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Isolation of Polyphenol Compounds from the Leaves of Korean Persimmon (Diospyrus kaki L. Folium) (한국산 감잎로부터 Polyphenol계 생리활성물질 분리)

  • An, Bong-Jeun;Bae, Man-Jong;Choi, Hee-Jin;Zhang, Yun-Bin;Sung, Tae-Soo;Choi, Cheong
    • Applied Biological Chemistry
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    • v.45 no.4
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    • pp.212-217
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    • 2002
  • We purified polyphenols from persimmon leaf and tested their biological activity. The 60% acetone extract was lyophilized and applied to test enzyme inhibition of glucosyltransferase and tyrosinase. GTase was 82.4% inhibited at $1.8{\times}10^{-1}$ mg/ml and tyrosinase 21.7% inhibited at 0.8 mg/ml. The acetone extract was fractionated into F-1, 2, 3, 4, 5 by Sephadex Q-50 gel filtration and the fraction-1 and 2 showed higher enzyme inhibition activity than the other fractions. To the Proteinase K treatment and autoclaving of the two fractions had no effect on the enzyme activity, but these results suggested that active fraction was not protein but phenol ring completed compounds. By Sephadex LH-20, MCI-gel and Bondapak $C_{18}$ column chromatographies, compouds 1, 2, 3 and 4 from F-1 fraction, compounds 5 and 6 from F-2 fraction and compounds 7 , 8 from F-3 fraction were purified and re-crystallized. The purified compounds was assumed to be condensed tannins of frame flavan-3-ol frame on the basis of color reagent reaction and to be a mixture of monomer, dimer and trimer according to TLC analysis.

Isolation, Expression Pattern, Polymorphism and Association Analysis of Porcine TIAF1 Gene

  • Wang, Y.;Xiong, Y.Z.;Ren, Z.Q.;Zuo, B.;Lei, M.G.;Deng, C.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.3
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    • pp.313-318
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    • 2009
  • TIAF1 is a TGF-${\beta}$1-induced anti-apoptotic factor that plays a critical role in blocking TNF (tumor necrosis factor) cytotoxicity in mouse fibroblasts and participates in TGF-${\beta}$-mediated growth regulation. In this study, we obtained the full-length cDNA sequence of the porcine TIAF1 gene. Real-time PCR further revealed that the TIAF1 gene was expressed at the highest level in liver and kidney with prominent expressions detected in uterus, and lower levels detected in heart, spleen, lung, stomach, small intestine, skeletal muscle and fat of Large White pigs. Sequence analysis indicated that a 6 base-pair deletion mutation existed in the exon of the TIAF1 gene between Meishan and Large White pigs. This mutation induced deletion of Gln and Val amino acids. PCR-RFLP was used to detect the polymorphism in 394 pigs of a "Large White${\times}$Meishan" $F_{2}$ resource population and four purebred pig populations. The frequencies of the A allele (with a 6 bp deletion) were dominant in Chinese Meishan and Bamei pigs, and the frequencies of the B allele (no 6 bp deletion) were dominant in Large White and Landrace pigs. Association analyses revealed that the deletion mutation had highly significant associations (p<0.01) with meat marbling score of the thorax-waist longissimus dorsi (LD) muscle (MM1) and intramuscular fat percentage (IMF), and significant associations (p<0.05) with carcass length (CL). The results presented here supply evidence that the 6 bp deletion mutation in the TIAF1 gene affects porcine meat quality and provides useful information for further porcine breeding.

Expression characterization and transcription regulation analysis of porcine Yip1 domain family member 3 gene

