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Induction of Midbrain Dopaminergic Phenotype in Nurr 1-Over expressing Human Neural Stem Cells (사람 신경 간세포에서 도파민 신경세포 분화유도에 대한 Nurr 1 유전자의 역할 규명)

  • Kim, Han-Jip;Lee, Haksup;Kim, Hyon-Chang;Min, Churl-Ki;Lee, Myung-Ae;Kim, Seung-Up;Han, Jin;Youm, Jae-Boum;Kim, Nari;Park, Won, Sun;Kim, Taeho;Kim, Euiyong;Han, Il-Yong
    • KSBB Journal
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    • v.20 no.5
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    • pp.363-370
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    • 2005
  • Neural stem cells (NSCs) of the central nervous system (CNS) have raised a great interest not only for their importance in basic neural development but also for their therapeutic potentials in neurologically degenerative diseases such as Parkinson's, Alzheimer and stroke. During the CNS development, two molecular cascades determine specification of midbrain dopamine system. In one pathway, FGF-8, sonic hedgehog and transcription factor Nurr1 specify dopamine neurotransmitter phenotype. In the other, transcription factors $Lm{\times}lb\;and\;Pt{\times}3$ are required for induction of dopaminergic neurons. In Nurr1 knockout mouse, tyrosine hydroxylase (TH) positive cells fail to appear in substantia nigra, indicating that Nurr1 is essential in specification of dopaminergic cell phenotype. In this study, we used the immortalized human NSCs retrovirally transduced with Nurr1 gene to probe the Nurr1 mediated mechanism to induce dopamine phenotype. While Nurr1 over-expression alone did not generate dopamine phenotype in NSCs, applications of retinoid and forskolin induced expression of TH and AADC mRNAs. In addition, co-cultures of Nurr1 expressing NSCs with human astrocytes induced a marked increase of TH expression. In this co-culture system, the addition of retinoid and forskolin dramatically increased expression of TH. These results indicate that the immortalized human NSCs with Nurr1 gene could have a clinical utility for cell replacement for the Parkinson patients.

Effects of Kale Juice Powder on Serum Lipids, Folate and Plasma Homocysteine Levels in Growing Rats (케일녹즙 분말식이가 흰쥐의 혈중 지질, 엽산 및 호모시스테인 수준에 미치는 효과)

  • Chung, Eun-Jung;Kim, Soo-Yeon;Nam, Young-Ju;Park, Jung-Hwa;Hwang, Hye-Jin;Lee, Yang-Cha
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1175-1181
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    • 2005
  • The purpose of this study was to investigate the effects of green vegetable (kale) juice powder supple-mentation on lipid profiles, plasma homocysteine, folate and vitamin $B_{12}$ in rats fed cholesterol (Chol. ) or Chol. free diet. 7-week old male Sprague Dawley rats (n=40) were divided into 4 groups, and experimental diets containing control diet group (CO), control diet plus 0.5 wt$ \% $ Chol. (CC), control diet added with 5 wt$ \% $ kale (KO), control diet added with 5 wt$ \% $ kale plus 0.5 wt$ \% $ Chol. (KC) were fed for 8 wks. Plasma homocysteine level was examined by amino acid analyzer and serum folate and vitamin $B_{12}$ level were measured by com-petitive radioimmunoassay methods. In various serum lipid profiles, TG level was lower in kale juice powder groups (KO, KC) compared to the corresponding groups (CO, CC) (p<0.001). In Chol. supplemented groups (CC, KC), HDL-Chol. level was lower (p<0.001) and LDL-Chol. level was higher (p<0.05) than Chol. free diet. HDL-Chol. level was higer (p<0.05) in kale juice powder groups. HDL/LDL ratio was lower in Chol. supplemented groups (CC, KC) and tended to be higher in kale juice powder groups (KO, KC) Serum folate and vitamin $B_{12}$ levels were not affected by dietary Chol. and kale juice powder supplementation. Plasma homocysteine level was not affected by dietary Chol. and kale juice powder supplementation, too. Serum folate level was positively correlated with serum vitamin $B_{12}$ level (r=0.5632, p<0.001), but plasma homocysteine level was not significantly correlated with any serum folate, vitamin $B_{12}$ and Chol. levels, respectively. In summary, kale juice powder supplementation have improved serum lipid profiles by increasing the HDL level and decreasing the TG level and have not altered homocysteine level under the sufficient supply of folate and vitamin B complex relating with the homocysteine metabolism.

