• Title/Summary/Keyword: pH dependent

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Optimization of the Production of a Thermostable Antifungal Antibiotic (내열성 항곰팡이 항생물질의 생산 최적화)

  • 신영준;정명주;정영기
    • KSBB Journal
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    • v.15 no.6
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    • pp.584-588
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    • 2000
  • The optimum conditions for the production of an antifungal antibiotic from Bacillus sp. YJ-63 were investigated. The oprimumized medium consisted of 1.5% soluble starch, 1% tryptone and 0.5% yeast extract, and temperature and initial medium pH for production were optimal at 35$^{\circ}C$ and pH 6.0, respectively. Production yield was significantly improved by shaking culture using 50 ml medium in 500 ml flasks. Under these conditions, the production of the antifungal antibiotic was growth-dependent, from 35hrs into cultivation to the stationary phase and endospore formation.

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Effect of Trolox C on CYP450 Isozymes Activity and Expression in Hepatic Ischemia/Reperfusion

  • Eum, Hyun-Ae;Kim, Sung-Ho;Lee, Sun-Mee
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.301.2-301.2
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    • 2002
  • The present study was done to determine the effect of trolox C. a hydrophilic analogue of vitamin E. on alteration in cytochrome P-450 (CYP)-dependent drug metabolism during ischemia and reperfusion. Rats were subjected to 60 min of hepatic ischemia and 5 h of reperfusion. Rats were treated intravenously with trolox C (2.5 mg/kg) or vehicle (PBS. pH 7.4), 5 min before reperfusion. Serum alanine aminotransferase and lipid peroxidation levels were markedly increased after ischemia and reperfusion. This increase was significantly suppressed by trolox C. (omitted)

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Resveratrol Exerts Dosage-Dependent Effects on the Self-Renewal and Neural Differentiation of hUC-MSCs

  • Wang, Xinxin;Ma, Shanshan;Meng, Nan;Yao, Ning;Zhang, Kun;Li, Qinghua;Zhang, Yanting;Xing, Qu;Han, Kang;Song, Jishi;Yang, Bo;Guan, Fangxia
    • Molecules and Cells
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    • v.39 no.5
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    • pp.418-425
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    • 2016
  • Resveratrol (RES) plays a critical role in the fate of cells and longevity of animals via activation of the sirtuins1 (SIRT1) gene. In the present study, we intend to investigate whether RES could promote the self-renewal and neural-lineage differentiation in human umbilical cord derived MSCs (hUC-MSCs) in vitro at concentrations ranging from 0.1 to $10{\mu}M$, and whether it exerts the effects by modulating the SIRT1 signaling. Herein, we demonstrated that RES at the concentrations of 0.1, 1 and $2.5{\mu}M$ could promote cell viability and proliferation, mitigate senescence and induce expression of SIRT1 and Proliferating Cell Nuclear Antigen (PCNA) while inhibit the expression of p53 and p16. However, the effects were reversed by 5 and $10{\mu}M$ of RES. Furthermore, RES could promote neural differentiation in a dose-dependent manner as evidenced by morphological changes and expression of neural markers (Nestin, ${\beta}III-tubulin$ and NSE), as well as pro-neural transcription factors Neurogenin (Ngn)1, Ngn2 and Mash1. Taken together, RES exerts a dosage-dependent effect on the self-renewal and neural differentiation of hUC-MSCs via SIRT1 signaling. The current study provides a new strategy to regulate the fate of hUC-MSCs and suggests a more favorable in vitro cell culture conditions for hUCMSCs-based therapies for some intractable neurological disorders.

Isolation and characterization of thioredoxin and NADPH-dependent thioredoxin reductase from tomato (Solanum lycopersicum)

  • Dai, Changbo;Wang, Myeong-Hyeon
    • BMB Reports
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    • v.44 no.10
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    • pp.692-697
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    • 2011
  • To investigate the pathways of oxidoreductases in plants, 2 key components in thioredox systems i.e. thioredoxin h (Trx h) and NADPH-dependent thioredoxin reductase (NTR) genes were first isolated from tomatoes (Solanum lycopersicum). Subsequently, the coding sequences of Trx h and NTR were inserted into pET expression vectors, and overexpressed in Escherichia coli. In the UV-Visible spectra of the purified proteins, tomato Trx h was shown to have a characteristic 'shoulder' at ~290 nm, while the NTR protein had the 3 typical peaks unique to flavoenzymes. The activities of both proteins were demonstrated by following insulin reduction, as well as DTNB reduction. Moreover, both NADPH and NADH could serve as substrates in the NTR reduction system, but the catalytic efficiency of NTR with NADPH was 2500-fold higher than with NADH. Additionally, our results reveal that the tomato Trx system might be involved in oxidative stress, but not in cold damage.

