• Title/Summary/Keyword: cell fractions

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Identification and Characterization of the Acid Phosphatase HppA in Helicobacter pylori

  • Ki, Mi-Ran;Yun, Soon-Kyu;Choi, Kyung-Min;Hwang, Se-Young
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.483-493
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    • 2011
  • An acid phosphatase (HppA) activated by $NH_4Cl$ was purified 192- and 34-fold from the periplasmic and membrane fractions of Helicobacter pylori, respectively. SDS-polyacrylamide gel electrophoresis revealed that HppA from the latter appears to be several kilodaltons larger in molecular mass than from the former by about 24 kDa. Under acidic conditions (pH${\leq}$4.5), the enzyme activity was entirely dependent on the presence of certain mono- and/or divalent metal cations (e.g., $K^+$,$ NH_4{^+}$, and/or $Ni^{2+}$). In particular, $Ni^{2+}$ appeared to lower the enzyme's $K_m$ for the substrates, without changing $V_{max}$. The purified enzyme showed differential specificity against nucleotide substrates with pH; for example, the enzyme hydrolyzed adenosine nucleotides more rapidly at pH 5.5 than at pH 6.0, and vice versa for CTP or TTP. Analyses of the enzyme's N-terminal sequence and of an $HppA^-$ H. pylori mutant revealed that the purified enzyme is identical to rHppA, a cloned H. pylori class C acid phosphatase, and shown to be the sole bacterial 5'-nucleotidase uniquely activated by $NH_4Cl$. In contrast to wild type, $HppA^-$ H. pylori cells grew more slowly. Strikingly, they imported $Mg^{2+}$ at a markedly lowered rate, but assimilated urea rapidly, with a subsequent increase in extracellular pH. Moreover, mutant cells were much more sensitive to extracellular potassium ions, as well as to metronidazole, omeprazole, or thiophenol, with considerably lowered MIC values, than wild-type cells. From these data, we suggest that the role of the acid phosphatase HppA in H. pylori may extend beyond 5'-nucleotidase function to include cation-flux as well as pH regulation on the cell envelope.

Hypofractionated radiotherapy for early glottic cancer: a retrospective interim analysis of a single institution

  • Lee, Jeong Won;Lee, Jeong Eun;Park, Junhee;Sohn, Jin Ho;Ahn, Dongbin
    • Radiation Oncology Journal
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    • v.37 no.2
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    • pp.82-90
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    • 2019
  • Purpose: To evaluate the results of hypofractionated radiotherapy (HFX) for early glottic cancer. Materials and Methods: Eighty-five patients with cT1-2N0M0 squamous cell carcinoma of the glottis who had undergone HFX, performed using intensity-modulated radiotherapy (IMRT, n = 66) and three-dimensional conformal radiotherapy (3D CRT, n = 19) were analyzed. For all patients, radiotherapy was administered at 60.75 Gy in 27 fractions. Forty-three patients received a simultaneous integrated boost (SIB) of 2.3-2.5 Gy per tumor fraction. Results: The median follow-up duration was 29.9 months (range, 5.5 to 76.5 months). All patients achieved complete remission at a median of 50 days after the end of radiotherapy (range, 14 to 206 days). The 5-year rates for locoregional recurrence-free survival was 88.1%, and the 5-year overall survival rate was 86.2%. T2 stage was a prognostic factor for locoregional recurrence-free survival after radiotherapy (p = 0.002). SIB for the tumor did not affect disease control and survival (p = 0.191 and p = 0.387, respectively). No patients experienced acute or chronic toxicities of ≥grade 3. IMRT significantly decreased the dose administered to the carotid artery as opposed to 3D CRT (V35, p < 0.001; V50, p < 0.001). Conclusions: Patients treated with HFX achieved acceptable locoregional disease control rates and overall survival rates compared with previous HFX studies. A fraction size of 2.25 Gy provided good disease control regardless of SIB administration.

Comparison of Anti-Adipogenesis Activity by Several Grape Extracts (포도 추출물로부터 C/EBP 전사인자 활성 비교)

  • Lee, Si-Rim;Park, Chul-Hong;Kuan, Eun-Young;Lu, Yan-Qing;Kim, Hong;Kim, Ki-Chan;Son, Hyeong-U;Lee, Hyun-Jin;Heo, Jin-Chul;Lee, Sang-Han
    • Food Science and Preservation
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    • v.18 no.1
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    • pp.119-123
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    • 2011
  • In order to compare what kinds of transcription factors are associated with the inhibition of preadipocyte cell proliferation, we prepared several grape extracts and tested the expression patterns by reverse transcription-polymerase chain reaction. As a result, 50% ethanol extract of Campbell early seed inhibited adipogenesis derived from the MDI solution. Extract of Campbell early seed was significantly inhibited lipid droplet formation and expression of molecular factors C/EBP-alpha and delta in 3T3-L1 cells. It is suggested that grape extracts of fractions would be a good candidate for the development of regional skin fat modulator.

