Owing to its diverse range of bioactive compounds, Ganoderma lucidumhas garnered significant research attention for health promotion and disease prevention. Ganodermanondiol, which has a triterpenoid structure, is one of the major active compounds of G. lucidum. In the present study, the anti-inflammatory effects of ganodermanondiol were investigated to evaluate its usefulness as a functional ingredient. Ganodermanondiol (0.5–2 g/mL) significantly inhibited the production of nitric oxide (NO), the expression of the cytokines tumor necrosis factor (TNF)??and interleukin 6 (IL-6), and the expression of cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) in lipopolysaccharide-induced RAW 264.7 (murine macrophage) cells. Ganodermanondiol (0.5–2 g/mL) also inhibited the phosphorylation of mitogen-activated protein kinase (MAPK) signal molecules, including p38 and c-Jun N-terminal protein kinase (JNK) in RAW 264.7 cells. Ganodermanondiol significantly inhibited the essential factors involved in the inflammatory responses of RAW 264.7 cells and would, therefore, serve as a potential prophylactic and therapeutic agent for immune-related diseases.
The deep geologic repository (DGR) concept is widely accepted as the most feasible option for the final disposal of spent nuclear fuels. In this concept, a series of engineered and natural barrier systems are combined to safely store spent nuclear fuel and to isolate it from the biosphere for a practically indefinite period of time. Due to the extremely long lifetime of the DGR, the performance of the DGR replies especially on the natural geologic barriers. Assessing the safety of the DGR is thus required to evaluate the impacts of a wide range of geological, hydrogeological, and physicochemical processes including rare geological events as well as present water cycles and deep groundwater flow systems. Due to the time scale and the complexity of the physicochemical processes and geologic media involved, the numerical models used for safety evaluation need to be comprehensive, robust, and efficient. This study describes the development of an accessible, transparent, and extensible integrated hydrologic models (IHM) which can be approved with confidence by the regulators as well as scientific community and thus suitable for current and future safety assessment of the DGR systems. The IHM under development can currently simulate overland flow, groundwater flow, near surface evapotranspiration in a modular manner. The IHM can also be considered as a framework as it can easily accommodate additional processes and requirements for the future as it is necessary. The IHM is capable of handling the atmospheric, land surface, and subsurface processes for simultaneously analyzing the regional groundwater driving force and deep subsurface flow, and repository scale safety features, providing an ultimate basis for seamless safety assessment in the DGR program. The applicability of the IHM to the DGR safety assessment is demonstrated using illustrative examples.
Deep geologic repositories (DGR) are designed to store spent nuclear fuel and to isolate it from the biosphere for an extended period of time as long as millions of years. The long-term performance of the DGR replies on the performance of the natural geologic barriers after the end of the lifetime for the engineered barrier systems. Typically, multiple analytical and numerical models are used to analyze and ensure the safety of the repositories along both engineered and natural barrier systems. Despite the immense advancement in computing power and modeling techniques over the last few decades, a series of models and their linkage often require many simplifying assumptions in this safety assessment. The degree of the reliability and confidence of the safety analysis is thus highly dependent on the validity of those tools used. Considering the significance of the DGR performance and public attention, the highest level of quality control is necessary for the models employed in the assessment. The performance of the ultimate long-term geologic barrier is determined by the expected travel time of the radioactive species of interest, the level of their dilution or radioactivity at compliance areas, and the uncertainty associated with them. As the species of interest can be carried away from the repository location by groundwater flow, the travel time is determined by groundwater velocity along the flow path from source to biosphere while the dilution is a function of the decay and production rates as well as the diffusion and dispersion. Due to the time scale and the complexity of the physicochemical processes and geologic media involved, the models used for safety evaluation will need to become more and more comprehensive, robust, and efficient which is difficult to achieve in principle. They will also need to be transparent and flexible to satisfy the regulatory quality control requirements. This study thus attempts to develop an accessible, transparent, and extensible integrated hydrologic models (IHM) which can be widely accepted by the regulators as well as scientific community and thus suitable for current and future safety assessment of the DGR systems. The IHM can be considered as a tool and a framework at the same time when it is designed to easily accommodate additional processes and requirements for the future as it is necessary. The IHM is capable of handling the atmospheric, land surface, and subsurface processes for simultaneously analyzing the regional groundwater driving force and deep subsurface flow, and repository scale safety features, providing an ultimate basis for seamless safety assessment in the DGR program. The applicability of the IHM to the DGR safety assessment is demonstrated using simple illustrative examples.
