• Animal cells and systems
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• 제8권4호
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• pp.329-333
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• 2004

Japanese flounder (Paralichthys olivaceus) head kidney (HK) leukocytes were incubated with $10^3$ to $10^{-3}$ ng levamisole/ml for 4, 24 or 48 h and then assayed for their natural cytotoxic activity against xenogeneic tumor cells. This activity was slightly increased after 24 h of incubation. In a second experiment, fish were fed 0, 75, 150 or 300 mg levamisole/kg diet for 10 consecutive days. The fish were then fed a commercial non-supplemented diet and sampled 0, 1, 2, 3, 4 or 6 weeks post-administration of levamisole. The cytotoxic activity was found to be increased with increasing levamisole dose and remained greatly enhanced until the end of the experiment. In conclusion, levamisole enhanced flounder natural cytotoxic cell activity both in vitro and in vivo and had a great lasting action when administered by feeding.

• Journal of Microbiology and Biotechnology
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• 제27권6호
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• pp.1098-1105
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• 2017

Vesicular stomatitis virus G glycoprotein (VSV-G) has been widely used for pseudotyping retroviral, lentiviral, and artificial viral vectors. The objective of this study was to establish a potential approach for large-scale production of VSV-G. To this end, VSV-G was cloned with an N-terminal His-tag into Pichia pastoris expression vector pPIC3.5K. Three clones ($Mut^s$) containing the VSV-G expression cassette were identified by PCR. All clones proliferated normally in expansion medium, whereas the proliferation was reduced significantly under induction conditions. VSV-G protein was detected in cell lysates by western blot analysis, and the highest expression level was observed at 96 h post induction. VSV-G could also be obtained from the condition medium of yeast protoplasts. Furthermore, VSV-G could be incorporated into Ad293 cells and was able to induce cell fusion, leading to the transfer of cytoplasmic protein. Finally, VSV-G-mediated DNA transfection was assayed by flow cytometry and luciferase measurement. Incubation of VSV-G lysate with the pGL3-control DNA complex increased the luciferase activity in Ad293 and HeLa cells by about 3-fold. Likewise, incubation of VSV-G lysate with the pCMV-DsRed DNA complex improved the transfection efficiency into Ad293 by 10% and into HeLa cells by about 1-fold. In conclusion, these results demonstrate that VSV-G could be produced from P. pastoris with biofunctionalities, demonstrating that large-scale production of the viral glycoprotein is feasible.

• 한국토양비료학회지
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• 제31권3호
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• pp.233-237
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• 1998

토양에 시용되는 각종 폐유기물들의 토양중 질소 무기화율을 알아보기 위하여 3종의 폐유기물(A:도시고형폐기물+하수슬러지, B:목재잔사, C:목재잔사+하수슬러지)를 토양에 처리한 뒤 12주 동안 $25^{\circ}C$ 호기조건에서 질소 무기화율을 조사한 결과, 초기 질산화율은 C 처리구가 다른 처리구에 비하여 높았다. 또한 무기화된 총질소량은 C 처리구에서 가장 많았으나 실험기간 동안의 질소 무기화율과 질산화율은 무처리구에서 높게 나타났다.

• 대한수의학회지
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• 제30권4호
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• pp.479-485
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• 1990

This study was carried out to observe in vitro effect of praziquantel on the viability and internal organ changes of Heterophyopsis continua with light microscopy. Metacercariae were collected from the perch, Lateolabrax japonicus, by artificial digestion technique and fed to 2-week old chickens. Adult worms were recovered from the small intestines of chickens 8 days after infection. For working solutions, praziquantel was diluted with TC199 medium at the concentration of 0.01, 0.1, 1 and $10{\mu}g/ml$. To each petri dish containing 10ml of solution, 5~10 worms were introduced and incubated at $37^{\circ}C$. Motlity of worms was observed at 5, 15, 30, 60 minutes, 1, 2, 4 and 6 hours after incubation. For light microscopy, worms were fixed in 10% formalin under cover glass pressure and stained with Semichon's acetocarmine. The results were as follows: 1. In $0.01{\mu}g/ml$ praziquantel, the worms had their mobility until 6 hours post treatment. However, worms in over $0.1{\mu}g/ml$ of praziquantel contracted within 5 minutes and immobilized. 2. Intestine of the worm incubated in $0.001{\mu}g/ml$ praziquantel for 5 minutes was dilated and intestinal wall was thickened. 3. In incubated over $0.1{\mu}g/ml$ praziquantel, pharynx of the worm protruded out from oral sucker. 4. The lowest effective lethal concentration of praziquantel on H. continua was $0.1{\mu}g/ml$. The worms exposed to the drug were observed to be immobilized immediately after incubation in solutions of over $0.1{\mu}g/ml$ concentration. All of the worms in early period showed severe contraction and those in late period showed severe dilation.

