• Title/Summary/Keyword: Co-Expression

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Cooperative Induction of HL-60 Cell Differentiation by Combined Treatment with Eugenol and 1α,25-Dihydroxyvitamin D3 (Eugenol과 1α,25-dihydroxyvitamin D3의 병합처리에 의한 HL-60 세포의 분화 유도)

  • Oh, Mi-Kyung;Park, Seon-Joo;Kim, Nam-Hoon;Cho, Jin-Kyung;Jin, Jong-Youl;Kim, In-Sook
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1191-1196
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    • 2007
  • Eugenol (4-allyl-2-methoxyphenol) is a main component of essential oils obtained from various spices. Recent reports have shown that eugenol induces growth inhibition and apoptosis of malignant tumor cells. In this study, the stimulatory effect of eugenol on cell differentiation was investigated in HL-60 promyelocytic leukemia cells. When HL-60 cells were treated in combination with 150 ${\mu}M$ of eugenol and 3 nM of $1{\alpha},25-dihydroxyvitamin$ $D_{3}$, cell growth was slower than that of cells treated with eugenol or $1{\alpha},25-dihydroxyvitamin$ $D_{3}$ alone. Eugenol enhanced low dose of $1{\alpha,25-dihydroxyvitamin }$ $D_{3}-induced$ a $G_{0}/G_{1}$ phase arrest in cell cycle. Consistent with this, combined treatment of eugenol and $1{\alpha},25-dihydroxyvitamin$ $D_{3}$ cooperatively increased p27 level and decreased cyclin A, cdk 2 and cdk 4 levels, which are cell cycle regulators related to $G_{0}/G_{1}$ arrest. According to flow cytometric analysis, the expression of CD14 (monocytic differentiation marker) was more increased in the cells co-treated with eugenol and $1{\alpha},25-dihydroxyvitamin$ $D_{3}$. These results indicate that eugenol potentiates cell differentiation mediated by $1{\alpha},25-dihydroxyvitamin$ $D_{3}$ of suboptimal concentration. The differentiation-inducing property of eugenol maybe contributes to chemopreventive activity of cancer.

Development and Expression of Porcine Embryos by Direct Injection of Sperm Treated with Exogenous DNA (외래유전자 도입정자를 이용한 돼지 체외성숙 난포란의 Intracytoplasmic Sperm Injection (ICSI) 후 후기 배로의 발달율과 외래유전자의 발현에 관한 연구)

  • 정기화;조성근
    • Journal of Embryo Transfer
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    • v.17 no.1
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    • pp.45-53
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    • 2002
  • The main goal of this study was to produce transgenic porcine embryos by direct injection of sperm-mediated exogenous DNA. Spermatozoa (6$\times$10$^{6}$ sperms of final concentration) were mixed with pcDNA LAC Z (20 ng/$\mu$l) and subjected into electroporation (300~750 volts, 25 $\mu$F, 0.4 cm electrode). After sperm injection, the oocytes were activated electrically (1.7 KV/cm, 30$\mu$sec, single pulse) in 0.3 M mannitol solution or not. The sperm injected eggs were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air fur 144 h. The rates of cleavage and development into blastocyst stage in activation group were significantly higher than those of non-activation group (79.6% and 24.1% vs. 46.3% and 14.4%, respectively, p<0.05). Control oocytes and shame injection were developed to blastocysts low (2.5%). Sixty five (27.1%) out of 240 embryos observed in activation and non-activation groups were showed positive by X-gal staining. However, all embryos in both groups were expressed partial or mosaic pattern. These results suggested that electrical stimulation far oocytes activation after sperm injection enhances the incidence of both fertilization and development fellowing sperm injection in the pig. Our study also suggested that sperm-mediated transfer of exogenous DNA by ICSI would be used as a valuable tool for the production of transgenic porcine embryos.

