The aim of present experiment was to examine commercial synthetic extender(AndroMed) for semen cryopreservation of Korean Black Bull. Semen was collected from a Korean Black Bull using an artificial vagina and transported to the laboratory. The semen was diluted 1:1 by AndroMed. The pellect was diluted to final sperm concentration of $5{\times}10^5/ml$ by doubling in every 10 minutes at $4^{\circ}C$ cold chamber. The semen was equilibrated for 1 hr at cold chamber and packed to 0.5 ml straw. The semen straws were located above 5 cm of liquid nitrogen for 5 minutes, above 5 cm for 10 minutes and above 10 cm for 10 min. And then the frozen straw was plunged to $LN_2$. The presented straws were examined the viability and motility after thawed at $37^{\circ}C$ water bath. Hanwoo semen was used as KPN (Korea Proven Bull Number) in this experiment. The survival rates was significantly higher in fresh semen than frozen semen ($80{\pm}14%\;and\;43{\pm}11%$). However, the motility rates was similar (80.7% and 66.4%). The survival and motility rates were higher in 5cm, 10 min treatment group than the other two groups in straw-located height and duration above $LN_2$ ($50{\pm}14%$ and 70.7% vs, 33.18% and $65{\pm}7%$ vs, 30.14% and 65.7%, respectively). The development rates to cleavage was higher in Black Cow than Hanwoo semen (62.2%, 64.4%), However, The development rates to blastocyst was higher in Hanwoo than Black cow semen (25.9%, 23.0%). In conclusion. The present results that acceptable fertilization and cryopreservation could be obtained by in vitro fertilization with frozen-thawed semen using a synthetic semen extender (AndroMed).
In woody species with a long life span, the studies on inheritance of any trait may be very time consuming and laborious. Chloroplast DNA(cpDNA) has been a valuable tool in such studies since it has several unique features such as limited genome size and cytoplasmic inheritance. In the present study, cpDNAs from five different species of Populus(P. alba, P. glandulosa, P. alba${\times}$P. glandulosa, P. davidiana, and P. nigra), and Nicotiana tabacum were compared with regard to restriction fragment length polymophism. The results showed that cpDNAs among the species were very conserved, although some polymorphisms were observed when the DNAs were digested with restriction enzyme EcoRI or KphI. The other enzymes (Bgl II, and PstI) tested produced identical restriction fragmentation pattern among the species. However, cpDNAs from all the five Populus species showed different restriction fragmentation pattern from that of tobacco with the four restriction enzymes tested. Southern hybridization with tobacco rbcL gene fragment as a probe also produced identical pattern among Populus species. The results indicate that cpDNAs in the genus are very well conserved during evolution.
By SDS-polyacrylamide gel elcctrophoresis (SDS-PAGE) and immunoblot analysis that a protein with 66,000 daltons in size was recognized by P. aeruginosa anti-exotoxin A from P. sp. PY002. The yields of exotoxin A in P. sp. PY002 culture were influenced by the concentration of iron in the culture media. Increasing of the exotoxin A production and siderophore production was made slight increasing in the MKB medium. On the other hand, to clone the gene encoding the exoloxin A genomic library of P. sp. PY002 was constructed in pBluescript SK(+). From this library a exotoxin A homologous gene was isolated by immunological hybridization method using P. aemginosa anti-exoloxin A as a probe. Two putative clones were isolated and designated pETA23 and pETA42. Thc restriction analysis ol pETA42 demonstrated that thc 1760 bp insert contained one NcoI, PvuII, SstI, Kpnl and EcoRI site and three SmaI and HaeD sites.
This study was to investigate pregnancy rate of IVM/IVF/IVC Korean cattle (registered in government) embryos according to transport time course. For the production of embryos, oocytes recovered from slaughtered excellent grade cow and highly motile frozen-thawed bull semen (purchased from LIMC, KPN#497) was used. In vitro produced embryos were cultured in CR1aa medium for 8 days and some of them were frozen. The rate of average cleavage (>2-cell) was 83.0% (308/371) and blastocyst rate at day 8 was 34.7% (107/308). Among in vitro produced blastocyst embryos at day 8, most healthy embryos were freshly transferred on production day and some frozen embryos were direct transferred on appropriate day. These embryos were produced in a laboratory, embryo transfer (ET) was planned in 10 areas of the remote island (Jeju) from the laboratory by airplane. Thus, we examined the pregnancy rate in recipient cow according to embryo of transport time course before ET. From embryo transferred 44 recipient cows, overall pregnancy was 40.9% (18/44), these 18 cows were all calved [single, 94% (17/18); twin, 6% (1/18)] and total embryo implantation rate was 26% (19/66). Comparing transport time in the base of 6 hr, pregnancy rate in ET group required less 4 hr (60%, 9/15) was significantly higher than that required more 6 hr (26.3%, 5/19). In direct ET of freezing embryos, the pregnancy rate was 40% (4/10). However, it was difficult to find the meaning of temperature, pH and corpus luteum quality of recipients on comparison of pregnancy rate. When the cell death level of embryos according to storage time in thermos (straw container) before ET was measured by TUNEL staining, apoptotic index was increased with storage time-dependent. These results demonstrated that long distance transfer of IVM/IVF/IVC embryos is possible and the time of embryo transport is very important for the pregnancy rate on field trial.