• Horticulture, Environment, and Biotechnology : HEB
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• 제59권6호
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• pp.847-856
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• 2018

This research aimed to evaluate the effectiveness of hot water ($43-53^{\circ}C{\cdot}5min^{-1}$; $47^{\circ}C{\cdot}2-6min^{-1}$), 1-methylcyclopropene (1-MCP) at $50-300nL\;L^{-1}$ and a combination of hot water ($47/49^{\circ}C{\cdot}5min^{-1}$) and phosphite $40%\;P_2O_5+20%\;K_2O$;$40%\;P_2O_5+10%\;Zn$) in anthracnose control and the effect on fruit quality [fresh weight loss (FWL-%); pH, total soluble solids ($TSS-^{\circ}Brix$), and titratable acidity (TA = % citric acid (CA)] of passion fruit ( Passiflora edulis f. flavicarpa ) at the postharvest stage. When the fruits were in the stage of 0% dehydration and fully yellow peels, they were disinfested and inoculated with Colletotrichum gloeosporioides. They were then subjected to the above mentioned treatments; this was followed by incubation for 120 h. The diameter of the disease lesions was monitored daily. After the incubation, a physico-chemical analysis was performed. Hot-water treatment resulted in disease reduction at 47 and $49^{\circ}C$ for 4 and 5 min. The combination of hot-water treatment at $47^{\circ}C$ (4 or 5 min) and application of the phosphite of K or Zn significantly reduced disease severity in fruits. The 1-MCP treatment reduced anthracnose severity in passion fruit mainly at $200nL\;L^{-1}{\cdot} 24h^{-1} $. None of the treatments significantly changed the physico-chemical characteristics of the fruit [FWL (2.6-4.1%); pH (3.2-3.5), TSS ($8.9-10.9^{\circ}Brix$), and TA (1.8-2.5% CA)].

• 한국초지조사료학회지
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• 제36권4호
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• pp.309-317
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• 2016

본 연구는 국내산 조사료인 IRG 사일리지의 이용성을 증진하기 위해 다양한 미생물제의 첨가 시 IRG 사일리지의 in vitro 반추위 발효특성 및 소화율에 미치는 영향을 조사하였으며, 미생물제의 접종 후 배양시간이 경과함에 따라 IRG 사일리지의 품질 및 in situ 반추위 소화율에 미치는 영향을 분석하였다. 미생물제로는 LC, BS 및 SC ($2.7{\times}10^7CFU/m{\ell}$)를 사용하였으며, IRG 사일리지에 $0.5{\times}10^4CFU/g$가 되도록 첨가하여 수행하였다. In vitro 실험 결과, 암모니아태 질소 함량은 12시간 배양 시 대조구보다 미생물제 처리구에서 높았고(p<0.05), 총 VFA 농도와 건물 분해율의 경우에도 유의적 차이는 없었지만, 대조구보다 미생물제 처리구에서 증가하였으며, 특히 L. casei에서 높게 나타났다. 미생물제를 접종한 후 5일간 배양한 결과, IRG 사일리지의 pH는 대조구보다 미생물제 처리구에서 낮았으며(p<0.05), 젖산 농도는 배양 1~5일 동안 대조구보다 미생물제 처리구에서 높았으며(p<0.05), 다른 처리구보다 LC-IRGS에서 접종 직후 가장 높았다(p<0.05). In situ 건물 분해율은 대조구보다 모든 처리구에서 증가하는 경향을 나타내었으며, 접종 직후에는 SC-IRGS에서 높았으나 이후 LC-IRGS에서 증가하였다. 본 연구결과를 보면, IRG 사일리지의 이용성을 제고하기 위해 LC, BS 및 SC을 활용한다면 IRG 사일리지의 반추위 내 소화율이 개선될 수 있으며, 특히 IRG 사일리지에 L. casei를 첨가하여 단시간 추가 발효하여 젖소에 급여한다면 IRG 사일리지 내 초산 및 낙산의 감소로 품질 및 소화율 개선에 효과가 있을 것으로 사료된다.

