• Title/Summary/Keyword: H17/28

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Analysis of domain required for aggregates formation of ALS (Amyotrophic lateral sclerosis)/FTD (Frontotemporal dementia)-linked FUS in mammalian cells (루게릭병 및 전측두엽성 치매 연관 단백질 Fused in Sarcoma (FUS)의 스트레스 응집체 형성에 관여하는 도메인 분석)

  • Jun, Mi-Hee;Lee, Jin-A
    • Analytical Science and Technology
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    • v.28 no.5
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    • pp.331-340
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    • 2015
  • Mutations in Fused in Sarcoma (FUS) have been identified in patients with amyotrophic lateral sclerosis (ALS) or Frontotemporal Dementia (FTD). Pathological FUS is mis-localized to cytosol and forms aggregates associated with stress granules (SG), while FUS is normally localized to nucleus. However, it is largely unknown how pathological FUS forms SG-aggregates and which domains are responsible for this process. In this study, we examined cellular localization and aggregation of ALS-linked FUS missense mutants (P525L, R521C, R521H, R521G), analyzed the domains responsible for cytosolic FUS aggregation in HEK293T cells, and confirmed this in cultured mouse neurons. To do this, we firstly generated missense mutants of FUS and then examined their cellular localization. We found that P525L was mostly mis-localized to cytosol and formed FUS-positive SG aggregates while R521C, R521H, or R521G was localized to both nucleus and cytosol. To further characterize the domains required for aggregate formation of cytosolic FUS, we generated different domain-deletion mutants using FUS-∆17 which has a deletion of nuclear localization signal. Interestingly, cytosolic FUS without SYGQ and RGG1 domain or cytosolic FUS without RGG2-ZnF-RGG3 domain did not form FUS-positive SG aggregates, while cytosolic FUS without RRM domain generated more aggregates compared to FUS-∆17. Taken together, these data suggest that SYGQ-RGG1 or RGG2-ZnF-RGG3 domain contributes to formation of cytosolic aggregate, while RRM domain might interfere with FUS aggregation. Therefore, our studies will provide important insight for understanding cellular pathogenesis of neurodegeneration associated with FUS aggregate as well as finding therapeutic targets for ALS or FTD.

Propofol protects human keratinocytes from oxidative stress via autophagy expression

  • Yoon, Ji-Young;Jeon, Hyun-Ook;Kim, Eun-Jung;Kim, Cheul-Hong;Yoon, Ji-Uk;Park, Bong-Soo;Yu, Su-Bin;Kwak, Jin-Won
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.17 no.1
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    • pp.21-28
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    • 2017
  • Background: The skin consists of tightly connected keratinocytes, and prevents extensive water loss while simultaneously protecting against the entry of microbial pathogens. Excessive cellular levels of reactive oxygen species can induce cell apoptosis and also damage skin integrity. Propofol (2,6-diisopropylphenol) has antioxidant properties. In this study, we investigated how propofol influences intracellular autophagy and apoptotic cell death induced by oxidative stress in human keratinocytes. Method: The following groups were used for experimentation: control, cells were incubated under normoxia (5% $CO_2$, 21% $O_2$, and 74% $N_2$) without propofol; hydrogen peroxide ($H_2O_2$), cells were exposed to $H_2O_2$ ($300{\mu}M$) for 2 h; propofol preconditioning (PPC)/$H_2O_2$, cells pretreated with propofol ($100{\mu}M$) for 2 h were exposed to $H_2O_2$; and 3-methyladenine $(3-MA)/PPC/H_2O_2$, cells pretreated with 3-MA (1 mM) for 1 h and propofol were exposed to $H_2O_2$. Cell viability, apoptosis, and migration capability were evaluated. Relation to autophagy was detected by western blot analysis. Results: Cell viability decreased significantly in the $H_2O_2$ group compared to that in the control group and was improved by propofol preconditioning. Propofol preconditioning effectively decreased $H_2O_2$-induced cell apoptosis and increased cell migration. However, pretreatment with 3-MA inhibited the protective effect of propofol on cell apoptosis. Autophagy was activated in the $PPC/H_2O_2$ group compared to that in the $H_2O_2$ group as demonstrated by western blot analysis and autophagosome staining. Conclusion: The results suggest that propofol preconditioning induces an endogenous cellular protective effect in human keratinocytes against oxidative stress through the activation of signaling pathways related to autophagy.