Characterization and Cloning of the Gene Encoding Autoregulator Receptor Protein from Streptomyces longwoodensis (Streptomyces longwoodensis로부터 Autoregulator Receptor Protein 유전자의 클로닝 및 특성)

  • Yeo Soo-Hwan;Lee Sung-Bong;Kim Hyun-Soo
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.96-105
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    • 2005
  • For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.

Intracullular Functions of the mas2+ Gene in the Fission Yeast, Schizosaccharomyces pombe (분열형 효모에서의 mas2+ 유전자의 세포 내 기능)

  • Sin, Sang-Min;Cha, Jae-Young;Ha, Se-Eun;Sim, Sun-Mi;Kim, Hyoung-Do;Lee, Jung-Sup;Park, Jong-Kun
    • Journal of Life Science
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    • v.19 no.1
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    • pp.101-110
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    • 2009
  • The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel $mas2^+$ (mitosis associated protein) gene, a homolog of human SMARCAD1 was isolated and characterized from a fission yeast Schizosaccharomyces pombe (S. pombe) using gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 922 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that an SNF2 domain is located, which is involved in the chromosome remodeling. The quantitative analysis of the $mas2^+$ transcript against $adh1^+$ showed that the expression level of $mas2^+$ is high before septum formation in S. pombe. When $mas2^+$ null mutant cells were grown at 27 and $35^{\circ}C$, the cytokinesis of $mas2^+$ null mutant was greatly delayed and a large number of multi-septate and mis-segregated cells were produced. In addition, the number of multi-septate cells significantly increased. When cells were cultured in YES rich medium to increase proliferation, the abnormal phenotypes $mas2^+$ null mutant dramatically increased. These phenotypes could be rescued by an over-expression of the mast gene. The Mas2 protein localized in the nuclei of S. pombe, as evidenced by Mas2-EGFP signals. These results suggest that the $mas2^+$ is homologous to human SMARCAD1 gene and involved in septum formation and chromosome remodeling control.

Characterization of Oszinc626, knock-out in zinc finger RING-H2 protein gene, in Ac/Ds mutant lines of rice(Oryza sativar L.) (Zinc finger RING-H2 protein관련 Ac/Ds전이인자 삽입 변이체 Oszinc626 유전자의 특성 분석)

  • Park, Seul-Ah;Jung, Yu-Jin;Ahn, Byung-Ohg;Yun, Doh-Won;Ji, Hyeon-So;Park, Yong-Hwan;Eun, Moo-Young;Suh, Seok-Cheol;Lee, Soon-Youl;Lee, Myung-Chul
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.177-183
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    • 2008
  • Ac/Ds mutant lines of this study were transgenic rice plants, each of which harbored the maize transposable element Ds together with a GUS coding sequence under the control of a promoterless(Ds-GUS). We selected the mutants that were GUS expressed lines, because the GUS positive lines will be useful for identifying gene function in rice. One of these mutants was identified knock-out at Oszinc626(NP_001049991) gene, encoding a RING-H2 zinc-finger protein, by Ds insertion. In this mutant, while primary root development is normal, secondary root development from lateral root was very poor and seed development was incomplete compare with normal plant. RING zinc-finger proteins play important roles in the regulation of development in a variety of organisms. In the plant kingdom, a few genes encoding RING zinc-finger proteins have been documented with visible effects on plant growth and development. The consensus of the RING-H2(C3-H2-C3 type) domain for this group of protein is $Cys-X_2-Cys-X_{28}-Cys-X-His-X_2-His-X_2-Cys-X_{14}-Cys-X_2-Cys$. Oszinc626 encodes a predicted protein product of 445 amino acids residues with a molecular mass of 49 kDa, with a RING-zinc-finger motif located at the extreme end of the C-terminus. RT-PCR analysis indicated that the expression of Oszinc626 gene was induced by IAA, cold, dehydration, high-salinity and abscisic acid, but not by 2,4-D, and the transcription of Oszinc626 gene accumulated primarily in rice immature seeds, root meristem and shoots. The gene accumulation patterns were corresponded with GUS expression.

