• Title/Summary/Keyword: K-PDB

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Rapid Induction of mRNA for Prostaglandin H Synthase in Ovine Meningeal Fibroblasts

  • Nam, Myeong-Jin;Thore, Clara;Busija, David
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.4
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    • pp.435-441
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    • 1998
  • We examined effects of interleukin $1{\alpha}$ ($IL1{\alpha}$) and phorbol 12, 13 dibutyrate (PDB), an activator of protein kinase C, on mRNA for Prostaglandin H synthase (PGHS) and prostanoid production in cultured ovine meningeal fibroblasts. Immuno- and morphologically-identified fibroblasts were derived from cerebral cortex and white matter from fetal lambs (approximately 120 days gestation) and grown to confluence on glass coverslips in 12 well plates. Levels of prostaglandin $F_{2{\alpha}}$ and the stable hydrolysis product of prostacyclin (i.e., $6-keto-PGF_{1{\alpha}}$) were determined using enzyme immunoassay. Relative amounts of mRNA were determined by in situ hybridization using ovine cDNA for PGHS1. $IL1{\alpha}$ (10 ng/ml) increased mRNA levels over baseline by $62{\pm}19%$ (p<0.05) at 60 min., $37{\pm}12%$ (NS) at 120 min., and $36{\pm}18%$ (NS) at 240 min (n=12). Levels of $6-keto-PGF_{1{\alpha}}$ were $148{\pm}18%$ pg/ml during baseline, $246{\pm}41%$ pg/ml at 60 min., $248{\pm}40%$ pg/ml at 120 min., and $259{\pm}62%$ pg/ml at 240 min (all p<0.05) (n=12). $PGF_{2{\alpha}}$ was increased although it wasn't statistically significant. However, $IL1{\alpha}$ decreased $PGE_2$ level significantly (all p<0.05). PDB $(10^{-6}M)$ increased mRNA levels over baseline by $25{\pm}6%$ after 30 min., $40{\pm}6%$ after 60 min., and $20{\pm}8%$ after 90 min. (n=9) (all p<0.05). Levels of $6-keto-PGF_{1{\alpha}}$ were $200{\pm}43%$ pg/ml during baseline, $202{\pm}43%$ pg/ml after 30 min. (NS), $268{\pm}58%$ pg/ml after 60 min. (p<0.05), and $296{\pm}60%$ pg/ml after 90 min. (p<0.05) (n=9). Levels of $PGF_{2{\alpha}}$ were $178{\pm}26%$ pg/ml during baseline, $300{\pm}30%$ pg/ml after 30 min., $299{\pm}35%$ pg/ml after 60 min., and $355{\pm}32%$ pg/ml after 90 min (all p<0.05) (n=6). Actinomycin-D (1 mg/ml) prevented increases in mRNA, $6-keto-PGF_{1{\alpha}}$, and $PGF_{2{\alpha}}$ at 60 min. for both $IL1{\alpha}$ and PDB. We conclude that cerebral fibroblasts are avid producers of prostanoids, and that enhanced production of PGHS is responsible for augmented $PGF_{2{\alpha}}$ and prostacyclin production in the presence of an activator of protein kinase C and for decreased $PGE_2$ and increased prostacyclin production in the presence of $IL1{\alpha}$.

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Effect of addition amino acids on the mycelial growth and the contents of β-glucan and γ-aminobutyric acid (GABA) in Sparassis latifolia (아미노산 첨가가 꽃송이버섯 균사체 성장 및 베타글루칸, GABA 함량 변화에 미치는 영향)

