• Parasites, Hosts and Diseases
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• 제42권1호
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• pp.27-34
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• 2004

We investigated the optimal culture conditions for Cryptosporidium muris in a human stomach adenocarcinoma (AGS) cell line by determining the effects of medium pH and of selected supplements on the development of C. muris. The optimum pH of the culture medium required for the development of C. muris was determined to be 6.6. The number of parasites significantly increased during cultivation for 72 hr (p < 0.05) at this level. On the other hand, numbers decreased linearly after 24 hr of incubation at pH 7.5. When cultured in different concentrations of serum, C. muris in media containing 5% FBS induced 4-7 times more parasites than in 1% or 10% serum. Of the six medium supplements examined, only 1 mM pyruvate enhanced the number of C. muris in vitro. Transmission electron microscopic observation showed the developmental stages of C. muris in the cytoplasm of the cells, not in an extracytoplasmic location. The growth of C. muris in AGS cells provides a means of investigating its biological characteristics and of testing its response to therapeutic agents. However, a more optimized culture system is needed for the recovery of oocysts on a large scale in vitro.

• 대한한방부인과학회지
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• 제19권1호
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• pp.14-30
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• 2006

Purpose : This study was conducted to investigate the inhibitory effects of Cortex ulmi pumilae on cell proliferation in HeLa cell. Methods : Human uterine cervical carcinoma HeLa cells were cultured in the 1%, 5% and 10% concentration of Cortex ulmi pumilae solution for 24 hours, 48 hours and 72 hours for the direct inhibitory effects of Cortex ulmi pumilae. Afterwards, we executed the analysis of the effect of Cortex ulmi pumilae solution on cell proliferation inhibition using XTT assay, DNA fragmentation, molecular biological method through MAP kinase activity and FACS analysis of caspase activity in the HeLa cells. Results : After 48 and 72 hours cultivation, the HeLa cells showed the concentration-dependently significant increase in all Cortex ulmi pumilae solution containing groups compared to the control. In the FACS analysis, all Cortex ulmi pumilae solution containing groups showed concentration-dependent increase compared to the control after 24 hours cultivation and the caspase-3 activities were decreased in all Cortex ulmi pumilae solution containing groups compared to the control after 24, 48 and 72 hours cultivation. After 48 and 72 hours cultivation, we could examined the apparent DNA fragmentation in all Cortex ulmi pumilae solution containing groups. In the XTT study, all Cortex ulmi pumilae solution containing groups showed concentration-dependent decrease compared to the control after 24 and 72 hours cultivation but 10% group after 48 hours and 5% and 10% groups after 72hours were presumed statistically significant differences. The expressions of MAP kinase were decreased in all Cortex ulmi pumilae solution containing groups compared to the control after 24, 48 and 72 hours cultivation. Conclusion : From this study we could suggest that Cortex ulmi pumilae be available to the inhibition of apoptosis of human cervical carcinoma cell line in vitro.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.28-28
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• 2001

