• Journal of Plant Biotechnology
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• v.49 no.3
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• pp.187-192
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• 2022

The repeated monthly or weekly subculture of plant callus is labor intensive and increases the risk of somaclonal variation from the parental callus line. The most effective method for preserving plant callus is cryopreservation, which involves storage in liquid nitrogen. However, this method cannot be applied to the callus of different plant species in the same manner, so it is difficult to develop a standardized cryopreservation method. In addition, the survival rate of the frozen callus after thawing and the regeneration rate after survival are uncertain. Therefore, it is necessary to develop a method to extend the subculture interval of plant callus in an active state. In this study, active plant calli of various species without freezing was incubated at 15℃ for 4 to 12 weeks without subculture. After 12 weeks, 8 lines of plant callus grew less than 2-fold when cultured at 25℃, but at least 2 times as much when cultured at 15℃. Moreover, total antioxidant activity did not differ significantly between plant callus recovered at 25℃ after culturing at 15℃ or at 25℃. These results show that the subculture interval can be extended at a temperature of 15℃ without need for modified medium composition or additional processes. In addition, positive results in all calli of several plant species are expected to reduce labor as well as somaclonal variation by increasing the subculture.

• Journal of Conservation Science
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• v.36 no.6
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• pp.494-504
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• 2020

Ten historic sites (denoted as A- J in this study) of a tomb were found during the construction of the east-west expressway in District 2 of Hyangnam, Hwaseong, which is implemented by the Gyeonggi-do headquarters of the Korea Land & Housing Corporation. Thetombswere first detected at siteH, and further investigations revealed various tombs from the Three Kingdoms period; artifacts such as gilt-bronze shoes and caps were excavated from wooden coffins in the tombs. The pottery examined in this study was the only pottery artifact excavated from the site. Its raw clay was soft and loose, reddish brown, and had quartz and feldspar particles of < 1 mm, which appeared to have been added as reinforcing agents. The firing temperature of the pottery was estimated to be under 800-870 ℃ as the mica remains and tridimite, which is the phase transition mineral of quartz, was not produced; a slight endothermic peak was also detected because of the hydration of sericite at 800 ℃. The condition of the artifact was severely weakened because of various factors, such as soil pressure from the stratum formed over the site and repeated freezing and thawing. The artifact could not be collected alone, and thus, surrounding soil that had attached to the artifact was also collected; the artifact was transported to the laboratory and conservation treatment was conducted in a safe and systematic manner.

• Animal Bioscience
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• v.35 no.12
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• pp.1827-1838
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• 2022

Objective: The semen quality of stallions including sperm motility is an important target of selection as it has a high level of individual variability. However, effects of the molecular architecture of the genome on the mechanisms of sperm formation and their preservation after thawing have been poorly investigated. Here, we conducted a genome-wide association study (GWAS) for the sperm motility of cryopreserved semen in stallions of various breeds. Methods: Semen samples were collected from the stallions of 23 horse breeds. The following semen characteristics were examined: progressive motility (PM), progressive motility after freezing (FPM), and the difference between PM and FPM. The respective DNA samples from these stallions were genotyped using Axiom Equine Genotyping Array. Results: We performed a GWAS search for single nucleotide polymorphism (SNP) markers and potential genes related to motility properties of frozen-thawed semen in the stallions of various breeds. As a result of the GWAS analysis, two SNP markers, rs1141327473 and rs1149048772, were identified that were associated with preservation of the frozen-thawed stallion sperm motility, the relevant putative candidate genes being NME/NM23 family member 8 (NME8), olfactory receptor family 2 subfamily AP member 1 (OR2AP1), and olfactory receptor family 6 subfamily C member 4 (OR6C4). Potential implications of effects of these genes on sperm motility are herein discussed. Conclusion: The GWAS results enabled us to localize novel SNPs and candidate genes for sperm motility in stallions. Implications of the study for horse breeding and genetics are a better understanding of genomic regions and candidate genes underlying stallion sperm quality, and improvement in horse reproduction and breeding techniques. The identified markers and genes for sperm cryotolerance and the respective genomic regions are promising candidates for further studying the biological processes in the formation and function of the stallion reproductive system.

• Journal of The Geomorphological Association of Korea
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• v.17 no.4
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• pp.71-83
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• 2010

In this study, the volume of shoreline retreat at sea-cliffs in the Taean peninsula(West Coast of Korea) was estimated and their erosion and seasonal landforms characteristics changes were investigated through multi-temporal aerial photographs and field survey. Based on the analysis of aerial photographs through ortho-correction, the results show that the length of shoreline and erosion area increase as erosion at sea-cliffs occurs in Pado-li and Dundu-li. To obtain the seasonal quantitative landforms changes and retreat of sea-cliffs, we marked top, middle, and bottom datum-points, from which the distance to the nearest bedrock was repeatedly measured. In these regions, the retreat of sea-cliffs gradually increases in spring to summer, but gradually decreases in autumn. In particular, the typhoon that has a great influence on the Korean peninsula in July to September in summer would drastically increase the retreat of sea-cliffs in comparison with other seasons. As the outcrop of sea-cliffs repeats freezing and thawing in winter, the retreat of sea-cliffs increases a little due to active mechanical weathering. To know the erosion and seasonal landforms changes of sea-cliffs, we took pictures of them in every month and then analyze their condition. The retreat of sea-cliffs was repeatedly occurred by the circulation of the erosion of sea-cliff base, landslides, the formation of slope sediment debris and their erosion, in that order.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.72-72
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• 2003