A gene coding for a novel putative $\sigma$ factor of RNA polymerase has been identified from Streptomyces coelicolor A3(2) using Escherichia coli rpoS gene fragment as a probe. The 486 bp rpoS gene fragment was amplified from E. coli genomic DNA by PCR with two synthetic oligonucleotides, the sequences of which were deduced from the amino acid sequences in the regions 2.3 and 4.2 conserved among various bacterial factors. When E. coli genomic DNA fragments were hybridized with cloned rpoS probe, only one band corresponding to rpoS gene (3.2 kb PvuII fragment or 2.3 kb KpnI fragment) was detected. In S. coelicolor, however, two bands were detected both in PvuII digested DNA and SalI digested DNA. 3.5 kb PvuII fragment which binds the rpoS gene probe was cloned (pMS1) from the sublibrary, and the nucleotide sequences of 1.0 kb BamH'/HincII subclone (pBH2) was partially determined. The nucleotide sequences revealed extensive similarity to other $\sigma$ factor genes of S. coelicolor (hrdA, hrdB, hrdC, hrdD), S. aureofaciens (hrdA, hrdB, hrdC, hrdD), Synechococcus species, Pseudomonas aeruginosa, Stigmatella aurantiaca, and Anabaena species. The nucleotide sequences in regions 1.2 and 4 were compared with the corresponding regions of 5 known ${\sigma}$ factor genes of S. coelicolor by multiple alignment. It turned out that the cloned gene is most closely related to hrdA showing 88% amino acid similarity in region 1.2 and 75% in region 4.
Diacylglycerol kinase (DGK) phosphorylates the second messenger diacylglycerol (DAG) to phosphatidic acid and it may play a role in signal transduction in Escherichia coli as well as in eukaryotic cells. In addition, DGK is important for microorganisms to adapt to several physiological stimuli. In Bacillus subtilis, the effect of stress on dgk transcription was examined by northern hybridization. The high level of dgk transcription was induced against high osmolarity, low pH value and low temperature. Transcriptional analysis revealed that the dgk gene and dgk upstream locus (ORF2, ORF3 and ORF4) were transcribed as a polycistronic mRNA to form an approximately 2.5 kb transcript.
Hyo Sang Lee;Yeongkuk Kim;Doo Ho Lee;Dongwon Seo;Dong Jae Lee;Chang Hee Do;Phuong Thanh N. Dinh;Waruni Ekanayake;Kil Hwan Lee;Duhak Yoon;Seung Hwan Lee;Yang Mo Koo
Journal of Animal Science and Technology
/
v.65
no.4
/
pp.720-734
/
2023
In Korea, Korea Proven Bulls (KPN) program has been well-developed. Breeding and evaluation of cows are also an essential factor to increase earnings and genetic gain. This study aimed to evaluate the accuracy of cow breeding value by using three methods (pedigree index [PI], pedigree-based best linear unbiased prediction [PBLUP], and genomic-BLUP [GBLUP]). The reference population (n = 16,971) was used to estimate breeding values for 481 females as a test population. The accuracy of GBLUP was 0.63, 0.66, 0.62 and 0.63 for carcass weight (CWT), eye muscle area (EMA), back-fat thickness (BFT), and marbling score (MS), respectively. As for the PBLUP method, accuracy of prediction was 0.43 for CWT, 0.45 for EMA, 0.43 for MS, and 0.44 for BFT. Accuracy of PI method was the lowest (0.28 to 0.29 for carcass traits). The increase by approximate 20% in accuracy of GBLUP method than other methods could be because genomic information may explain Mendelian sampling error that pedigree information cannot detect. Bias can cause reducing accuracy of estimated breeding value (EBV) for selected animals. Regression coefficient between true breeding value (TBV) and GBLUP EBV, PBLUP EBV, and PI EBV were 0.78, 0.625, and 0.35, respectively for CWT. This showed that genomic EBV (GEBV) is less biased than PBLUP and PI EBV in this study. In addition, number of effective chromosome segments (Me) statistic that indicates the independent loci is one of the important factors affecting the accuracy of BLUP. The correlation between Me and the accuracy of GBLUP is related to the genetic relationship between reference and test population. The correlations between Me and accuracy were -0.74 in CWT, -0.75 in EMA, -0.73 in MS, and -0.75 in BF, which were strongly negative. These results proved that the estimation of genetic ability using genomic data is the most effective, and the smaller the Me, the higher the accuracy of EBV.