본 연구에서는 Flammulina velutipes var. lupinicola의 laccase 유전자를 동정하고 최적 활성 pH, 온도, 시간을 분석하고 하였다. F. velutipes var. lupinicola 유전체에서 선별된 laccase 유전자 서열을 바탕으로 구리 결합 부위 및 신호 펩타이드 분석을 수행한 결과 5종의 laccase 유전자(g1934, g1937, g2415, g2539, g5858)를 동정하였다. 5종의 선별된 laccase 유전자 크기는 1,488~1,662 bp로 확인되었고, cDNA 염기서열 분석 결과 14~17개의 인트론이 확인되었다. Laccase 유전자의 신호펩타이드로 예측된 절단 부위는 N-말단으로부터 20~34 bp 사이에 위치하는 것으로 확인되었다. F. velutipes var. lupinicola laccase의 활성 특성을 규명하기 위해 분리 정제를 수행하였으며, Zymogram을 수행하여 0.2 M 및 0.3 M NaCl과 1.6 M 및 1.7 M의 ammonium sulfate로 정제된 단백질에서 5개의 laccsae 활성 밴드를 확인하였다. pH, 온도 및 시간별로 분리 정제된 단백질의 최적 활성을 분석한 결과, 반응의 최적 pH는 5.5이고 최적 온도는 40 ̊C로 확인되었다. 따라서 본 연구를 통하여 확인된 F. velutipes var. lupinicola 유전체의 laccase 유전자 구조 및 활성에 대한 특성은 laccase를 이해하는 데 도움이 될 것이며 추가 연구를 통하여 향후 다양한 산업적 활용이 가능할 것으로 사료된다.
최근 Flammulina 종들에 대한 유전체 염기서열 분석 결과가 보고되었고, 그로 인해 다양한 유전자 정보가 밝혀지고 있다. 본 연구에서는 Flammulina elastica 전체 유전체 서열의 laccase 유전자를 동정하고 구조적 특징 분석을 수행하고자 하였다. 유전체 분석 및 생물정보분석을 통하여 F. elastica 유전체 내 3개의 laccase 유전자(Felac1, Fe-lac2, Fe-lac3)를 확인하였고, 이들 유전자 내에는 10개의 히스티딘 잔기와 1개의 시스테인 잔기를 가지는 구리 이온 결합 영역과 4개의 시스테인 잔기를 가지는 이황화결합 형성 부위가 존재하는 것을 확인하였다. 1,548~1,602 bp의 laccase 유전자에 대한 전장 cDNA 염기 서열 분석을 통하여 12~16개의 인트론이 존재하는 것을 확인되었으며, N-말단으로부터 17~22 bp의 사이에 신호 펩타이드가 존재하는 것이 확인되었다. 본 연구를 통하여 F. elastica의 laccase 유전자를 최초로 동정하여 구조적 특징을 분석하였고, 이러한 결과는 F. elastica의 바이오매스 분해에 대한 이해를 돕는데 활용될 것으로 사료된다.
Immortalization is an essential process of the transformation of cells to a neoplastic growth. High risk human papillomavirus (hrHPV) infection has been the major cause of head and neck squamous cell carcinoma (HNSCC). The aim of this study was to search for a novel pathway causing immortalization in HPV16 E6/E7 transfected immortalized oral keratinocytes (IHOK). hrHPV integration sites were identified through DNA sequencing. HPV16 E6/E7 genes were integrated into 1q32.2, 12q21.2, 15q15.2, and 19q13.43 in IHOKs. Array-CGH was conducted to examine the deranged sites of the genes of IHOK. Of the 587 amplification genes, 70 genes were resided on chromosome 20. We selected PLAGL2 and MAPRE1 as the most amplified genes. PLAGL2 and MAPRE1 mRNA showed higher expression in IHOK than in normal keratinocytes. Knockdown of MAPRE1 significantly reduced telomerase activity. The analysis using a public database substantiated our data, showing the amplification of chromosome 20 and MAPRE1. In conclusion, our results suggest that MAPRE1 could play a crucial role in activating telomerase activity in hrHPV-infected cells. This finding may provide basic data to develop a novel target therapy for hrHPV-related HNSCC.