Effect of One- or Two-Week Triple Therapy with Omeprazole, Amoxicillin, and Clarithromycin on Eradication of Helicobacter pylori Infection in Children (소아에서 Omeprazole, Amoxicillin, Clarithromycin 삼제 병합 요법의 치료기간별 Helicobacter pylori 제균율)

  • Choi, In-Kyoung;Lee, Seung-Yun;Chung, Ki-Sup
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.5 no.1
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    • pp.19-25
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    • 2002
  • Purpose: The triple therapy with proton pump inhibitor (PPI) has been recognized as the treatment of choice in Helicobacter pylori (H. pylori) infection in adults. However, the effect of triple therapy with omeprazole, amoxicillin and clarithromycin (OAC) on eradication of H. pylori infection in children has not been established yet. This study was performed to evaluate the efficacy of OAC triple therapy and to compare the effect of one-week with two-week therapy on H. pylori eradication. Methods: From July 1998 to July 2000, 34 children with upper gastrointestinal symptoms, who underwent upper gastrointestinal endoscopy with biopsy at entry and 4 or more weeks after therapy, were enrolled in this study. H. pylori infection was assessed by CLO test and histologic examination (Hematoxylin-Eosin stain or Alcian yellow stain) with biopsy specimens. The regimen consisted of omeprazole (0.7 mg/kg/day), amoxicillin (50 mg/kg/day), and clarithromycin (25 mg/kg/day) for 1 week (n=21) or 2 weeks (n=13). Eradication of H. pylori was determined after the termination of treatment by the CLO test and histologic examination. Results: One-week treatment group consisted of 21 children (11 male, 10 female) with a mean age of $9.5{\pm}3.0$ years. Two-week group consisted of 13 children (4 male, 9 female) with a mean age of $9.9{\pm}4.0$ years. The endoscopic diagnoses included nodular gastritis in 19 cases, superficial gastritis in 7 cases, gastric ulcer in 4 cases, purpuric duodenitis in 2 cases, and normal in 2 cases. H. pylori was eradicated in 28 of total 34 children (82.4%). In 1-week group, H. pylori was eradicated in 17 of 21 children (81%). In 2-week group, H. pylori was eradicated in 11 of 13 children (84.6%). In remaining 6 cases in whom H. pylori had not been eradicated with OAC regimen, H. pylori infection persisted despite of the treatment with additional drugs such as colloidal bismuth subcitrate ($Denol^{(R)}$) and metronidazole. Conclusion: In this study, eradication rate of H. pylori with OAC regimen was 82.4%, and the triple therapy would be highly effective as primary treatment. However, there was no significant difference in the eradication rate between the 1-week and 2-week treatment group (P=0.785).

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Shelf Life Extension of Wasabi Paste Products by Addition of Citric Acid (구연산 첨가에 의한 와사비 페이스트 제품의 저장성 향상)