The objective of this study was to the effect on subsequent development of EGF present in defined medium during bovine 1)oocyte maturation or 2)embryo culture. The presence of EGF during IVM, irrespective of concentration(1, 10, 100ng/$m\ell$), stimulated cumulus expansion and significantly increased the proportion of oocytes attaining metaphaseII, the rate of cleavage, and develop to blastocyst. 1. In the group of EGF-added medium(1, 10, 100ng/$m\ell$), nuclear maturation rate for in vitro maturation was 91% to 92% but was not significantly higher than control group(87%). 2. For in vitro maturation, in the group of EGF-added medium(1, 10, 100ng/$m\ell$)the rate of cumulus cell expansion degree 2 ranged from 81% to 87%, which was significantly higher than the control group(medium with EGF not added). The rate of in vitro fertilization, developing to 2-to 4- cell stage, was 76% to 80%, which was also significantly higher(p<0.05)than control group(62%). 3. For in vitro maturation, in the group of EGF added in medium(1, 10, 100ng/$m\ell$)the development rate to blastocyst was 24.3% to 27%, which was significantly higher than control group(13.7%). The total cleavage rate in the group of EGF-added medium was 77% to 82%, which was higher than control group. 4. The development rate to blastocyst for 6 days of cultivation and the hatching blastocyst were 30.6% and 59.1%, respectively, in the group of 100ng/$m\ell$ of EGF, which were significantly higher(p<0.05)than control group(14.0% and 24%, respectively), The numbers of cells in blastocyst were 140.2 and 148, respectively, in 10ng/$m\ell$ and 100ng/$m\ell$ of EGF-added medium, which were higher than 108.5 in control group. 5. The development rate of in vitro fertilized embryos to blastocyst in 10ng/$m\ell$ of EGF-added medium co-cultured with somatic cell was 28%, which was significantly higher(p<0.05)than control group(11.8%). The numbers of cells in blastocyst were 141.6 for EGF-added medium and 145 for EGF+co-culture group, which were higher than control(101.6)and medium co-cultured with somatic cells(110.6). These results showed that in vitro maturation and fertilization, EGF was found a significant effect of increase of development rate to blastocyst and cell number.

• 방사선산업학회지
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• 제4권4호
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• pp.297-306
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• 2010

Due to the increasing incidence of diabetes, obesity and hypertensive, stevia has been placed great attentions as the sweetener to substitute sucrose in the world. Stevia was introduced to Korea in 1970's, but it has not been an attractive crop in that time. However, recently it has more attention for the natural food sweet additives. Because stevia have many problems for cultivation especially cultivar, seed germination, fertility, uniformity and glycoside quality, the sport mutation was attempted to in vitro plants for the improvement of some characteristics. The young in vitro plants was nursed on MS medium supplemented with $1mg\;l^{-1}\;GA_3$. Shoots of 10 cm height were irradiated with 0~200 Gy of gamma ray and the every node was separated and inoculated on MS basic medium. The lethality, number and length of shoot, numbers of node and branch were investigated for the evaluation of radiosensitivity. The optimum dose of gamma ray seemed to be around 80 Gy for the sport mutation induction in stevia. The lower node was more sensitive than higher node to radiation.

• 한국자원식물학회지
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• 제32권6호
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• pp.735-742
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• 2019

This study investigated a suitable method that could be applied for Asian chain fern [Woodwardia japonica (L. f.) Sm.] to propagate gametophytes and promote sporophyte formation. The gametophytes used in all experiments were obtained from germinated spores in vitro and were subcultured at 8-week intervals. The most appropriate media for gametophyte propagation was identified by culturing 300 mg of gametophyte in Murashige and Skoog (MS) basal medium (1/8, 1/4, 1/2, 1, 2), and Knop medium for 8 weeks. As a result, fresh weight of the gametophyte was increased by 56.7-fold on MS medium. Moreover, antheridium formation as well as gametophyte growth was improved on MS medium, especially. To improve the sporophyte formation ex vitro, 1.0 g of gametophyte was ground with distilled water and spread on eight combinations onto four different culture mediums, such as bed soil, peat moss, perlite and decomposed granite. Then generation and growth of sporophytes were investigated after cultivation for 10 weeks. As a result of this experiment, peat moss had a promotive effect of sporophyte formation at single-use and mixed culture soils. In particular, a mixture of bed soil, peat moss and perlite in a 1:1:1 ratio (v/v/v) led to the accelerated formation (782.5 ea/pot) and the frond growth of sporophytes. This included increases in length and width of fronds. However, promotive effect of gametophyte growth and sporophyte formation was not found at single-use and treatment with high ratio of bed soil.

