The present study was carried out to investigate the effects of maturation media on penetrability of pig oocytes by liquid boar sperm coincubated with different sperm concentrations in a modified Trisbuffered medium (mTBM). Follicular oocytes collected from ovaries of prepubertal gilts were matured in a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. The spermich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ far 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes in 5% $CO_2$in air at 38.5$^{\circ}C$, cumulus cells were removed and oocytes (30~40) were coincubated for 6 h in 0.5 $m\ell$ mTBM fertilization medium with five different (1$\times$10$^{6}$ , 2$\times$10$^{6}$ , 4$\times$10$^{6}$ , 6$\times$10$^{6}$, 10$\times$10$^{6}$$m\ell$) sperm concentrations. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ NCSU-23 culture medium fur further culture of 6 h. At 12 h after IVF, sperm penetration, polyspermy and male pronuclear formation of oocytes were evaluated. Oocytes of NCSU-23 maturation medium decreased polyspermy and increased male pronuclear formation compared to those of mTCM199 and mWaymouth MB 752/1 maturation media. Of oocytes matured in NCSU-23 medium and inseminated in mTBM medium with 2~4$\times$10$^{6}$$m\ell$ sperm concentrations, 50.8~50.9% showed sperm penetration, 13.3~19.5% polyspermy and 43.9~45.4% male pronuclear formation. In conclusion, we found out that oocytes matured in NCSU23 medium and inseminated in mTBM medium showed superior invitro fertilization compared to those matured in mTCM199 and mWaymouth MB 752/1 maturation media and inseminated in mTBM medium. The optimum sperm concentrations for in-vitro fertilization of oocytes matured in NCSU-23 medium by liquid boar semen stored at 17$^{\circ}C$ for 5 days were 2~4$\times$10$^{6}$$m\ell$.
The present study was carried out to investigate the effects of the maturation media such as a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium on penetrability of pig oocytes by liquid boar sperm. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. When immature pig oocytes were cultured in mTCM-199, mWaymouth MB 752/1 and NCSU-23 maturation media for 44 h in 5% $CO_2$, in air at 38.5$^{\circ}C$, the germinal vesicle breakdown (CVBD) rates of the oocytes were 95.6, 94.1 and 94.9%, respectively, and the maturation rates (metaphase II) of oocytes were 92.5, 90.1 and 91.1%, respectively. No differences were observed among the maturation media. The spermrich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ for 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes, cumulus cells were removed and oocytes (30~40) coincubated far 6 h in 0.5 $m\ell$ mTCM-199 and mTBM fertilization media with 2$\times$1061$m\ell$ sperm concentration. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ mTCM-199 and NCSU-23 culture media for further culture 6 or 42 h. Sperm penetration, polyspermy and male pronuclear formation of oocytes at 12 h after IVF, and developmental ability of oocytes at 48 h after IVF were evaluated. The oocytes in combination with NCSU-23 medium for maturation and mTBM medium for IVF increased male pronuclear formation (48.0%) compared to those in combination with mTCM-199 media for maturation and IVF, and mWaymouth MB 752il medium for maturation and mTCM-199 medium far IVF. The rates of cleaved embryos (2~4 cell stage) at 48 h after IVF were 24.1% in combination with mTCM-199 media for maturation, IVF and culture, 43.6% in combination with mWaymouth MB 75211 medium fur maturation and mTCM-199 media for IVF and culture, and 71.2% in combination with NCSU-23 medium for maturation, mTBM medium for IVF and NCSU-23 medium for culture. In conclusion, we found out the oocytes matured in vitro were fertilized by liquid boar sperm stored in BTS extender at 17$^{\circ}C$ for 5 days. We recommend the simple defined NCSU-23 medium for nuclear maturation, mTBM medium and liquid boar sperm for IVF, and NCSU-23 medium for embryo culture.