Journal of the korean academy of Pediatric Dentistry
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v.30
no.2
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pp.196-203
/
2003
The application of sealants is a highly technique-sensitive procedure, requiring an extremely dry field prior to placement. Moisture contamination of the etched enamel surface before sealant placement is cited as the main reason for sealant failure. The purpose of this study was to evaluate the effects of different methods of sealant application on the shear bond strength of sealants to enamel. In groups 1, 2, 3, 4 Teethmate(unfilled sealant) was used, while Ultraseal XTplus(filled sealant) was used in groups 5, 6, 7, 8. Groups 1 and 5(control) were acid etched for 15 seconds using 35% phosphoric acid, washed and then dried. In groups 2, 6 drying agents were applied, and in groups 3, 7 bonding agents were applied and light cured. In groups 4 and 8 both drying agent and bonding agent were applied. Then sealant was cured to the specimen using molds 3mm in diameter and 2mm in height. Thermocycling was performed and shear bond strength was finally measured. The following results were obtained : 1. Groups using filled sealant(groups 5, 6, 7, 8) showed higher shear bond strengths compared to groups using unfilled sealant(groups 1, 2, 3, 4). 2. Among groups using unfilled sealant(groups 1, 2, 3, 4), groups 2, 3, 4 showed significantly higher shear bond strength compared to group 1(p<0.05). There were no significant differences among groups 2, 3 and 4. 3. There were no significant differences(p>0.05) among groups using filled sealant(groups 5, 6, 7, 8). 4. When modes of fracture were examined, cohesive failure was observed in groups 2, 3 and 4.
Hanwoo young bulls are selected based on performance test using the weight at 12 months and pedigree index comprising marbling score. Pedigree index was not based on the progeny tested data but the breeding value of the proven bulls; resulting a lower accuracy. The progeny testing of the young bulls was categorized into testing at farm and at the test station. The farm tested data was difficult to compare with those from test station data. Farm tested bulls had different slaughter ages than those for test station bulls. Therefore, this study had considered a different age at slaughter for respective records on ultrasound traits. Records on body weight at 12 months, ultrasound measures at 12 and 24 months(uIMF, uEMA, uBFT, and uRFT), and carcass traits(CWT, EMA, BFT, and MS) were collected from steers and bulls of Hanwoo national improvement scheme between 2008 and 2013. Fixed effects of batch, test date, test station, personnel for measurement, personnel for judging, and a linear covariate of weight at measurement were fitted in the animal models for ultrasound traits. The ranges of heritability estimates of the ultrasound traits at 12 and 24 months were 0.21-0.43 and 0.32-0.47, respectively. Ultrasound traits at 12 and 24 months between similar carcass traits was genetically correlated at 0.52-0.75 and 0.86-0.89, respectively.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.27
no.3
/
pp.239-249
/
2001
The purpose of this study is to evaluate the relationship of the factors which could be influenced by orthognathic surgery especillay SSRO. We measured the amounts of the maximum opening, lateral movements, maximum velocity and pattern of mandibular path during the opening and closing of mandible at the following times ; preoperative, 1 month after operation, 6 months after operation respectively using MKG. And the results were compared according to the categorized subgroups. Following results were obtained : 1. The change of the amounts of mandibular lateral movement and maximum opening velocity were statistically different between male and female (p<0.05), but the others were not. 2. According to the method of operation, there was no difference in the change of the mandibular movements between the group of SSRO and SSRO plus LeFort I osteotomy (p>0.05). 3. According to the amounts of mandibular movement, the recovery of left lateral movement of the group of $6{\sim}10mm$ was better than the other groups (p<0.05). 4. In the frontal pattern of the opening and closing of the mandible, the complex deflected type (F5), simple deflected type (F4), complex deviated type (F3), simple deviated type (F2), straight type (F1) were obtained in order at the time of preoperative, simple deflected type, simple deviated type, complex deviated type, straight type, complex deflected type in order at the time of 1 month after surgery, and the result at the time of 6 months after surgery was the same with that of the time of preoperative. In the sagittal pattern, non-coincident type (S2) was predominant at the time of preoperative, and coincident type (S1) was predominant at the time of 1 month after surgery. After 6 months, the result was also the same with that of the preoperative in sagittal pattern. 5. There was not a statistical difference in the change of the mandibular movement between group of presence of the preoperative TMJ symptoms and non-presence group (p>0.05). 6. There was not a statistical difference in the change of the mandibular movement between repositioning device applied group and non-applied group (p>0.05). 7. Sixty three percents of the patients who had preoperative TMJ symptoms were improved after surgery and preoperative TMJ symptoms were more improved after operation in the repositioning device non-applied group statistically (p<0.05).
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