• Title/Summary/Keyword: Co-incubated

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Studies on the Immunomodulating Effects of Polysaccharide Extracted from Ganoderma lucidum on Macrophage (영지버섯 다당체의 마우스 대식세포 면역증강 효과)

  • 김성환;김을상
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.1
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    • pp.148-153
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    • 1997
  • The immunomodulating effects of polysaccharide extracted from Ganoderma lucidum(PSG) on macrophage were evaluated using murine macrophage cell line ATCC TIB 71 cells or peritoneal exudate cells of BALB/c mice. The cell were incubated with various content of PSG for 24 hours at 0.5% $CO_2$ incubator under varying experimental conditions. PSG stimulated the non-specific activites of macrophage such as mitotic activity and expression of surface interleukin-2 receptors by dose-dependent pattern with statistic significance(p<0.001): however, PSG had little immunoregulatory effects on cytokines derived from peritoneal macrophages of BALB/c mice. There were no significant changes in the se-cretion of interleukin-6, interleukin-6, or tumor necrosis factors(Tn) of PSG treated cells compared to the control group. But PSG increased secretion of cytokines(IL-1 and TNF) when the cells were primed and trigged with BCG and IFN-${\gamma}$.

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Fertilizing Ability of Bovine Spermatozoa Following Oviduct Epithelial Cell Co-culture In Vitro (난관상피세포와 공배양한 소 정자의 체외수정능)

  • 황우석;노상호;이병천
    • Journal of Embryo Transfer
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    • v.13 no.3
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    • pp.227-233
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    • 1998
  • The aim of these experiments was to investigate the effects of oviduct epithelial cells on bovine in vitro fertilization. Oviduct epithelial cell monolayers (OEC) on the 4-well dish were prepared according to general procedures. Monolayers were formed within 5days. The medium for OEC culture (TCM199 with 10% FBS) was replaced with IVF-TALP 2h before each experiment. Macromolecules/proteins from oviductal conditioned medium (OM) were recovered by ultrafiltration, which desalted and concentrated macromolecules greater than 5kDa, and this OM were added to W medium (experiment 1). The cleavage rate in OM+OEC group was significantly higher than in OM group (p〈0.01). In this experiment 2, oocytes were inseminated on OEC with sperm which had been pre-incubated with OEC for 0 or 4h before insemination. In this experiment, oocytes were exposed to sperm only 8 h for clarifying the effect. After insemination, oocytes were cultured in CRlaa. At 42 h post insemination, oocytes were denuded and examined for evidence of cleavage. The cleavage rates of oocytes which were inseminated with OEC treated sperm for 4 h were significantly higher than those of the other group (p〈0.01). In conclusion, sperm released from OEC have more fertilizing ability than those before attachment.

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Effect of Red ginseng extract on immune function of Israeli carp, Cyprinus carpio. (홍삼추출물이 이스라엘잉어의 면역반응에 미치는 영향)

  • Choi, Min-Soon;Park, Sung-Woo;Park, Kwan-Ha
    • Journal of fish pathology
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    • v.18 no.3
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    • pp.277-285
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    • 2005
  • Israeli carps were intraperitoneally injected with an aqueous extract of red ginseng (RG) at five different concentrations (0, 1, 10, 25 and 50 mg/kg of body weight). The fish were sacrificed after 3 days and measured the neutrophil activity and plasma components. The activity of lysozyme, Ht, TIC and NBT reduction were significantly enhanced in the concentrations of more than 10 mg/kg. Further attempts were made to measure the effects of RG on leucocyte function at five different concentrations (1-100 ${\mu}g/m{\ell}$), in vitro. The proliferation of lymphocytes co-incubated with phytohaemagglutin (PHA) and concanavalin A (Con A) was significantly increased at 10 ${\mu}g/m{\ell}$ concentration, but suppressed at the highest concentration (100 ${\mu}g/m{\ell}$). RG alone or with lipopolysaccharide (LPS) had no effect on proliferation at any concentrations tested. Migration and reactive oxygen intermediates (ROI) production of leucocytes were enhanced in all concentration higher than 10 ${\mu}g/m{\ell}$. These results suggest that the RG activates the leucocyte function and may be able to be used as a stimulant of fish innate defences.

