• 제목/요약/키워드: Co-incubation

검색결과 508건 처리시간 0.023초

Inhibition of Lipopolysaccaride-induced Inducible Nitric Oxide (iNOS) mRNA Expression and Nitric Oxide Production by Higenamine in Murine Peritoneal Macrophages

  • Lee, Hoi-Young;Lee, Jang-Soon;Kim, Eun-Ju;Han, Jeung-Whan;Lee, Hyang-Woo;Kang, Young-Jin;Chang, Ki-Churl
    • Archives of Pharmacal Research
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    • 제22권1호
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    • pp.55-59
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    • 1999
  • Nitric oxide synthesized by inducible nitric oxide synthase (iNOS) has been implicated as a mediator of inflammation in rheumatic and autoimmune diseases. The effects of higenamine, a tetrahydroisoquinoline compound, on induction of NOS by bacterial lipopolysaccaride (LPS) were examined in murine peritoneal macrophages. LPS-induced nitrite/nitrate production was markedly inhibited by higenamine which at 0.01 mM, decreased nitrite/nitrate levels by $48.7{\pm}4.4%$This was comparable to the inhibition of LPS-induced nitrite/nitrate production by tetrandrin ($49.51{\pm}2.02%$). at the same concentration. Northern and Western blot analysis of iNOS expression demonstrated that iNOS expression was significantly attenuated following co-incubation of peritoneal macrophages with LPS (10 $\mu\textrm{g}$/m;; 18hrs) and higenamine (0.001, 0.,01 mM; 18hrs). These results suggest that higenamine can inhibit LPS-induced expression of iNOS mRNA in murine peritoneal macrophages. The clinical implications of these findings remain to be established.

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The role and characterization of .betha.-1, 3-glucanase in biocontrol of fusarium solani by pseudomonas stutzeri YPL-1

  • Lim, Ho-Seong;KiM, Sang-Dal
    • Journal of Microbiology
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    • 제33권4호
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    • pp.295-301
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    • 1995
  • An antifungal Pseudomonas stutzeri YPL-1 produced extracellular chitinase and .betha.-1, 3-glucanase that were key enzymes in the decomposition of fungal hyphal walls. These lytic extracellular enzymes markedly inhibited mycelial growth of the phytopathogenic fungus Fusarium solani. A chitinase from P. stutzeri YPL-1 inhibited fungal mycelial growth by 87%, whereas a .betha.-1, 3-glucanase from the bacterium inhibited growth by 53%. Furthermore, co-operative action of the enzymes synergistically inhibited 95% of the fungal growth. The lytic enzymes caused absnormal swelling and retreating on the fungal hyphal walls in a dual cultures. Scanning electron microscopy clearly showed hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. In an in vivo pot test, P. stutzeri YPL-1 proved to have biocontrol ability as a powerful agent in controlling plant disease. Planting of kidney bean (Phaseolus vulgaris L.) seedlings with the bacterial suspension in F. solani-infested soil significantly suppressed the development of fusarial root-rot. The characteristics of a crude preparation of .betha.-1, 3-glucanase produced from P. stutzeri YPL-1 were investigated. The bacterium detected after 2 hr of incubation. The enzyme had optimum temperature and pH of 40.deg.C and pH 5.5, respectively. The enzyme was stable in the pH range of 4.5 to 7.0 and at temperatures below 40.deg.C, with a half-life of 40 min at 60.deg.C.

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Inhibitory Effect of 7-O-butyl Naringenin on Growth of Helicobacter pylori ATCC 26695

  • Kim, Kee-Tae;Moon, Sun-Hee;Yeo, Eun-Ju;Park, Yong-Sun;Han, Ye-Sun;Nah, Seung-Yeol;Lee, Na-Gyong;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • 제15권3호
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    • pp.466-468
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    • 2006
  • The antimicrobial effect of a novel flavonoid (7-O-butyl naringenin) on Helicobacter pylori ATCC 26695 and its inhibitory effects on the urease activity of the strain were evaluated by comparing with quercetin and naringenin. H. pylori was cultured with brain heart infusion supplemented with 5% horse serum at $37^{\circ}C$ under 10% $CO_2$ atmosphere and the inhibitory effects of flavonoids against the strain were detected using micro-plate methods. During 12 hr of incubation time, the optical densities of phenol red reduced (pink color) in the urea broth by producing ammonia were detected at 560 nm with a spectrophotometer. The results indicated that both quercetin and 7-O-butyl naringenin were effective against the growth of H. pylori. Moreover, inhibitory effect of 7-O-butyl naringenin on the growth of H. pylori was about two-fold higher than quercetin at the same concentration. With regard to H. pylori urease activity, 7-O-butyl naringenin had a greater inhibitory effect than did naringenin or quercetin at the same concentration.