  • Ni, Dongjiao;Huang, Xiang;Wang, Zhibo;Deng, Lin;Zeng, Li;Zhang, Yiwei;Lu, Dongdong;Zou, Xinhua
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.3
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    • pp.398-407
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    • 2020
  • Objective: The Yip1 domain family (YIPF) proteins were proposed to function in endoplasmic reticulum (ER) to Golgi transport and maintenance of the morphology of the Golgi, which were homologues of yeast Yip1p and Yif1p. YIPF3, the member 3 of YIPF family was a homolog of Yif1p. The aim of present study was to investigate the expression and regulation mechanism of porcine YIPF3. Methods: Quantitative realtime polymerase chain reaction (qPCR) was used to analyze porcine YIPF3 mRNA expression pattern in different tissues and pig kidney epithelial (PK15) cells stimulated by polyinosine-polycytidylic acid (poly [I:C]). Site-directed mutations combined with dual luciferase reporter assays and electrophoretic mobility shift assay (EMSA) were employed to reveal transcription regulation mechanism of porcine YIPF3. Results: Results showed that the mRNA of porcine YIPF3 (pYIPF3) was widely expressed with the highest levels in lymph and lung followed by spleen and liver, while weak in heart and skeletal muscle. Subcellular localization results indicated that it expressed in Golgi apparatus and plasma membranes. Upon stimulation with poly (I:C), the level of this gene was dramatically up-regulated in a time- and concentration-dependent manner. pYIPF3 core promoter region harbored three cis-acting elements which were bound by ETS proto-oncogene 2 (ETS2), zinc finger and BTB domain containing 4 (ZBTB4), and zinc finger and BTB domain containing 14 (ZBTB14), respectively. In which, ETS2 and ZBTB4 both promoted pYIPF3 transcription activity while ZBTB14 inhibited it, and these three transcription factors all played important regulation roles in tumorigenesis and apoptosis. Conclusion: The pYIPF3 mRNA expression was regulated by ETS2, ZBTB4, and ZBTB14, and its higher expression in immune organs might contribute to enhancing ER to Golgi transport of proteins, thus adapting to the immune response.

Long Non-coding RNAs are Differentially Expressed in Hepatocellular Carcinoma Cell Lines with Differing Metastatic Potential

  • Fang, Ting-Ting;Sun, Xiao-Jing;Chen, Jie;Zhao, Yan;Sun, Rui-Xia;Ren, Ning;Liu, Bin-Bin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10513-10524
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    • 2015
  • Background: Metastasis is a major reason for poor prognosis in patients with cancer, including hepatocellular carcinoma (HCC). A salient feature is the ability of cancer cells to colonize different organs. Long non-coding RNAs (lncRNAs) play important roles in numerous cellular processes, including metastasis. Materials and Methods: In this study, the lncRNA expression profiles of two HCC cell lines, one with high potential for metastasis to the lung (HCCLM3) and the other to lymph nodes (HCCLYM-H2) were assessed using the Arraystar Human LncRNA Array v2.0, which contains 33,045 lncRNAs and 30,215 mRNAs. Coding-non-coding gene co-expression (CNC) networks were constructed and gene set enrichment analysis (GSEA) was performed to identify lncRNAs with potential functions in organ-specific metastasis. Levels of two representative lncRNAs and one representative mRNA, RP5-1014O16.1, lincRNA-TSPAN8 and TSPAN8, were further detected in HCC cell lines with differing metastasis potential by qRT-PCR. Results: Using microarray data, we identified 1,482 lncRNAs and 1,629 mRNAs that were differentially expressed (${\geq}1.5$ fold-change) between the two HCC cell lines. The most upregulated lncRNAs in H2 were RP11-672F9.1, RP5-1014O16.1, and RP11-501G6.1, while the most downregulated ones were lincRNA-TSPAN8, lincRNA-CALCA, C14orf132, NCRNA00173, and CR613944. The most upregulated mRNAs in H2 were C15orf48, PSG2, and PSG8, while the most downregulated ones were CALCB, CD81, CD24, TSPAN8, and SOST. Among them, lincRNA-TSPAN8 and TSPAN8 were found highly expressed in high lung metastatic potential HCC cells, while lowly expressed in no or low lung metastatic potential HCC cells. RP5-1014O16.1 was highly expressed in high lymphatic metastatic potential HCC cell lines, while lowly expressed in no lymphatic metastatic potential HCC cell lines. Conclusions: We provide the first detailed description of lncRNA expression profiles related to organ-specific metastasis in HCC. We demonstrated that a large number of lncRNAs may play important roles in driving HCC cells to metastasize to different sites; these lncRNAs may provide novel molecular biomarkers and offer a new basis for combating metastasis in HCC cases.