At the Flower Research Institute of Chungcheongnam-do agricultural research and extension service, ‘Shusin’ and ‘ST06-005-11’ were crossed and a candidate descendant, ‘ST09-173-06’, was selected based on disease resistance and other morphological traits. We intensively tested the candidate line on various characteristics, including the floral qualities through shading and retarding culture, starting from year 2009. In 2011, we registered this line as the standard chrysanthemum (Dendranthema grandiflorum Ramat.) cultivar ‘Baekseol’. The natural flowering time of 'Baekseol' starts the 24th of October, which is earlier than ‘Jinba’ (29th of October). ‘Baekseol’ is favorable at winter growing season and the flower vase life is 17 days. The cut flower length of ‘Baekseol’ is 87.2 cm, which is shorter than ‘Jinba’ (97.4 cm) and the lateral branch number of ‘Baekseol’ (15.6 per plant) is less than ‘Jinba’ (21.8 per plant). The flower size of ‘Baekseol’ is 14.6 cm (‘Jinba’, 13.4 cm) and the petal number per flower is 430.4 (316.3 per flower for ‘Jinba’). Careful nutrient management is required at t he e arly g rowing s tage. T he s hort d ay t reatment i s appropriate when the plant is approximately 40 - 45 cm. In addition, flower bud differentiation temperature is approximately 16 - 17℃.
In our previous study, active-single compound (ASC), inhibitor of melanogenesis in B16F10 melanoma cell, was identified from Ganoderma lucidum. Thus, the aim of this study is to analyze the change of the ASC contents in G. lucidum when lacquer tree (LT) was added in cultivation of G. lucidum. In HPLC analysis, significantly increased ASC peak was observed in G. lucidum extract supplemented with 15% (v/v) of LT. In addition, melanogenesis inhibitory effect of G. lucidum extract supplemented with 15% of LT significantly increased when compared to G. lucidum extract without LT supplementation. Furthermore, LT supplementation increased mycelial growth of G. lucidum in both solid and liquid cultivation. These results suggest that is useful as natural ingredient for increasing bioactivity including skin-whitening effect and mycelial growth of G. lucidum.
Ganoderma lucidum, a species of the class Basidiomycetes, attracts international attention due to the wide variety of its biological activities and great potential as cosmetic ingredient, such as skin care cosmetics including ‘skin-whitening’. However, there is little information available regarding the tyrosinase inhibitory activity. Therefore, the objectives of this study were to investigate the chemical composition and tyrosinase inhibitory activity of the G. lucidum. To isolate active single compound from G. lucidum (GASC), we conducted ethanol extraction and chloroform fractionation. In addition, we assayed the inhibitoty effect of tyrosinase activity and melanin biosynthesis in B16F10 mouse melanoma cells. In the present study, we identified a GASC, which exhibited inhibitory effects of cellular tyrosinase activity, the protein expression of cellular tyrosinase and tyrosinase-related protein-1, 2. In additional, microphthalmia-associated transcription factor (MITF), as known as crucial regulator of melanogenesis-related genes, was down-regulated by treatment with GASC in a concentration-dependent manner. GASC exhibited significant inhibition of tyrosinase activity and melanin synthesis in B16F10. The finding that melanogenesis inhibitory effect of GASC will contribute to facilitate various approaches of this mushroom for use in skin whitening products.
In our previous study, active-single compound (ASC), inhibitor of melanogenesis in B16F10 melanoma cell, was identified from Ganoderma lucidum. Thus, the aim of this study is to analyze the change of the ASC contents in G. lucidum when oriental raisin tree (ORT) was added in cultivation of G. lucidum. In HPLC analysis, significantly increased ASC peak was observed in G. lucidum extract supplemented with 15% (v/v) of ORT. In addition, melanogenesis inhibitory effect of G. lucidum extract supplemented with 15% of ORT significantly increased when compared to G. lucidum extract without ORT supplementation. Furthermore, ORT supplementation increased mycelial growth of G. lucidum in both solid and liquid cultivation. These results suggest that oriental raisin tree is useful as natural ingredient for increasing bioactivity including skin-whitening effect and mycelial growth of G. lucidum.
