• 한국수정란이식학회지
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• 제32권2호
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• pp.53-58
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• 2017

It is very difficult to get the information about semen quality analysis in transgenic pigs because of limited numbers and research facilities. Therefore, in the present study, we analyzed the semen quality of transgenic boars generated for xenotransplantation research. Briefly, the semen samples were collected from 5 homozygous ${\alpha}1,3$-Galactosyltransferase knock-out ($GalT^{-/-}$) transgenic boars and immediately transported to the laboratory. These semen samples were decupled with DPBS and conducted to analyze semen parameters by a computer-assisted semen analysis (CASA) system. The boar semen were examined all 12 parameters such as total motility (TM), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), and hyperactivated (HYP), etc. In results, among the 5 $GalT^{-/-}$ boars, three boars (#134, 144, and 170) showed normal range of semen parameters, but #199 and 171 boars showed abnormal ranges of semen parameters according to standard ranges of semen parameters. Unfortunately, #171 boar showed azoospermia symptom with rare sperm counts in the original semen. Conclusively, assessment of semen parameters by CASA system is useful to pre-screening of reproductively healthy boar prior to natural mating and artificial insemination for multiplication and breeding.

• 한국지반공학회논문집
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• 제16권4호
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• pp.157-170
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• 2000

본 연구에서는 자연점토에 대한 구리 흡착 및 구리와 아연의 경쟁 흡착 시 온도변화에 따른 흙의 각 구성성분별 흡착거동을 살펴보기 위해 혼합 흡착-연속추출법(combined adsorption-sequential extraction analysis, CASA)을 사용하였다. 실험결과 중금속 아연의 경우 농도에 따른 차이는 있지만 온도증가에 따라 주로 이온교환 형태로 흡착되는 것을 알 수 있고, 구리와 경쟁흡착의 경우 이러한 경향은 더욱 커져서 약 80~90%정도가 이온교환형태로 흡착되어진다. 반면 구리의 경우 실험을 수행한 거의 모든 농도 범위에서 약 50%이상이 탄산염 형태로 흡착되어진다. 탄산염형태로의 흡착 양상은 단일 흡착의 경우 온도증가에 따라 약 5% 증가하는 경향을 보이고, 경쟁흡착의 경우에는 약 10%정도 증가하는 경향을 나타내었다. 그리고 자연점토에서의 아연과 구리의 각 구성성분 별 흡착거동은 이온 교환 형태로 분배되는 경우를 제외하고는 온도증가에 따라 흡착량이 증가하는 흡열반응$(\DeltaH0>0)$인 것으로 나타났다.

• 건축역사연구
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• 제13권2호
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• pp.21-38
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• 2004

This study provides a descriptive and analytical account of major aspects of urban development and transformation of housing types of Italian Firenze from the 13th century to the 19th century. It is a typo-morphological depiction of urban spatial structure of the extraordinary city, Firenze, the center of Italian Renaissance. And this study has proceeded on the assumption that the evolving form of the urban structure and housing types cannot be understood without reference to the larger context of political, economic, and social life. Based on these backgrounds, the purpose of this study is threefold: to provide a comprehensive discussion of general characteristics of urban spatial structure of Firenze, and to explain the process of formation of working-class neighborhoods by constructing new city wall in later 13th century, and to discuss transformation of housing types of the working-class neighborhood with understanding the mechanism of existence of housing in the newly formed residential neighborhoods. The development of residential neighborhoods was pursued by 'planned' manner through forming square-shaped blocks, and characterized by the subdivision of larger properties into standardized building lots for the construction of houses. On the bases of documentary evidences, several ecclesiastical institutions are identified as the agents of a distinctive type of development. While the institutions did the major role for developing lands, the construction of houses was done by small scale construction agents with moderate amount of properties. The major housing type of working-class neighborhoods of Firenze has been the 'casa a schiera' characterized by the form of narrow front and long depth. The type was generalized by the newly formed middle and working-class of Firenze which grew their body very rapidly, Even though the type assumed very uniform in its fen there were many variations. And through passing time, the casa a schiera developed to be multi-family housing, and the level of variation became deepen. Eventually, transformation of housing type of Firenze was ended by appearance of the 'casa in linea', which was very similar to modern apartment in its spatial organization.

• Journal of Plant Biotechnology
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• 제30권3호
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• pp.251-255
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• 2003

An efficient system for in vitro bulblet formation of Lilium oriental hybrids(cvs). Casa Blanca and Siberia is described. Transverse thin cell layer(tTCL)(1mm thick) explants of 'Casa Blanca' formed bulblets at a frequency of 97.7％ when cultured on Murashige and Skoog (MS) medium supplemented with 1mg/L 2,4 dichlorophenoxyacetic acid(2,4-D) (On average 15.6 bulblets were formed per explant). The frequency of bulblet formation was drastically reduced when the explant ghickness was thinner than 1 mm. Explants from the outermost layer of bulb scale produced greater frequency of bulblet formation than middle or innermost layer. Among auxins supplemented to culture medium at 1 mg/L, 2,4-D led to greater frequency of bulblet formation on explants than dicamba, picamba, or phenylacetic acid(PAA). tTCL explants from the middle region of the outermost layer bulb scale yielded greater frequency of bulblet formation than the upper or lower region. tTCL stem explants of 'Siberia' formed bulblets at a frequency of 95.3% when cultured on MS medium with 1 mg/L 2,4-D(On average 9.1 bulblets were formed per explant). The system estabilished in this study will be useful for in vitro rapid propagation and genetic transformation of Lilium Oriental hybrids.

