Pyung-Gyun Shin

The regulatory effect of veratric acid on NO production in LPS-stimulated RAW264.7 macrophage cells.

Veratric acid, a simple benzoic acid derived from plants and fruits, has been reported to have anti-oxidant, anti-inflammation, and blood pressure-lowering effects. This study was designed to evaluate the inhibitory effects of veratric acid on nitric oxide (NO) production in LPS-stimulated RAW264.7 cells. It was found that veratric acid inhibited NO production and inducible nitric oxide synthase (iNOS) expression in LPS-stimulated RAW264.7 cells. The inhibitory effects of veratric acid on the generation of interleukin-6 (IL-6) and interferon-γ (IFN-γ) was determined. Furthermore, veratric acid facilitated the inactivation of glycogen synthase kinase-3β (GSK-3β), STAT-1, and STAT-3 in dose-dependent manner. Notably, NF-κB and members of the mitogen activated protein kinase (MAPK) family including p38, ERK, and JNK were dephosphorylated by veratric acid. These findings suggest that the treatment of veratric acid might be effective in neutralizing the over-expression of NO in inflammatory disorders.

Veratric acid inhibits iNOS expression through the regulation of PI3K activation and histone acetylation in LPS-stimulated RAW264.7 cells

In the present study, we investigated regulatory effects of veratric acid on the production of nitric oxide (NO) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. NO production was significantly decreased by veratric acid in the LPS-stimulated RAW264.7 cells in a dose-dependent manner. The reduction in nitric oxide production was induced by the downregulation of inducible NO synthase (iNOS) expression. Veratric acid suppressed the LPS-induced effects on the regulatory and catalytic subunits of phosphoinositide 3-kinase (PI3K), comprised of p85, p110α, p110β and Akt. The acetylation of p300 and the phosphorylation of activating transcription factor 2 (ATF-2) induced by LPS were downregulated following treatment with veratric acid; similar effects were observed following treatment with LY294002, a specific inhibitor of PI3K/Akt. The LPS-induced expression of histone deacetylase (HDAC)3 decreased to basal levels following treatment with veratric acid, and its expression was also downregulated by LY294002. In the measurement of histone acetylation levels, the LPS-stimulated acetylation of histone H4 was significantly attenuated by veratric acid, and was also reduced following the inhibition of PI3K/Akt with LY294002. From our data, it can be concluded that veratric acid exerts a regulatory effect on LPS-induced iNOS expression. Our results suggest that veratric acid impedes the PI3K/Akt-mediated histone acetyl-transferase (HAT) activation and HDAC expression induced by LPS, thereby abrogating iNOS expression.

Breeding progress and characterization of a Korean white variety `Baek-A` in Flammulina velutipes

We made the first Korean white commercial strain `Baek-a` developed by crossing between monokaryons derived from brown strains. This variety can be estimated as the Korea`s indigenous one different from the origin of Japanese white ones. The optimum temperature of mycelial growth was but it needed to adjust to when incubated at the bottle cultivation. The optimum temperatures of fruiting body initiation and development were and , respectively. Fruiting body of `Baek-a` was pure white even developed from crossing with brown strains. `Baek-a` was a good variety with high quality and high productivity characterized as quite even budding habit, long stipes and hemi-spherical pilei. The days for the fruiting was 7 days and the productivity was g per 850 ml. This variety needed high concentration of carbon dioxide and it had to be adjusted up to 4,000 ppm for the good quality.

Anti-inflammatory effects of Sparassis crispa extracts

Sparassis crispa is a medicinal mushroom, which has been reported to have anti-cancer effect. In this study, we designed to investigate the effects of Sparassis crispa extracts on the production of nitric oxide (NO) in LPS-stimulated RAW264.7 cells. The pre-treatment of the extracts prior to add LPS in RAW264.7 cells suppressed NO production and iNOS expression at protein and mRNA levels. The phosphorylation of was inhibited by the extracts, which was induced through suppressing the activation of . Sparassis crispa extracts showed the effect on the down-regulation of STAT-1 activation in a dose-dependent manner. In LPS-stimulated RAW264.7 cells, was translocated into the nucleus, while the treatment of Sparassis crispa extracts induced to sequestered in the cytosol. These experimental results determined that Sparassis crispa extracts play a inhibitory role in inflammatory reactions via regulating NO production, which suggests potential as a component of inflammatory drugs.