• 한국수정란이식학회지
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• 제11권3호
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• pp.259-269
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• 1996

The present study was undertaken to examine the critical effect of $Ca^2$+ concentration on electrostimulation and post-electrostimulation media for electric activation of in vitro matured oocytes of Korean Native Cattle. Oocytes collected from slaughterhouse ovaries were matured in TCM 199 containing FSH, estradiol-17$\beta$ and FBS with granulosa cell monolayer for 24 hours and denuded with hyaluronidase. And then cumulus-free oocytes were submitted to a DC field of 1.0 kV/cm for 60 $\mu$sec in electroporation media(0.28 M mannito' and PBS) with different $Ca^2$+ concentations (0.00, 0.05, 0.10 and 0.15 mM). Stimulated oocytes were stained and examined for pronuclear formation after incuhation in SOF for 12 hours. The rates of pronuclear formation in hovine oocytes electrically stimulated in 0.28 M mannitol with 0.05, 0.10 and 0.15 mM $Ca^2$+(60.3, 82.2 and 75.0%) were significantly higher than without $Ca^2$+(6.3%) at 12 hours after an electric pulse(p<0.005). The activation rates of Korean Native Cattle oocytes stimulated in PBS supplemented with 0.05, 0.10 and 0.15 mM $Ca^2$+(71.0, 75.8 and 75.4%) were significantly higher than without $Ca^2$+(23.5%) after post-stimulation incubation(p<0.005). After incubation of oocytes in SOF with and without $Ca^2$+ following electric stimulation in 0.28 M mannitol with 0.10 mM $Ca^2$+, the rates of pronuclear formation of bovine oocytes in $Ca^2$+-free SOF(85.7%) was significantly higher than in SOF with 1.71 mM $Ca^2$+(62.5%, p<0.05). When oocytes were stimulated in two electrostimulation media supplemented with $Ca^2$+ and incubated in $Ca^2$+-free SOF, there were no significant differences in the rates of pronuclear formation hetween 0.28 M mannitol and PBS. These results indicate that a single electric pulse could induce activation of Korea Native Cattle oocytes in 0.28 M mannitol and PBS supplemented with $Ca^2$+. Furthermore, to improve the activation rates, it was hetter that stimulated oocytes were incubated in $Ca^2$+-free SOF after electric stimulation than in SOF with $Ca^2$+.

• 한국미생물·생명공학회지
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• 제16권6호
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• pp.517-521
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• 1988