• 한국수정란이식학회지
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• 제28권1호
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• pp.63-71
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• 2013

In this study, we used flow a cytometric assay to evaluate plasma membrane integrity and mitochondrial activity in post-thawed sperm that was supplemented with ginsenoside-$Rg_1$. Varying concentrations of ginsenoside-$Rg_1$ (0, 25, 50 and $100{\mu}M/ml$) were used in the extender during cryopreservation to protect the DNA of thawed sperm, thereby increasing the viability and motility rate as evaluated using a computer-assisted sperm analysis (CASA) method. The results derived from CASA were used to compare the fresh, control, and ginsenoside-$Rg_1$ groups. Sperm motility and the number of progressively motile sperm were significantly (p<0.05) higher in the $50{\mu}M/ml$ ginsenoside-Rg1 group ($61.0{\pm}4.65%$) than in the control ($46.6{\pm}7.02%$), $25{\mu}M/ml$ ($46.2{\pm}4.76%$), and $100{\mu}M/ml$ ginsenoside-$Rg_1$ ($52.0{\pm}1.90%$) groups. However, the velocity distribution of post-thawed sperm did not differ significantly. Membrane integrity and MMP staining as revealed using flow cytometry were significantly (p<0.05) higher ($91.6{\pm}0.82%$) in the $50{\mu}M/ml$ ginsenoside-$Rg_1$ group than in the other groups. Here, we report that ginsenoside-$Rg_1$ affects the motility and viability of boar spermatozoa. Moreover, ginsenoside-$Rg_1$ can be used as a protective additive for the suppression of intracellular mitochondrial oxidative stress caused by cryopreservation.

• Animal Bioscience
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• 제34권12호
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• pp.1912-1920
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• 2021

Objective: This study investigated the effects of 5-aminolevulinic acid (5-ALA) on the motility parameters, mitochondrial membrane depolarization, and ATP levels in chicken sperm. Methods: The pooled semen from Barred Plymouth Rock males was used. In the first experiment, the semen was diluted 4-times with phosphate-buffered saline (PBS (-)) containing various concentrations (0, 0.01, 0.05, and 0.1 mM) of 5-ALA, and then the sperm motility parameters after incubation were evaluated by computer-assisted sperm analysis (CASA). In the second experiment, the semen was diluted 4-times with PBS (-) containing 0.05 mM 5-ALA, and then sperm mitochondrial membrane depolarization and ATP levels after 1.5 h of incubation were analyzed with the MitoPT^® JC-1 Assay and ATP Assay kits, respectively. In the third experiment, the semen was removed from the seminal plasma and resuspended with the mediums of PBS (-), PBS (-) supplemented with CaCl₂ and MgCl₂ (PBS (+)) + 5-ALA, PBS (+) + caffeine, and PBS (+) + caffeine + 5-ALA. Then, the sperm motility parameters after incubation were evaluated by CASA. In the last experiment, the semen was treated with the mediums of PBS (-), PBS (-) + 5-ALA, 5.7% glucose, 5.7% glucose + 5-ALA after removing the seminal plasma, and then the sperm motility parameters were evaluated by CASA. Results: The addition of 0.05 mM 5-ALA significantly increased the chicken sperm motility, progressive motility, linearity, average path velocity, curvilinear velocity, straight-line velocity, and the wobble. The sperm mitochondrial membrane depolarization was also increased by the 5-ALA treatment. The 5-ALA treatment decreased the sperm ATP levels. Both the caffeine treatment and glucose treatment decreased the sperm motility during incubation period. Conclusion: 5-ALA might increase sperm mitochondrial membrane depolarization to utilize the ATP for enhancing sperm movement.

• Animal Bioscience
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• 제34권8호
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• pp.1253-1270
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• 2021

Assessment of male fertility is based on the evaluation of sperm. Semen evaluation measures various sperm quality parameters as fertility indicators. However, semen evaluation has limitations, and it requires the advancement and application of strict quality control methods to interpret the results. This article reviews the recent advances in evaluating various sperm-specific quality characteristics and methodologies, with the help of different assays to assess sperm-fertility status. Sperm evaluation methods that include conventional microscopic methods, computer-assisted sperm analyzers (CASA), and flow cytometric analysis, provide precise information related to sperm morphology and function. Moreover, profiling fertility-related biomarkers in sperm or seminal plasma can be helpful in predicting fertility. Identification of different sperm proteins and diagnosis of DNA damage has positively contributed to the existing pool of knowledge about sperm physiology and molecular anomalies associated with different infertility issues in males. Advances in methods and sperm-specific evaluation has subsequently resulted in a better understanding of sperm biology that has improved the diagnosis and clinical management of male factor infertility. Accurate sperm evaluation is of paramount importance in the application of artificial insemination and assisted reproductive technology. However, no single test can precisely determine fertility; the selection of an appropriate test or a set of tests and parameters is required to accurately determine the fertility of specific animal species. Therefore, a need to further calibrate the CASA and advance the gene expression tests is recommended for faster and field-level applications.

