• Korean Journal of Poultry Science
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• v.45 no.4
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• pp.237-244
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• 2018

In this study, to increase the survival rate of frozen/thaw rooster semen, standard protocols of semen thawing procedures were tested by computer-assisted sperm assay (CASA). We tested 4 different thawing protocols for frozen semen, $5^{\circ}C$ for 2 min, $35^{\circ}C$ for 30 s, $54^{\circ}C$ for 13 s, and $70^{\circ}C$ for 7 s. The pooled semen from 5 to 8 Ogye rooster line was diluted in the HS-1 diluent and frozen in 8% methylacetamide (MA) in liquid nitrogen vapors. To determine standard thawing method, straws were plunged into different temperatures and times. The resulting motilities were recorded by the CASA system. The results of this study showed that the best viability of the spermatozoa was shown by exposure at $5^{\circ}C$ for 2 min. Moreover, the longevity test of thawed sperm at $5^{\circ}C$ for 2 min also supported the higher viability under low temperature preservation of $17^{\circ}C$ for 1 hr. Further research is needed to increase the motility of thawed rooster semen for field application. In addition, the in vivo tests for different rooster lines are also needed for the establishment of avian genetic resource bank.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.63-63
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• 2003

This study was carried out to investigate the effects of liquid boar sperm stored at 4$^{\circ}C$ on sperm motility, normal acrosome, and in-vitro fertilization and culture of pig oocytes matured in-vitro. The sperm-rich fraction (30~60 ml) of ejaculate was collected into an insulated vacuum bottle. Semen was slowly cooled to room temperature (20~23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min at 800$\times$g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of lactose, egg yolk and N-acetyl-D-glucosamine (LEN) diluent to provide 1.0$\times$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$ and preserved for 5 days to examine sperm motility and normal acrosome. The medium used for oocyte maturation was modified tissue culture medium (TCM) 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 6 h in 500 ${mu}ell$ mTBM fertilization media with 0.2, 1, 5 and 10$\times$10$^{6}$ /ml sperm concentration, respectively. At 6 h after IVF, oocytes were transferred into 500 ${mu}ell$ Hepes-buffered NCSU-23 culture medium for further culture of 6, 48 and 144 h. There were significant differences in sperm motility and normal acrosome among preservation days and incubation times, respectively. The rates of sperm penetration and polyspermy were higher in 5 and 10$\times$10$^{6}$ sperm/ml than in 0.2 and 1$\times$10$^{6}$ sperm/ml. Male pronuclear formation was lower in 0.2$\times$10$^{6}$ sperm/ml than in 1, 5 and 10$\times$10$^{6}$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in 10$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. The rate of blastocysts from the cleaved oocytes (2~4 cell stage) was highest in 1$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend 1$\times$10$^{6}$ ml sperm concentration for in-vitro fertilization of pig oocytes.

• Applied Chemistry for Engineering
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• v.35 no.3
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• pp.214-221
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• 2024

Polyether ketone (PEEK) has been widely used in membranes because of its excellent thermal stability, chemical resistance, and significant mechanical strength. However, the melting temperature is very high, making it difficult to find suitable solvents. Therefore, in this study, PEEK and benzophenone (DPK) were used as diluents to prepare a membrane with excellent mechanical strength and chemical stability using the thermally induced phase separation (TIPS) method to compensate for the shortcomings of PEEK membrane preparation and achieving the highest performances. The optimal membrane manufacturing conditions were confirmed through the crystallization temperature and cloud point according to the polymer content through the phase diagram. Subsequently, the morphological changes of the membrane, influenced by the polymer and diluent content, were confirmed through scanning electron microscopy (SEM). Additionally, the membrane thickness tended to increase with higher polymer content. Tensile strength and DI-water permeability tests were conducted to confirm the mechanical strength and permeability of the membrane. Through the previous characteristic evaluation, it was confirmed that the membrane using PEEK had excellent mechanical strength and permeability.