Treatment of Food Garbage Using a Treatment Reactor and Microbial Consortium (발효소멸기를 이용한 음식물 쓰레기의 감량 및 악취제거)

  • Koh, Rae-Hyun;Lee, Kang-Hyoung;Yoo, Jin-Soo;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.306-312
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    • 2006
  • Disposal of food garbage in most large cities is very troublesome task. To date, microbiological treatment has been received an attention as a garbage decomposition process. In this study, the inoculation effect of some cellulase, amylase and protease-producing bacteria and photosynthetic bacteria on food garbage treatment was examined. They were added into a treatment reactor specially designed in this study together with food garbage and incubated in various conditions for 15 days and the removals of food garbage and foul smell produced during the treatment were analyzed. Average decomposition percentages of the inoculated food garbage in treatment reactor were 11 and 18.8% under intermittent aeration (once in a day) and continuous aeration conditions (2 L/min), respectively, and these were higher than removal percentages in the corresponding uninoculated reactors,3.4 and 13.8%. Optimal pH and temperature for food garbage decomposition by inoculated bacteria were pH 7.0 and $30^{\circ}C$. Maximal decomposition percentage in the inoculated food garbage was 35% under the optimal condition (pH 7, $30^{\circ}C$, and continuous aeration). The malodor compounds generated from food garbage treatment such as complex foul smell and sulfur compounds were effectively reduced about 84% and 25.5%, respectively, with a biofilter composed of purple nonsulfur bacteria trapped in sponge. This decomposing capability of food garbage by these bacteria can be utilized for the rapid and efficient treatment of food garbage.

Characterization of microbial communities and soil organic carbon degradation associated with the depth and thawing effects on tundra soil in Alaska (Alaska 툰드라 토양의 깊이 및 해동 영향에 따른 미생물 군집과 토양 유기 탄소 분해 특성)

  • Park, Ha Ju;Kim, Dockyu;Park, Hyun;Lee, Bang Yong;Lee, Yoo Kyung
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.365-374
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    • 2016
  • In high-latitude regions, temperature has risen ($0.6^{\circ}C$ per decade) and this leads to the increase in microbial degradability against soil organic carbon (SOC). Furthermore, the decomposed SOC is converted into green-house gases ($CO_2$ and $CH_4$) and their release could further increase the rate of climate change. Thus, understanding the microbial diversity and their functions linked with SOC degradation in soil-thawing model is necessary. In this study, we divided tundra soil from Council, Alaska into two depth regions (30-40 cm and 50-60 cm of depth, designated as SPF and PF, respectively) and incubated that for 108 days at $0^{\circ}C$. A total of 111,804 reads were obtained through a pyrosequencing-based metagenomic study during the microcosm experiments, and 574-1,128 of bacterial operational taxonomic units (OTUs) and 30-57 of archaeal OTUs were observed. Taxonomic analysis showed that the distribution of bacterial taxa was significantly different between two samples. In detail, the relative abundance of phyla Actinobacteria and Firmicutes largely increased in SPF and PF soil, respectively, while phyla Crenarchaeota was increased in both soil samples. Weight measurement and gel permeation chromatography of the SOC extracts demonstrated that polymerization of humic acids, main component of SOC, occurred during the microcosm experiments. Taken together our results indicate that these bacterial and archaeal phyla could play a key function in SOC degradation and utilization in cold tundra soil.

$In$ $Vitro$ Development of Goat Parthenogenetic Oocytes Derived from Different Activation Methods (도축장 유래 산양난자의 단위 발생 유기 방법에 따른 체외 발달)

  • Yun, Yun-Jin;Park, Kyeong-Jin;Park, Hee-Sung
    • Journal of Embryo Transfer
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    • v.27 no.1
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    • pp.57-62
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    • 2012
  • Efficient oocyte activation is a key step for the success of nuclear transfer in cloning. Ionomycin sequentially combined with 6-DMAP is now widely used to activate normal oocytes for analytical studies of oocyte activation and to activate reconstructed oocytes after nuclear transfer. The present study investigated sources of oocytes, duration of ionomycin and 6-DMAP, laser and electric stimulation in goat oocyte activation in order to optimize the protocols. Goat ovaries were collected in individual abattoirs during the breeding season and were delivered to the laboratory within 6 h in saline with 100 IU/ml streptomycin and 0.05 mg/ml penicillin. The oocytes were denuded from the cumulus cell by pipetting with 0.2% hyaluronidase in PBS at 20~22 hr post maturation. Oocytes with the polar body were selected and assigned to four groups for parthenogenetic activation. To examine the effect of duration of ionomycin treatment, oocytes after 20~22 hr of maturation were treated with 2.5 uM ionomycin for 1 or 5 min times and then cultured in 2 mM 6-DMAP for 2 or 4 hr. The activated oocytes were cultured in mSOF at $38.5^{\circ}C$ in $CO_2$ 5%, $O_2$ 5% and $N_2$ 90% multi incubator. Cleavage and blastocyst development was observed at 48 hr and day 8 of culture $in$ $vitro$, respectively. Activation rates of oocytes exposed to ionomycin for 1 min(86.4%) were significantly higher than those treated for 5 min(74.3%) duration. This indicated that 1 min ionomycin treatment was most suitable for activation of goat oocytes. The duration of 6-DMAP treat duration was in 2 mM 6-DMAP for 2 hr after 1 min exposure to 2.5 uM ionomycin. The activation rate of oocytes incubated in 6-DMAP for 2 hour(82.5%) was significantly higher than those in oocytes treated with 4 hr(75.5%).