Quality Characteristics on Enzyme Treatment of Brown Rice(Goami) Alcohol Fermentation By-Product (현미(고아미) 알코올발효 부산물의 효소처리에 따른 품질특성)

  • Jang, Se-Young;Woo, Seung-Mi;Kim, Tae-Young;Yeo, Soo-Hwan;Kim, Sang-Burm;Hong, Ju-Yeon;Jeong, Yong-Jin
    • Korean Journal of Food Preservation
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    • v.15 no.3
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    • pp.477-482
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    • 2008
  • The quality characteristics of Goami by-product under the mixed enzyme treatment condition of $\alpha$-amylase and cellulase have been compared, and found the highest amount of soluble solids and reducing sugars at the $\alpha$-amylase treated group (A), and the contents revealed to show gradual decrease with the increase of cellulase content. The amounts of total dietary fiber and total sugars did not show large difference by both of enzyme concentration. The result of sugar analysis revealed the presence of all $G{\sim}G5$ in all treatment groups, and the content of malto-oligosaccharide recorded the highest content of 2,200 mg% at the $\alpha$-amylase treatment group (A). When the quality characteristic of the hydrolyzed powders manufactured by the optimum hydrolysis condition was compared, no significant color difference was found between samples. Among the contents of dietary fibers, insoluble dietary fiber was found to present in the lowest content of 6.95% at the Goami flour (GF) and the Goami by-product powder (GBPP) and Goami by-product hydrolysate powder (GBPHP) resulted the similar content around 14% and the highest soluble dietary fibers content was found in Goami by-product hydrolysate powder (GBPHP), which was followed by in the order of Goami by-product powder (GBPP) and Goami flour (GF), but the content variation was not large. The free amino acid was found to be highest in Goami by-product hydrolysate powder (GBPHP) followed by in the order of Goami by-product powder (GBPP) and Goami flour (GF). In the sugar analysis, the Goami by-product hydrolysate powder (GBPHP) was found with all $G{\sim}G5$ sugars by showing the highest amount of 1,800 mg% At the Goami by-product powder (GBPP), $G{\sim}G2$ sugars were detected with about 66 mg% and malto-oligosaccharides were not detected at the Goami flour (GF). Based upon the results, the functionality of Goami by-product hydrolysate powder (GBPHP) was found to be enforced compared to Goami flour (GF) and Goami by-product powder (GBPP), which allow us to expect it to be used as the various rice processing food source.

The Growth Characteristics and Germanium Uptake by Water Celery in Soil Treated with Germanium (게르마늄 처리 토양에서 미나리 생육 특성과 게르마늄 흡수)

  • Lee, Seong-Tae;Lee, Young-Han;Heo, Jae-Young;Hong, Kwang-Pyo;Dahlgren, Randy A.;Heo, Jong-Soo
    • Korean Journal of Environmental Agriculture
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    • v.27 no.2
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    • pp.185-190
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    • 2008
  • In order to obtain the basic information for agricultural utilization of Germanium(Ge), the growth characteristics and the germanium uptake by water celery were investigated at different concentration of germanium in soil. This experiment was carried out in the Wagner pot(1 $5,000^{-1}a$). Germanium concentrations in soil for water celery cultivation were maintained at 0.26, 25.0, 62.5, and 125.0 mg $kg^{-1}$, respectively. The treatment of over Ge 25.0 mg $kg^{-1}$ in the soil led to germanium phytotoxicity such as reduction of plant height and fresh weight. The contents of germanium in water celery were increased with the increase of germanium concentration in the soil. When water celery was cultivated from soil maintained with Ge 25.0 and 62.5 mg $kg^{-1}$, its germanium contents in plant were 89.9 and 371.6 mg $kg^{-1}$, respectively. Then, the efficiency of germanium uptake of water celery in Ge 25.0 and 62.5 mg $kg^{-1}$ maintained plots was 1.7 and 2.4%, respectively. When water celery was cultivated from soil maintained with Ge 25.0, 62.5 and 125.0 mg $kg^{-1}$, its content of amino acid was found to be 89.8, 198.4, and 318.2 mg $g^{-1}$, respectively. To investigate the effect of N fertilizer application in uptake of germanium by water celery, these were treated with nontreatment, 1.0, 1.5 and 2.0 times of N application based on soil testing for cultivation of water celery. However, the amount of the N fertilizer application did not affect the contents of germanium in the water celery. When water celery was cultivated from soil maintained with two kinds of inorganic and organic germanium 50 mg $kg^{-1}$, respectively, the content of germanium were 24.2 mg $kg^{-1}$ in the Ge-132 treatment and 11.8 mg $kg^{-1}$ in the $GeO_2$ treatment.