The present study examined the possibility of cryopreservation of the D-shaped and umbo larvae of arkshell (Scapharca broughtonii), in terms of the survival rates after freezing and thawing. D-shaped and umbo larvae of arkshells were obtained from a shellfish farming on Yosu city. The average shell lengths were $93.3 \pm 10.1 \mu$m and $201.7 \pm 13.5 \mu$, respectively. Five cryoprotectants (CPAs), dimethyl sulfoxide (DMSO), glycerol, ethylene glycol (EG), propylene glycol (PG), and methanol, were tested at the concentrations of 1.5, 2.0 and 2.5 M. After larvae suspended in CPAs, cryoprotectants were loaded in 0.5 ml straws at a larval density of 50-100 larvae per straw, and epuilibrated for 10 and 20 minute at room temperature ($23^{\circ}C$), repectively. Straws were cooled at a rate of $1^{\circ}C$/min from $0^{\circ}C$ to $-12^{\circ}C$, held for 5 min at $-12^{\circ}C$, and then cooled at $2^{\circ}C$/min to $-35^{\circ}C$ and equilibrated for 5 min followed by plunging in liquid nitrogen. After storage in liquid nitrogen for 1 day, straws were thawed in a $30^{\circ}C$ water. As soon as straws were observed to melt, larvae were diluted with an equal volume of ASW and then washed twice with a large volume of ASW at an interval of 2 min to unload the CPAs. The results showed that after equilibration for 10 and 20 minute at room temperature, no larvae survived using methanol as CPAs, and it was observed that larval shells all open slightly, and larval flesh broke down and slopped over the shells. The highest survival rates (D-shaped larvae: 77.6%, umbo larvae: 59.3%) were obtained with 2M DMSO, and 1.5M glycerol yielded survival rates of 53.8% for D-shaped larvae and 37.5% for umbo larvae. The surviving D-shaped larvae showed active rotary motion and perfect membrane integrity and cytoplasmic normality, and the vigorous movement of veliger cilia was observed inside the closed shells. The breakdown of tissue occurred in the abnormal larvae, and the isolated cell often run out of shells.

• Journal of the Korean Geotechnical Society
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• v.22 no.10
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• pp.77-91
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• 2006

Domestic practices in shotcrete use have developed in many respects even now, but it still has issues about material, construction, quality standard and so on. In overseas, the construction using high strength shotcrete with $39.2{\sim}58.8 MPa$ of compressive strength is becoming common based on the shotcrete technology of high strength and durability. However, domestic shotcrete design strength is low at around 20.6 MPa of compressive strength and a long term durability is also insufficient. In this paper, field tests using high-quality additives and accelerators were performed to obtain the improvement of shotcrete strength and EFNARC standard was used to evaluate the field test results. In addition, deterioration test combined with the freezing-thawing and carbonation was also performed in order to investigate a long-term durability of high-strength shotcrete. As a result of the field test, the promotion ratio of early strength was $90{\sim}97%$ in case of using alkali-free accelerators. And the compressive strength of the shotcrete using Micro-silica fume was $45.2{\sim}55.8MPa$ and flexible strength was $5.01{\sim}6.66MPa$, so the promotion ratio of strength was $37{\sim}79%$ and $17{\sim}61%$ respectively. The promotion effect of strength by silica fine additives ratio of $7.5{\sim}10%$ for cement mass was much superior to the other cases. It was especially examined that using Micro-silica fume reduced deterioration due to mixed steel fiber and improved a long-term durability of shotcrete.

• Journal of the Korean Geotechnical Society
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• v.40 no.3
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• pp.93-100
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• 2024

Pore water freezes in low-temperature compaction, which leads to different compaction characteristics compared to room temperature conditions. In regions like Alberta, Canada, where organic soils are prevalent, compaction performance is influenced by the high water retention and compressibility of organic soils, as well as their sensitivity to freezing and thawing. Alberta's strict environmental regulations demand the reuse of excavated soil for backfill, and the long winter season creates challenging conditions for civil engineering projects. In this study, a laboratory compaction test was conducted to evaluate the low-temperature compaction characteristics of organic soils with varying organic content. The results indicate that the optimum moisture content increases as the organic content increases, and the maximum dry unit weight decreases by up to 21.9%. In addition, under temperature conditions below -4℃, no optimum moisture content was observed, and the dry unit weight decreased as the moisture content increased.

