• Food Science and Biotechnology
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• v.18 no.4
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• pp.965-970
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• 2009

Oleuropein content of olive leaf extracts (OLE; ethanol extract) was evaluated by high performance liquid chromatography analysis. Oleuropein contents were $4.21{\pm}0.57$, $3.92{\pm}0.43$, $0.32{\pm}0.03$, $5.76{\pm}0.32$, and $32.47{\pm}0.25$ mg/100 g for ethanol extract, and hexane, chloroform, ethyl acetate, and butanol fraction, respectively. The removal of DPPH free radical increased in OLE and all 5 fractions of OLE in a concentration dependent manner. In order to investigate the antioxidant effect of OLE in vitro, 80%(v/v) ethanol OLE, $H_2O_2$, or combined treatment of 80%(v/v) ethanol OLE and $H_2O_2$ were applied on mouse embryonic fibroblast (MEF) cells. Cells were damaged by oxidative stress decreased their viability followed by increasing concentration of $H_2O_2$, but co-treatment of OLE and $H_2O_2$ showed an increase in cell growth about 20% compare to the cells treated with $H_2O_2$. OLE suppresses cytotoxicity induced by $H_2O_2$ in dose dependent manner. OLE treatment on MEF cells was also examined by analyzing cell cycle and apoptotic rate using flow cytometry. Apoptotic and necrotic cell accumulation was decreased in addition of OLE to $H_2O_2$ compare to the oxidative damaged cells. Taken together, these results demonstrated that OLE suppresses cytotoxicity induced by $H_2O_2$ and protect cells against oxidative stress on MEF cells.

• The Journal of Pediatrics of Korean Medicine
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• v.27 no.4
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• pp.17-30
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• 2013

Objectives This experimental study was designed to investigate the effects of Mulberry leaves contained herbal mixture (MLHM) on body weight, serum lipid level and adipocyte differentiation in high fat diet-fed obese mice. Methods Four-week old mice (wild-type C57/BL6) were used for all experiments. Cells were incubated with MLHM at the indicated concentration (0.04-4mg/ml) for 24h, and growth rate was assessed by MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. 3T3-L1 preadipocytes were incubated in DMEM for 2 days with the indicated concentrations of MLHM, and on Day 6, the cells were fixed and the cellular lipid contents were assessed by Oil-Red-O staining. The expression of peroxisome proliferator-activated receptor ${\gamma}$ (PPAR ${\gamma}$) and cytidine-cytidine-adenosine-adenosine-thymine (CCAAT)/enhancer-binding proteins ${\alpha}$ (C/EBP ${\alpha}$) as adipocyte-specific proteins were determined by real time RT-PCR and western blotting. In addition, body weight gain and serum lipid levels were measured in the mice with obesity induced by the high fat-diet for four weeks. Results Though MLHM did not show toxicity even at the concentration of 4mg/ml, MLHM significantly inhibited the differentiation of 3T3-L1 preadipocites in a dose-dependent manner. Also, MLHM significantly reduced the expressions of PPAR ${\gamma}$ and C/EBP ${\alpha}$ in a dose-dependent manner. Furthermore, MLHM significantly reduced body weight gain and LDL-cholesterol contents in high fat diet-fed obese mice. Conclusions These results demonstrate that MLHM exerts anti-obesity effect in 3T3-L1 cells and mice with obesity by high-fat diet.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.986-993
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• 2001

Translation Initiation in prokaryotes involves the formation of a 30 S preinitiation complex, in which translation initiation factors play a role in the stimulation of fMet-tRNA (fMet) binding. However, the specific function and precise mechanism of initiation factor IF1 are still unclear. One a functionally active factor with a high purity. In the present study a large quantity of active IF was rapidly purified, obtained by the overexpression of the infA gene, and then used for a functional study. The induction of infA did not appreciably affect the growth rate of the protease-deficient strain E. coli AR68 harboring the IF1 overproducing plasmid. The level of IF1 obtained was approximately $1-2\%$ of the total cell protein, which enabled the yield of highly purified IF1 (>$98\%$ pure) to be increased to 0.15 mg of IF1/g of cells. The IF1 was isolated within one day by the centrifugatioin of the ribosomal washed fraction, by ammonium sulfate fractionation, chromatography on batch of phosphocellulose, and FPLC Mono S. The overexpressed IF1 was found to be comparable to the factor isolated from normal cells, as determined by migration in NEPHGE/SDS 2-D gels. For binding of fMet-tRNA(fMet) to the 30 S ribosomal subunitis, relatively high levels of binding were obtained when IF2 was present. The addition of IF1 up to 110 pmol proportionally stimulated the binding to a variable extent. This IF1-dependent stimulation of the 30 S preinitiation complex formation demonstrated that IF1 would appear to be exclusively essential for promoting the initiation phase of protein synthesis.

• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.208-216
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• 2010

Fusarium verticillioides is an important pathogen of maize, being responsible for ear rots, stalk rots, and seedling blight worldwide. During the past decade, F. verticillioides has caused several severe epidemics of maize seedling blight in many areas of China, which lead to significant losses. In order to understand the molecular mechanisms regulating fungal development and pathogenicity in this pathogen, we isolated and characterized the gene fpk1 (GenBank Accession No. EF405959) encoding a homolog of the cAMP-dependent protein kinase catalytic subunit, which included a 1,854-bp DNA sequence from ATG to TAA, with a 1,680-bp coding region, and three introns (lengths: 66 bp, 54 bp, and 54 bp), and the predicated protein precursor had 559 aa. The mutant ${\Delta}fpk1$, which was disrupted of the fpkl gene, showed reduced vegetative growth, fewer and shorter aerial mycelia, strongly impaired conidiation, and reduced spore germination rate. After germinating, the fresh hypha was stubby and lacking of branch. When inoculated in susceptible maize varieties, the infection of the mutant ${\Delta}fpk1$ was delayed and the infection efficiency was reduced compared with that of the wild-type strain. AU this indicated that gene fpk1 participated in hyphal growth, conidiophore production, spore germination, and virulence in F. verticillioides.

• Nutrition Research and Practice
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• v.8 no.2
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• pp.132-137
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• 2014

BACKGROUND/OBJECTIVES: In this study, the apoptogenic activity and mechanisms of cell death induced by hexane extract of aged black garlic (HEABG) were investigated in human leukemic U937 cells. MATERIALS/METHODS: Cytotoxicity was evaluated by MTT (3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyl tetrazoliumbromide) assay. Apoptosis was detected using 4,6-diamidino-2-phenyllindile (DAPI) staining, agarose gel electrophoresis and flow cytometry. The protein levels were determined by Western blot analysis. Caspase activity was measured using a colorimetric assay. RESULTS: Exposure to HEABG was found to result in a concentration- and time-dependent growth inhibition by induction of apoptosis, which was associated with an up-regulation of death receptor 4 and Fas legend, and an increase in the ratio of Bax/Bcl-2 protein expression. Apoptosis-inducing concentrations of HEABG induced the activation of caspase-9, an initiator caspase of the mitochodrial mediated intrinsic pathway, and caspase-3, accompanied by proteolytic degradation of poly(ADP-ribose)-polymerase. HEABG also induced apoptosis via a death receptor mediated extrinsic pathway by caspase-8 activation, resulting in the truncation of Bid, and suggesting the existence of cross-talk between the extrinsic and intrinsic pathways. However, pre-treatment of U937 cells with the caspase-3 inhibitor, z-DEVD-fmk, significantly blocked the HEABG-induced apoptosis of these cells, and increased the survival rate of HEABG-treated cells, confirming that HEABG-induced apoptosis is mediated through activation of caspase cascade. CONCLUSIONS: Based on the overall results, we suggest that HEABG reduces leukemic cell growth by inducing caspase-dependent apoptosis through both intrinsic and extrinsic pathways, implying its potential therapeutic value in the treatment of leukemia.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.22
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• pp.10015-10020
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• 2014

