• Title/Summary/Keyword: suspension cell

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Oocyte-sperm Binding Assay (OSBA) Technique for Rapid Q/C of IVF Culture Condition (체외수정용 배양조건의 신속한 Q/C를 위한 정자-난자 결합분석법(OSBA) 개발)

  • 정구민;신영수
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.163-169
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    • 2001
  • OSBA(oocytes-sperm binding assay) is a tool developed for rapid test of optimal condition of IVF medium and protein source by binding ability of mouse sperm and egg. Mouse oocyte-cumulus complexes were prepared by removing of the cumulus cells with 0.1% hyaluronidase. 10$\pm$2 oocytes per 30 ${mu}ell$ medium drop were inseminated with 3 ${mu}ell$ sperm suspension and were cultured f3r 3 hours and 24 hours, respectively. And the oocytes were recovered gently and the No. of sperm bound on oocytes were counted. In the Exp. 1, the ratio of oocytes bound with one sperm at least were 60.2%(50/83), 2%(2/77) and 100%(79/79) in the medium with no protein, FBS(15%, v/v) and BSA(0.4%. w/v), respectively, Fetal bovine serum(FBS) seriously inhibited sperm binding on oocyte, although bovine serum albumin(BSA) promoted the binding ability. The inhibiting effect of FBS was dependent on the concentration of FBS. The sperm binding ability according to oocyte maturity was tested in the Exp. 2. There was no significant difference between Met. II (mature) and Met. I (intermediate mature) oocytes in the number of oocytes bound with sperm and the number of sperm bound on oocytes. Finally, in Exp. 3, two batches of Ham's F10 medium with good and poor quality by OSBA were tested (The ratios of embryos developed from PN 1-cell stage to hatched blastocyst; 25% vs. 70%). In the medium with good quality, sperm binding ability was significantly increased (P < 0.05). The ratio of oocytes bound with one sperm at least was 66% and 90% in the medium with poor and good quality, respectively. Conclusively, It was possible to test IVF medium condition rapidly and easily by OSBA.

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Combination of Hydrophobic Filtration and Enrichment Methods for Detecting Bacillus cereus in Fresh-Cut Cabbage

  • Lee, Sujung;Choi, Yukyung;Lee, Heeyoung;Kim, Sejeong;Lee, Jeeyeon;Ha, Jimyeong;Oh, Hyemin;Lee, Yewon;Kim, Yujin;Yoon, Yohan;Lee, Soomin
    • Journal of Food Hygiene and Safety
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    • v.33 no.5
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    • pp.325-329
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    • 2018
  • This study developed a rapid detection method for Bacillus cereus in fresh-cut cabbages. Fresh-cut cabbage samples were inoculated at 1-, 2- and 3-Log CFU/g, and pathogens were enriched in tryptic soy broth containing 0.15% polymyxin B at $30^{\circ}C$, $37^{\circ}C$, and $42^{\circ}C$ to determine the detection limit and appropriate enrichment temperature for multiplex PCR detection. Enriched bacterial cells in enrichment broth were collected in a hydrophobic filter prior to DNA extraction for multiplex PCR. Filters were resuspended in distilled water, and DNA was extracted from the suspension. DNA samples were further analyzed by multiplex PCR. Detection limit of multiplex PCR was 5-Log CFU/mL. B. cereus cell counts were higher (P < 0.05) at $42^{\circ}C$ than other temperatures. Detection rate of 1-, 2-, and 3-Log CFU/g inoculated samples were 60%, 80%, and 100% after enrichment respectively. However, when enriched samples were filtered with hydrophobic membrane filter, detection rates became 100%, regardless of inoculation level. Results indicate a combination of enrichment with hydrophobic filtration improves rapid detection efficiency of B. cereus in fresh-cut cabbage by multiplex PCR.

Influence of $TiO_2$ Concentrations and Irradiation Lights on the Photocatalytic Reaction for Inhibiting Growth of Streptococcus mutans ($TiO_2$ 농도 및 여기광에 따른 광촉매 반응이 Streptococcus mutans의 생장에 미치는 영향)

  • Kang, Si-Mook;Lee, Hae-Na;Kim, Hee-Eun;Kim, Baek-Il
    • Journal of dental hygiene science
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    • v.14 no.3
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    • pp.319-324
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    • 2014
  • The aim of this study was to evaluate influences of titanium dioxide ($TiO_2$) concentrations and irradiation times on growth of Streptococcus mutans when irradiated by visible light (405 nm wavelength) and by ultraviolet light (254 nm wavelength). To find the optimal antibacterial concentration of $TiO_2$, 0.01, 0.1, 1.0, and 10.0 mg/ml $TiO_2$ suspension was prepared with sterilized distilled water. S. mutans cultured media was added to $TiO_2$ solution to set the final cell count to $10^4CFU/ml$. The photocatalytic reaction was induced by irradiating 254 nm and 405 nm lights for 10 minutes. To compare the bactericidal activities according to irradiation times, all photocatalytic reaction was carried out with 0.1 mg/ml $TiO_2$ for 0, 10, 20, 30, and 40 minutes with both lights. After the photocatalytic reaction, $100{\mu}m$ of the reaction mixture was immediately plated on brain heart infusion agar. These plates were placed at 5% $CO_2$, $37^{\circ}C$, for 24 hours and the bacterial colonies were counted. All experiments were performed in quintuplicate. One-way ANOVA was used to determine whether there were any significant differences between the $TiO_2$ concentrations or the irradiation times. The most effective concentration of $TiO_2$ for its photocatalytic bactericidal effect on S. mutans was 0.1 mg/ml when irradiated with 254 nm and 405 nm lights. The longer the irradiation time, the bigger the bactericidal effect for both wavelengths. Over 99% of bacteria in the inoculum were killed after irradiation with 254 nm for 20 minutes and with 405 nm for 40 minutes. In conclusion, a photocatalytic reaction of $TiO_2$ induced by visible light of 405 nm constitutes the bactericidal effect on S. mutans.