Nitric Oxide Production and Elastase Inhibitory Activities of Extract and Its Fraction from Phellodendri Cortex (황백추출물 및 분획물의 Nitric Oxide 생성 억제 효과 및 Elastase 억제 효과)

  • Um, Ji Na;Min, Jin Woo;Joo, Kwang Sik;Kang, Hee Cheol
    • Korean Journal of Medicinal Crop Science
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    • v.25 no.1
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    • pp.22-28
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    • 2017
  • Background: To date, the anti-wrinkle efficacy of phellodendri cortex has not been defined. In this study, we investigated the nitric oxide (NO) production and elastase inhibitory activities of 80% methanol extract of Phellodendri cortex and its ethyl acetate fraction. Methods and Results: We prepared 80% methanol extract, and its fractions from phellodendri cortex. The treatment of RAW 264.7 cell with $25{\mu}g/m{\ell}$ 80% methanol extract and ethyl acetate fraction resulted in no toxicity. We conducted assays of nitric oxide (NO) production and elastase inhibition. In the NO production assay, the ethyl acetate fraction showed an inhibitory effect approximately 17 times stronger than the 80% methanol extract. In elastase inhibitory assay, the ethyl acetate fraction also showed a stronger effect than the 80% methanol extract. In order to standardize the extract and fraction, we used TLC to separate the extract and observed the plate under UV light. We confirmed that the known pharmacological ingredients berberine, and palmatine in the 80% methanol extract and the ethyl acetate fraction. Conclusions: These results indicated that phellodendri cortex extract and its ethyl acetate fraction produced strong inhibitory effect on elastase and NO production.

Purification and Properties of a Membrane-bound Alcohol Dehydrogenase from Acetobacter sp. HA (Acetobacter sp. HA로부터 Membrane-bound Alcohol Dehydrogenase의 정제 및 특성)

  • Yoo, Jin-Cheol;Sim, Jung-Bo;Kim, Heung-Keun;Chun, Hong-Sung;Kim, Sung-Jin
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.78-83
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    • 1994
  • Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneous state fron an acetic acid producing bacteria, Acetobacter sp. HA. The enzyme was purified about 153-fold with an overall yield of 35% from the crude cell extract by solubilization and extraction of the enzyme with Triton X-100 and subsequent fractions by column chromatography. Upon sodium dodecyl sulphate-PAGE, the enzyme showed the presence of three subunits with a molecular mass of 79,000 daltons, 49,000, and 45,000 daltons, respectively. Absorption oxidized aliphatic alcohols with a straight carbon chain except for methanol. Formaldehyde, acetaldehyde and glutaraldehyde were also oxidizable substrates. The apparent $K_m$ for ethanol was 1.38mM. The optimun pH and temperature were 5.0~6.0 and 32${\circ}C$, respectively. $V_2O_5$ and heavy metals such as $ZnCl_2\;and\; NiCl_2$ were inhibitory to the enzyme activity.

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Cellular Responses of Salmonella typhimurium Exposed to Green Tea Polyphenols (녹차폴리페놀에 노출된 Salmonella typhimurium의 세포반응)

  • Choi, Hyo-Kyung;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.87-92
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    • 2012
  • The purpose of this study was to examine the cellular response of Salmonella typhimurium exposed to tea polyphenols (TPP) extracted from Korean green tea (Camellia sinensis L.). TPP showed a dose-dependent bactericidal effect on S. typhimurium. Analysis of cell membrane fatty acids of S. typhimurium cultures treated with TPP identified unique changes in saturated and unsaturated fatty acids, while scanning electron microscopic analysis demonstrated the presence of perforations and irregular rod forms with wrinkled surfaces in cells treated with TPP. Two-dimensional polyacrylamide gel electrophoresis of soluble protein fractions from S. typhimurium cultures showed 16 protein spots increased by TPP. These up-regulated proteins including proteins involved in antioxidants and chaperons, transcript and binding proteins, energy and DNA metabolism were identified by peptide mass fingerprinting using MALDI-TOF. These results provide clues for understanding the mechanism of TPP induced stress and cytotoxicity on S. typhimurium.

Characterization of Hemolytic Aeromonas sp. MH-8 Responding to the Exposure of Green Tea Catechin, EGCG (녹차 카테킨 EGCG의 노출에 따른 식중독 세균인 용혈성 Aeromonas sp. MH-8의 특성조사)

  • Kim, Dong-Min;Oh, Kye-Heon
    • KSBB Journal
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    • v.31 no.4
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    • pp.228-236
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    • 2016
  • The aim of this study was to characterize the hemolytic Aeromonas sp. MH-8 exposed to green tea catechin, epigallocatechin gallate (EGCG). Initially, the hemolytic Aeromonas sp. MH-8 was enriched and isolated from stale fish. Bactericidal effects of MH-8 exposed to EGCG ranging from 1 mg/mL to 4 mg/mL were monitored, and complete bactericidal effects were achieved within 3 h at 3 mg/mL and higher concentrations. SDS-PAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain MH-8 treated to different concentrations and exposing periods of EGCG in exponentially growing cultures. The stress shock proteins (70-kDa DnaK and 60-kDa GroEL), which might contribute to enhancing the cellular resistance to the cytotoxic effect of EGCG, were induced at different concentrations of EGCG exposed to cell culture of MH-8. Scanning electron microscopic analysis demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with EGCG. 2-DE of soluble protein fractions from MH-8 cultures showed 18 protein spots changed by EGCG exposure. These proteins involved in chaperons (e.g., DnaK, GroEL and trigger factor), enterotoxins (e.g., aerolysin and phospholipase C precursor), LPS synthesis (e.g., LPS biosynthesis protein and outer membrane protein A precursor), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. In consequence, EGCG was found to have substantial antibacterial effects against food-poisoning causing bacterium, hemolytic Aeromonas sp. MH-8. Also the results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on Aeromonas sp. MH-8.