  • Jo, Han-Gyo;Shin, Hyun-Jae
    • Journal of Mushroom
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    • v.15 no.1
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    • pp.38-44
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    • 2017
  • Sparassis latifolia (formerly S. crispa) is used in food and nutraceuticals or dietary supplements, as rich in flavor compounds and ${\beta}-glucan$. Some previous studies have reported the effects of mushroom on brain function, including its neuroprotective effect. Thus, for this mushroom to be used as an effective nutraceutical for brain function, it would be desirable for it to contain other compounds such as ${\gamma}-aminobutyric$ acid (GABA) in addition to ${\beta}-glucan$. In this study, the enhancement of growth and GABA production in the mycelium of medicinal and edible mushroom S. latifolia was investigated. Amino acids were added externally as the main source of nutrition, and the effects of amino acids were investigated using liquid medium, specifically amino acid-free potato dextrose broth (PDB). The amino acids added were L-glutamic acid (named PDBG medium) and L-ornithine (named PDBO medium). The growth of mycelia was determined to be $0.9{\pm}0.00g/L$, $2.2{\pm}0.16g/L$, and $1.93{\pm}0.34g/L$ PDBG respectively. The GABA content was $21.3{\pm}0.9mg/100g$ in PDB medium, and it in PDBG 1.4% medium, at $115.4{\pm}30.2mg/100g$. However, the PDBO medium was not effective in increasing the GABA content of mycelia. Amino acids had little effect on the ${\beta}-glucan$ content of mycelia. The ${\beta}-glucan$ content was $39.7{\pm}1.4mg/100mg$, $34.4{\pm}0.2mg/100mg$, and $35.2{\pm}9.2mg/100mg$ in PDB, PDBG 1.8% and PDBO 1.4% media, respectively. Addition of glutamic acid and ornithine positively affected the growth of S. latifolia mycelia, and glutamic acid positively affected GABA production; no degradation of GABA was observed with addition of glutamic acid.

Purification and Characterization of Extracellular Laccase from Trametes versicolor (Trametes versicolor 배양액으로부터 단리 정제된 Laccase의 효소적 특성)

  • Kim, Hyun Joo;Bae, Hyeun Jong
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.4
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    • pp.61-66
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    • 2007
  • The study was performed to purify and characterize laccase in culture of Trametes versicolor. The fungus was grown in liquid culture media of PDB and added 2,5-xylidine (0.2 mM) after 5 days to enhance the production of laccase. The fungal culture was incubated at $25^{\circ}C$ on a rotary shaker (120 rpm) for 7days, and the culture broth was clarified through Glass filter (GF/C). The aqueous solution was concentrated by ultramicrofiltration (Viva flow 50, GE Healthcare Bioscience, USA) and loaded onto a Hitrap Q FF column. Laccase activity could be detected at one peak, and this enzyme has a molecular mass of approximately 53kDa as determined by SDS-PAGE The optimum pH and temperature for syringaldazine were 5.0 and $60^{\circ}C$, respectively. The specific activity of crude, concentrated and purified laccase were 32, 409, and 1,243 U/mg, respectively.

A Study on Design of Schema Integration based Biological Information Retrieval System (스키마 통합 기반 생명정보 검색시스템(BIRS) 설계에 관한 연구)

  • Han, Keon;Lee, Sang-Ho;Ahn, Bu-Young
    • Journal of Information Management
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    • v.40 no.1
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    • pp.217-234
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    • 2009
  • In computer-based virtual lab, a bioscience researcher who wants to obtain bio information first uses a biodiversity-related database to retrieve information on species, ecology and distribution of an organism. The researcher also needs to access gene/protein databases such as GenBank or PDB to find information on the organism's genetic sequence and protein structure. Furthermore, the researcher should search for academic papers containing the information on the organism so that his research is based on comprehensive and accurate information. This series of activities often undermines research efficiency as it takes a lot of time and causes inconvenience on the part of researchers. To solve such inconvenience, we analyzed various methods for integrated search and chosen schema integration. In addition, we analyzed each databases and extracted metadata for designing schema integration. This paper introduces a biological information retrieval system(BIRS) using schema integration and it's interface that will increase research efficiency for bioscience.