저온성 미생물의 살균후 오염에 관하여 HTST 살균법에 의하여 제조된 음용유를 대상으로 연구하였다. 살균후 제조공정중의 시료와 충병제품의 시료론 시유와 탈지유별로 대형 음용유 공장에서 채취하였다. 제조공정중의 시료를 시유의 경우 아홉개의 각각 다른 시료채취 장소에서 각각 채취하였다. 각각 의 제조공정중의 시료는 21$^{\circ}C$에서 16시간 동안 예비배양(PI)을 한 후 표준평판검사(SPC)바 그람음성균 검사(CVT)를 실시하였다. 보존성검사를 위하여 각시료를 7.2$^{\circ}C$에서 7일간 보존한 후 풍미검사와 SPC 및 저온미생물검사(PBC)를 실시하였다. 제조공정중의 시료 이외에 실제품의 보존성을 검사하기 인하여 1000ml용 종이용기에 충병된 완제품을 각 제품(시유 및 탈지유)의 생산이 시작될 때마다 일련적으로 연속하여10개씩 충병기로부터 채취하였다. 충병제품 시료들에 대해 예비배양(PI)을 거친 후 SPC와 CVT를 실시하였다. 보존성 검사를 인하여 각 충병제품 시료를 7.2$^{\circ}C$에서 7, 10 및 14일간 보존 후 풍미 검사와 SPC 및 PBC를 실시하였다. 제조공정중의 시료의 예비배양 후 CVT(PI-CVT) 평균치는 시료채취 장소에 따라 차이가 있었다. 살균유 저장탱고로부터 충병기까지의 제조공정에서 오염이 주로 발생하였다. 충병제품 시료의 PI-CVT 값은 10 및 14일간 보존 후 풍미와 밀접한 상관관계를 나타내었다. 제조공정중의 시료의 PI-CVT 값은 10일간 보존후 양호한 풍미를 유지한 충병제품 시로의 전체 충병제품 시료에 대한 백분율과 좋은 상관관계를 보였다.

• 생명과학회지
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• 제17권1호
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• pp.64-69
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• 2007

본 연구에서는 UVB에 조사된 NIH3T3 세포에서 세포고사와 DNA 단사절단에 미치는 fisetin후처리의 효과에 대해서 연구하였다. 세포에 UVB $(200J/m^2)$를 조사하고 정상배지에서 48시간 배양한 세포의 세포고사에 수반되는 핵분절은 50% 정도의 세포에서 관찰되었다. 흥미롭게도 배양배지에 fisetin이 첨가될 경우 핵분절을 보이는 세포의 빈도는 상당한 감소를 보였다. 알칼리 아가로스 겔에 의한 DNA 단사절단 분석에서 자외선 조사 후 fisetin처리는 정상배지 배양시보다 단사절단의 빈도를 감소시켜 DNA크기의 증가를 유도하였는데 이는 fisetin이 UVB에 의한 DNA 상해의 회복에 긍정적 효과를 나타냄을 시사한다 Western blot 분석에 의해 fisetin은 자외선 조사에 의해 활성화되는 p53의 수준을 유의한 수준으로 감소시키며 자외선 상해의 결과 세포주기의 정지에 수반되는 PCNA의 감소 경향을 다소 완화시켰다. 이러한 결과들은 fisetin이 DNA 회복의 활성을 통해 세포고사의 감소에 기여하며 이 과정에서 p53 및 PCNA의 수준변화와 관련하여 행동함을 시사한다.

• 농업과학연구
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• 제44권4호
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• pp.558-565
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• 2017

This study was conducted to determine the effect of inoculation of microorganism isolated from pig feces on nutrient contents of fermented hulless barley. The microbial flora in feces of a total of four crossbred piglets ($Landrace{\times}Yorkshire{\times}Duroc$) was analyzed by 16s rRNA sequencing. The most abundant strain was then selected for fermentation of hulless barley. Lactobacillus plantarum (L. plantarum) was dominant (64.56%) in intestinal microbial flora in the pig feces. The selected candidate strain showed significantly higher survival rate at pH 7 than at pH 2.5 and 3.0 (p < 0.05). Incubated culture containing the candidate strain showed an increased growth rate with lower pH levels after 7.5 h incubation compared to initial incubation period (p < 0.05). When compared with commercial multiple probiotics which were used as control, the selected strain showed faster growth rate at 5 h post-incubation (p < 0.05). During the fermentation period, neither inoculated nor non-inoculated control hulless barley showed any change in pH value. Crude fat, fiber and ash contents were lower (p < 0.05) in hulless barley inoculated by the selected strain compared to control. However, moisture, energy, NDF and ADF were not affected by the inoculation of strain or fermentation period. Lactic acid was increased and acetic acid was decreased in the hulless barley inoculated with the selected strain during the fermentation period (p < 0.05). Taken together, our results suggest that L. plantarum derived from the pigs could be utilized as a new microorganism for manufacturing fermented feed stuffs.