  • Jeong, Eun-Jeong;Lee, Hyo-Kyung;Kim, Yong-Suk
    • Journal of Food Hygiene and Safety
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    • v.34 no.4
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    • pp.354-360
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    • 2019
  • In order to extend the shelf life of Wasabi paste, the effects of citric acid were confirmed at $35^{\circ}C$ for 28 days. Citric acid-treated groups contained citric acid in amounts of 0.05, 0.10, 0.17, 0.30, and 0.40%, respectively. Quality characteristics of Wasabi pastes were determined in pH, titratable acidity, soluble solid content, color values, microbial analysis (aerobic bacteria, yeast), gas production, and content of allyl isothiocyanate. The pH and titratable acidities of Wasabi pastes added with citric acid were indicated as 4.03-5.19 and 4.23-4.82%, respectively. Soluble solid content was significantly different according to concentrations of citric acid. L values showed the highest at $50.05{\pm}0.46$. a and b values were increased during the storage period. Total aerobic bacteria and yeast counts of Wasabi pastes were decreased in a dose-dependent manner. Gas production from Wasabi pastes showed at 19.55-19.80 mL/tube after 28 days of storage. The addition of citric acid (0.3% or more) to the Wasabi paste resulted in increased storage stability.

Effects of Different Dietary Acidifier Sources of Calcium and Phosphorus on Ammonia, Methane and Odorant Emission from Growing-finishing Pigs

  • Kim, I.B.;Ferke, P.R.;Powers, W.J.;Stein, H.H.;Van Kempe, T.A.T.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.8
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    • pp.1131-1138
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    • 2004
  • The objective of this study was to investigate the effects of different sources of Ca and P on urine and ileal digesta pH, and ammonia ($NH_{3}$), methane ($CH_{4}$), and odor emission. In experiment 1, eight pigs (commercial three-way cross; initial BW 67$\pm$3 kg) were arranged in a repeated 4$\times$4 Latin Square design. All pigs were equipped with a T-cannula in the distal ileum. Four corn-soybean meal based diets were formulated. Diet 1 was the control in which dicalcium phosphate (DCP) and limestone ($CaCO_{3}$) were used as the sources of inorganic P and Ca. In Diets 2 and 3, ${H_{3}}{PO_{4}}$, monocalcium phosphate (MCP), and $CaSO_{4}$replaced DCP and $CaCO_{3}$ as the inorganic sources of P and Ca. Diet 4 was similar to Diet 1 except that it was fortified with HCl to provide an acid load similar to that of diet 2. Urine and ileal digesta pH were determined in pigs fed each of these diets. In Exp. 1, urine pH decreased (p<0.05) in animals consuming diets containing ${H_{3}}{PO_{4}}$-$CaSO_{4}$ (5.85$\pm$0.38) and MCP-$CaSO_{4}$(5.73$\pm$0.30) compared with the DCP-$CaCO_{3}$ diet (6.89$\pm$0.24). In the pigs consuming ${H_{3}}{PO_{4}}$-$CaSO_{4}$, ileal digesta pH decreased compared with the control (5.52$\pm$0.28 vs. 6.66$\pm$0.17; p<0.05). Based on the results of Exp. 1, a total of four trials were performed in environmental chambers for determining how $NH_{3}$, $NH_{4}$, and odor were affected by the different dietary Ca and P sources (Exp. 2). In Exp. 2, pigs fed the ${H_{3}}{PO_{4}}$-$CaSO_{4}$ diet had decreased (30%) $NH_{3}$ emissions compared with the control (p<0.05). Also, a combination of MCP-$CaCO_{3}$-$CaCl_{12}$ decreased $NH_{3}$ emission by 15% (p<0.05). Emission of $CH_{4}$ was decreased only with the ${H_{3}}{PO_{4}}$-$CaSO_{4}$ diet with 14% (p<0.05). Odorant emission of phenolics and volatile fatty acids increased roughly three-fold with the DCP-$CaSO_{4}$ diet but was not affected by other test diets. In conclusion, acidogenic Ca and P sources in swine diets can decrease the urinary pH and reduce $NH_{3}$ and $CH_{4}$ emission from swine facilities.