The design and fabricationt for ion fraction measurement of plasma generator (플라즈마발생기의 이온분율 측정 장치 설계 및 제작)

  • Lee, Chan-Young
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.11a
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    • pp.368-368
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    • 2008
  • Ion implantation has been widely developed during the past decades to become a standard industrial tool. To comply with the growing needs in ion implantation, innovative technology for the control of ion beam parameters is required. Beam current, beam profile, ion fractions are of great interest when uniformity of the implant is an issue. Especially, it is important to measure the spatial distribution of beam power and also the energy distribution of accelerated ions. This energy distribution is influenced by the proportion of mass for ion in the plasma generator(ion source) and by charge exchange and dissociation within the accelerator structure and also by possible collective effects in the neutralizer which may affect the energy and divergence of ions. Hydrogen atom has been the object of a good study to investigate the energy distribution. Hydrogen ion sources typically produce multi-momentum beams consisting of atomic ion ($H^+$) and molecular ion ($H_2^+$ and $H_3^+$). In the beam injector, the molecular ions pass through a charge-exchanges gas cell and break up into atomic with one-half (from $H_2^+$) or one-third (from $H_3^+$) according to their accelerated energy. Burrell et al. have observed the Doppler shifted lines from incident $H^+$, $H_2^+$, and $H_3^+$ using a Doppler shift spectroscopy. Several authors have measured the proportion of mass for hydrogen ion and deuterium using an ion source equipped with a magnetic dipole filter. We developed an ion implanter with 50-KeV and 20-mA ion source and 100-keV accelerator tube, aiming at commercial uses. In order to measure the proportion of mass for ions, we designed a filter system which can be used to measure the ion fraction in any type of ion source. The hydrogen and helium ion species compositions are used a filter system with the two magnets configurations.

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Response Evaluation after Stereotactic Ablative Radiotherapy for Lung Cancer (초기 폐암의 정위방사선치료후 반응평가 분석)

  • Choi, Ji Hoon
    • Progress in Medical Physics
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    • v.26 no.4
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    • pp.229-233
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    • 2015
  • We retrospectively reviewed lung cancer patients who were treated with stereotactic ablative radiotherapy (SABR). We investigated the value of response evaluation after treatment by measuring the volume change of tumors on serial chest computed tomography (CT) examinations. The study included 11 consecutive patients with early-stage (T1-T2aN0M0) non-small cell lung cancer (NSCLC) who were treated with SABR. The median dose of SABR was 6,000 cGy (range 5,000~6,400) in five fractions. Sequential follow-up was performed with chest CT scans. Median follow-up time was 28 months. Radiologic measurement was performed on 51 CT scans with a median of 3 CT scans per patient. The median time to partial response ($T_{PR}$) was 3 months and median time to complete remission ($T_{CR}$) was 5 months. Overall response rate was 90.9% (10/11). Five patients had complete remission, five had partial response, and one patient developed progressive disease without response. On follow-up, three patients (27.2%) developed progressive disease after treatment. We evaluated the the response after SABR. Our data also showed the timing of response after SABR.

Screening of Monocyte Chemoattractant Protein-1-Induced Chemotaxis Inhibitors from Medicinal Herbs (생약자원으로부터 Monocyte Chemoattractant Protein-1에 의한 Chemotaxis 저해활성 검색)

  • Lee, Seung-Woong;Kwon, Oh-Eok;Chung, Mi-Yeon;Kim, Young-Ho;Lee, Hyun-Sun;Kim, Young-Kook;Rho, Mun-Chual
    • Korean Journal of Pharmacognosy
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    • v.33 no.4 s.131
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    • pp.352-358
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    • 2002
  • Blood monocytes are the precursors for the lipid-laden foam cells of early atherosclerotic lesions. Monocyte chemoattractant protein-1(MCP-1), a CC chemokine, and chemokine receptor 2 (CCR2) play a crucial role in the recruitment of monocytes to the vascular lesion. Using the human monocyte THP-1 cell line, we investigate the inhibitory effects of methanol extracts of 127 medicinal herbs on MCP-1 induced chemotaxis. Seven kinds of methanol extracts of medicinal herbs showed above 40% inhibitory effect with the concentration of $25{\mu}g/ml$. They were divide three fractions of $CHCI_3$, BuOH, $H_2O$ to use solvent partition. Among them, butanol extract of Junci Medulla and $CHCI_3$ extract of Clematidis Radix are showed significant inhibitory activities (above 50% inhibition) at the same concentration.