Potential and Significance of Ammonium Production from Helicobacter pylori

  • KI, Mi-Ran;Yun, Soon-Kyu;Choi, Kyung-Min;Hwang, Se-Young
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.673-679
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    • 2003
  • Glutamine and urea, abundant in body fluids or plasma, yield net ammonium ions upon hydrolysis by ${\gamma}-glutamyl$ transpeptidase (${\gamma}-GTP$) and urease, respectively, and these two enzymes are largely produced from Helicobacter pylori. To investigate bacterial potential of ammonium production, we first quantified those in whole-cell systems and found that the relative ratio of their amounts varied greatly, especially with pH values and the cell's aging. During the H. pylori cultivation, the ratio appeared to be inversely proportional to each other, showing a progressive increase of the ${\gamma}-GTP$ with decreasing of the urease. Under the urease-defective conditions due to low pH or coccoids, the bacterial cells still possessed a considerable amount of ${\gamma}-GTP$, which was found exclusively in the external compartment, therefore, the cell's ammonium production was found to be solely dependent upon glutamine, and the external ammonium concentration was constant without any contribution of urea concentration. Such ammonium constancy would definitely have an adverse effect on the host, because of its absolute requirement for vacuolar degeneration by H. pylori VacA, maximized at approximately 10 mM $NH_4Cl$. It was also found that, by using the metal-saturated membrane vesicles, ammonium ions were likely to be involved in the pH-dependent cation-flux across the H. pylori membrane, where the role of ${\gamma}-GTP$ in ammonium homeostasis around cells was suggested, especially under the hostile conditions against H. pylori.

Characterization of a cysteine proteinase from adult worms of Paragonimus westermani (폐흡충(Parnonimr westemani)성충에서 정제한 cysteine proteinase의 특성)

  • 송철용;김동수
    • Parasites, Hosts and Diseases
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    • v.32 no.4
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    • pp.231-242
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    • 1994
  • Pnragonimus westermnni, the lung fluke, is known to migrate to the pulmonary tissue of mammalian hosts and causes pathological changes in the lungs. An acidic thiol-dependent proteinase with a molecular weight of approximately 20,000 daltons was purified to homogeneity using ion-exchange chromatography and gel filtration chromatography. On SDS-PAGE, the molecular weight of the enzyme was 17,500 daltons. Isoelectric point was 6.45. The enzyme was similar to the acidic cysteine proteinase of vertebrates in the properties of pH optimum, substrate specificity and inhibitor sensitivity. Enzymatic activity was stable at pH 5.5 for at least two days when stored at 4℃. The cysteine proteinase was capable of degrading collagen and hemoglobin. Sera of patients with paragonimiasis and mice infected with R westermani reacted in immunoblots with the partially purified proteinase. This result suggested that the cysteine proteinase of P. westermnni may play a role in migration in tissues, and in acquisition of nutrients by parasites from the host. It is also potentially an antigen for the serodiagnosis of paragonimiasis.

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Effect of estradiol-$17{\beta}$ on proliferation in primary cultured chicken hepatocytes (초대 배양한 닭 간세포 증식에 대한 estradiol-$17{\beta}$의 효과)

  • Baek, Gyul;Kang, Ju-Won
    • Korean Journal of Veterinary Service
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    • v.31 no.4
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    • pp.457-463
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    • 2008
  • The sex steroid hormone estradiol-$17{\beta}(E_2)$ mediate their biological effects on development, differentiation and maintenance of reproductive tract and other target tissue through gene regulation by nuclear steroid receptors. Although the importance of $E_2$ in many physiological process has been reported, but little is known about the effects of $E_2$ on primary cultured chicken hepatocyte. therefore, in the present study, we have examined the effect of $E_2$ on cell proliferation and it's related signal cascades. $E_2$ increase $[^3H]$-thymidine incorporation in time-(${\leq}8hr$) and dose-($10^{-10}M$)dependent manner and treatment of $E_2$ increased the phosphorylation of p44/43 MAPKs(p44/42 mitogen-activated protein kinase) and JNK(c-Jun N-terminal kinase) in a time dependent manner. In addition, PD98059(p44/42 blocker, $10^{-5}M$), SP600125(JNK blocker, $10^{-6}M$) blocked the estrogen-induced increase in $[^3H]$-thymidine incorporation. In conclusion, $E_2$ stimulates the proliferation of primary cultured chicken hepatocytes and this action is mediated by p44/42 MAPKs and JNK signal transduction pathway.