Effects of Medium, Temperature and pH on Mycelial Growth and Cellulase Activity of Ectomycorrhizal Fungi from Korean Forests (우리나라 산림에서 분리한 외생균근균의 균사생장에 있어 배지, 온도, pH의 영향과 셀룰라아제 활성)

  • Jeon, Sung-Min;Kim, Min-Soo;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.40 no.4
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    • pp.191-203
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    • 2012
  • Mycelial growth of ectomycorrhizal fungi (27 strains of 8 species) collected from Korean forests was observed on various culture conditions (media, temperature, pHs). After 60 days of incubation, all strains grown on potato dextrose agar (PDA) and modified Melin-Norkran's agar (MMNA), whereas no mycelial growth was observed on malt extract agar (MEA) or sabouraud dextrose agar (SDA) in some strains including Tricholoma matsutake. Mycelial growth on PDA was poor at high temperature ($30^{\circ}C$) than the low temperature ($10^{\circ}C$). The optimal temperature on PDA and pH in potato dextrose broth (PDB) for mycelial growth in most strains were $20-25^{\circ}C$ and pH 4-5, respectively. All strains tested showed the carboxymethyl cellulase (CM-cellulase) activity and the maximal cellullase activity was expressed by the mycelium of T. matsutake (KFRI 1266) on the CMC agar plate with pH 5.0.

Mycelial Culture and Fruiting Analysis of Panellus edulis Strains Collected in Korea (Panellus edulis 수집 균주의 균사배양 및 자실체 특성 분석)

  • Woo, Sung-I;Ryoo, Rhim;Jang, Yeongseon;Park, Youngae;Jeong, Yeun Sug;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.46 no.3
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    • pp.281-294
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    • 2018
  • Molecular analysis using the internal transcribed spacer region sequences revealed that the strains used in this study, which were formerly identified as Panellus serotinus, are Panellus edullis. After Universal Fungal PCR Fingerprinting (UFPF) analysis, eight strains of P. edulis were divided into two groups. We conducted fundamental research on mycelial growth and sawdust cultivation to understand the cultural characteristics of eight wild P. edulis strains collected from Korean forests. All strains showed faster and denser mycelial growth on potato dextrose agar (PDA) than on other media (malt extract agar, Sabouraud dextrose agar). Optimal conditions for mycelial growth were: $20^{\circ}C$ on PDA, $25^{\circ}C$ on potato dextrose broth (PDB), and pH 5~8 on PDB at $25^{\circ}C$. Two strains (NIFoS 2407, 3993) were selected as excellent strains based on mycelial growth and density on PDA. NIFoS 2792 showed high cellulase activities on carboxymethyl cellulose (CMC) agar, and NIFoS 2387 and 2804 exhibited high laccase activities on ABTS-containing agar media. The mycelial growth of P. edulis was the fastest on Quercus acutissima and Q. mongolica sawdust media, and mycelial density was the highest on Quercus spp. sawdust-containing media. Sawdust cultivation of P. edulis was successful. The conditions were 80~85 days of cultivation period after spawn inoculation, 10~11 days for primordial formation at $17{\sim}18^{\circ}C$, and 15~20 days for fruiting growth. NIFoS 2804 and 3993 were selected as good strains in terms of cultivation period and mushroom production. These results could be useful for the artificial cultivation of P. edulis.

Leaf Blight of Perilla Caused by Alternaria alternata (Alternaria alternata에 의한 들깨 잎마름병)

  • 차외진;김철승;송주희;김현주;이영병;문병주
    • Journal of Life Science
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    • v.12 no.6
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    • pp.708-714
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    • 2002
  • A new leaf blight was found on the perilla leaves at the major perilla-cultivating areas such as Kangdong in Busan and Miryang in Kyungnam province. Symptoms of the disease initially appeared on the edge of perilla leaves showing black necrosis and drying, and the infected leaves were finally fell down. The SD1 isolate showing strong pathogenicity and forming abundant conidial spores on the diseased lesions was isolated. Among the tested media, mycelial growth was abundant on PDA (Potato Dextrose Agar) medium at $25^{\circ}C$ under dark condition, but conidial formation was greater on V8A (V-8 juice A8ar) medium than that on PDA medium. Optimal temperatures for mycelial growth and conidial formation on PDA medium were respectively $25^{\circ}C$ and 3$0^{\circ}C$. The rate of conidial germination and the elongation of germ tube were more effective in 10% tomato juice than those in PDB (Potato Dextrose Broth) and sterile water. In 10% tomato juice, the rate of conidial germination and the length of germ tube were 100% after incubation for 24k and 535.2${\mu}{\textrm}{m}$ after incubation for 36hr, respectively. According to the result of pathogenicity, it revealed that conidial suspention with 10% tomato juice was the most effective for pathogenicity test showing as 100% of disease incidence, and the symptoms caused by artificial inoculum were same as those of naturally infected perilla. In this study, the SD1 isolate according to the results of morphological characteristics, the incubation characteristics and pathogenicity was firstly identified A. alternata, and named as leaf blight of perilla.