Enhanced Production of Galactooligosaccharides Enriched Skim Milk and Applied to Potentially Synbiotic Fermented Milk with Lactobacillus rhamnosus 4B15

  • Oh, Nam Su;Kim, Kyeongmu;Oh, Sangnam;Kim, Younghoon
    • 한국축산식품학회지
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    • 제39권5호
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    • pp.725-741
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    • 2019
  • In the current study, we first investigated a method for directly transforming lactose into galacto-oligosaccharides (GOS) for manufacturing low-lactose and GOS-enriched skim milk (GSM) and then evaluated its prebiotic potential by inoculating five strains of Bifidobacterium spp. In addition, fermented GSM (FGSM) was prepared using a potentially probiotic Lactobacillus strain and its fermentation characteristics and antioxidant capacities were determined. We found that GOS in GSM were metabolized by all five Bifidobacterium strains after incubation and promoted their growth. The levels of antioxidant activities including radical scavenging activities and 3-hydroxy-3-methylglutaryl-CoA reductase inhibition rate in GSM were significantly increased by fermentation with the probiotic Lactobacillus strain. Moreover, thirty-nine featured peptides in FGSM was detected. In particular, six peptides derived from ${\beta}$-casein, two peptides originated from ${\alpha}s_1$-casein and ${\kappa}$-casein were newly identified, respectively. Our findings indicate that GSM can potentially be used as a prebiotic substrate and FGSM can potentially prevent oxidative stress during the production of synbiotic fermented milk in the food industry.

The Effect of Oligosaccharides on Ethylene Production in Mung Bean (Vigna radiata W.) Hypocotyl Segments

  • Choy, Yoon-Hi;Lee, Dong-Hee;Lee, June-Seung
    • Journal of Plant Biology
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    • 제39권4호
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    • pp.295-300
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    • 1996
  • The physiological effects of oligogalacturonic acid (OGA:D. P. 6-7), a product of acid hydrolysis of polygalacturonic acid (PGA), on ethylene biosynthesis in mung bean (Vigna radiata W.) hypocotyl segments was studied. Among PGA, OGA and monogalacturomic acid (MGA), only OGA stimulated ethylene production in mung bean hypocotyl segments, and the most effective concentraton of OGA was 50$\mu\textrm{g}$/mL. Time course data indicated that this stimulatiion effect of OGA appeared after 90 min incubation period and continued until 24 h. When indol-3-acetic acid (IAA) and 1-aminocyclopropane-1-carboxylic acid (ACC) were treated with OGA to investigate the mechanism of OGA on ethylene production, they did not show synergistic effects on ethylene production. The stimulation of ethylene production by OGA was due to the increase of in vivo ACC synthase activity, but OGA treatment had no effect of in vivo ACC oxidase activity. The effect of aminoethoxy vinyl glycine (AVG) and Co2+, the inhibitor of ethylene synthesis, was siminished a little by the OGA, but the treatment of Ca2+, known to increase ACC, with OGA did not increase the ethylene production, this effect seems to be specific for Ca2+ because other divalent cation, Mg2+, did not show the inhibition of OGA-indyuced ethylene production. It is possible that the OGA adopts a different signal transduction pathway to the ethylene bioxynthesis.

