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Apoptotic Effect of Co-Treatment with Chios Gum Mastic and Eugenol on SCC25 Human Tongue Squamous Cell Carcinoma Cell Line (사람혀편평세포암종세포에서 Chios gum mastic과 eugenol의 병용처리가 미치는 세포자멸사 효과에 관한 연구)

  • Sohn, Hyeon-Jin;Yea, Byeong-Ho;Kim, In-Ryoung;Park, Bong-Soo;Jeong, Sung-Hee;Ahn, Yong-Woo;Ko, Myung-Yun
    • Journal of Oral Medicine and Pain
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    • v.36 no.3
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    • pp.147-160
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    • 2011
  • Eugenol (4-allyl-2-methoxyphenol) is a natural phenolic constituent extensively used in dentistry as a component of zinc oxide eugenol cement and is applied to the mouth environment. Chios gum mastic (CGM) is a resinous exudate obtained from the stem and the main leaves of Pistacia lenticulus tree native to Mediterranean areas. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with a natural product, CGM and natural phenolic compound, eugenol on SCC25 human tongue squamous cell carcinoma cell line. To investigate whether the co-treatment with eugenol and CGM compared to each single treatment efficiently reduces the viability of SCC25 cells, MTT assay was conducted. Induction and augmentation of apoptosis were confirmed by Hoechst staining, TUNEL staining and DNA hypoploidy. Westen blot analysis and immunofluorescent staining were performed to study the alterations of the expression level and the translocation of apoptosis-related proteins in co-treatment. In this study, co-treatment of with eugenol and CGM on SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensations, DNA fragmentation, the increase and decrease of Bax and Bcl-2, decrease of DNA content, the release of cytochrome c into cytosol, translocation of AIF and DFF40 (CAD) onto nuclei, and activation of caspase-3, caspase-6 caspase-7, caspase-9, PARP, Lamin A/C and DFF45 (ICAD) whereas each single treated SCC25 cells did not show or very slightly these patterns. Although the single treatment of 40 ${\mu}g$/ml CGM and 0.5 mM eugenol for 24 h did not induce apoptosis, the co-treatment of these reagents prominently induced apoptosis. Therefore our data provide the possibility that combination therapy with CGM and eugenol could be considered as a novel therapeutic strategy for human oral squamous cell carcinoma.

Potential Antitumor $\alpha$-Methylene-$\gamma$-butyrolactone-Bearing Nucleic Acid Base. 3. Synthesis of $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans

  • Kim, Jack-C.;Kim, Si-Hwan;Kim, Ji-A;Choi, Soon-Kyu;Park, Won-Woo
    • Archives of Pharmacal Research
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    • v.21 no.4
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    • pp.458-464
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    • 1998
  • Search for a new $\alpha$-methylene-$\gamma$-butyrolactone-bearing 6-substituted purine as a potental antitumor agent has led to synthesize seven, hitherto unreported, $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$- methylenetetrahydrofurans (H, Cl, l, $CH_3$, $NH_2$, SH, >C=O) (6a-g). These include $5^1$-Methyl-$5^1$-[(9H-purin-9-yl)methyll-$2^1$-oxo-$3^1$ -methylenetetrahydrofurans (6a), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydr ofurans (6b), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6c), $5^1$-Methyl-$5^1$-[(6-methyl-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6d), $5^1$-Methyl-$5^1$-[(9H-adenin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6e), $5^1$-Methyl-$5^1$-[(6-mercapto-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6f) and $5^1$-Methyl-$5^1$-[(9H-hypoxanthin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrof urans (6g) which were made by the Reformatsky-type reaction of ethyl $\alpha$-(bromomethyl) acrylate with the corresponding (6-substituted-9H-purin-9-yl)-2-propanone intermediates (5a-g). These ketone intermediates 5a-g, 1-(9H-purin-9-yl)-2-propanone (5a), 1-(6-chloro-9H-purin-9-yl)-2-propanone (5b), 1-(6-iodo-9H-purin-9-yi)-2-propanone (5c), 1-(6-methyl-9H-purin-9-yl)-2-propanone (5d), 1-(9H-adenin-9-yl)-2-propanone (Se), 1-(6-mercapto-9H-purin-9-yl)-2-propanone (5f), and 1-(9H-hypoxanthin-9-yl)-2-propanone (5g) were directly obtained by the alkylation of the 6-substituted purine bases with the chloroacetone in the presence of $K_2$$CO_3$ (or NaH) under DMF (or DMSO). The preliminary in vitro cytotoxcity assay for the synthetic .alpha.-methylene-y-butyro-lactone compounds (6a-g) were determined against three cell lines (PM-3A, P-388, and K-562) and showed the moderate antitumor activity ($IC_50$ ranged from 1.4 to 4.3 $\mu\textrm{g}$/ml) with the compound $5^1$-methyl-$5^1$ -[(9H-hypoxanthin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofuran (6g) showing the least antitumor activity.

