1 gene has already been identified and characterized based on its homology with mouse Mx1; the full-length coding region of the pig Mx1 gene spans 2,545 bp (M65087) and is organized into 17 exons compared with the human ortholog mRNA. In this study, the exons 9, 10 and 11 and introns 6 and 9 of the porcine Mx1 gene were cloned and sequenced. Two SNPs were identified in exons 9, 10 and 11 but none of the SNPs led to an amino acid exchange, and the other eleven variants were detected in introns 6 and 9, respectively. Differences in allele frequency between Meishan and other pig breeds were observed within intron 6, of which an $A{\rightarrow}G$ substitution at position 371 was detected as an SnaBI PCR-RFLP. The association analysis using the Large White${\times}$Meishan $F_2$ offspring suggested that the Mx1 genotype was associated with variation in several immunity traits that are of interest in pig breeding. However, further investigations in more populations are needed to confirm the above result.

• Journal of Embryo Transfer
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• v.32 no.1
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• pp.9-15
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• 2017

The present study was aimed to determine the effect of green tea extract (GTE) and beta-mercaptoethanol (${\beta}$-ME) supplementation in boar sperm freezing extender on sperm motility, viability and reactive oxygen species (ROS) level. Experimental groups were allocated into Lactose-egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/L GTE in LEY) and ${\beta}$-ME ($50{\mu}M$ ${\beta}$-ME in LEY). Spermatozoa extended with LEY were cooled to $5^{\circ}C$ for 3 h and then kept at $5^{\circ}C$ for 30 min following dilution with LEY containing 9% glycerol and 1.5% Equex STM (final sperm concentration: $1{\times}10^8/mL$). Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. Following thawing at $37^{\circ}C$ for 25 sec, sperm viability and ROS level were measured using fluorescent double stain Fertility(R) and cytometry, respectively. Motility and viability of GTE supplemented-group were higher than those of control and ${\beta}$-ME without significance. ROS level in GTE group showed significantly lower than control (P < 0.05). In conclusion, GTE supplementation in boar sperm freezing extender can reduce ROS generation during freezing.

• The Journal of the Korean bone and joint tumor society
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• v.1 no.1
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• pp.7-16
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• 1995

In countries where confucianism is popular, it is extremely hard to get fresh cadaver bone for allograft. Therefore in Korea, the reimplantation of resected autoclaved autogenous bone segments has been increasingly performed for limb reconstruction of extremities with malignancies. To preserve the bone morphogenetic protein and mechanical strength of heated bone, many studies have reported that pasteurization of bone is far better than autoclaving over $100^{\circ}C$. Based on this assumption, replacement with a pasteurized autogenous bone graft after resection of a malignant bone tumor was deemed feasible for reconstruction. However, little is known about how high a temperature and how much time for pasteurization is needed to make tumors completely necrotic and to maintain the mechanical strength of bone. Consequantly, experimental studies were carried out to test heat conductivity of human bone and torsional strength of porcine tibia after pasteurization. First, two pairs of human proximal tibia and distal femur were used. We used T-type thermocoples to check core temperature of the bone and a computerized data acquisition system to record results. Without reaming of the medullary cavity, in a $60^{\circ}C$-thermostatic saline tub, it took 32 minutes and 50 seconds to raise the core temperature of human proximal tibia from $20^{\circ}C$ to $58^{\circ}C$, and 30 minutes for distal femur. In a $80^{\circ}C$ saline tub, it took 12 minutes and 50 seconds for proximal tibia, and 11 minutes and 10 seconds for distal femur. In contrast, using porcine tibia whose cortical thickness is similar to that of human tibia, after reaming of the medullary canal, it took less than 3 minutes and 30 seconds in a $60^{\circ}C$ saline tub, less than 1 minute and 45 seconds in a $70^{\circ}C$ tub, and less than 1 minute in a $80^{\circ}C$ tub to elevate core temperature from $20^{\circ}C$ to $58^{\circ}C$. Second, based on data of the heat conductivity test, we compared the torsional strength before and after pasteurization. Twenty matched pairs of porcine tibia were used, The left one was used as a non-heated control group and the right one as a pasteurized experimental group. Wighout reaming of the medullary cavity, there was no statistical difference in torsional strength between the pasteurization of the $60^{\circ}C$-35minute and of $80^{\circ}C$-15minute. With reaming, we also found no statistical difference among pasteurization of $60^{\circ}C$-15 minute, of $70^{\circ}C$-15 minute, and of $80^{\circ}C$-15 minute groups. In conclusion, reaming of the medullary canal is very helpful in saving pasteurization time. And, in a $60^{\circ}C$ saline tub, no significant weakness in torsional strength occurs with pasteurization of the bone for up to 35 minutes. Also no significant weakness in torsional strength occurs with an exposure of 15 minutes to the $80^{\circ}C$ saline tub.