• Restorative Dentistry and Endodontics
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• v.33 no.1
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• pp.1-8
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• 2008

The purpose of this study was to evaluate the effect of a new resin monomer, filler size and polishing technique on the surface roughness of composite resin restorations using confocal laser scanning microscopy. By adding new methoxylated Bis-GMA (Bis-M-GMA, 2,2-bis[4-(2-methoxy-3-methacryloyloxy propoxy) phenyl] propane) having low viscosity, the content of TEGDMA might be decreased. Three experimental composite resins were made: EX1 (Bis-M-GMA/TEGDMA = 95/5 wt%, 40 nm nanofillers); EX2 (Bis-M-GMA/TEGDMA = 95/5 wt%, 20 nm nanofillers); EX3 (Bis-GMA/TEGDMA = 70/30 wt%, 40 nm nanofillers). Filtek Z250 was used as a reference. Nine specimens (6 mm in diameter and 2 mm in thickness) for each experimental composite resin and Filtek Z250 were fabricated in a teflon mold and assigned to three groups. In Mylar strip group, specimens were left undisturbed. In Sof-lex group, specimens were ground with #1000 SiC paper and polished with Sof-lex discs. In DiaPolisher group, specimens were ground with #1000 SiC paper and polished with DiaPolisher polishing points. The Ra (Average roughness), Rq (Root mean square roughness), Rv (Valley roughness), Rp (Peak roughness), Rc (2D roughness) and Sc (3D roughness) values were determined using confocal laser scanning microscopy. The data were statistically analyzed by Two-way ANOVA and Tukey multiple comparisons test (p = 0.05). The type of composite resin and polishing technique significantly affected the surface roughness of the composite resin restorations (p < 0.001). EX3 showed the smoothest surface compared to the other composite resins (p < 0.05). Mylar strip resulted in smoother surface than other polishing techniques (p < 0.05). Bis-M-GMA. a new resin monomer having low viscosity, might reduce the amount of diluent, but showed adverse effect on the surface roughness of composite resin restorations.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.6
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• pp.1114-1119
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• 2012

This study was conducted to assess the microbial toxicity of ionic silver solution ($Ag^+N$) and silver nanoparticle suspension ($Ag^0NP$) based on the Microtox bioassay. In this test, the light inhibition of luminescent bacteria was measured after 15 and 30 min exposure to aqueous solutions and soils spiked with a dilution series of $Ag^+N$ and $Ag^0NP$. The resulting dose-response curves were used to derive effective concentration (EC25, $EC_{50}$, EC75) and effective dose ($ED_{25}$, $ED_{50}$, $ED_{75}$) that caused a 25, 50 and 75% inhibition of luminescence. In aqueous solutions, $EC_{50}$ value of $Ag^+N$ after 15 min exposure was determined to be < $2mg\;L^{-1}$ and remarkably lower than $EC_{50}$ value of $Ag^0NP$ with $251mg\;L^{-1}$. This revealed that $Ag^+N$ was more toxic to luminescent bacteria than $Ag^0NP$. In soil extracts, however, $ED_{50}$ value of $Ag^+N$ with 196 mg kg-1 was higher than $ED_{50}$ value of $Ag^0NP$ with $104mg\;kg^{-1}$, indicating less toxicity of $Ag^+N$ in soils. The reduced toxicity of $Ag^+N$ in soils can be attributed to a partial adsorption of ionic $Ag^+$ on soil colloids and humic acid as well as a partial formation of insoluble AgCl with NaCl of Microtox diluent. This resulted in lower concentration of active Ag in soil extracts obtained after 1 hour shaking with $Ag^+N$ than that spiked with $Ag^0NP$. With longer exposure time, EC and ED values of both $Ag^+N$ and $Ag^0NP$ decreased, so their toxicity increased. The toxic characteristics of silver nanomaterials were different depending on existing form of Ag ($Ag^+$, $Ag^0$), reaction medium (aqueous solution, soil), and exposure time.