Characterization and Expression in Escherichi coli of Streptococcus pneumoniae FtsH

  • Kim, Hee-Soo;Lee, Jae-Jung
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.109-115
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    • 2000
  • FtsH is a membrane-bound, ATP-dependent metalloprotease that is involved in a variety of cellular functions including the regulation of responses to heat and stress shock. Previously, we had cloned and sequenced pneumococcal ftsH gene whose deduced amino acid sequence was very similar to those of several gram-positive bacteria and Escherichia coli, except for the N-terminal domain that was responsible for membrane anchoring. In order to better understand the role of Streptococcus pneumoniae FtsH, we expressed pneumococcal ftsH gene in Escherichia coli. When it was expressed from a strong promoter, $P_{tac}$, a considerable amount of the recombinant FtsH was produced, although the prolonged induction resulted in not only accumulation of breakdown products but also ceasing of the further growth of E. coli host. This indicated that the expression of the exogenous ftsH gene was tightly regulated since the excessive FtsH appeared detrimental to bacterial cells. In Western blotting, the pneumococcal FtsH protein, whether native or recombinant, was reactive to anti-E. coli FtsH serum. The observation that FtsH proteins were well conserved throughout the bacterial kingdom and its expression level was fine-tuned suggests an important role for this protein in the stress adaptation which may be related to infecting process by pneumococci.

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pH-Dependent Dye Adsorption and Release Behaviors of Poly(ethylene-alt-maleic anhydride)/poly(4-vinyl pyridine) Multiplayer Films (pH 의존 특성을 갖는 Poly(ethylene-alt-maleic anhydride)/Poly(4-vinyl pyridine) 다층막의 염료 흡착 및 방출 거동 연구)

  • Hong, Sook-Young;Lee, Joon-Youl
    • Polymer(Korea)
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    • v.29 no.6
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    • pp.593-598
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    • 2005
  • This work studied the loading capabilities and release behaviors of poly(ethylene-alt-maleic anhydride) (PEMAh)/poly(4-vinyl pyridine) (P4VP) multilayer films formed by the layer-by-layer(LbL) sequential self-assembly method, using Rodamine 6G(R6G) as an indicator. Thickness of the multilayer, and loading and subsequent release behavior of R6G from the multilayer were studied using UV-visible spectroscopy. The amount of R6G loaded in multilayer film increased linearly with increasing film thickness. pH-Sensitive permeability was observed, where lower pH environments increased both release rate and release amount. By additional assembling of PEMAh/poly(ethyleneimine) (PEI) capping layers on top of (PEMAh/P4VP)n multilayers, the release of R6G was better controlled.

Schedule-Dependent Effect of Epigallocatechin-3-Gallate (EGCG) with Paclitaxel on H460 Cells

  • Park, Sunghoon;Kim, Joo-Hee;Hwang, Yong Il;Jung, Ki-Suck;Jang, Young Sook;Jang, Seung Hun
    • Tuberculosis and Respiratory Diseases
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    • v.76 no.3
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    • pp.114-119
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    • 2014
  • Background: Epigallocatechin-3-gallate (EGCG), a major biologically active component of green tea, has anti-cancer activity in human and animal models. We investigated the schedule-dependent effect of EGCG and paclitaxel on growth of NCI-H460 non-small cell lung cancer cells. Methods: To investigate the combined effect of EGCG (E) and paclitaxel (P), combination indices (CIs) were calculated, and cell cycle analysis was performed. For the effect on cell apoptosis, western blot analysis was also performed. Results: CI analysis demonstrated that both concurrent and sequential E ${\rightarrow}$ P treatments had antagonistic effects (CIs >1.0), but sequential P ${\rightarrow}$ E had synergistic effects (CIs <1.0), on the growth inhibition of NCI-H460 cells. In the cell cycle analysis, although paclitaxel induced $G_2/M$ cell cycle arrest and increased the sub-G1 fraction, concurrent EGCG and paclitaxel treatments did not have any additive or synergistic effects compared with the paclitaxel treatment alone. However, western blot analysis demonstrated that sequential P ${\rightarrow}$ E treatment decreased the expression of Bcl-2 and procaspase-3 and increased poly(ADP-ribose) polymerase (PARP) cleavage; while minimal effects were seen with concurrent or sequential E ${\rightarrow}$ P treatments. Conclusion: Concurrent or sequential E ${\rightarrow}$ P treatment had opposite effects to P ${\rightarrow}$ E treatment, where P ${\rightarrow}$ E treatment showed a synergistic effect on growth inhibition of NCI-H460 cells by inducing apoptosis. Thus, the efficacy of EGCG and paclitaxel combination treatment seems to be schedule-dependent.