Effects of Dietary Silicate Based Complex Mineral on Performance, Meat Quality and Immunological Competence in Broiler (규산염 복합광물질의 급여가 육계의 생산능력, 계육품질 및 면역능력에 미치는 영향)

  • Lim, Chun Ik;Park, Jin Ern;Kim, Sang Eun;Choe, Ho Sung;Ryu, Kyeong Seon
    • Korean Journal of Poultry Science
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    • v.44 no.4
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    • pp.275-282
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    • 2017
  • This study was conducted to investigate the effect of dietary silicate based complex mineral (SCM) on the performance of broiler chicks. Four hundred fifty one day old Cobb ${\times}$ Cobb broiler chicks were fed with commercial diets at 0%, 0.05%, 0.10%, 0.15% and 0.20% SCM with five replicates for five weeks. Weight gain, feed intake and feed conversion were measured weekly, and blood composition, immunity and meat quality were evaluated at the end of experiment. During overall period weight gain in chicks fed diet containing 0.1% SCM was significantly increased as compared with that of control (p<0.05). Feed intake showed no consistency among the treatments. Feed conversion appeared to increase in the chickens fed with SCM addition diets during prestarter period. Albumin, glucose and other blood parameters related to chicken health tended to improve at the level of 0.05% SCM addition treatments. Drip loss in breast meat was significantly decreased in more than 0.05% SCM addition (p<0.05). The expression of IL-2 (Interleukin-2) in blood increased significantly in the chickens fed with SCM of 0.05% or 0.10% level than other treatments (p<0.05). The optimum SCM concentration for commercial dietary supplementation for improving broiler performance and other health-related parameters was 0.10%.

Evaluation of Scab Resistance and Effect of Photosynthetic Rates on Fruit Characteristics among Elite Pear Seedlings (배 우량계통의 검은별무늬병 저항성 평가 및 광합성률이 과실 품질에 미치는 영향 구명)

  • Won, Kyung-Ho;Kang, Sam-Seok;Kim, Yoon-Kyeong;Sherzod, Rajametov;Lim, Kyeong-Ho;Lee, Han-Chan
    • Korean Journal of Environmental Agriculture
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    • v.32 no.2
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    • pp.117-122
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    • 2013
  • BACKGROUND: The scab, which is caused by Venturia nashicola, gives serious damages to pear trees. 'Niitaka' accounts for 82% of areas in pear cultivation. However 'Niitaka' is a scab susceptible cultivar. So, most of Korean farmers who growing pear trees have suffered by economic losses with the scab. In this research, we evaluated the scab resistance among elite pear seedlings to clarify genetics about the scab resistance. And we analyzed photosynthetic features with these seedlings to develop suitable cultivar which is advantageous for producing quality fruits during the growth and development of plants. METHODS AND RESULTS: We measured the rates of scab incidence among seedlings in a field experiment condition and an in-vitro test. An in-vitro test has been done with field experiment-based results. We made plant materials by grafting branches of each seedlings with 'Kongbae' rootstocks. And they had been grown for one month. Then, scab conidia suspension is sprayed to seedlings and sustained for 40 days under the controlled environment. As the results, 6 seedlings displayed lower incidence rates than other seedlings and 'Niitaka'. We also measured instant photosynthetic rates of each seedlings to determine the correlation between photosynthetic rates and fruit characteristics. However, it seemed that there is no correlation between them. CONCLUSION(S): Among the seedlings, 6 seedlings displayed the higher resistance to scab than other seedlings and 'Niitaka'. This characteristics is considered to be come from the gene expression of European pear. And we found that photosynthetic rate in trees rarely does not influence the fruit characteristics. It is considered to be affected by cultivar's own characteristics.

Substrate chain-length specificities of polyhydroxyalkanoate synthases PhaC1 and PhaC2 from Pseudomonas aeruginosa P-5 (Pseudomonas aeruginosa P-5에 존재하는 polyhydroxyalkanoate synthase PhaC1과 PhaC2의 기질특이성)