• Journal of the Korean Chemical Society
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• v.42 no.6
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• pp.618-628
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• 1998

Pure CaO, Mn-doped CaO, Mn/CaO, and K/CaO catalysts were prepared and tested as catalysts for the oxidative coupling of methane in the temperature range of 600 to 800$^{\circ}C$ to investigate the effects of Mn- and K-addition on the catalytic activity of calcium oxide. To characterize the catalysts, X-ray powder diffraction(XRD), XPS, SEM, DSC, and TG analyses were performed. The catalytic reaction was carried out in a single-pass flow reactor using on-line gas chromatography system. Normalized reaction conditions were generally $p(CH_4)/p(O_2)=250$ Torr/50 Torr, total feed flow rate=30 mL/min, and 1 atm of total pressure with He being used as diluent gas. Among the catalysts tested, 6.3 mol% Mn-doped CaO catalyst showed the best $C_2$ yield of 8.0% with a selectivity of 43.2% at 775$^{\circ}C$. The $C_2$ selectivity increased on lightly doped CaO catalysts, while decreased on heavily doped CaO([Mn] > 6.3 mol%) catalysts. 6 wt.% Mn/CaO and 6 wt.% K/CaO catalysts showed the $C_2$ selectivities of 13.2% and 30.9%, respectively, for the reaction. Electrical conductivities of CaO and Mn-doped CaO were measured in the temperature range of 500 to 1000$^{\circ}C$ at Po2's of $10^{-3}\; to\;10^{-1}\;atm.$ The electrical conductivity was decreased with Mn-doping and increased with increasing $P0_2$in the range of $10^{-3}\;to\;10^{-1}\;atm,$ indicating the specimens to be p-type semiconductors. It was suggested that the interstitial oxygen ions formed near the surface can activate methane and the formation of interstitial oxygen ions was discussed on the basis of solid-state chemistry.

• Food Science and Preservation
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• v.23 no.6
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• pp.797-803
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• 2016

In this study, we investigated the effect of fermentation conditions on the amylolytic and proteolytic activities of Aspergillus luchuensis strain 74-5 and Aspergillus oryzae strain 75-2, which are used in the preparation of the starter culture, for Takju (Korean traditional rice wine). The starter culture was optimized using different conditions, such as inoculum size, inoculation temperature, and incubation time. The enzyme activities under each condition were measured. In the A. luchuensis strain 74-5 starter culture, the ${\alpha}-amylase$ and glucoamylase activities increased, however the activity of acidic protease decreased as the diluent to starter culture ratio increased. In the A. oryzae 75-2 starter culture, all enzyme activities were maintained at a higher level even at 5% inoculation ratio. Higher enzyme activities were observed in the middle range of inoculation temperature (35, $40^{\circ}C$), than in the lower range (20, $30^{\circ}C$). Enzyme activity in the starter culture varied with incubation time, however it was the highest at 144 and 120 hr, respectively, for A. luchuensis strain 74-5 and A. oryzae strain 75-2. The spore count of the starter culture was approximately $2{\times}10^7$ during fermentation, out of which contamination by aerobic bacteria was about $3{\times}10^3$. The results suggested that the starter culture of each strain could be used as an inoculum for fermentation. However, we needs to conduct further research for the selection of suitable diluting agents as well as drying methods to reduce the contamination by aerobic bacteria, while retaining the enzyme activity.

• Research in Plant Disease
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• v.21 no.2
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• pp.50-57
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• 2015

Control of nematode has become difficult owing to the restricted use of effective soil fumigant, methyl bromide, and other non-fumigant nematicides. Therefore, it is urgently necessary to develop microbial nematicide to replace chemical nematicides. In this study, the 50% aqueous methanol extraction solution of fermentation broths of 2,700 actinomycete strains were tested for their nematicidal activity against second stage of juveniles (J2s) of Meloidogyne incognita. As the results, only the 50% aqueous methanol extraction solution of AN110065, at 20% equivalent to 10% fermentation broth, showed strong nematicidal activity with 78.9% of mortality 24 h after treatment and 94.1% of mortality at 72 h. The 16S rRNA gene sequencing showed that the strain sequence was 99.78% identical to Streptomyces netropsis. The extract of S. netropsis AN110065 fermentation broth was successively partitioned with ethyl acetate and butanol and then the ethyl acetate, butanol and water layers were investigated for their nematicidal activity against the M. incognita. At $1000{\mu}g/ml$, ethyl acetate layer showed the strongest activity of 83.5% of juvenile mortality 72 h after treatment. The pot experiment using the fermentation broth of AN110065 on tomato plant against M. incognita displayed that it evidently suppressed gall formation at a 10-fold diluent treatment. The tomato plants treated with the fermentation broth of S. netropsis AN110065 did not show any phytotoxicity. The results suggest that S. netropsis AN110065 has a potential to serve as microbial nematicide in organic agriculture.