Novel Alkali-Tolerant GH10 Endo-${\beta}$-1,4-Xylanase with Broad Substrate Specificity from Microbacterium trichothecenolyticum HY-17, a Gut Bacterium of the Mole Cricket Gryllotalpa orientalis

  • Kim, Do Young;Shin, Dong-Ha;Jung, Sora;Kim, Hyangmi;Lee, Jong Suk;Cho, Han-Young;Bae, Kyung Sook;Sung, Chang-Keun;Rhee, Young Ha;Son, Kwang-Hee;Park, Ho-Yong
    • Journal of Microbiology and Biotechnology
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    • v.24 no.7
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    • pp.943-953
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    • 2014
  • The XylH gene (1,167-bp) encoding a novel hemicellulase (41,584 Da) was identified from the genome of Microbacterium trichothecenolyticum HY-17, a gastrointestinal bacterium of Gryllotalpa orientalis. The enzyme consisted of a single catalytic domain, which is 74% identical to that of an endo-${\beta}$-1,4-xylanase (GH10) from Isoptericola variabilis 225. Unlike other endo-${\beta}$-1,4-xylanases from invertebrate-symbiotic bacteria, rXylH was an alkali-tolerant multifunctional enzyme possessing endo-${\beta}$-1,4-xylanase activity together with ${\beta}$-1,3/${\beta}$-1,4-glucanase activity, which exhibited its highest xylanolytic activity at pH 9.0 and 60oC, and was relatively stable within a broad pH range of 5.0-10.0. The susceptibilities of different xylosebased polysaccharides to the XylH were assessed to be as follows: oat spelts xylan > beechwood xylan > birchwood xylan > wheat arabinoxylan. rXylH was also able to readily cleave p-nitrophenyl (pNP) cellobioside and pNP-xylopyranoside, but did not hydrolyze other pNP-sugar derivatives, xylobiose, or hexose-based materials. Enzymatic hydrolysis of birchwood xylan resulted in the product composition of xylobiose (71.2%) and xylotriose (28.8%) as end products.

Screening of Wood-Rot Fungi Based on RBBR Decolorization and Its Laccase Activity (RBBR 탈색능을 이용한 목재부후균의 선발 및 이들 균의 Laccase 효소활성)

  • Choi, Yun-Jeong;Shin, Yoo-Su;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.4
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    • pp.46-53
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    • 2006
  • This study was to screen white-rot fungi possesing strong lignin degrading enzymes, glucose-1 oxidase (GOD), laccase (LAC) and Mn-peroxidase (MnP), based on their decolorization activity of Remazol Brilliant Blue R (RBBR). In the midst of 20 tested fungi, 9 isolates were shown 4 kinds of activities such as RBBR decolorization, GOD, LAC and MnP. Relatively high active strains were identified as Phlebia radiata, Trametes versicolor, Abortiporus biennis, Gleophyllum odoratum and Cerrena unicolor. In particular, T. versicolor, G. odoratum, and C. unicolor, which have high activities of LAC, were used to confirm the optimal temperature and pH and to evaluate the effect of inducer, 2,5-xylidine on their LAC activity. The optimum temperatures for mycelial growth were $28^{\circ}C$ for T. versicolor and G. odoratum, and $25^{\circ}C$ for C. unicolor. The optimum pH for mycelial growth was 5.5. Three strains showed the increase of LAC enzyme activity by the addition of 2,5-xylidine. T. versicolor had the highest LAC activity of $22,700nkat/{\ell}$, corresponding to 11.3 times, G. odoratum $15,400nkat/{\ell}$, 9 times and C. unicolor $17,330nkat/{\ell}$, 5.5 times higher than those of the control.

Changes of Physicochemical Properties and Microbial during Storage of Commercial Makgeolli (시판막걸리의 저장기간에 따른 품질 특성 및 미생물의 변화)