  • Woo, Sang Hee;Lee, Sun Hee;Rhee, Young Ha
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.455-462
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    • 2016
  • Pseudomonas aeruginosa P-5 is an unusual organism capable of synthesizing polyhydroxyalkanoates (PHAs) consisting of 3-hydroxyvalerate (3HV) and medium-chain-length (MCL) 3-hydroxyalkanoate (3HA) monomer units when C-odd alkanoic acids are fed as the sole carbon source. Evaluation of the substrate chain-length specificity of two P. aeruginosa P-5 PHA synthases ($PhaC1_{P-5}$ and $PhaC2_{P-5}$) by heterologous expression of $PhaC1_{P-5}$ and $PhaC2_{P-5}$ genes in Pseudomonas putida GPp104 revealed that $PhaC2_{P-5}$ incorporates both 3HV and MCL 3HAs into PHA, whereas $PhaC1_{P-5}$ favors only MCL 3HAs for polymerization. In order to obtain $PhaC2_{P-5}$ mutants with altered substrate specificity, site-specific mutagenesis for $PhaC2_{P-5}$ was conducted. Amino acid substitutions of $PhaC2_{P-5}$ at two positions (Ser326Thr and Gln482Lys) were very effective for synthesizing copolymers with a higher 3HV fraction. When recombinant P. putida GPp104 harboring double mutated $phaC2_{P-5}$ gene ($phaC2_{P-5}QKST$) was grown on nonanoic acid, 2.5-fold increase of copolymer content with 3.8-fold increase of 3HV fraction was observed. The $phaC2_{P-5}QKST$-containing Ralstonia eutropha PHB-4 supplemented with valeric acid also produced copolymers consisting of 3HV and 3-hydroxyheptanoate with a high 3HV fraction. These results suggest that recombinants containing $phaC2_{P-5}QKST$ could be useful for production of new PHA copolymers with improved material properties.

The Effect of Antihistamine on Endotoxin-induced Acute Lung Injury (내독소 유도 급성폐손상에서 항히스타민의 역할)

  • Jung, Bock-Hyun;Koh, Youn-Suck;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.3
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    • pp.219-229
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    • 2002
  • Background : Sepsis-induced acute lung injury (ALI) is caused by many cellular and humoral mediators induced by an endotoxin. Histamine, which is widely distributed in the lungs and has been considered as an important mediator of sepsis. It increases P-selectin expression on the endothelial cell surfaces and induces IL-8 secretion. Therefore, an endotoxin-induced histamine may be related to neutrophil-mediated ALI by inducing the migration and activation of neutrophils in the lung tissue. However, the role of endogenous histamine in endotoxin ALI has not been clarified. The purpose of this study was to investigate how endotoxin-induced ALI is influenced by endogenous histamine and to identify the possible mechanism of action. Materials and Methods : The study consisted of 4 groups using Sprague-Dawley rats : 1) control group, where the rats were infused intratracheally by normal saline, 2) an endotoxin group, where lipopolysaccharide (LPS) was administered intratracheally 3) the $H_2$ receptor antagonist-treated group ($H_2$ group) and 4) the $H_1$ receptor antagonist-treated group ($H_1$ group), where $H_2$-receptor blocker (ranitidine) and $H_1$-receptor blocker(pyrilamine) were co-treated intravenously with the intratracheal administration of an endotoxin. The lung leak index using $I^{125}$-BSA, the total protein and LDH concentration in the lung lavage fluid, myeloperoxidase(MPO) activity in the lung tissue, the pathologic score and the total number of neutrophils, TNF-$\alpha$, IL-$1{\beta}$ and IL-10 in lung lavage (BAL) fluid were measured in each group as the indices of lung injury. Results : Compared to the control group, the endotoxin group exhibited significant increases in all lung injury indices. Significant reductions in the endotoxin-mediated increases in lung leak index (p<0.05) were observed in both the $H_1$ and $H_2$ groups. In addition the total protein (p<0.05) and LDH concentration (p<0.05) in the BAL fluid were also lower in the $H_2$ group compared to the endotoxin group. However, there was no change in the MPO activity in the lung tissue, the pathologic score and the total number of neutrophils in the BAL fluid in both the $H_2$ and $H_1$ groups compared to the endotoxin group. The increases in TNF-$\alpha$ IL-$1{\beta}$ and IL-10 concentrations in the BAL fluid observed in the endotoxin group were not reduced in the $H_2$ and $H_1$ groups. Conclusion : Antihistamine attenuated the enhanced alveolar-capillary permeability induced by the endotoxin via the $H_2$ receptor. However the attenuating mechanism may not be related to the pathogenesis of neutrophil dependent lung injury.