Investigation and Analysis of Allergy-related SNPs for Allergy Affected Students in a high school. (과학영재학교 학생들이 알러지 관련 SNP 탐색고 분석)

  • 김경원;이호경;김현근;김수영;안정훈
    • Journal of Life Science
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    • v.14 no.5
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    • pp.847-854
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    • 2004
  • Allergy is a multi-factorial disease influenced by genetic and environmental factors. As the number of allergy-affected people is increasing in developed countries, there is an increasing interest in genetic predisposition to the allergy. A number of genes and chromosomal region have been identified to be linked to allergy including rhinitis, asthma and atopy. In order to understand the genetic background for the allergy-affected people, we investigated genetic predisposition among students enrolled in Busan Science Academy. Among 138 students, about 30% students had some allergy-related disorder including rhinitis, asthma and atopy. We analyzed several single nucleotide polymorphisms (SNPs) within two genes, Inter-leukin-4(IL-4) and Interleukin-4 receptor(IL-4R), which are involved in the induction of allergy reaction with the Th2 immunity. For 96 samples obtained from students, we analyzed 9 SNPs including -590 C/T and -34 C/T in IL-4, and I75V, Q576R, E375A, e406R, 5411L, S761P and S727A in IL-4R. From the analysis, these SNPs showed slight differences among normal and allergy-affected students, but these differences was not enough to predict the predisposition to the allergy. In contrast to previous reports, we could not find SNP(s) related with allergy. These results suggest that genetic tests recently performed in Korea widely have to be reassessed for its validity of genetic predisposition. [Supported by grants from MOST]

Current Trends and Recent Advances in Diagnosis, Therapy, and Prevention of Hepatocellular Carcinoma

  • Wang, Chun-Hsiang;Wey, Keh-Cherng;Mo, Lein-Ray;Chang, Kuo-Kwan;Lin, Ruey-Chang;Kuo, Jen-Juan
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3595-3604
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    • 2015
  • Hepatocellular carcinoma (HCC) has been one of the most fatal malignant tumors worldwide and its associated morbidity and mortality remain of significant concern. Based on in-depth reviews of serological diagnosis of HCC, in addition to AFP, there are other biomarkers: Lens culinaris agglutinin-reactive AFP (AFP-L3), descarboxyprothrombin (DCP), tyrosine kinase with Ig and eprdermal growth factor (EGF) homology domains 2 (TIE2)-espressing monocytes (TEMs), glypican-3 (GPC3), Golgi protein 73 (GP73), interleukin-6 (IL-6), and squamous cell carcinoma antigen (SCCA) have been proposed as biomarkers for the early detection of HCC. The diagnosis of HCC is primarily based on noninvasive standard imaging methods, such as ultrasound (US), dynamic multiphasic multidetector-row CT (MDCT) and magnetic resonance imaging (MRI). Some experts advocate gadolinium diethyl-enetriamine pentaacetic acid (Gd-EOB-DTPA) MRI and contrast-enhanced US as the promising imaging madalities of choice. With regard to recent advancements in tissue markers, many cuting-edge technologies using genome-wide DNA microarrays, qRT-PCR, and proteomic and inmunostaining studies have been implemented in an attempt to identify markers for early diagnosis of HCC. Only less than half of HCC patients at initial diagnosis are at an early stage treatable with curative options: local ablation, surgical resection, or liver transplant. Transarterial chemoembolization (TACE) is considered the standard of care with palliation for intermediate stage HCC. Recent innovative procedures using drug-eluting-beads and radioembolization using Yttrium-90 may exhibit beneficial effects in HCC treatment. During the past few years, several molecular targeted agents have been evaluated in clinical trials in advanced HCC. Sorafenib is currently the only approved systemic treatment for HCC. It has been approved for the therapy of asymptomatic HCC patients with well-preserved liver function who are not candidates for potentially curative treatments, such as surgical resection or liver transplantation. In the USA, Europe and particularly Japan, hepatitis C virus (HCV) related HCC accounts for most liver cancer, as compared with Asia-Pacific regions, where hepatitis B virus (HBV) may play a more important role in HCC development. HBV vaccination, while a vaccine is not yet available against HCV, has been recognized as a best primary prevention method for HBV-related HCC, although in patients already infected with HBV or HCV, secondary prevention with antiviral therapy is still a reasonable strategy. In addition to HBV and HCV, attention should be paid to other relevant HCC risk factors, including nonalcoholic fatty liver disease due to obesity and diabetes, heavy alcohol consumption, and prolonged aflatoxin exposure. Interestingly, coffee and vitamin K2 have been proven to provide protective effects against HCC. Regarding tertiary prevention of HCC recurrence after surgical resection, addition of antiviral treatment has proven to be a rational strategy.