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Organic-Inorganic Hybrid Nanoflowers as Potent Materials for Biosensing and Biocatalytic Applications

  • Tran, Tai Duc;Kim, Moon Il
    • BioChip Journal
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    • 제12권4호
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    • pp.268-279
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    • 2018
  • Flower-shaped organic-inorganic hybrid nanostructures, termed nanoflowers, have received considerable recent attention as they possess greatly enhanced activity, stability, durability, and even selectivity of entrapped organic biomolecules, which are much better than those from the conventional methods. They can be synthesized simply via co-incubation of organic and inorganic components in aqueous buffer at room temperature and yield hierarchical nanostructures with large surface-to-volume ratios, allowing for low-cost production by easy scale-up, as well as the high loading capacity of biomolecules without severe mass transfer limitations. Since a pioneering study reported on hybrid nanoflowers prepared with protein and copper sulfate, many other organic and inorganic components, which endow nanoflowers with diverse functionalities, have been employed. Thanks to these features, they have been applied in a diverse range of areas, including biosensors and biocatalysis. To highlight the progress of research on organic-inorganic hybrid nanoflowers, this review discusses their synthetic methods and mechanisms, structural and biological characteristics, as well as recent representative applications. Current challenges and future directions toward the design and development of multi-functional nanoflowers for their widespread utilization in biotechnology are also discussed.

Process Optimization of Nypa fruticans Wurmb Extract using Mixed Solvent and its Functional Component Analysis

  • Kim, Jin;Kim, Han Sung;Son, Jang Wan;Moon, Seong Yong;Lee, Sook-Young
    • 대한의생명과학회지
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    • 제28권1호
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    • pp.34-41
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    • 2022
  • In this study, conditions for extraction of functional component from Nypa fruticans wurmb was optimized. The yield by extraction with 30% ethanol (LE30, 42.12%) was higher than those hot water extraction (LDW 33.32%), 50% ethanol (LE50, 40.12%) and 70% ethanol (LE70, 34.5%). The extract was purified and analyzed by GC MS. The prevailing compounds found in extract were Cyclodecasiloxane-, pentadecanoic acid, -eicosane, undecanal and tridecanoic acid. The presence of saturated and unsaturated fatty acids in ethanolic extract vindicate the use of this plant to treat many diseases in traditional medicine. The total phenolic contents in the LDW, LE30, LE50, LE70 extract were 128±1.65 mg/g, 205±2.3 mg/g, 210±4.23 mg/g and 180±5.6 mg/g, respectively. The DPPH was highest in LE70 extract (1,000 ㎍/mL, 81.14%), ABTS was highest in LE50 extract (1,000 ㎍/mL, 84.14%). The protective effects against oxidative stress in raw 264.7 cell imparted by the LE50 extract was better than those imparted by the other extracts. The findings of the present study suggest that 50% ethanol is best solvent for extraction of Nypa fruticans Wurmb, considering yield, polyphenol content, and antioxidant activities with extraction cost.

Kinesin-13, a Motor Protein, is Regulated by Polo-like Kinase in Giardia lamblia

  • Park, Eun-Ah;Kim, Juri;Shin, Mee Young;Park, Soon-Jung
    • Parasites, Hosts and Diseases
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    • 제60권3호
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    • pp.163-172
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    • 2022
  • Kinesin-13 (Kin-13), a depolymerizer of microtubule (MT), has been known to affect the length of Giardia. Giardia Kin-13 (GlKin-13) was localized to axoneme, flagellar tips, and centrosomes, where phosphorylated forms of Giardia polo-like kinase (GlPLK) were distributed. We observed the interaction between GlKin-13 and GlPLK via co-immunoprecipitation using transgenic Giardia cells expressing Myc-tagged GlKin-13, hemagglutinin-tagged GlPLK, and in vitro-synthesized GlKin-13 and GlPLK proteins. In vitro-synthesized GlPLK was demonstrated to auto-phosphorylate and phosphorylate GlKin-13 upon incubation with [γ-32P]ATP. Morpholino-mediated depletion of both GlKin-13 and GlPLK caused an extension of flagella and a decreased volume of median bodies in Giardia trophozoites. Our results suggest that GlPLK plays a pertinent role in formation of flagella and median bodies by modulating MT depolymerizing activity of GlKin-13.