• 미생물학회지
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• 제24권2호
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• pp.147-153
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• 1986

나균(M. leprae)을 비롯한 항산균(mycobacterium)의 시험관내 생리력 또는 생명력평가법(in vitro assessment of physiological potential or viability)으로 형광분광측정법(fluorospectrophotometry)를 고안하였다. 균액을 비형광성의 fluorescein diacetate(FDA)로 처리, 균체의 생대사능에 의해 FDA로부터 유리된 체내 fluorescein 량을 Aminco-Bowman 형광분광기로 측정함으로 균체의 생리능을 fluorounit로 표기해 보았다. Fluorounit로 표기된 균체의 생명력을 균배양의 광학밀도(optical density, colony forming unit, 체내 ATP 량 intracellular ATP content)등으로 표기되는 항산균의 생명력고 비교 검토함으로 형광분광법에 의한 시험관내 항산균의 생명력 평가법의 적합성을 살펴보았다. 형광분광측정에 의한 생명도의 평가는 객관성을 띤 기기정량법으로 조작이 간단하고 신속하게 결과를 얻을 수 있어 시험관내 배양의 속도가 완만한 균이나(slow growing mycobacteria, I.e.M. lerpae, Listeriae sp)등의 생명력의 상대적 평가에 활용될 수 있다고 사료된다.

• 미생물학회지
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• 제44권4호
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• pp.358-361
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• 2008

미나리(Oenanthe stolonifera) 발효액이 장내 병원성 미생물과 유익균의 생육, 그리고 장내 세균효소에 미치는 영향을 in vitro 에서 조사하였다. 고상 배지 (Agar plate) 에서의 clear zone 형성에 의한 생육저해를 측정한 결과, Vibro, Salmonella 등의 유해 미생물에 대해서 강한 생육저해 효과를 보였으며 Bifidobacterium longum 에 대해서는 생육저해 효과가 크지 않았다. 액체배지에서의 최소저해농도(MIC) 측정에서도 고장배지에서와 같은 경향을 보여 상대적으로 B. longum 에 대한 생육저해가 가장 적었다. 발효 기간에 따른 영향을 보면 발효 기간이 길수록 적은 양으로도 유해한 균들의 생육을 잘 저해할 수 있는 것으로 나타났다. 장내 세균 효소인 ${\beta}$-glucuronidase와 tryptophanase의 활성에 대해 미나리 발효액은 발효하지 않은 액에 비해 저해효과가 컸으며 발효기간이 길수록 저해효과도 증가하였다. 이상의 실험 결과로서 미나리 발효액은 유해세균에 대한 생육저해능이 크고 상대적으로 유익균인 B. longum에 대한 저해는 적어, 음용 시 정장 효과가 있을 것으로 추정된다.

• 대한한의학회지
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• 제27권1호
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• pp.23-35
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• 2006

Objectives : This study was conducted to investigate the inhibitory effects of Gaejibokryunghwan on cell proliferation in HeLa cells. Methods : Human uterine cervical carcinoma HeLa cells were cultured in the 1%, 5% and 10% concentration of Gaejibokryunghwan extract solution. All three were cultured for 24 hours, 48 hours and 72 hours each, to examine the inhibitory effects of Gaejibokryunghwan. Afterwards, we drew out the effect of Gaejibokryunghwan extract solution by making 5 analysis. First analysis was to measure the proliferation rate of cells. Second was FACS analysis. Third was to estimate the activity or caspase-3. Fourth, we used XTT assay to analyze the activation or cells. Ana lastly, a molecular biological method was used to determine activation of MAP kinase in the HeLa cells. Results : After 24, 48 and 72 hours cultivation, the proliferation of HeLa cells showed the dose-dependent decrease in all Gaejibokryunghwan extract solution groups compared to the control group. In the FACS analysis, Gaejibokryunghwan extract solution groups showed increased caspase expression compared to the control group, except for the group for 48 and 72 hours in 1 % concentrate. Caspase-3 activities were increased in all, except tile group cultured for 24 hours in 5% concentrate and the groups cultured for 48 hours in 1% and 5% concentrate. In the XTT study, 1% Gaejibokryunghwan extract solution groups showed increase compared to the control group, but other Gaejibokryunghwan extract solution containing groups showed significant decrease compared to the control after 24, 48 and 72 hours of cultivation. The expressions of MAP kinase were decreased in all Gaejibokryunghwan extract solution containing groups compared to the control group after 24, 48 and 72 hours of cultivation. Conclusions : From this study, we could suggest that Gaejibokryunghwan be available to the inhibition of proliferation of human cervical carcinoma cell line, HeLa cells in vitro.