Bioactive and Chemical Properties by Silkworm (Bombyx mori L.) Powder Degradation with Kiwifruit, Papaya, Pineapple and Pear Juice (키위, 파파야, 파인애플 및 배 과즙 처리에 의한 누에분말의 이화학적 특성과 생리활성)

  • Cha, Jae-Young;Kim, Yong-Soon;Ahn, Hee-Young;Eom, Kyung-Eun;Heo, Su-Jin;Cho, Young-Su
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1718-1724
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    • 2010
  • Bioactive and chemical properties of silkworm powder (SP) degradation by fruit extract containing the proteolytic enzymes of kiwifruit, papaya, pineapple and pear were investigated. Silkworm powder was incubated with extracts from each fruit at $60^{\circ}C$ for 24 hr. Protein content was slightly higher in the SP treated with fruit extract than that in the control SP. Major minerals were K, Ca, Mg, and Zn. Major fatty acids were linolenic acid, oleic acid, and palmitic acid. When total protein patterns were analyzed by SDS-polyacrylamide gel electrophoresis (PAGE), silkworm protein was strongly degraded by the treatment of fruit extract from pineapple, papaya, and pear, but little silkworm degradation was observed in kiwifruit extract treatment. Fibriolytic activity was only detected in the SP by the fruit extract treatments from papaya and pear. DPPH radical scavenging activity was slightly stronger in the SP treated with fruit extract than that in silkworm powder. However, all these samples exhibiteda relatively low activity compared with the butylated hydroxytoluene (BHT). These results may provide the basic data for understanding the biological activities and chemical characteristics of SP treated with fruit extract for development of functional foods.

Effect of Glucose and Insulin on Human Gingival Fibroblasts and Periodontal Ligament Cells (포도당 및 인슐린이 인체 치은섬유모세포와 치주인대세포에 미치는 영향)

  • Han, Hee-Ran;Kim, Eung-Tea;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.28 no.1
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    • pp.133-143
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    • 1998
  • Diabetes mellitus is a systemic disease with profound effects on oral health and periodontal wound healing. Uncontrolled diabetes adversely affects surgical wound healing and is often associated with abnormal proliferation of fibroblasts. Human gingival fibroblasts and PDL cells were chosen because they are intimately involved in periodontal therapy and are important for the success of surgical procedure such as guided tissue regeneration. The aim of the present study was to elucidate whether cellular activity and collagen synthesis by glucose pre-treated human gingival fibroblasts and PDL cells are influenced by insulin, and whether healthy cells differ from glucose treated cells. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidified incubator. To evaluate the effect of glucose on gingival fibroblasts and periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4\;cells/well$ culture plates and treated with 20 and 50mM of glucose for 5 days. Then MTT assay was carried out. To evaluate the effect of insulin on glucose-pretreated cells, the cells were seeded at a cell density of $1{\times}10^4\;cells/well$ culture plates and treated with 20 and 50mM of glucose for 5 days. After incubation, $10^3$, $10^4$ and $10^5mU/l$ of insulin were also added to the each well and incubated for 2 days, respectively. Then, MTT assay and collagen synthesis assay were carried out. The results indicate that cellular activity of gingival fibroblasts significantly increased by glucose while periodontal ligament cells were unaffected and cellular activity of gingival fibroblasts and periodontal ligament cells were unaffected by insulin. Collagen synthesis of gingival fibroblast with 20mM glucose and insulin unaffected, but 50mM glucose and insulin increased than control. Collagen synthesis of periodontal ligament cell with 20mM glucose and $10^5mU/l$ insulin significantly increased than other groups and 50mM glucose pretreated PDL cells significantly increased at $10^3mU/l$ insulin but decreased at $10^4mU/l$ insulin. Our findings indicated that these cell types differed in their growth response to glucose, and the increase in collagen synthesis was significantly raised at insulin level of $10^3mU/l$ in gingival fibroblasts and periodontal ligament cells except 20mM glucose pretreated periodontal ligament cells.