The purpose of this experiment is to measure the change in the growth rate and the amount of bioactive compounds when additives are added to mushrooms. According to the additive content, Ganoderma lucidum ASI7004, Ganoderma lucidum ASI7071, Ganoderma. meredithae KACC42868 and Ganoderma lucidum ASI7013 mycelial growth rate variation analysis, oriental raisin tree, Siberian ginseng, chestnut shell, apple pomace, and Korean cabbage additive increased mycelial growth by approximately 2.5 times when compared to the control group(MCM medium). The results of analyzing the total amount of polyphenol in mycelia of G. lucidum based on different kinds of additives have shown that the amount of polyphenol increased only in G. lucidum ASI7004 with 2% Korean cabbage additive. However, when other additives are added, the amount of polyphenol turned out to be lower than original G. lucidum and other individual additives. The results of analyzing the total amount of triterpenoid in mycelia of G. lucidum based on different kinds of additives have shown that the amount of triterpenoid increased in G. lucidum mycelia with apple pomace and Korean cabbage additives. Siberian ginseng additive was effective in increasing the amount of triterpenoid only when 4% Siberian ginseng was added to G. lucidum ASI7004, yet the increased amount was not significant. Although the addition of additives turned out to be effective to the growth rate, it was not effective enough towards the amount of bioactive compounds. Further experiments are required.
The aim of this study was to identify and characterize new Flammulina velutipes laccases from its whole-genome sequence. Of the 15 putative laccase genes detected in the F. velutipes genome, four new laccase genes (fvLac-1, fvLac-2, fvLac3, and fvLac-4) were found to contain four complete copper-binding regions (ten histidine residues and one cysteine residue) and four cysteine residues involved in forming disulfide bridges. fvLac-1, fvLac-2, fvLac3, and fvLac-4, encoding proteins consisting of 516, 518, 515, and 533 amino acid residues, respectively. Potential N-glycosylationsites(Asn-Xaa-Ser/Thr) were identified in the cDNA sequence of fvLac-1(Asn-454), fvLac-2(Asn- 437andAsn-455), fvLac-3(Asn-111andAsn-237), and fvLac4 (Asn-402andAsn-457). In addition, the first 19–20 amino acid residues of these proteins were predicted to comprise signal peptides. Laccase activity assays and reverse transcription polymerase chain reaction(RT-PCR) analyses clearly reveal that CuSO4 affects the induction and the transcription level of these laccase genes.
The 35.6-mb genome contained 12,218 predicted protein-encoding genes and 287 tRNA genes assembled into 11 scaffolds corresponding with the 11 chromosomes of strain KACC 42780. Based on presence of copper binding region and signal peptide, 8 laccase genes (Fv Lac-1, Fv Lac-2, Fv Lac-3, Fv Lac-4, Fv Lac-5, Fv Lac-6, Fv Lac-7, Fv Lac-8) were selected in F.velutipes genome. The laccase genes of F.velutipes ranged from 1,300 to 1,700 base pair in their size. In addition, the molecular mass and PI values of laccase genes ranged from 50 to 62 kDa and from 3 to 11, respectively.
In laccase activity assays, the highest laccase activity (3unit/ml)was shown in medium with 0.5mM of CuSO4 on 9th day. However, laccase activity was severely decreased in medium with 1mM and 2mM of CuSO4 was drastically decreased from day 3 through day 9. Interestingly, laccase activity in the medium without CuSO4 showed higher level than in the medium with 0.25, 1 and 2mM on both 6th and 9th day.
RT_PCR showed the highest transcription level with 0.25mM and 0.5mM of CuSO4 supplementation on day 3 and on day 9, respectively. In addition, the transcription level of Lac-1 increased depending on concentration of CuSO4 supplementation.
Flammulina velutipes is a popular edible basidiomycete mushroom found in East Asia and is commonly known as winter mushroom. Mushroom development showing dramatic morphological changes by different environmental factors is scientifically and commercially interesting. We have sequenced the 35.6-megabase pair genome of F. velutipes KACC42780 using a next generation sequencing (NGS) approach. Then, HiSeq2000 sequencing system was used for comparing gene expression of F. velutipes genome at three different developmental stages (mycelium, primordium, and fruit body). Ninety one percents (10,116 genes) of the 10,999 predicted genes are expressed in at least one developmental stage. In addition, analysis of expressed genes in three different developmental stages showed that only 0.25–1.5% (28–165) of the total expressed genes (10,013) were uniquely expressed in a single developmental stage, whereas 83.6% (9,198) was shared in all stages.