Proceedings of the Korean Society of Plant Pathology Conference
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2003.10a
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pp.84.2-85
/
2003
Magnaporthee grisea causes the devastating blast disease of rice. Entensive research has been conducted on infection mechanisms, particularly on appressorium formation and penetration, of this fungus during the last decade. However, the role(s) of cell-wall-degrading enzymes (CWDEs) on pathogenesis is not clearly demonstrated at molecular level. Many CWDES in plant pathogenic fungi including M. grisea are redundant; that is, there are multiple genes encoding enzymes with a similar or overlapping spectrum of activities. It is laborious to isolate all of the genes encoding related enzymes and to construct mutants lacking all 9f them. Thus, we considered alternative strategies to address the role of CWDEs in pathogenesis. Since expression of CWDE genes Is repressed by a simple sugar, as the first step, we cloned a Snfl (sucrose non-fermenting) gene (MgSnf1) from M. grisea. The predicted amino acid sequence showed a high identity with other Snf1 genes from various fungi. To elucidate molecular function of MgSnf1, a transformant lacking MgSnf1 was created by targeted gene replacement. En glucose, sucrose, and xylan the MgSnf1 mutant grew normally but in pectin and complex media, it grew slower than wild type. Expression of various CWDEs in MgSnf1 mutant was investigated and found that expression of some CWDEs is repressed. However, no significant difference was observed in conidial germination, appressorium formation, and pathogenicity in MgSnf1 mutant. However, MgSnf1 functionally complemented a yeast MgSnf1 mutant. These results suggest that MgSnf1 is involved in regulation of CWDEs and MgSnf1 is dispensable in pathogenicity of M. grisea.
Chromium nitride (CrN) samples with two different layer structures (multilayer and single layer) were coated on bipolar plates of polymer electrolyte membrane fuel cells (PEMFC) using the reactive sputtering method. The effects with respect to layer structure on corrosion resistance and overall cell performance were investigated. A continuous and thin chromium nitride layer ($Cr_{0.48}\;N_{0.52}$) was formed on the surface of the SUS 316L when the nitrogen flow rate was 10 sccm. The electrochemical stability of the coated layers was examined using the potentiodynamic and potentiostatic methods in the simulated corrosive circumstances of the PEMFC under $80^{\circ}C$. Interfacial contact resistance (ICR) between the CrN coated sample and the gas diffusion layer was measured by using Wang's method. A single cell performance test was also conducted. The test results showed that CrN coated SUS316L with multilayer structure had excellent corrosion resistance compared to single layer structures and single cell performance results with $25\;cm^2$ in effective area also showed the same tendency. The difference of the electrochemical properties between the single and multilayer samples was attributed to the Cr interlayer layer, which improved the corrosion resistance. Because the coating layer was damaged by pinholes, the Cr layer prevented the penetration of corrosive media into the substrate. Therefore, the CrN with a multilayer structure is an effective coating method to increase the corrosion resistance and to decrease the ICR for metallic bipolar plates in PEMFC.
Infrared (IR) wavelength is out of visible range and thus usually cut by hot filters in general commercial cameras. However, some information from the near-IR (NIR) range is known to improve the overall visibility of scene in many cases. For example when there is fog or haze in the scene, NIR image has clearer visibility than visible image because of its stronger penetration property. In this paper, we propose an algorithm for fusing the RGB and NIR images to obtain the enhanced images of the outdoor scenes. First, we construct a weight map by comparing the contrast of the RGB and NIR images, and then fuse the two images based on the weight map. Experimental results show that the proposed method is effective in enhancing visible image and removing the haze.
Kim, Taehyun;Lee, Song Hee;Oh, Young Taek;Jeon, Junhyun
The Plant Pathology Journal
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v.36
no.4
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pp.314-322
/
2020
Interplay between histone acetylation and deacetylation is one of the key components in epigenetic regulation of transcription. Here we report the requirement of MoHDA1-mediated histone deacetylation during asexual development and pathogenesis for the rice blast fungus, Magnaporthe oryzae. Structural similarity and phylogenetic analysis suggested that MoHDA1 is an ortholog of Saccharomyces cerevisiae Hda1, which is a representative member of class II histone deacetylases. Targeted deletion of MoHDA1 caused a little decrease in radial growth and large reduction in asexual sporulation. Comparison of acetylation levels for H3K9 and H3K14 showed that lack of MoHDA1 gene led to significant increase in H3K9 and H3K14 acetylation level, compared to the wild-type and complementation strain, confirming that it is a bona fide histone deacetylase. Expression analysis on some of the key genes involved in asexual reproduction under sporulation-promoting condition showed almost no differences among strains, except for MoCON6 gene, which was up-regulated more than 6-fold in the mutant than wild-type. Although the deletion mutant displayed little defects in germination and subsequent appressorium formation, the mutant was compromised in its ability to cause disease. Wound-inoculation showed that the mutant is impaired in invasive growth as well. We found that the mutant was defective in appressorium-mediated penetration of host, but did not lose the ability to grow on the media containing H2O2. Taken together, our data suggest that MoHDA1-dependent histone deacetylation is important for efficient asexual development and infection of host plants in M. oryzae.