• 한국동물생명공학회지
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• 제37권3호
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• pp.193-201
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• 2022

Subgroups of sperm which share similar motility features documented in mammals indicate between-subject variations that might be related to fertilizing potential of the respective ejaculates. The objectives of this study were to define subpopulations of motile sperm in Gyr falcon semen using kinematic parameters driven by Computer Assisted Semen Analysis (CASA) and to investigate the subject-related variations in these subpopulations. A total of 24 fresh ejaculates from 6 falcons were used to assign each of the 20473 sperms into 3 subpopulations by a multivariate cluster analysis. The proportion of sperms in different sub-populations were compared among subjects by a generalized linear model and repeatability of sperm frequency in different subpopulations was investigated by corelation analysis. The resulting 3 categories of sperm indicated significant differences in all kinematic parameters (p < 0.05). Subpopulation 1 (15.91%) contained sperms with the highest velocity and progressiveness of movement trajectory while subpopulation 3 (6.4%) included the least progressively motile sperms. Proportion of rapid and medium progressive sperm were consistently higher in the ejaculate of three falcons compared to the two other birds which also had the highest proportion of slow non-progressive sperms (p < 0.05). Respective proportion of sperms in each subpopulations indicated significant repeatability over multiple measurements (p < 0.05). In conclusion, subpopulations of motile sperm in Gyr falcon can be identified using kinematic parameters generated by CASA. Individual differences in the proportion of these subpopulations might have potential application for identifying the males with higher fertilizing capacity.

• 식물조직배양학회지
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• 제28권3호
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• pp.135-140
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• 2001

본 연구는 오리엔탈 나리 'Casa Blanca' 의 기내인편에서 저반부가 비대된 자구를 형성하고, 형성된 자구절편을 배양하여 저반부가 비대된 자구의 증식 및 자구를 비대시키는, 5 L의 생물반응기를 이용한 일련의 대량생산 체계를 확립하기 위하여 실시하였다. 5 L airlift 생물반응기를 이용한 액체배양에서는 저반부가 비대된 자구의 형성 및 증식은 전혀 이루어 지지 않았으며, 인편엽만 무성하게 자랐다. 재료가 배지에 일시적으로 침지되는 ebb & flood형의 생물반응기에서 저반부가 비대된 자구 절편체를 배양한 결과, 저반부가 비대된 자구들이 증식하였다. 광은 자구의 증식에 영향을 미치지 않았으나, 명배양에서는 인편엽의 발생률이 높았으며, 저반부가 비대된 자구 증식은 MS 배지에 BA 2.0 mg/L와 IAA 0.3 mg/L가 첨가된 배지에서 양호하였다. 5L airlift 생물반응기에서 저반부가 비대된 자구 절편체로부터 자구비대는 MS 배지에 sucrose 70 g/L가 첨가된 배지에서 액체배양하는 것이 양호하였으며, 16주 배양기간 동안 배양 8주 후에 배지를 교체하는 것이 자구비대에 효과적이었다. 또한 5 L의 생물반응기에 3 L 액체배지에는 절편체를 15 g을 주입하는 것이 자구비대에 적절하였다.

• 한국환경복원기술학회지
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• 제20권4호
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• pp.29-42
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• 2017

As the "Guidelines for GHG Environmental Assessment" was revised, it pointed out that the developers should evaluate GHG sequestration and storage of the developing site. However, the current guidelines only taking into account the quantitative reduction lost within the development site, and did not consider the qualitative decrease in the carbon sequestration capacity of forest edge produced by developments. In order to assess the quantitative and qualitative effects of vegetation carbon uptake, the CASA-NPP model and satellite image spatial-temporal fusion were used to estimate the annual net primary production in 2005 and 2015. The development projects between 2006 and 2014 were examined for evaluate quantitative changes in development site and qualitative changes in surroundings by development types. The RMSE value of the satellite image fusion results is less than 0.1 and approaches 0, and the correlation coefficient is more than 0.6, which shows relatively high prediction accuracy. The NPP estimation results range from 0 to $1335.53g\;C/m^2$ year before development and from 0 to $1333.77g\;C/m^2$ year after development. As a result of analyzing NPP reduction amount within the development area by type of forest development, the difference is not significant by type of development but it shows the lowest change in the sports facilities development. It was also found that the vegetation was most affected by the edge vegetation of industrial development. This suggests that the industrial development causes additional development in the surrounding area and indirectly influences the carbon sequestration function of edge vegetaion due to the increase of the edge and influx of disturbed species. The NPP calculation method and results presented in this study can be applied to quantitative and qualitative impact assessment of before and after development, and it can be applied to policies related to greenhouse gas in environmental impact assessment.