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Recommendation Rate of Slow Release Nitrogen Fertilizer Application for Early Dry Seeding Culture of Rice in Honam Plain Area (호남평야지에서 벼 건답직파 조기파종재배에 알맞은 완효성 질소비료 시비량)

  • Kim, Sang-Su;Choi, Weon-Yeong;Back, Nam-Hyun;Choi, Min-Gyu;Park, Hong-Kyu;Nam, Jeong-Kweon
    • Korean Journal of Soil Science and Fertilizer
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    • v.39 no.5
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    • pp.292-297
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    • 2006
  • A research was carried out to investigate the release pattern of slow release nitrogen fertilizer compound(SRNC) that is latex coated urea(LCU) and to recommend the application rate of the fertilizer at dry seeding rice culture in Honam plain area. The experiment was conducted at experimental field(Jeonbug series, fine silty, mixed, nonacid, mesic family of Fluvaquentic Endoaquepts) of National Honam Agricultural Research Institute. A medium late maturing rice variety (Dongjinbyeo) was seeded at the rate of $60kg\;ha^{-1}$ by drill seeder on April 1, 1997 and March 30, 1998. Fertilizer application rate was $160-90-110kg\;ha^{-1}$ for $N-P_2O_5-K_2O$. Soil ammonium nitrate($NH_4-N$) contents remained higher in all SRNC plots than conventional ones and higher with the increase of SRNC application rate until panicle formation stage, but the contents was higher in the conventional than NRNC plots at the heading stage. The plant hight was taller in SRNC than conventional plots until maximum tiller stage, but became similar in both conventional and SRNC plots at heading stage. The culm and panicle number was greater in SRNC than conventional plots throughout all growth stage, but the effective tiller rate was higher in conventional ones. Nitrogen efficiency was higher in the SRNC than conventional plots, but the efficiency was decreasing with the higher SRNC level. The spikelet number per unit area was greater in SRNC than conventional plots, and increased with higher SRNC level. The more spikelet number with higher NRNC level resulted in lower 1,000-grain weight. The rice yield in conventional plot was similar with only 60% SRNC level, but lower than 80% and 100% SRNC levels. However, slight lodging was observed in 100% SRNC level. In conclusions, we recommend NRNC application level as 80% of standard nitrogen application for early dry seeding culture of rice Honam plain area.

The Effect of the Addition of Carbohydrate Sources on the Concentration of Odorous Compounds for Recycling of Pig Slurry to Grassland (발효탄수화물 첨가 비육돈사료가 초지환원용 슬러리의 악취물질 농도에 미치는 영향)

  • Cho, Sung Back;Hwang, Ok Hwa;Park, Kyu Hyeun;Choi, Dong Yun;Yang, Seung Bong;Kim, Do Hwan;Park, Sung Kwon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.4
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    • pp.257-262
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    • 2013
  • This study investigates the effect of fermentable carbohydrate on the concentration of odorous compounds in pig slurry. Four types of pig diet were studied: control, peanut hull (crude fiber 29.87, NDF 44.02%), golden fiber (crude fiber 48.77, NDF 65.88%), and almond hull (crude fiber 44.30, NDF 64.44%). Pigs (average BW 37.0 kg) were fed diets that met the Korean Feeding Standard (2012) and their excreta samples were collected from the slurry pits. Levels of volatile organic compounds (phenols and indoles) and volatile fatty acids were analyzed by gas chromatography. Phenol level was the lowest (p<0.05) in golden fiber (33.26 ppm) group and the highest in control (97.29 ppm). The concentration of indoles in the peanut hull (1.27 ppm), almond hull (1.20 ppm), and golden fiber (1.02 ppm) groups was lower (p < 0.05) than that of control (1.79 ppm). Levels of short chain fatty acid (SCFA) were lower (p < 0.05) in golden fiber (1,319 ppm) and almond hull (1,433 ppm) groups than in control (1,893 ppm). Concentration of branched chain fatty acid (BCFA) in the golden fiber group (74 ppm) was lower (p < 0.05) than that of control (98 ppm). Taken together, the concentration levels of phenols, indoles, and VFAs decreased on addition of peanut hull, golden fiber or almond hull to the diet, suggesting that fermentable carbohydrate may contribute to reducing odorous compounds in pig slurry.