Analysis on the Risk-Based Screening Levels Determined by Various Risk Assessment Tools (II): Derivation of Particulate Emission Factor at Former Janghang Smelter Site (다양한 위해성평가 방법에 따라 도출한 토양오염 판정기준의 차이에 관한 연구 (II): (구)장항제련소부지의 기상 및 부지 특성을 반영한 비산계수 결정)

  • Jung, Jae-Woong;Yang, Kyung;Lee, Gwang-Hun;Ryu, Hye-Rim;Nam, Kyoung-Phile
    • Journal of Soil and Groundwater Environment
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    • v.17 no.3
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    • pp.21-31
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    • 2012
  • This paper presents the short-term and long-term measures to determine the fugitive dust concentration in a contaminated site, which is a crucial step for the determination of particulate emission factor (PEF) for risk assessment. As a long-term measure, USEPA method employing Q/C value (inverse of the ratio of the geometric mean air concentration to the emission flux at center of a 0.5-acre square source) seems to be suitable as it reflects regional-specific meteorological conditions. However, it requires nation-wide database collection and interpretation. Use of ASTM method is an alternative as a short-term measure. The method is readily field-applicable as PEF calculation equation is simple and input parameters can be easily derived at the site of interest as well without the nation-wide efforts. Using ASTM method, PEF at the Former Janghang Smelter Site was determined. According to various mode of aggregate size distribution and fractions of vegetative cover, which are the most important factors in PEF calculation, PEF values at the Former Janghang Smelter Site varied greatly. When the mode of aggregate size distribution was set at 0.25 mm, PEF values at the Former Janghang Smelter Site was 5~20 times higher than the default PEF value (i.e., 35 ${\mu}g/m^3$) shown in the current Korean Soil Contamination Risk Assessment Guidance. On contrast, when the mode was set at 2 mm, PEF values at the Former Janghang Smelter Site was 160~640 times lower than the default PEF value in the Korean Guidance.

Studies on the Ethanol Production by Clostridium thermosaccharolyticum (Clostridium thermosaccharolyticum에 의한 에탄올생산에 관한 연구)

  • 조은경;이윤광;변유량;유주현
    • Microbiology and Biotechnology Letters
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    • v.13 no.4
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    • pp.397-402
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    • 1985
  • The fermentation of various sugars by C. thermosaccharolyticum was examined under pH controlled, anaerobic condition. The kinetic model for Product formation at various sugars was the combination of growth and non-growth associated mode. In the utilization of a single sugar, glucose was the best carbon source for growth. The specific growth rate of glucose, xylose and cellobiose were 0.363 h$^{-1}$, 0.242 h$^{-1}$ and 0.144 h$^{-1}$ respectively. The production of ethanol from glucose showed a negatively growth associated mode, so the higher growth rate decreased the productivity of ethanol. The maximum concentrations of the produced ethanol were 2.42 g/l, 3.76 g/l, and 3.4 g/l on glucose, xylose, and cellobiose. No glucose was detected during cellobiose fermentation. Sequential utilization of sugars was observed in the mixtures of glucose, xylose and cellobiose. It preferred glucose, followed by xylose and then cellobiose. The presence of other sugars had little or no effect on the rate of another sugar utilization. Cell lysis at the end of fermentation occured more slowly in the mixtures of sugars than a single sugar.