  • Kwon, Young-Hee;Lee, Ae-Ran;Kim, Jae-Ho;Kim, Hye-Ryun;Ahn, Byung-Hak
    • The Korean Journal of Mycology
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    • v.40 no.4
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    • pp.210-214
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    • 2012
  • The aim of this study was to determine changes of physicochemical properties and microorganisms of commercial Makgeolli during storage at $10^{\circ}C$ and $25^{\circ}C$ During storage at $10^{\circ}C$ for 11 days, ethanol contents did not increased up to 7.0%, and pH and total acidity were ranged with 3.12-3.99 and 0.22-0.28%, respectively. Makgeolli which was storaged at $25^{\circ}C$ for 11 days showed more than 7.0% of alcohol contents, pH of 3.96-4.17 and total acidity of 0.27-0.30%. Yeast cell counts showed maximal $2.90-16.00{\times}10^7$ CFU/mL at $10^{\circ}C$ for 3 days and $1.12-29.40{\times}10^7$ CFU/mL at $25^{\circ}C$ for 1 day. Lactic acid bacteria and total bacteria cell counts of Makgeolli which was storaged at $25^{\circ}C$ for 11 days were higher than those $10^{\circ}C$ storage. In preference test, Makgeolli from storage at $10^{\circ}C$ for 11 days was showed the level of 4.9-5.2 points however, Makgeolli from storage at $25^{\circ}C$ for one day showed below 5.0 point.

Effects of Evaporation on the Weathering Rate and Chemical Composition of Iranian Heavy Crude Oil (이란산 원유의 증발에 따른 풍화율 및 화학적 성상 변화)

  • Kim, Beom;Kim, Gi-Beum;Sim, Won-Joon;Yim, Un-Hyuk
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.15 no.3
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    • pp.238-246
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    • 2012
  • Once oil is spilled into marine environment, it experiences various weathering processes among which evaporation is the most dominant process in the initial stage of weathering. This study aimed to elucidate the effects of evaporation on the physicochemical properties of spilled oil using standardized laboratory experiments. Laboratory evaporation process was successfully reproduced using controlled rotary evaporation method. In case of Iranian Heavy crude (IHC), evaporation rate after 48 hours was $29.3{\pm}0.4%$ (n=40, p<0.001). Evaporation was simulated using ADIOS2 weathering model and the result was in agreement with laboratory experiment. Chemical composition changes of petroleum hydrocarbons including alkanes, polycyclic aromatic hydrocarbons (PAHs) and biomarkers by evaporation rate were also analyzed. As oil evaporated, low molecular weight alkanes and PAHs decreased, while biomakers showed conservative characteristics. Among biomarkers, $17{\alpha}(H)$, $21{\beta}(H)$-hopane was used for calculation of weathering rates, which matched with evaporative mass losses. Weathering rate calculation using hopane showed that stranded oils of weathering stage I (28.9%) and mesocosm oil weathering experiment till 5 days (26.5%) were mainly affected by evaporation process.

The Chemical Characteristics of Korean Red Wine with Different Grape Varieties (포도품종을 달리한 적포도주의 이화학적 성분변화 (I))

  • Lee, Jang-Eun;Won, Yoo-Dong;Kim, Sung-Soo;Koh, Kyung-Hee
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.151-156
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    • 2002
  • Five kinds of red wine from three different grape varieties, Gerbong (G), Campbell (C), Moru (M), Gerbong+Moru (70:30, GM) and Gerbong+Campbell (70:30, GC), were prepared. Their chemical and microbiological changes were monitored during alcohol fermentation and aging. The changes of pH, brix and total acidity were $3.2{\sim}3.6,\;17.9{\sim}6.0$ and $2.4{\sim}4.6\;g/L$, respectively. The viable cell numbers of initial, stationary and death phases were $6.0{\times}10^6,\;1.0{\times}10^8$ and $7.0{\times}10^5\;cfu/mL$ during alcohol fermentation. The sugar fermentability, glucose and fructose contents were greatly decreased less than 0.2 g/L, and the final contents of ethanol and sulfur dioxide were $11.4{\sim}12.3%$ and $40{\sim}62\;mg/L$, respectively. The conversion ratios of malic acid to lactic acid were 23% (G), 67% (M), 28% (C), 33% (GM) and 39% (GC). The chemical characteristics of five red wine were significantly different in pH, total acidity, sulfur dioxide and lactic acid contents (p<0.05).