Inhibitory Effects of Myelophycus simplex Papenfuss Methanol Extract on Melanogenesis in B16F10 Melanoma Cells (바위수염 메탄올 추출물이 B16F10 흑색종 세포에서의 멜라닌합성에 미치는 영향)

  • Kim, Hyang Suk;Cheon, Ji Min;Kwon, Da Hye;Choi, Eun Ok;Kim, Min Ju;Choi, Yung Hyun;Kim, Byung Woo;Hwang, Hye Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.1
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    • pp.34-38
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    • 2017
  • Myelophycus simplex Papenfuss, a type of brown algae, is known to be majorly distributed in along the southern coast of Korea and Japan. The purpose of this study was to investigate the effects of M. simplex Papenfuss methanol extract (MSPME) on melanogenesis in ${\alpha}$-melanocyte-stimulating hormone-stimulated B16F10 melanoma cells. Melanin contents of B16F10 melanoma cells were decreased by 27, 41, and 59% in a dose-dependent manner, upon MSPME treatment at 100, 300, and $500{\mu}g/mL$, respectively. Tyrosinase activities in B16F10 melanoma cells were decreased by 18, 49, and 61% in a dose-dependent manner, upon MSPME treatment at 100, 300, and $500{\mu}g/mL$, respectively. MSPME suppressed expression of tyrosinase, tyrosinase-related protein-1, tyrosinase-related protein-2, and melanocyte-inducing transcription factor in B16F10 melanoma cells. Concentration of $50{\mu}g/mL$ of MSPME especially induced greater decreases in tyrosinase activity, melanin contents, and melanogenic enzyme protein expressions. This results indicate that MSPME inhibits melanin synthesis and tyrosinase activity, and M. simplex Papenfuss extract may be an ideal candidate as a skin whitening agent.

Cyanobacteria and Secondary Metabolites (시아노박테리아의 이차대사물질에 대한 연구)

  • Kim, Gi-Eun;Kwon, Jong-Hee
    • KSBB Journal
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    • v.22 no.5
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    • pp.356-361
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    • 2007
  • Cyanobacteria are a very old group of prokaryotic organisms that produce very diverse secondary metabolites, especially non-ribosomal peptide and polyketide structures. Although some cyanobacteria produce lethal toxins such as microcystins and anatoxins, some may be useful either for development into commercial drugs or as biochemical tools. Detection of unknown secondary metabolites was carried in the present study by a screening of 98 cyanobacterial strains from Cyanobiotech GmbH in order to establish a screening process, isolate pure substances and determine their bioactivities. A degenerated polymerase chain reaction technique as molecular approaches has been used for general screening of NRPS gene and PKS gene in cyanobacteria. A putative PKS gene was detected by DKF/DKR primer in 38 strains (38.8%) and PCR amplicons resulted from a presence of NRPS gene were showed by MTF2/MTR2 primer in 30 strains (30.6%), respectively. A screening of interesting strains was performed by comparing PCR screening results with HPLC analyses of extracts. HPLC analysis for a detection of natural products was performed in extracts from biomass. 5 strains were screened for further scale-up processing. 7 pure substances were isolated from the scale-up cultures and tested for bioactivities under consideration to purity, amount and molecular weight of substances. One substance isolated from CBT 635 showed cytotoxic activity. This substance may be regarded as Microcystin LR.

The Effect of Insulin-Like Growth Factor-I(IGF-I) and IGF Binding Protein-3(IGFBP-3) on Cellular Proliferation in Mouse 3T3 Fibroblast Cells (마우스 섬유아세포(3T3 fibroblast cells)에서 Insulin-like Growth Factor-I(IGF-I) 및 IGF Binding Protein-3 (IGFBP-3)이 세포증식에 미치는 영향)