Seo, Young-Rok;Han, Sung-Sik;Yu, Yong-Man;Lee, Jun-Jae;Ryu, Jae-Chun
Proceedings of the Korea Society of Environmental Toocicology Conference
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1996.12a
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pp.70-70
/
1996
The cytotoxicological effect of Bt 1-endotoxin, well-known as a bioinsecticide, was investigated on ion channel of insect cell lines. This study attempted to evaluted the specificity by simple experiment to measure the cell swelling using lepidopteran cell lines in isotonic solution containing only one cation. Cell swelling was stimulated in KCI-sucrose isotonic solution as well as TC-100 media containg in solubilized crystal 5-endotoxin. It suggested that the cell swelling by Bt toxin have a relation to K+ channel. The cell swelling may be due to the stimulation K+ influx and simultaneously the penetration of H2O induced by Bt toxin, because the stimulation of swelling was observed with the solubilized toxin in KCI-sucrose isotonic solution, but not in sucrose isotonic solution. Moreover the specific K+ channel blocker, such as 4-arnjnopyrimidine(4-AP) and ouabain, showed the significant effect on the cell swelling induced by Bt toxin. The increasement of the cell swelling induced by 4-AP suggested to be caused by the block of K+ efflux through K+ leak channels. The inhibition of cell swelling by ouabain, which is the well-known inhibitor of Na+, K+-ATPase, suggested to be due to decreasement of K+ influx following diminishment of Na+, K+-ATPase activities.
Recently smartphone has propagated all age groups rapidly due to the fact that the advance of media and the increase of smartphone penetration. In South Korea, high-school students' smartphone retention rate is 90.2%. This study proceed an analysis of high school students' smartphone usage by a feasibility study, and also by recording the log data of actual usage patterns of smartphone applications. The feasibility study investigates subject's smartphone usage, and the log data analysis measures the accurate usage recorded for about three months. We compared the feasibility study and the log data for the daily smartphone usage, and investigated the change of usage pattern during the weekdays, weekends, and the before, during and after exams. High-school students are unique group in capital area. As a result, we found that high-school students' smartphone usage pattern in capital area has not affected by weekday or weekend but has affected by the before, during and after exams.
This study is a user experience study based on location branding at mobile experience stores. As mobile devices have recently become saturated beyond the penetration, the IT industry is encouraging users to buy through experience and can be competitive by increasing brand loyalty through experience stores. The purpose of this study is to analyze the effect of place branding among experience stores through qualitative and quantitative measurements and analyze the user experience factors related to the location branding to present a future direction. A total of eight subjects were given tasks and then experimented with participatory observations and conducted in-depth interviews. The experiment showed that the experience had a significant effect on users, but the difference between each brand was obvious. Since this study has identified the experience-oriented elements, we expect to provide better service and experience for future users.
Sen, Nazmiye;Sermet, Ibrahim Bulent;Gurler, Nezahat
The Journal of Advanced Prosthodontics
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v.11
no.2
/
pp.105-111
/
2019
PURPOSE. Limited data is available regarding the differences for possible microleakage problems and fitting accuracy of zirconia versus titanium abutments with various connection designs. The purpose of this in vitro study was to investigate the effect of connection design and abutment material on the sealing capability and fitting accuracy of abutments. MATERIALS AND METHODS. A total of 42 abutments with different connection designs [internal conical (IC), internal tri-channel (IT), and external hexagonal (EH)] and abutment materials [titanium (Ti) and zirconia (Zr)] were evaluated. The inner parts of implants were inoculated with $0.7{\mu}L$ of polymicrobial culture (P. gingivalis, T. forsythia, T. denticola and F. nucleatum) and connected with their respective abutments under sterile conditions. The penetration of bacteria into the surrounding media was assessed by the visual evaluation of turbidity at each time point and the number of colony forming units (CFUs) was counted. The marginal gap at the implant- abutment interface (IAI) was measured by scanning electron microscope. The data sets were statistically analyzed using Kruskal-Wallis followed by Mann-Whitney U tests with the Bonferroni-Holm correction (${\alpha}=.05$). RESULTS. Statistically significant difference was found among the groups based on the results of leaked colonies (P<.05). The EH-Ti group characterized by an external hexagonal connection were less resistant to bacterial leakage than the groups EH-Zr, IT-Zr, IT-Ti, IC-Zr, and IC-Ti (P<.05). The marginal misfit (in ${\mu}m$) of the groups were in the range of 2.7-4.0 (IC-Zr), 1.8-5.3 (IC-Ti), 6.5-17.1 (IT-Zr), 5.4-12.0 (IT-Ti), 16.8-22.7 (EH-Zr), and 10.3-15.4 (EH-Ti). CONCLUSION. The sealing capability and marginal fit of abutments were affected by the type of abutment material and connection design.
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