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Effects of Inclusion Levels of Wheat Bran and Body Weight on Ileal and Fecal Digestibility in Growing Pigs

  • Huang, Q.;Su, Y.B.;Li, D.F.;Liu, L.;Huang, C.F.;Zhu, Z.P.;Lai, C.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.6
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    • pp.847-854
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    • 2015
  • The objective of this study was to determine the effects of graded inclusions of wheat bran (0%, 9.65%, 48.25% wheat bran) and two growth stages (from 32.5 to 47.2 kg and 59.4 to 78.7 kg, respectively) on the apparent ileal digestibility (AID), apparent total tract digestibility (ATTD) and hindgut fermentation of nutrients and energy in growing pigs. Six light pigs (initial body weight [BW] $32.5{\pm}2.1kg$) and six heavy pigs (initial BW $59.4{\pm}3.2kg$) were surgically prepared with a T-cannula in the distal ileum. A difference method was used to calculate the nutrient and energy digestibility of wheat bran by means of comparison with a basal diet consisting of corn-soybean meal (0% wheat bran). Two additional diets were formulated by replacing 9.65% and 48.25% wheat bran by the basal diet, respectively. Each group of pigs was allotted to a $6{\times}3$ Youden square design, and pigs were fed to three experimental diets during three 11-d periods. Hindgut fermentation values were calculated as the differences between ATTD and AID values. For the wheat bran diets, the AID and ATTD of dry matter (DM), ash, organic matter (OM), carbohydrates (CHO), gross energy (GE), and digestible energy (DE) decreased with increasing inclusion levels of wheat bran (p<0.05). While only AID of CHO and ATTD of DM, ash, OM, CHO, GE, and DE content differed (p<0.05) when considering the BW effect. For the wheat bran ingredient, there was a wider variation effect (p<0.01) on the nutrient and energy digestibility of wheat bran in 9.65% inclusion level due to the coefficient of variation (CV) of the nutrient and energy digestibility being higher at 9.65% compared to 48.25% inclusion level of wheat bran. Digestible energy content of wheat bran at 48.25% inclusion level (4.8 and 6.7 MJ/kg of DM, respectively) fermented by hindgut was significantly higher (p<0.05) than that in 9.65% wheat bran inclusion level (2.56 and 2.12 MJ/kg of DM, respectively), which was also affected (p<0.05) by two growth stages. This increase in hindgut fermentation caused the difference in ileal DE (p<0.05) to disappear at total tract level. All in all, increasing wheat bran levels in diets negatively influences the digestibility of some nutrients in pigs, while it positively affects the DE fermentation in the hindgut.

The Nutritive Values in Different Varieties of Corn Planted in One Location Fed to Growing Pigs over Three Consecutive Years

  • Zhang, L.;Li, Y.K.;Li, Z.C.;Li, Q.F.;Lyu, M.B.;Li, D.F.;Lai, C.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.12
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    • pp.1768-1773
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    • 2016
  • This experiment was conducted to determine the effect of variety and planting year on the nutritive values of corn fed to growing pigs. Four corn varieties examined in this experiment were planted in the same village located in Longhua County, Heibei Province, China, in 2012, 2013, and 2014, respectively. During each year, corn was hand-harvested in early October and sun dried to about 14% moisture content. Three batches of twenty-four barrows ($33.27{\pm}4.30$, $31.88{\pm}2.93$, $34.21{\pm}3.81kg$ body wight [BW] in 2012, 2013, and 2014, respectively) were used and allotted to a complete block design with 4 diets and 6 replicate pigs per diet. Pigs were individually placed in metabolic crates. The four experimental diets were formulated by mixing each variety of corn and vitamins and minerals, respectively. A five-day collection period followed a seven-day diet acclimation period. The results indicated that variety of corn significantly influenced the available energy content (digestible energy [DE] on dry matter basis, p<0.05; metabolizable energy (ME) on dry matter basis, p<0.05, respectively), and the apparent total tract digestibility (ATTD) of organic matter (p<0.01), dry matter (p<0.05), gross energy (p<0.05), neutral detergent fiber (p<0.01), acid detergent fiber and ether extract (p<0.05). The planting year also significantly influenced the available energy contents (DE on dry matter basis, p<0.05; ME on dry matter basis, p<0.01, respectively) and the ATTD of neutral detergent fiber (p<0.01), acid detergent fiber (p<0.01), crude protein (p<0.01), and ether extract (p<0.01). No interaction was observed between the variety and planting year in DE and ME contents in corn. In conclusion, the variety and planting year significantly influenced the available energy and nutrient digestibility of corn fed to growing pigs.