Background: Fentanyl is used as an analgesic to treat pain in a variety of patients with cancer and recently it has become considered to also act as an antitumor agent. The study present was designed to investigate the effects of fentanyl on colorectal cancer cell growth and plausible mechanisms. Materials and Methods: The human colorectal carcinoma cell line HCT116 was subcutaneously injected into nude mice. The viability of HCT116 was tested by MTT assay, and apoptosis by flow cytometry and caspase-3 activity. The expression of Sirt1 and NF-${\kappa}B$ were evaluated by Western blotting and the levels of Sirt1 and NF-${\kappa}B$ by fluorescence method. SiRNA was used to silence and Ad-Sirt1 to overexpress Sirt1. Results: Our data showed that fentanyl could inhibit tumor growth, with increased expression of Sirt1 and down-regulation of Ac-p65 in tumors. Compared with control cells without treatment, HCT116 cells that were incubated with fentanyl had a higher apoptotic rate. Moreover, fentanyl could increase expression and activity of Sirt1 and inhibitor expression and activity of NF-${\kappa}B$, which might be mechanisms of fentanyl action. Conclusions: Fentanyl increased colorectal carcinoma cell apoptosis by inhibition of NF-${\kappa}B$ activation in a Sirt1-dependent manner.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.23
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• pp.10281-10287
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• 2015

Background: Development of a nanosized polymeric delivery system for erlotinib was the main objective of this research. Materials and Methods: Poly caprolactone-polyethylene glycol-polycaprolactone (PCEC) copolymers with different compositions were synthesized via ring opening polymerization. Formation of triblock copolymers was confirmed by HNMR as well as FT-IR. Erlotinib loaded nanoparticles were prepared by means of synthesized copolymers with solvent displacement method. Results: Physicochemical properties of obtained polymeric nanoparticles were dependent on composition of used copolymers. Size of particles was decreased with decreasing the PCL/PEG molar ratio in used copolymers. Encapsulation efficiency of prepared formulations was declined by decreasing their particle size. Drug release behavior from the prepared nanoparticles exhibited a sustained pattern without a burst release. From the release profiles, it can be found that erlotinib release rate from polymeric nanoparticles is decreased by increase of CL/PEG molar ratio of prepared block copolymers. Based on MTT assay results, cell growth inhibition of erlotinib has a dose and time dependent pattern. After 72 hours of exposure, the 50% inhibitory concentration (IC50) of erlotinib hydrochloride was appeared to be $14.8{\mu}M$. Conclusions: From the obtained results, it can be concluded that the prepared PCEC nanoparticles in this study might have the potential to be considered as delivery system for erlotinib.

• The Korean Journal of Physiology
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• v.25 no.2
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• pp.147-158
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• 1991

The effects of noradrenaline on the spontaneous contraction recorded from a strip of mucosa-free antral circular muscle were studied in the guinea-pig stomach, and the changes in slow waves and membrane resistance were analyzed in order to elucidate the mechanism for the excitatory response to noradrenaline. Electrical responses of circular muscle cells were recorded using glass microelectrodes filled with 3 M KCI. Electrotonic potentials were produced to estimate membrane resistance by the partition stimulating method. All experiments were performed in tris-buffered Tyrode solution which was aerated with 100% $O_2$ and kept at $35^{\circ}C$. The results obtained were as follows: 1) The spontaneous contractions were potentiated dose-dependently by the application of noradrenaline. 2) Through the experiments using adrenoceptor-blockers, the strong excitatory effect via $[\alpha}-adrenoceptors$ and the weak inhibitory efffect via ${\beta}-adrenoceptors$ were noted. 3) Noradrenaline produced hyperpolarization of membrane potential, and increases in the amplitude and the maximum rate of rise of slow waves. 4) In the presence of apamin, Ca-dependent K channel blocker, the characteristic hyperpolarization was not developed. However, the excitatory effect of noradrenaline on spontaneous contraction remained. 5) Membrane resistance was reduced during the hyperpolarized state by the application of noradrenaline, and the change of membrane resistance and the hyperpolarized state were completely abolished by apamin. From the above results, following conclusions could be made: Excitatory responses to noradrenaline result from the dominant ${\alpha}-excitatory$, and the weak ${\beta}-inhibitory$ action of noradrenaline. Hyperpolarization of membrane potential by noradrenaline is due to the activation of Ca-dependent K channel.