• Journal of the Korean Geographical Society
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• v.43 no.3
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• pp.296-311
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• 2008

In order to examine the rates and factors of path widening in Mount Halla, the retreat of path sidewalls was monitored at 32 sites of Seongpanak Hiking Trail located between 875 m and 1,400 m in elevation. The mean rate of sidewall retreat for the period 2002-2008 is 50.6 mm, equivalent to 10.0 mm/yr. The retreat rate of frozen period is 19.3 mm/yr, while the rate of unfrozen period is 4.3 mm/yr. The latter is divided into the rainy and dry periods that exhibit the retreat rates of 5.9 mm/yr and 2.9 mm/yr, respectively. The retreat rate of sidewalls is also varied with seasons; winter shows the maximum rate of 42.2 mm/yr, while summer exhibits the minimum rate of 1.3 mm/yr. Spring and fall show the intermediate rates of 13.9 mm/yr and 6.4 mm/yr, respectively. Soil hardness and elevation are not closely related to the retreat rate of sidewalls, even though the retreat rate is larger at the north-faced sidewalls than the south-faced sidewalls during the frozen period. Pipkrake is likely to be the most important factor contributing to the path widening in that the retreat of winter months accounts for 76.7% of the total retreat. The hiking trail is placed under the climatic conditions which develop pipkrake in 85 days annually. In addition, it is usual to observe the path sidewall covered with pipkrake in the freezing month of December and the thawing months of March and April. On the other hand, deflation and rainsplash erosion are not important due to the weak wind speed and the forested trail. Rainwash is also insignificant in that the path has been almost paved to mitigate trampling effects. Although biological activity is not dominant, hikers cause a large retreat of sidewalls in the thawing months since they would walk on the sidewalls to avoid snow-melting pools on the path.

• Journal of Veterinary Clinics
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• v.30 no.6
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• pp.442-448
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• 2013

The aim of the present study was to develop glycerol-free TRIS extender using glucose for dog sperm cryopreservation. We determined the appropriate concentration of glucose in glycerol-free TRIS and the exposure time in glycerol-free TRIS containing 0.3 M glucose at $4^{\circ}C$. Ejaculates of six dog sperm were cooled in glycerol-free TRIS through $4^{\circ}C$ for 100 min, cooled at $4^{\circ}C$ in TRIS with different glucose concentrations 0 M, 0.04 M, 0.1 M, 0.2 M and 0.3 M, respectively for 30 min followed by cryopreservation. After thawing at $37^{\circ}C$ for 25 sec, membrane and acrosome integrities of dog sperm were evaluated. In addition, the effect of exposure time (10, 30, 50 and 70 min) of sperm to glycerol-free TRIS containing 0.3 M glucose at $4^{\circ}C$ on progressive motility, viability, and DNA integrity following sperm cryopreservation was studied. Membrane integrity and acrosome integrity were assessed by 6-carboxyfluoresceindiacetate (6-CFDA)/propidium iodide (PI) fluorescent staining and Pisum sativum agglutinin conjugated to fluorescein isothiocyanate, respectively. DNA integrity was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling, using flow cytometry. Sperm frozen in glycerol-free TRIS supplemented with 0.2 M or 0.3 M glucose have an intact plasma membrane (CFDA+/PI-) after cryopreservation than sperm frozen in the extenders with lower glucose concentrations (p<0.05). Acrosome integrity was significantly higher in the 0.3 M group than less than 0.1 M groups (p<0.05). The sperm DNA fragmentation index did not differ according to exposure time, although progressive motility was significantly higher in the 50 min exposure group than the other groups (p<0.05). These results indicate that cryopreservation of dog sperm is feasible and yields more motile sperm following freezing and thawing in glycerol-free TRIS containing 0.3 M glucose with the exposure time for 50 min at $4^{\circ}C$.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.3
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• pp.199-205
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• 2020

Cryopreservation of semen is useful for animal breeding via artificial insemination (AI). However, the use of frozen-thawed boar semen is limited due to cryodamage. The aim of this study was to investigate the effects of different concentrations of MitoTEMPO (a mitochondria-targeted antioxidant) in lactose-egg yolk (LEY) extenders on kinetic characteristics of frozen-thawed boar sperms. Semen samples were collected from mature Duroc boars (2~3 years old) and cryopreserved in LEY extenders containing 0, 0.5, 5, 50, and 500 μM MitoTEMPO. The kinetic characteristics of frozen-thawed sperms were determined 0 and 30 min after thawing using computer-assisted sperm analysis (CASA). Results indicated that sperm motility immediately after thawing was significantly higher with 5 and 50 μM (50.46±2.71% and 46.96±2.66%, respectively) than with 500 μM MitoTEMPO (35.40±2.95%) (P<0.05). However, there were no significant differences in other kinetic characteristics except motility. In conclusion, the addition of MitoTEMPO to the sperm freezing extender may have a beneficial effect on motility of post-thawed boar semen.