• 대한한방부인과학회지
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• 제19권2호
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• pp.34-48
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• 2006

Purpose : This study was conducted to investigate the inhibitory effects of Dangguijakyaksan on cell proliferation in HeLa cells. Methods : Human uterine cervical carcinoma HeLa cells were cultured in the 1%, 5% and 10% concentration of Dangguijakyaksan extract solution for 24 hours, 48 hours and 72 hours for the direct inhibitory effects of Dangguijakyaksan. Afterwards, we executed the analysis of the effect of Dangguijakyaksan extract solution on cell proliferation inhibition using XTT assay, molecular biological method through MAP kinase activity and FACS analysis of caspase activity in the HeLa cells. Results : After 24, 48 and 72 hours cultivation, Dangguijakyaksan extract solution group showed significant decrease of HeLa cells except 1% solution after 24 hours compared with the control group. In the FACS analysis, Dangguijakyaksan extract solution groups showed increase of caspase activity except 1% solution after 48 hours compared with the control group. In the XTT assay, the caspase-3 activities were increased in Dangguijakyaksan extract solution groups except 1% solution after 24 hours in a dose-dependent manner. In the XTT study, cell activities were significantly decreased in 10% Dangguijakyaksan extract solution groups after 48 and 72 hours cultivation compared with the control group. In all Dangguijakyaksan extract solution groups, The activities of MAP kinase were decreased after 24, 48 and 72 hours cultivation compared with the control group. Conclusion : It could be concluded that Dangguijakyaksan is available to the inhibition of proliferation of human cervical carcinoma cell line in vitro.

• 한국유기농업학회지
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• 제6권1호
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• pp.133-149
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• 1997

Recently, the importance of management and cultivation of grasses has been increased in Korea. Among these cultural practices, the appropriate control of diseases is considered more important than other cultivation techniques such as fertilization and irrigation. The damages of brown patch and large patch caused by Rhizoctonia spp. and Pythium blight caused by Pythium spp. are serious in the major cultivation area of turfgrass in Korea. Since these diseases are difficult to control by agrochemicals, the damages are very serious if these are occured. The periodic spray of agrochemicals, to protect and control these diseases could make some problems of toxicity and environmental pollution as well as rising of non-target diseases. Therefore, the biological methods to control diseases have been required to decrease problems resulted from overuse of agrochemicals, to conserve natural ecosystem, and to control effectively diseases of grasses in the long period. The number of studies about biological control using antagonistic microorganisms have been increased for last half century. However, the application of biological control method has been very limited. In this study, thirteen isolates of R. cerealis, 8 isolates of R. solani and 3 isolates of Phthyn spp. have been isolated from diseased turfgrass in golf course and grass-culture area that have patch and wilting symptoms of zoysia grass and creeping bentgrass. Isolation frequency of R. cerealis and R. solani was high in especially zoysiagrass, while Pythym spp. was isolated from bent grass at low frequency but showed high pathogenicity. Totally, 205 isolates of soil microorganisms were isolated in this study as primary antagonistic microorganism by Herr's triple agar layer plate and dual culture method using rhizosphere of grasses, soil of crop field as the source of antagonistic microorganisms. Among the 205 isolates, 23 isolates were actinomycetes and 182 isolates were bacteria. All of the actinomycetes were isolated by Herr's method. Antagonistic effect of primary isolated microorganisms was tested for in vitro mycelial growth inhibition against pathogenic fungi isolated from grasses and for inhibition of disease occurrence in 24 well tissue culture plate and pot experiment. Then, four isolated of bacteria which are BG23, BG74, BG136 and BG171 were selected as antagonistic microorganisms against soil-born pathogenic fungi of bentgrass.