Healthy Functional Food Properties of Phenolic Compounds Isolated from Ulmus pumila (유근피(Ulmus pumila)로부터 분리한 phenol성 물질의 건강기능식품 활성)

  • Kim, Kyung-Bum;Jo, Bun-Sung;Park, Hye-Jin;Park, Ki-Tae;An, Bong-Jeun;Ahn, Dong-Hyun;Kim, Myung-Uk;Chae, Jung-Woo;Cho, Young-Je
    • Food Science and Preservation
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    • v.19 no.6
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    • pp.909-918
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    • 2012
  • The phenolic compounds which were extracted with 70% ethanol from Ulmus pumila for 12 hr were the highest as $17.9{\pm}1.0\;mg/g$. DPPH scavenging activity of 70% ethanol extracts was also the highest as $89.5{\pm}1.9%$ and it was confirmed to be high as 80% over in both of water and 70% ethanol extracts containing $50{\mu}g/mL$ over phenolic concentration. ABTS radical cation decolorization activities of water and 70% ethanol extracts were higher as $96.8{\pm}2.9%$, antioxidant protection factor (PF) was 2.0 PF in 70% ethanol and showed higher activities in both of water and 70% ethanol extracts containing $200{\mu}g/mL$ phenolic concentration as 2.5 PF than BHA. TBARs of 70% ethanol extracts was $86.5{\pm}4.6%$, it showed high anti-oxidative activity in $50{\sim}200{\mu}g/mL$ phenolic concentrations of water and 70% ethanol extracts as 80% over. The angiotensin converting enzyme (ACE) inhibitory activity of Ulmus pumila extracts against hypertension was 77.4% and 90.6% in water and 70% ethanol extracts of $200{\mu}g/mL$ phenolic concentration. Xanthine oxidase inhibitory activity of Ulmus pumila extracts for anti-gout effect was not observed in water extracts, but it showed 30% inhibitory activity in 70% ethanol extracts, and 48.1% at $200{\mu}g/mL$ phenolics concentration.

Discrimination of African Yams Containing High Functional Compounds Using FT-IR Fingerprinting Combined by Multivariate Analysis and Quantitative Prediction of Functional Compounds by PLS Regression Modeling (FT-IR 스펙트럼 데이터의 다변량 통계분석을 이용한 고기능성 아프리칸 얌 식별 및 기능성 성분 함량 예측 모델링)

  • Song, Seung Yeob;Jie, Eun Yee;Ahn, Myung Suk;Kim, Dong Jin;Kim, In Jung;Kim, Suk Weon
    • Horticultural Science & Technology
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    • v.32 no.1
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    • pp.105-114
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    • 2014
  • We established a high throughput screening system of African yam tuber lines which contain high contents of total carotenoids, flavonoids, and phenolic compounds using ultraviolet-visible (UV-VIS) spectroscopy and Fourier transform infrared (FT-IR) spectroscopy in combination with multivariate analysis. The total carotenoids contents from 62 African yam tubers varied from 0.01 to $0.91{\mu}g{\cdot}g^{-1}$ dry weight (wt). The total flavonoids and phenolic compounds also varied from 12.9 to $229{\mu}g{\cdot}g^{-1}$ and from 0.29 to $5.2mg{\cdot}g^{-1}$dry wt. FT-IR spectra confirmed typical spectral differences between the frequency regions of 1,700-1,500, 1,500-1,300 and $1,100-950cm^{-1}$, respectively. These spectral regions were reflecting the quantitative and qualitative variations of amide I, II from amino acids and proteins ($1,700-1,500cm^{-1}$), phosphodiester groups from nucleic acid and phospholipid ($1,500-1,300cm^{-1}$) and carbohydrate compounds ($1,100-950cm^{-1}$). Principal component analysis (PCA) and subsequent partial least square-discriminant analysis (PLS-DA) were able to discriminate the 62 African yam tuber lines into three separate clusters corresponding to their taxonomic relationship. The quantitative prediction modeling of total carotenoids, flavonoids, and phenolic compounds from African yam tuber lines were established using partial least square regression algorithm from FT-IR spectra. The regression coefficients ($R^2$) between predicted values and estimated values of total carotenoids, flavonoids and phenolic compounds were 0.83, 0.86, and 0.72, respectively. These results showed that quantitative predictions of total carotenoids, flavonoids, and phenolic compounds were possible from FT-IR spectra of African yam tuber lines with higher accuracy. Therefore we suggested that quantitative prediction system established in this study could be applied as a rapid selection tool for high yielding African yam lines.