본 연구는 다양한 영지버섯 균주의 균사체 메탄올추출물을 이용한 항염, 항산화 및 항알러지 효능을비교 분석하기 위해 수행되었다. G. species ASI-7150 (흑영지), G. lucidum ATCC46755 (Canada), G.species ASI-7151 (흑룡), G. lucidum ATCC64251(Taiwan), G. neo-japonicum ASI-7032, 및 G. lucidumASI-7071 (영지2호)의 균사체 추출물 항산화 효과를 분석한 결과 G. species ASI-7150 (흑영지) 의 균사체 추출물에서 DPPH radical scavenging 활성이 가장 높았다.항염활성 분석에서는 G. lucidum ATCC64251 (Taiwan)의 균사체 추출물을 처리하였때 NO 생성 저해능이 가장 좋았다. 또한 G. lucidum ATCC64251 (Taiwan)의 균사체 추출물을 처리하였을 때, -hexosaminidase생성억제율이 가장 좋았다. MTT assay를 통하여 각 영지버섯 균사체 추출물은 세포 생존율에는 영향이 없는것으로 확인 되었다. 이러한 선행 연구결과는 추후에국내 및 국외 품종 등 다양한 영지버섯 균사체의 약리효과 분석에 기초자료로 사용될 것이다.
Ganoderma have been used in traditional medicine for thousands of years in Korea, China and Japan as well as in other regions of the world. Ganoderma species is known to grow on hardwoods, conifers, palms, old and declining trees, decay dead wood and stumps. Several species is cause serious diseases of plantation and other economically important tropical crops. It also has been reported to prevent and treat immunological diseases, including tumorigenesis, control of blood glucose levels, modulation of the immune system, hepatoprotection, bacteriostasis, and more. In the present study, we examined the effects of various Ganoderma species, including Ganoderma lucidum and Ganoderma neo-japonicum on nitric oxide inhibition activities, β-hexosaminidase secretions, and DPPH free radical scavenging capacities to compare the anti-inflammatory, anti-allergic, and antioxidant effects.
1. 세계 26개국에서 도입된 녹두 유전자원 705점에 대하여16개의 형태적 형질 특성을 이용하여 각각 두 가지의 군집방법, 핵심집단 자원수 결정방법 및 표본추출 방법을 조합하여 총8개의 핵심집단(SUNPR, SUNPPr, UNPR, UNPPr, SUNLR, SUNLPr, UNLR, UNLPr)을 작성하여 원수집단과 비교한바 8개핵심집단 모두 원수집단과는 분산과 평균치에서 차이가 없었다.
2. Nei의 다양성지수는 8개 핵심집단 모두 원수집단 보다높았고, 특히 UNLPr이 가장 높았으며, UNPR은 다른 핵심집단에 비하여 상대적으로 낮은 지수를 보였다.
3. 원수집단과 핵심집단간 자원분포의 동질성을 파악하기 위하여 카이자승 검증을 한 결과 12개 양적형질과 1개의 질적형질(종피색)은 모든 핵심집단이 동질분포를 보였으나 3개의질적 형질 (배축색, 종피광택, 생육습성)은 핵심집단에 따라서동질분포를 하는 것과 그렇지 않은 것이 있었으며, 8개 핵심집단 중 UNPPr은 동질성이 가장 높았다. 평균차율(MD%)과일치율(CR%)은 8개 핵심집단 모두 유의한 수준을 보였다.
4. 8개의 모든 핵심집단이 원수집단과 평균치에서 차이가 없고, 높은 다양성을 과 동질한 분포를 보이며, 평균차율(MD%)과 일치율(CR%)이 유의한 수준을 보여 원수집단의 다양성을잘 나타내는 것으로 해석되었다.
5. 핵심수집단 크기(10%와 15%)에 따른 다양성은 유의한차이가 없었으며, 8개의 핵심집단 중 평균치, 동질분포, 일치율 및 도복내성을 고려할 때 UNPPr이 가장 좋은 핵심집단으로 사료되었다.