Involvement of NOX2-derived ROS in human hepatoma HepG2 cell death induced by Entamoeba histolytica

  • Young Ah Lee ;Myeong Heon Shin
    • Parasites, Hosts and Diseases
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    • 제61권4호
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    • pp.388-396
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    • 2023
  • Entamoeba histolytica is an enteric tissue-invasive protozoan parasite causing amoebic colitis and liver abscesses in humans. Amoebic contact with host cells activates intracellular signaling pathways that lead to host cell death via generation of caspase-3, calpain, Ca2+ elevation, and reactive oxygen species (ROS). We previously reported that various NADPH oxidases (NOXs) are responsible for ROS-dependent death of various host cells induced by amoeba. In the present study, we investigated the specific NOX isoform involved in ROS-dependent death of hepatocytes induced by amoebas. Co-incubation of hepatoma HepG2 cells with live amoebic trophozoites resulted in remarkably increased DNA fragmentation compared to cells incubated with medium alone. HepG2 cells that adhered to amoebic trophozoites showed strong dichlorodihydrofluorescein diacetate (DCF-DA) fluorescence, suggesting intracellular ROS accumulation within host cells stimulated by amoebic trophozoites. Pretreatment of HepG2 cells with the general NOX inhibitor DPI or NOX2-specific inhibitor GSK 2795039 reduced Entamoeba-induced ROS generation. Similarly, Entamoeba-induced LDH release from HepG2 cells was effectively inhibited by pretreatment with DPI or GSK 2795039. In NOX2-silenced HepG2 cells, Entamoeba-induced LDH release was also significantly inhibited compared with controls. Taken together, the results support an important role of NOX2-derived ROS in hepatocyte death induced by E. histolytica.

섬유소 분해균을 이용한 건조 청보리 발효사료가 돼지의 In vitro 발효 특성에 미치는 영향 (Effects of Dried Whole Crop Barley Treated with Cellulolytic Microorganisms on In Vitro Fermentation Characteristics in Swine)

  • 박도연;박중국;조성백;김창현
    • 한국초지조사료학회지
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    • 제30권2호
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    • pp.179-190
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    • 2010
  • 본 연구는 높은 섬유소 분해력을 검증 받은 Aspergillus niger (KCCM 60357)와 Bacillus licheniformis (KCCM 40934)를 단독 및 혼합 배양한 미생물제제로 양돈용 청보리 발효사료를 제조하였을 때 사료 성상변화, in vitro 대장발효 및 전장소화율에 미치는 영향을 평가하였다. 실험 설계는 건조 청보리 (control), A. niger (control + A. niger), B. licheniformis (control + B. licheniformis) 및 Mixture (control + Aspergillus niger + Bacillus licheniformis)구로, 균주를 첨가시킨 발효사료에 따라 네 처리군으로 나누었다. 사료 성분변화는 발효사료의 일반 성분분석을 통해 확인하였으며, 그 결과 A. niger 및 B. licheniformis구에서 조단백질과 조지방 함량이 유의하게 증가하였다 (p<0.05). In vitro 대장발효성상의 변화는 배양 후 24시간에 건물 분해율이 A. niger와 Mixture구에서 각각 55 및 57.42%로 대조구 및 B. licheniformis구와 비교하여 유의하게 높았다 (p<0.05). 가스 발생량은 대조구에서 유의적으로 가장 높았으며, 메탄가스 농도는 모든 처리구가 대조구에 비해 증가되었다. pH는 대조구를 비롯한 모든 처리구가 6.8 로 확인되었으며, 암모니아 질소는 모든 발효사료에서 유의하게 증가되었다. Xylanase 활력은 A. niger구가 유의적으로 높았으며 모든 처리구의 활력이 증가하였다 (p<0.05). 총 VFA 농도는 배양 후 24시간에 B. licheniformis구에서 높은 농도 (12.61mM)를 보여주며 VFA 개선효과를 나타냈다 (p<0.05). In vitro 전장 소화율은 B. licheniformis구에서 49.61%로 대조구의 45.65% 보다 소화율이 유의적으로 증진되었다(p<0.05). 따라서 본 실험에 사용된 미생물을 이용하여 청보리 발효사료를 제조하였을 때 전장 소화율 및 대장발효 환경 개선에 충분한 효과가 있을 것으로 기대된다.