Inhibitory Effects of Various Mulberry Fruits (Morus alba L.) on Related Enzymes to Adult Disease (품종이 다른 오디(Morus alba L.)의 성인병 관련 효소 억제효과)

  • Chae, Jung-Woo;Park, Hye-Jin;Kang, Sun-Ae;Cha, Won-Seup;Ahn, Dong-Hyun;Cho, Young-Je
    • Journal of Life Science
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    • v.22 no.7
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    • pp.920-927
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    • 2012
  • The objective of this research was to develop the functional material of water and 60% ethanol extracts from nine kinds of mulberry fruits (Morus alba L.) that influence the inhibitory activity on angiotensin-converting enzyme, xanthine oxidase, ${\alpha}$-amylase, and ${\alpha}$-glucosidase. The total phenolic contents in the water extracts were over 2 mg/g in two species (Cheongilppong and Kangwon III) and five species (Daeyoupchosaeng, Cheongilppong, Kangwon III, Hihak, and Cataneo) of 60% ethanol extracts. The inhibitory activity against the angiotensin-converting enzyme was determined with them. Baekwoon III was $90.9{\pm}4.5%$ in the water extracts, and Hihak was $81.8{\pm}4.5%$ in the 60% ethanol extracts. The inhibitory activity of Kuksang 20 against xanthin oxidase was about 10% in the water extracts, and Cataneo was $21.4{\pm}2.3%$ in the 60% ethanol extracts. Six of the species (Daeyoupchosaeng, Suwonppong, Cheongilppong, Kangwon III, Hihak, and Kuksang 20) in the water extracts showed inhibitory activities against ${\alpha}$-amylase, as 100%, respectively. The inhibitory activity of ${\alpha}$-glucosidase was determined for these nine species. Four species (Baekwoon III, Daeyoupchosaeng, Cheongilppong, Kangwon III, Hihak, and Kuksang 20) in the water extracts and three species (Daechoukmyeun, Kangwon III, and Kuksang 20) in the 60% ethanol extracts showed inhibition of over 20%. The results revealed strong biological activity in spite of little total phenolic contents. These water and 60% ethanol extracts with high-quality biological activity from various mulberry fruits (Morus alba L.) are expected to represent good candidates for the development of antihypertentive and antidiabetes sources.

Antifungal and Antioxidant Activities of Extracts from Leaves and Flowers of Camellia japonica L. (동백나무 잎과 꽃 추출물의 항미생물 활성 및 항산화 효과)

  • Lee, Sook-Young;Hwang, Eun-Ju;Kim, Gi-Hae;Choi, Young-Bok;Lim, Chae-Young;Kim, Sun-Min
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.3
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    • pp.93-100
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    • 2005
  • This research was performed to investigate the possibilities of industrial usage of camellia (Camellia japonica L.) by examining the antioxidant and antimicrobial effects of methanol extract with different sections. Content of total phenolics, DPPH radical scavenging activities and antibacterial activity of young leaf, mature leaf, flower bud, flower, bark, and seed of camellia were compared in vitro experimental models. Total phenolics was contained the higher in young leaf (74.62 mg), flower bud (65.02 mg) and flower (62.42 mg) but less than 20.95 mg per 100 g of dry weight in other parts of Camellia japonica L. And effects of antioxidant measured by DPPH radical scavenger activity ($RC_{50}$, reduce concentration 50%), was shown higher $7.16{\sim}18.14\;{\mu}g/m{\ell}$ in methanol extract of young leaf, flower bud and flower than $61.23\;{\mu}g/m{\ell}$ of BHT as a chemical oxidant. Also, the antimicrobial activity of Camellia japonica L. extracts determined using a paper disc method against food-borne pathogen and food spoilage bacteria, the young leaves extracts showed the most active antimicrobial activity against 7 kinds of harmful microorganisms. Flower bud extracts showed the highest antibacterial activity against P. aeruginosa and Enterobacter spp. C1036. In addition, the minimum inhibitory concentration (MIC) of young leaf extract against B. subtillis,S. fradiae,S. aureus,E. coli,P. aeruginosa, Enterobacter spp. C1036, and S. typhimurium were revealed 1 to 15 ${\mu}g/m{\ell}$. As a result, antimicrobial activity of camellia extracts was shown higher gram positive bacteria than gram negative bacteria.