Quality changes in Agaricus bisporus varieties due to period and temperature during their storage

This study was carried out to investigate storage stability of harvested mushroom in developed varieties of button mushroom through identification of quality change during 35 days. The mushroom harvested up to flush mushroom was stored on different storage temperature(4, 7, ) and weight, length, thickness, color of pileus and stipe in fruit body was tested every 7 days. The morphological trait and color of stipe in fruit body were influenced by harvesting period and storage temperature. On the morphological traits of fruit body, mushroom harvested at flush showed smaller difference than those at flush and mushroom stored at indicated more difference than ones at the other temperature. Mostly color of pileus in brown button mushroom had lower (color difference) than white mushroom during storage period. The color of pileus of fruit body in white mushroom had low at whereas that of brown mushroom was low at . The safe period of storage based on the L value is 7 days under a specified environmental condition. `Seolgang` was stored for 14 days as good quality of mushroom, `Saedo` had best quality after harvesting and `Seojeong` was most stable during storage change in white mushroom varieties.

Agrobacterium-mediated transformation using gill tissue of Flammulina velutipes

Agrobacterium-mediated transformation was conducted in order to generate DNA insertional mutants of Flammulina velutipes. Agrobacterium tumefaciens AGL-1 harboring pBGgHg was transformed into gill tissues of Flammulina velutipes strain KACC42777. The transformants resistant on hygromycine (30 µg/ml) were confirmed by PCR. The targeted insertional sites were amplified by inverse PCR and sequenced. To find the phenotype variation of all generated transformants, bottle cultivation which followed by the standard cultivation protocol were conducted. Color variation was observed on the cultivated fruiting bodies. Futhermore, the transformant pool will be used as a good genetic resources for studying gene function.

Comparison of amino acid and free amino acid contents between cap and stipe in Flammulina velutipes and Pleurotus ostreatus

팽이버섯과 느타리버섯의 색깔이 다른 계통과 부위별로 갓과 대로 나누어 아미노산 및 유리아미노산 분석을 실시하여 성분 조성과 함량을 비교하였다. 그 결과 버섯 종류와 계통, 그리고 부위에 따라 서로 다른 아미노산 종류가 검출되었다. 팽이, 느타리 모두에서 tryptophane은 검출되지 않았으며, 느타리에서는 특이적으로 alanine이 검출되지 않았다. 또한 팽이에서는 glutamic acid 함량이 가장 높았으며 methionine과 phenylalanine은 극히 적게 검출되었다. 느타리버섯에서는 glutamic acid 함량이 가장 높았고 phenylalanine과 cysteine이 낮았으며 특이하게 흑색변이계통에서 leucine이 검출되지 않았다. 또한 부위별로는 팽이와 느타리 모두 아미노산 종류별 비율은 동일한 경향을 보였으며, 일반적으로 대보다는 갓에서 아미노산 함량이 더 높은 경향을 보였다. 유리아미노산 분석 결과에서는 버섯계통별로 검출되는 아미노산 함량이 서로 달랐으며, 버섯의 갓과 대에서도 검출되는 아미노산 조성이 일치하지 않는 것으로 나타났다. 【The composition and content of amino acid and free amino acid were separately compared in cap and stipe of fruitbody obtained from different color strains in winter mushroom and oyster mushroom strains. The result showed different kinds and amount of amino acids according to the mushroom parts, strains and kinds. Tryptophane was not detected in any mushrooms. Alanine was also not detected in all oyster mushroom strains. The content of glutamic acid was the highest and that of methionine, phenylalanine and cysteine were very low in all mushroom strains. When compared by parts, the ratio of amono acids showed similar tendency in composition. Generally the content of amono acids showed higher in caps than in stipes. The profile and content of free amino acid showed a lot of variation between cap and strip in the intra and inter mushrooms.】

Genetic Diversity and Phylogenetic Relationship in Korean Strains of Lentinus lepideus Based on PCR Polymorphism