Effect of Silk in Silk/PLGA Hybrid Films on Attachment and Proliferation of Human Aortic Endothelial Cells (실크/PLGA 하이브리드 필름에서 실크가 인간 대동맥 내피세포의 부착과 증식에 미치는 효과)

  • Lee, Jihye;Lee, Sojin;Kim, Seulji;Kim, Kyounghee;Kim, Younglae;Song, Jeongeun;Lee, Dongwon;Khang, Gilson
    • Polymer Korea
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    • v.37 no.2
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    • pp.127-134
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    • 2013
  • The vascular endothelial cells are the inner layers of blood vessels. It regulates the function of blood vessels and proliferation of vascular smooth muscle cells. Poly(lactide-co-glycolic acid) (PLGA) is a biodegradable synthetic polymer with a well-controlled degradation rate and an acceptable mechanical strength. It can be easily fabricated into many shapes. Silk consists of 18 amino acids. It found important for attaching cells cultured in vitro, and maintaining cell functions. In this study, we fabricated silk/PLGA biomaterial hybrid films of 0, 10, 20, 40 and 80 wt% silk. We performed MTT, SEM, ELISA, and immunocytochemistry analyses. We confirmed the adhesion and the proliferation of HAECs on silk/PLGA according to the content of silk, and 40 wt% silk/PLGA hybrid films have superior adhesion and proliferation properties. These results demonstrate that silk/PLGA hybrid films provide suitable surfaces for HAECs, and there is the effect of silk on cell growth and proliferation.

Genetic Variations of Chicken MC1R Gene and Associations with Feather Color of Korean Native Chicken (KNC) 'Woorimatdag' (토종 '우리맛닭' 부계 및 실용계에서 MC1R 유전자 변이 및 모색과의 연관성 분석)

  • Park, Mi Na;Kim, Tae-Hun;Lee, Hyun-Jeong;Choi, Jin Ae;Heo, Kang-Nyeong;Kim, Chong-Dae;Choo, Hyo-Jun;Han, Jae-Yong;Lee, Taeheon;Lee, Jun-Heon;Lee, Kyung-Tai
    • Korean Journal of Poultry Science
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    • v.40 no.2
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    • pp.139-145
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    • 2013
  • There are several loci controlling the feather color of birds, of which one of the most studied is Extended black (E) encoding the melanocortin 1-receptor (MC1R). Mutations in this gene affect the relative distribution of eumelanin, phaeomelanin. The association of feather color and sequence polymorphism in the melanocortin 1-receptor (MC1R) gene was investigated using Korean native chicken H breed (H_PL) and 'Woorimatdag' commercial chickens (Woorimatdag_CC). In order to correlate gene mutation to Korean native chicken feather color, single nucleotide polymorphism (SNP) from MC1R gene sequence were investigated. A total of 307 birds from H_PL and Woorimatdag_CC were used. H_PL have black, black-brown feather color and Woorimatdag_CC have black with brown spots or brown with black spots. There are 6 SNPs in MC1R gene, locus T69C, C212T, A274G, G376A, G636A, T637C. 3 SNPs are nonsynonymous that change amino acid. But it is difficult to find correlation of feather color and polymorphisms. It will be needed to increase the population of Korean native chicken H breed and correlation analysis of genetic variation with feather colors.

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.