Antioxidant Activities of Methanol Extracts from Prunella vulgaris (하고초 메탄올 추출물의 항산화 활성)

  • Lee, Soo-Jung;Sung, Nak-Ju;Jeong, Hey-Gwang;Shin, Jung-Hye;Chung, Young-Chul;Seo, Jong-Kwon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.12
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    • pp.1535-1541
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    • 2008
  • The aim of the present study was to investigate the antioxidant activities of methanol extracts from whole plant, flower stalk and stem of Hagocho (Prunella vulgaris). Content of total phenolic compound was the highest in flower stalk (77.1 mg/100 g) and those of others were below 54.0 mg/100 g. Flavonoid contents was the highest in stem (36.1 mg/100 g) compared to other samples. Electron donating ability of Prunella vulgaris was activated at over 70% in all samples at $500{\mu}g/mL$ concentration, especially, the activity was the highest (92.1%) in flower stalk extracts. Reducing power showed similar tendency to electron donating ability, which was significantly higher flower stalk ($0.3{\sim}1.9$), whole plant ($0.2{\sim}1.6$) and stem ($0.2{\sim}1.5$). Hydroxyl radical was scavenged over 80% in $100{\mu}g/mL$ concentration and was not significantly different between parts. Antioxidant activity in $\beta$-carotene-linoleic acid system was $47.5{\sim}84.6%$ when $1,000{\mu}g/mL$ methanol extracts was added to reaction mixtures, and flower stalk showed the highest activity. Ability of ABTs cation decolorization from Prunella vulgaris was activated over 50% in all samples when $250{\mu}g/mL$ of methanol extracts was added to reaction mixtures and $500{\mu}g/mL$ were the most suitable concentration for its activation. Nitric oxide scavenging activity was lower under 20%, but its activity was significantly higher in flower stalk than other parts. The results indicate that flower stalk from Prunella vulgaris has potent antioxidant activities.

Reconstruction of Metabolic Pathway for the Chicken Genome (닭 특이 대사 경로 재확립)

  • Kim, Woon-Su;Lee, Se-Young;Park, Hye-Sun;Baik, Woon-Kee;Lee, Jun-Heon;Seo, Seong-Won
    • Korean Journal of Poultry Science
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    • v.37 no.3
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    • pp.275-282
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    • 2010
  • Chicken is an important livestock as a valuable biomedical model as well as food for human, and there is a strong rationale for improving our understanding on metabolism and physiology of this organism. The first draft of chicken genome assembly was released in 2004, which enables elaboration on the linkage between genetic and metabolic traits of chicken. The objectives of this study were thus to reconstruct metabolic pathway of the chicken genome and to construct a chicken specific pathway genome database (PGDB). We developed a comprehensive genome database for chicken by integrating all the known annotations for chicken genes and proteins using a pipeline written in Perl. Based on the comprehensive genome annotations, metabolic pathways of the chicken genome were reconstructed using the PathoLogic algorithm in Pathway Tools software. We identified a total of 212 metabolic pathways, 2,709 enzymes, 71 transporters, 1,698 enzymatic reactions, 8 transport reactions, and 1,360 compounds in the current chicken genome build, Gallus_gallus-2.1. Comparative metabolic analysis with the human, mouse and cattle genomes revealed that core metabolic pathways are highly conserved in the chicken genome. It was indicated the quality of assembly and annotations of the chicken genome need to be improved and more researches are required for improving our understanding on function of genes and metabolic pathways of avian species. We conclude that the chicken PGDB is useful for studies on avian and chicken metabolism and provides a platform for comparative genomic and metabolic analysis of animal biology and biomedicine.