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Desorption Kinetics and Structural Changes of Tetracycline Treated Barrier Membranes for Guided Tissue Regeneration (Tetracycline 처리된 조직유도재생술용 차폐막의 약제유리양상 및 구조적 변화)

  • Lee, Sung-Mi;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.27 no.1
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    • pp.1-17
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    • 1997
  • Tetracycline is known to be effective in eliminating periodontopathogens and have collagenolytic activity. This study was performed to observe the desorption kinetics and structural changes of tetracycline-treated barrier membranes for guided tissue regeneration. Four kinds of barrier membranes were tested : $Tefgen^{(R)}$(American Custom Medical, USA) and $Gore-Tex^{(R)}$(W.L. Gore & Associates Inc., USA) as nonresorbable membranes ; Resolut(polyglycolide & polylactide copolymer, W.L. Gore & Associates Inc., USA) and $Biomend^{(R)}$(collagen, Collatec Co., USA) as resorbable membranes. The membranes were cut into discs(diameter : 4mm) and were immersed in 5% tridodecylmethylammonium chloride(TIMAC) ethanol and air-dried. The membrane discs were absorbed with $100{\mu}g/ml tetracycline solution(pH8) for one minute and dried. For desorption kinetics, TC treated discs were immersed in phosphate buffered saline solution (PBS, pH 7.4). PBS was exchanged daily and TC concentration was measured by absorbance at 276nm on UV spectrophotometer. To measure remaining antibacterial activity, discs of 1 day to 4 weeks after desorption were placed on Mueller Hinton agar containing Bacillus cereus and incubated aerobically in $37^{\circ}C$ for twelve hours and the inhibition diameters were measured. To observe the structural change of membranes after TIMAC treatment or immersion in PBS, the membrane discs were examined under SEM. The results were as follows : 1. Total amounts of TC absorbed into membrane discs($0.7536mm^2$) were $2000{\mu}g$, $1800{\mu}g$, $2625{\mu}g$ and $2499{\mu}g$ for $Tefgen^{(R)}$, $Gore-Tex^{(R)}$, $Biomend^{(R)}$ and $Resolut^{(R)}$. 2. The concentration of TC released from barrier membrane discs was maintained over $4{\mu}g/ml$ until the fifth day in nonresorbable membranes and $Resolut^{(R)}$, but until the fourth day in $Biomend^{(R)}$, Until the ninth day in nonresorbable membranes and until the seventh day in resorbable membranes, the TC concentration was maintained over $1{mu}g/ml$. 3. The four membrane discs in the first day showed similar size of inhibition zone. One to four weeks later, the inhibition zone was much smaller in resorbable membrane discs than nonresorbable membrane discs. 4. Any structural change due to treatment of TIMAC was not observed on the nonresorbable membranes. $Resolut^{(R)}$ did not show any structural change except fibrillar loosening during immersion period, but Biomend showed destruction of membrane structure from the first week of immersion. This study indicates that tetracycline treated barrier membranes lead to the sustained release of tetracycline for over 7 days. This slow release pattern of tetracycline may contribute to the favorable clinical outcome of guided tissue regeneration.

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Effects of Extract of Hyperici Japonici Herba on Lung Cells in Asthma-indused Mice by OVA Exposure (지이초(地耳草) 추출물이 OVA로 천식이 유발된 생쥐의 폐세포에 미치는 영향)

  • Lee, Young-Yong;Seo, Young-Bae;Lee, Young-Cheol;Seo, Bu-Il;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.23 no.1
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    • pp.75-83
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    • 2008
  • Objectives : The present study was carried out to investigate the effect of Hyperici Japonici Herba on the proliferation and activation of eosinophils which were prepared from lung cells of asthma-induced mice by ovalbumin(OVA) treatment. Methods : C57BL/6 mouse was exposed to OVA three times a week for 6 weeks. The mouse lung tissues were dissected out, chopped and dessiciated with collagenase(1${\mu}g$/ml). Eosinophils were activated by rIL-3/rmIL-5 co-treatments. The lung cells were treated with extract of Hyperici Japonici Herba(EHH), incubated for 48 hr at $37^{\circ}C$, and analyzed by flow cytometer. ELBA, RT-PCR, immunocytochemistry stain. Results : The cell number ratio of granulocyte, $CD3e^-$/$CCR3^+$, $CD3e^+$/$CD69^+$, $CD4^+$, $CD23^+$/$B220^+$ cells was increased in rmIL-5/rIL-3 treated control group compared to the normal group. Cells numbers in the experimental animal group treated with EHH was all decreased. In ELISA analysis, IL-4, IL-5, IL-13 protein levels and histamine release level were greatly increased in the control group compared to the normal animal group, then significantly decreased in the experimental group with 100 ${\mu}g$/ml of EHH treatment. In RT-PCR analysis, the HT value of IL-4, IL-5, IL-13, CCR3, Eotaxin were increased in the control group compared to the normal animal group, then decreased in the experimental group with 100 ${\mu}g$/ml of EHH treatment. And eosinophil proliferation levels were 18847${\pm}$1527(cpm) in the control group, 4676${\pm}$972(cpm) in the positive control group, and 8675${\pm}$159(cpm), 11352${\pm}$1005(cpm), 14325${\pm}$677(cpm) in the experimental group with 100 ${\mu}g$/ml, 10 ${\mu}g$/ml, 1 ${\mu}g$/ml of EHH treatment. Conclusions : The present data suggested that Hyperici Japonici Herba may have an effects on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of EHH.

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