The Roles of Hydroxyl Substituents in Tyrosinase Inhibitory Activation of Flavone Analogues (Flavone 유도체들의 Tyrosinase 저해활성화 반응에서 Hydroxyl 치환기들의 역할)

  • Park, Joon-Ho;Sung, Nack-Do
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.56-62
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    • 2011
  • Molecular docking of polyhydroxy substituted flavone analogues (1-25) as substrate molecules to the active site of tyrosinase (PDB ID: Deoxy-form (2ZMX) & Oxy-form (1WX2)) and Free-Wilson analysis were studied to understand the roles of hydroxyl substituents ($R_1-R_9$) in substrate molecules for the tyrosinase inhibitory activation. It is founded from Free-Wilson analysis that the $R_1$=hydroxyl among $R_1-R_9$ substituents had the strongest influence on the tyrosinase inhibitory activity. H-bonds between the hydroxyl substituents of substrate molecules and amino acid residues in the active site of tyrosinase were contributed to make a stable substrate-receptor complex compound. Particularly, it is proposed from the findings that the noncompetitive inhibitory activation would take place via H-bonding between peroxide oxygen (Per404) atom in the active site of tyrosinase and the hydroxyl substituents in substrate molecule.

Identification of Auxin from Pseudomonas sp. P7014 for the Rapid Growth of Pleurotus eryngii Mycelium (큰느타리버섯 균사체의 생육촉진을 위한 Pseudomonas sp. P7014으로부터 옥신 확인)

  • Kang, Young Min;Cho, Kye Man
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.15-21
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    • 2014
  • The promoting effect of Pseudomonas sp. P7014 on the mycelia growth of Pleurotus eryngii was investigated. An ethyl acetate fraction (F5) from the culture supernatant of the bacteria was confirmed to contain the growth promoting compound (GPC). The GPC was identified to be indole acetic acid (IAA) by TLC, HPLC, MS/MS, and NMR analyses. P. eryngii mycelia grew rapidly both on PDA and in PDB after the treatment of GPC. The promoting concentration of GPC was as low as 1.0 nM. Tryptophan, the aminated form of IAA, was confirmed to be the precursor of IAA. These results suggested that bacterial secreted compound was IAA and plays an important role in promoting growth of mushroom mycelia.

Characterization of the Bacterial Cell Wall Lytic Enzyme Produced by Aspergillus sp. HCLF-4 (Aspergillus sp. HCLF-4에 의해 생성되는 세균세포벽 분해효소의 특성)

  • 임진하;민병례;최영길
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.15-20
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    • 2001
  • In this study, we have isolated bacterial cell wall lytic enzyme in the culture supernatant of Aspergillus sp. HCLF-4. This hydrolase showed cell wall lytic activity against Anabaena cylindrica. The extracellular enzyme was produced by Aspergillus sp. HCLF-4 when it was grown in a PDB media containing 0.05% heat killed Micrococcus luteus cells. The molecular weight of lytic enzyme was about 14.3 kDa. The optimal pH and temperature for the activity of this enzyme were 3.0~4.0 and $30^{\circ}C$, respectively. This hydrolase activity was reduced by $Na^{+}$, $Li^{+}$, $Ca^{2+}$, $Cu^{2+}$, $Fe^{3+}$, EDTA, and PMSF, whereas it was increased by $Mg^{2+}$, $Mn^{2+}$>. The enzyme has N-acetylmuramyl-L-amidase or endopeptidase activity.

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