Lentinus lepideus, known as train wrecker fungus, has been used for nutritional and medicinal purposes. Recently, commercial cultivation technique and a new cultivar of the mushroom were developed. To investigate the genetic diversity and phylogenetic relationship for identifying the mushroom strains and cultivar, one commercial and 13 strains of Lentinus lepideus from different geographical regions of Korea were analyzed by ITS regions of rDNA and RAPD of genomic DNA. Three strains of Lentinus edodes were also used for the analysis. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 173 to 179 bp and 203 to 205 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical with 156 base pairs. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into four clusters, while 3 strains of L. edodes was divided into a new cluster. Ten primers out of 20 arbitrary primers used in the RAPD-PCR efficiently amplified the genomic DNA. The numbers of amplified DNA bands varied with the primers and strains, with polymorphic DNA fragments in the range from 0.2 to 2.6 kb. The results showed that phylogenetic relationship among Korean strains of Lentnus lepideus is high, but genetic diversity is low.

Toluhydroquinone from Aspergillus sp. suppress inflammatory mediators via nuclear factor-κB and mitogen-activated protein kinases pathways in lipopolysaccharide-induced RAW264.7 cells

The purpose of this study is to investigate anti‐inflammatory effects of toluhydroquinone in lipopolysaccharide (LPS)‐stimulated RAW264.7 cells.

Development of a new brown button mushroom cultivar 'Hogam'

ABSTRACT: A new brown button mushroom cultivar, ‘Hogam’, C34 line, was made by crossing homokaryons, ASI1164-37 andASI1175-66, selected by RAPD analysis and by cultivating three times. Mycelium of ‘Hogam’ on CDA (compost dextrose agar)media grew well at 25 o C. The optimum pin-heading temperature of new variety and optimum growing temperature was 14-18 o C. The thickness of the mature cap and stipe were thicker than a control, ‘Dahyang’ that developed in 2010. The color ofpileus was light brown and lighter than ‘Dahyang’. Days required from casing to first harvesting were three days longer thancontrol strain, but the weight of harvested fruiting body increased by 1.35 times. ‘Hogam‘ cultivar are expected to contribute tothe diversification of domestic mushroom cultivars.KEYWORDS: Agaricus bisporus, Brown button mushroom cultivar, Morphological and cultural characteristics 서론 양송이(Agaricus bisporus)는 생긴 모양을 빗대어 buttonmushroom으로 불리며, 식용버섯으로 맛과 향기가 뛰어나서 세계적으로 가장 널리 소비되는 버섯 중 하나이다(Jzsefet al. 2004). 최근 우리나라의 양송이 생산량은 6,678MT이고, 생산액은 433억이다(Special crop production in 2013).양송이에는 일반적으로 많이 소비되는 백색양송이 와 갓이갈색을 띄는 갈색양송이로 구성된다. 특히 갈색양송이는크래미버섯, 베이비 포토벨로, 베이비벨라, 미니벨라, 포터벨리니, 로먼버섯, 이탈리아버섯 또는 갈색버섯등과 같은여러 이름으로 불리며 일반 버섯보다 크게 키워, 갓이 약간벌어진 형태의 버섯은 potobello라고 불린다. 우리나라에서는 갈색양송이는 병충해에 강해 과거 705 등을 재배하였으나 가격이 낮고 재배규모는 적어 학교급식을 납품하는 농가에서 소규모로 재배를 하고 있다.양송이는 다른 버섯과 달리 이차 자웅동주성(secondaryhomothalism)으로 한 담포자내에 두 개의 반수체핵(haploidnuclei) 을가지는다핵성(multinucleate) 지닌다. 이에따라 교잡을 하지 않아도 포자내에 화합성을 가진 두 개의 핵이 임성을 띄어 대다수의 담자포자가 자실체를 형성한다.이에교잡을위한동핵균주확보가어렵고, 교잡후꺽쇠연결체(clamp connection)가 형성되지 않아 교잡을 확인하기가 어렵다(Rapar and Raper, 1972). 현재까지 육종연구는돌연변이(Fritsche, 1965; Stoller and Stauffer, 1953), 교배(Elliott, 1978; Kneebone, 1972; Moessner, 1962), 원형질체분리(Horgen, 1991), 형질전환(Rhee et al, 1996; Chen etal, 2000; Tomas et al, 2001) 등이 이루어졌다. 동핵균주의