Study on Ti-doped LiNi0.6Co0.2Mn0.2O2 Cathode Materials for High Stability Lithium Ion Batteries (고안정성 리튬이온전지 양극활물질용 Ti 치환형 LiNi0.6Co0.2Mn0.2O2 연구)

  • Jeon, Young Hee;Lim, Soo A
    • Journal of the Korean Electrochemical Society
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    • v.24 no.4
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    • pp.120-132
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    • 2021
  • Although the development of high-Nickel is being actively carried out to solve the capacity limitation and the high price of raw cobalt due to the limitation of high voltage use of the existing LiCoO2, the deterioration of the battery characteristics due to the decrease in structural stability and increase of the Ni content. It is an important cause of delaying commercialization. Therefore, in order to increase the high stability of the Ni-rich ternary cathod material LiNi0.6Co0.2Mn0.2O2, precursor Ni0.6Co0.2Mn0.2-x(OH)2/xTiO2 was prepared using a nanosized TiO2 suspension type source for uniform Ti substitution in the precursor. It was mixed with Li2CO3, and after heating, the cathode active material LiNi0.6Co0.2Mn0.2-xTixO2 was synthesized, and the physical properties according to the Ti content were compared. Through FE-SEM and EDS mapping analysis, it was confirmed that a positive electrode active material having a uniform particle size was prepared through Ti-substituted spherical precursor and Particle Size Analyzer and internal density and strength were increased, XRD structure analysis and ICP-MS quantitative analysis confirmed that the capacity was effectively maintained even when the Ti-substituted positive electrode active material was manufactured and charging and discharging were continued at high temperature and high voltage.

Biological Control of Tomato and Red Pepper Powdery Mildew using Paenibacillus polymyxa CW (Paenibacillus polymyxa CW를 이용한 고추 및 토마토 흰가루병 방제)

  • Kim, Yong-Ki;Choi, Eun-Jung;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Jee, Hyeong-Jin;Park, Jong-Ho;Han, Eun-Jung;Jang, Bo-Kyung;Yun, Jong-Cheul
    • The Korean Journal of Pesticide Science
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    • v.17 no.4
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    • pp.379-387
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    • 2013
  • In order to improve practical utility of agro-microorganisms (AMs) which had been cultured and disseminated to promote plant growth and to control crop diseases, 51 isolates of AMs were collected from 18 agricultural extension centers in local government and screened for multi-functions such as antifungal activity, activities of phosphorus solubilization, IAA and siderophore production, nitrogen fixation, and hydrolytic enzyme activity. Finally we selected one isolate showing good antifungal activity and multi-functions related to plant growth and disease control. The selected isolate, Paenibacillus polymyxa CW, showed good inhibitory effect against plant pathogens, Pyricularia gresea, Colletotrichum acutatum, Fusarium oxysporum, Phomopsis sp., Aspergillus niger, Rhizoctonia solani and Phytophthora capsici. Suppressive effect of P. polymyxa CW against the used plant pathogens except for R. solani was much higher than that of P. polymyxa AC-1 storing in National Academy of Agricultural Science. We found P. polymyxa CW isolate showed good activity in siderophore and IAA formation, and nitrogen fixation. With P. polymyxa CW isolate, siderophore formation activity was similar to that of P. polymyxa AC-1, but IAA formation and nitrogen fixation activity was much higher than that of P. polymyxa AC-1. However neither P. polymyxa CW nor P. polymyxa AC-1 showed hydrolytic enzyme (chitinase, pectinase and cellulase) activity. The treatment of P. polymyxa CW with culture suspension of different cell density ($10^8$, $10^7$. $10^6$ cfu/ml) showed that the highest density reduced incidence of red pepper powdery mildew by 68.3% after 10 days of application. As application density of P. polymyxa CW was decreased, its control efficacy was proportionally decreased. In addition, when P. polymyxa CW was treated to control tomato powdery mildew at the same concentrations and their control effects were investigated after 7 days of inoculation, disease incidence was 0.03, 19.5, 45.7%, respectively, compared to 56.3% that of untreated check. Like red pepper powdery mildew, increase of application density of P. polymyxa CW resulted in increase of its control efficacy proportionally. P. polymyxa CW showed a density-dependent control efficacy against red pepper and tomato powdery mildews. Therefore we think that mode of action of the antagonist for suppressing two powdery mildew diseases might be antibiosis and density of more than $10^8cfu/ml$ was needed to control effectively the two diseases. On this basis, we think that P. polymyxa CW can be a promising control agent for suppressing powdery mildews of red pepper and tomato.

Protoplast Fusion of Nicotiana glauca and Solanum tuberosum Using Selectable Marker Genes (표식유전자를 이용한 담배와 감자의 원형질체 융합)

  • Park, Tae-Eun;Chung, Hae-Joun
    • The Journal of Natural Sciences
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    • v.4
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    • pp.103-142
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    • 1991
  • These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

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