• The Korean Journal of Nuclear Medicine Technology
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• v.26 no.1
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• pp.42-46
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• 2022

Purpose In the meantime, there have been not many samples that require dilution, and it has been difficult for the examiner to set an appropriate dilution multiple for RIA test item and report the results. Accordingly, it was judged that it was necessary to set the maximum dilution multiple for each test and to verify the upper limit of the clinical reportable range. Therefore, in this study, the maximum dilution multiple for each RIA test was set and the upper limit of the clinical reportable range was verified accordingly Materials and Methods Among all RIA tests conducted at Asan Medical Center, the study treated on 30 types of tests which also conduct the dilution test. Data from March to July 2021 were collected and analyzed. The study was conducted on samples subjected to serial dilutions such as X2, X4 or X10, X10², X10³, X10⁴, X10⁵. Results Among a total of 30 test types, 18 test types have more than 5 N values in the tolerance range of 80~120%. As a result of the verification of maximum dilution multiples, the test set to 10⁴ is 𝛼-fetoprotein and thyroglobulin, and the test set to 10³ is CA-125, CEA, and 𝛽-hCG, and the test set to 10² is Free PSA, PSA, CA15-3, SCC, Ferritin, PTH, Cortisol, and Calcitonin. Tests set to 10 include three categories: 𝛽2-Microglobulin, C-peptide, and Testosterone. Conclusion It is expected that it will contribute to improving the quality of nuclear medicine blood tests as the results of dilution experiments can be reported quickly and accurately through the verification of the clinical reportable range.

• Korean Journal of Animal Reproduction
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• v.27 no.3
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• pp.215-223
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• 2003

This study was to investigate the effects of fertilization time and culture medium of pig oocytes matured in-vitro by liquid boar sperm. The sperm rich fraction (30∼60 ml) was slowly cooled to room temperature (20∼23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min 800 ${\times}$ g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of the LEN diluent to provide 1.0${\times}$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$. The medium used for oocyte maturation was TCM-199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10 $\mu\textrm{g}$/ml insulin, 2 $\mu\textrm{g}$/ml vitamin B$_{12}$ , 25 mM HEPES, 10 $\mu\textrm{g}$/ml bovine apotransferrin, 150 $\mu$M cysteamine, 10 IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75 $\mu\textrm{g}$/ml sodium penicillin G, 50 $\mu\textrm{g}$/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9 h in 500 ${mu}ell$ mTBM fertilization media with 1.0${\times}$10$^{6}$ sperm/ml concentration, respectively. Thereafter, oocytes were transferred into 500 ${mu}ell$ NCSU-23, HEPES buffered NCSU-23, PZM-3 and PZM-4 culture media, respectively, for further culture of 6, 48 and 144 h. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times for 6 and 9 h than in those for 1 and 3 h. The rates of cleaved oocytes were higher in the fertilization times for 6 and 9 h (85.0 and 84.6%) than in those for 1 and 3 h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time for 6 h (33.6%) than in that for 1, 3 and 9 h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9, 27.6, 26.3 and 24.4 in the fertilization times for 6, 9, 3 and 1 h, respectively. The rate of blastocyst from the cleaved oocytes and the number of cells per blastocyst were higher in HEPES buffered NCSU-23 culture medium than in NCSU-23, PZM-3 and PZM-4 culture media. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend the coincubation time of 6 h in 500 ${mu}ell$ TBM fertilization medium with 1${\times}$10$^{6}$ sperm/ml concentration and the HEPES buffered NCSU-23 culture medium for in-vitro fertilization of pig oocytes matured in-vitro.