• The Korean Journal of Physiology
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• v.21 no.2
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• pp.201-210
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• 1987

수온변화에 따른 심맥관계 및 신장기능의 변화와 생체실험을 통한 온도에 의한 adrenoceptor의 변형을 알아보기 위해, 무마취 자라에서 $18^{\circ}C$에서 $25^{\circ}C$로 수온을 증가시 나타나는 혈압, 심박동수 및 신장기능의 변화를 관찰하고, epinephrine 1 ug/kg과 10 ug/kg을 상이한 온도에 노출된 자라의 정맥내 투여하여 나타나는 효과를 비교하였다. 1) $18^{\circ}C$에서 $25^{\circ}C$로 수온을 증가시킴에 따라 심박동수는 현저히 증가하여 일정하게 유지되었으나, 혈압 및 혈장 renin 활성도는 변화하지 않았다. 온도증가에 의해 뇨량, 사구체여과율 및 전해질 배설량의 현저한 증가를 보였으나 90분부터는 서서히 감소하기 시작하였다. 2) 수온 $18^{\circ}C$에 노출된 자라에서 epinephrine은 dose-dependent한 양상으로 혈압 및 심박동수를 증가시켰으며, 다량의 epinephrine 투여시 작용시간은 현저히 연장되어 있었다. $25^{\circ}C$에 노출된 자라에서는 epinephrine에 의한 혈압상승 효과 및 심박동수 증가는 나타났으나, dose dependency나 작용시간의 차이는 발견할 수 없었다. 3) 동량의 epinephrine에 의한 혈압 및 심박동수의 증가효과는 $18^{\circ}C$와 $25^{\circ}C$에 노출된 자라에서 유의한 차이를 발견할 수 없었으나, $18^{\circ}C$에 노출된 자라에서 epinephrine의 작용시간 및 반감기가 현저히 연장되어 있었다. 4) Epinephrine 투여에 의해 뇨량, 사구체여과율 및 전해질 배설량의 증가를 관찰하였으며, 이는 dose-dependent 양상이었다. 그러나, 신장효과의 유의한 차이는 상이한 온도에 노출된 두 군에서 발견할 수 없었다. 이상의 결과로, 온도증가에 의한 이뇨 및 sodium 배설효과는 혈관이완에 의한 사구체여과율의 증가에 기인한 것으로 사료되며, 상이한 온도에 노출된 자라에서 epinephrine 효과의 차이를 발견할 수 없었던 것은 본 실험에서 가한 좁은 범위의 온도의 변화 내에서는 adrenoceptor의 변형이 나타나지 않을 것이라고 추론하였다. 그러나 저온에서의 epinephrine의 작용시간의 연장은 아마도 epinephrine의 파괴 효소의 활성도의 감소인 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• v.26 no.7
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• pp.1333-1340
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• 2016

The main objective of this study was to investigate whether Lactobacillus rhamnosus GG (LGG) ameliorated the effects of Citrobactor rodentium infection in Toll-like receptor 2 (TLR2) knockout (KO) and TLR4 KO mice, as well as in wild-type C57BL/6 (B6) mice. TLR2 KO, TLR4 KO, and B6 mice were divided into three groups per each strain. Each group had an uninfected control group (n = 5), C. rodentium-infected group (n = 8), and LGG-pretreated C. rodentium-infected group (n = 8). The survival rate of B6 mice infected with C. rodentium was higher when pretreated with LGG. Pretreatment with LGG ameliorated C. rodentium-induced mucosal hyperplasia in B6 and TLR4 KO mice. However, in C-rodentium-infected TLR2 KO mice, mucosal hyperplasia persisted, regardless of pretreatment with LGG. In addition, LGG-pretreated B6 and TLR4 KO mice showed a decrease in spleen weight and downregulation of tumor necrosis factor alpha, interferon gamma, and monocyte chemotactic protein 1 mRNA expression compared with the non-pretreated group. In contrast, such changes were not observed in TLR2 KO mice, regardless of pretreatment with LGG. From the above results, we conclude that pretreatment with LGG ameliorates C. rodentium-induced colitis in B6 and TLR4 KO mice, but not in TLR2 KO mice. Therefore, LGG protects mice from C. rodentium-induced colitis in a TLR2-dependent manner.