  • Cho, Chul-Ho;Kwak, Seung-Min;Moon, Tae-Hun;Cho, Jae-Hwa;Ryu, Jeong-Seon;Lee, Hyong-Lyeol
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.5
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    • pp.618-628
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    • 1999
  • Background: Cell growth is a balance between cell proliferation and cell death. Insulin-like growth factor-I(IGF-I), which binds IGF-I receptor(IGF-IR), mediates cellular proliferation as a potent mitogen. IGF binding protein-3(IGFBP-3) as a circulating major IGFBP can inhibit or enhance the effects of IGF-I on cellular growth by binding IGFs. Methods: We investigated the expressions of mRNA of IGF-I and IGF-IR by northern blot and phosphorylation of IGF-IR with the treatment of IGF-I by western blot in 3T3 fibroblast cells. The cellular proliferations of 3T3 cells with the treatments of IGF-I were evaluated using $^3H$-thymidine incorporation and MTT assay. Also to observe the effect of IGFBP-3 on cellular proliferation, 3T3 cells were treated with anti-IGFBP-3 and ${\alpha}IR_3$(monoclonal antibody to IGF-IR) alone or in combination. Results: Our results demonstrated that 3T3 cells showed mRNA expressions of IGF-I and IGF-IR and the IGF-I increased phosphorylation of IGF-IR. The treatments of 3T3 cells with IGF-I increased cellular proliferation in 5 % and 1 % seruma-containing media, not in serum-free media. The addition of anti-IGFBP-3 to neutralize IGFBP-3 showed 2-fold increase of cellular proliferation, and also co-incubation of anti-IGFBP-3 and ${\alpha}IR_3$ together showed similar increase of cellular proliferation in 3T3 cells. Interestingly, when the cells were pretreated with ${\alpha}IR_3$ for 4 hr, prior to the simultaneous addition of ${\alpha}IR_3$ and anti-IGFBP-3, anti-IGFBP-3-mediated cellular proliferation was decreased to control level. All of these results suggest that free IGF-I released from IGF-I/IGFBP-3 complex would be involved in the cellular proliferation. Conclusion: IGF-I is a mitogen through the activation of IGF-IR in 3T3 cells, and IGFBP-3 could be a potent inhibitor for IGF-I action by binding IGF-I.

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Effect of Hericium erinaceus Mycelia Supplementation on the Oxidative Stress and Inflammation Processes Stimulated by LPS and Their Mechanisms in BALB/C Mice (BALB/C Mice에서 노루궁뎅이버섯 균사체 보충이 LPS로 인한 산화적 스트레스와 염증 반응에 미치는 효과 및 기전)

  • Jang, Ji-Hyun;Noh, Kyung-Hee;Choi, Ji-Na;Jin, Kyong-Suk;Shin, Jin-Hyuk;On, Joon-Ho;Cho, Chung-Won;Jeong, Woo-Sik;Kim, Myo-Jeong;Song, Young-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.2
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    • pp.227-236
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    • 2010
  • This study was carried out to investigate the anti-oxidative and anti-inflammatory actions of Hericium erinaceus mycelia in BALB/C mice injected with lopopolysaccharide (LPS), called endotoxin. Mice (6 weeks of age) weighing approximately $24.73\pm0.11$ g were divided into 5 groups and were fed on the experimental diets containing Hericium erinaceus mycelia powder (HMP) for 1 week. Experimental groups were NC (normal control), HMP-C (HMP control), LC (LPS control), HMP 3%, and HMP 10%. Endotoxin shock was induced by intraperitoneal injection of LPS (100 mg/kg BW). NC and HMP-C groups were injected with saline solution (100 mg/kg BW). Food efficiency ratio were significantly (p<0.05) decreased in the HMP supplementation groups. Total fat and $\beta$-glucan excretion were higher in HMP supplementation groups than NC and LC groups, while plasma TG level was not different among groups. Plasma ALT levels were significantly (p<0.05) lower in the HMP supplementation groups than in LC group at 8 hr after LPS injection, while tumor necrosis factor-$\alpha$ and interleukine-6 levels of plasma were not different among groups. Hepatic superoxide dismutase, glutathione-reductase (GSH-red), and glutathione-peroxidase activities were higher in the HMP supplementation groups than in LC group at 4 hr after intraperitoneal injection of LPS. Hepatic GSH levels and protein expression of GSH-red was significantly (p<0.05) higher in the HMP supplemented groups than in LC group at 1 hr, 4 hr and 8 hr after LPS injection. From the above results, it is concluded that Hericium erinaceus mycelia may ameliorate hepatic oxidative stress by LPS through the elevation of hepatic glutathione level and antioxidant enzyme activities, which support the hepatoprotective effect of Hericium erinaceus mycelia.