• Title/Summary/Keyword: Amino Acid Metabolism

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Regulation of hPTH Expression In Virto Using the Tetracycline Inducible Retrovirus Vector System (Tetracycline Inducible Retrovirus Vector System을 이용한 In Vitro에서의 인간 부갑상선 호르몬의 발현 조절)

  • Koo, Bon-Chul;Kwon, Mo-Sun;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.30 no.3
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    • pp.157-162
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    • 2006
  • Endogenous 84 amino acid parathyroid hormone(PTH) is synthesized as a pre-pro hormone by the chief cells of the parathyroid glands. Physiological actions of PTH include regulation of bone metabolism, renal tubular reabsorption of calcium and phosphate, and intestinal calcium absorption. In addition, PTH stimulates new bone formation by extraordinary stimulation of osteoblastic activity and decreasing calcium excretion by the kidney. In this study, we constructed and tested retrovirus vectors designed to express the human parathyroid hormone(hPTH) gene under the control of the tetracycline-inducible promoters. To increase the hPTH gene expression at turn-on state, woodchuck hepatitis virus posttranscriptional regulatory element(WPRE) sequence was also introduced into retrovirus vector at downstream region of either the hPTH gene or the sequence encoding reverse tetracycline-controlled transactivator(rtTA). Transformed primary culture cells(porcine fetal fibroblast, PFF, chicken embryonic fibroblast, CEF) were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the hPTH gene expression level using two step RT-PCR and ELISA Higher hPTH expression($3{\tims}10^4\;pg/ml,\;5.3{\times}10^4\;pg/ml$) and tighter expression control(up to 8 fold) were observed from the vector in which the WPRE sequence was placed at downstream of the hPTH gene. The resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

Effect of Semisulcospira libertina Extracts from Different Extraction Processes on Liver Cell Toxicity and Ethanol Metabolism (간세포 독성과 에탄올 대사에서 추출 조건에 따른 다슬기 추출물의 효과)

  • Cho, Kyoung Hwan;Choo, Ho Jin;Seo, Min Gyun;Kim, Jong Cheol;Shin, Yu Jin;Ryu, Gi Hyung;Cho, Hee Young;Jeong, Chi-Young;Hah, Young-Sool
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.158-166
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    • 2017
  • Although Semisulcospira libertina is generally regarded as a supplement for the alleviation of alcohol hangover, little is known about its effects on cell metabolism. Therefore, this study was conducted to analyze the constituents of the extracts prepared using different extraction methods and to compare their biochemical properties. The amino acid contents were found to be much higher in acidic and enzymatic hydrolysates than hot water extracts from S. libertina. DPPH radical scavenging activities in acidic and enzymatic hydrolysates were higher than those of hot water extracts. Three types of S. libertina hydrolysate was added to HepG2 cells damaged by acetaminophen (AAP), after which the survival rate of HepG2 cell were measured. In addition, lactate dehydrogenase (LDH) activities in the culture media were evaluated. The survival rates of HepG2 cells were $77.0{\pm}4.3%$ and $81.5{\pm}1.3%$ at 3 h and 5h enzymatic hydrolysates, respectively. These cell survival rates were higher compared to those of the negative control group ($67.8{\pm}4.3%$) treated only with acetaminophen. Cellular toxicities induced by treatment with AAP were also significantly alleviated in response to treatment with the extracts of S. libertina. In addition, the activities of 2 key enzymes that metabolize ethanol, alcohol dehydrogenase and aldehyde dehydrogenase, were upregulated by 4.7- and 2.7-fold respectively in response to treatment with a 3 h enzymatic hydrolysate of S. libertina. Taken together, these results provide biochemical evidence of the method by which S. libertina exerts its biological functions, including the alleviation of alcohol hangover and the protection of liver cells against toxic insults.

Molecular Characterization of Hanwoo Glucose Transporter 4 Gene (한우 Glucose Transporter 4 유전자의 분자생물학적 해석)

  • Lee, S.M.;Jeong, Y.H.;Kim, H.M.;Park, H.Y.;Yoon, D.H.;Moon, S.J.;Chung, E.R.;Kang, M.J.
    • Journal of Animal Science and Technology
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    • v.47 no.6
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    • pp.1087-1094
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    • 2005
  • The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose transport protein. In the glucose transport protein family, GLUT4 plays a key role in cellular glucose uptake stimulated by insulin in skeletal muscles and adipose tissue in rodents and human. In this studies, we reported the identification, characterization, and expression of Hanwoo GLUT4 gene. The Hanwoo GLUT4 cDNA includes a 1527 bp open reading frame encoding a protein of 509 amino acids. The GLUT4 amino acid sequences of the Hanwoo show strong conservation with the corresponding sequences reported in other species. The highest mRNA expression of GLUT4 was detected in heart and lower expression was detected in rib meat, sirloin, and colon. We confirmed the expression of GLUT4 in the subcutaneous and small intestinal adipose tissue using RT-PCR. To investigate the expression of GLUT4 in the bovine intramuscular adipose differentiation, fibroblast-like cells were isolated from the sirloin of Hanwoo bull aged 12 months by collagenase digestion of minced tissue and cultured with activators of PPAR gamma. We identified that GLUT4 mRNA expression decreased during differentiation of preadipocytes into adipocyte in Korean cattle. These results indicated that function of GLUT4 in bovine adipose tissue was different from that of mouse and human.

Studies on the amino acid metabolism of young rice root (Part 3) - Effects of nitrogen, phosphorus, potassium and respiratory inhibitor on the enzyme activities of rice root - (수도근(水稻根)의 Amino산(酸) 대사(代謝)에 관한 연구 -제(第) 3 보(報) 수도근(水稻根)의 몇가지 효소(酵素)의 활성(活性)에 미치는 삼요소(三要素)및 호흡저해제(呼吸沮害劑)의 영향(影響))

  • Kim, Young Ung
    • Korean Journal of Soil Science and Fertilizer
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    • v.7 no.4
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    • pp.201-207
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    • 1974
  • Some effects of nitrogen, phosphorus, potassium and respiratory inhibitor on growth of rice plant and activity of GOT, GPT and peroxidase for the rice root were investigated. Obtained results were summarized as follows: 1. Growth of rice root and plant applied with $NO_3$-N in culture solution was generally increased in the length and weight compared with that of $NH_4$-N plot. On the other hand, the GOT, GPT and peroxidase activity was more increased in the $NH_4$-N plot than in the $NO_3$-N plot. 2. Oxidative power of ${\alpha}$-naphthylamine in rice root was stronger in the $NO_3$-N plot than in the $NH_4$-N plot. 3. When rice plant was cultured in the medium which did not supplied nitrogen, phosphorus or potassium, respectively, GOT activity was more decreased than GPT activity, while peroxidase activity was increased mostly in the potassium-free plot. 4. When rice plant was cultivated in the culture solution added respiratory inhibitor, NaF, plant height was shortened in the order of nitrogen-free > $NH_4$-H > urea-N > $NO_3$-N plot, and GOT and GPT activity was also decreased in the order of nitrogen-free > $NH_4$-N > urea-N > $NO_3$-N plot.

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Establishment of Reference Value of Insulin Using the Statistical Analysis (통계적 분석을 통한 Insulin의 정상 참고치 설정)

  • Kim, Whe-Jung;Yoon, Pil-Young;Shin, Young-Goon;Yoo, Seon-Hee;Cho, Shee-Man
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.1
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    • pp.143-146
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    • 2010
  • Purpose: Insulin is involved in carbohydrate metabolism and also it's very important because it increases storage of glycogen, synthesis of fatty acids, absorption of amino acid, synthesis of protein. Insulin is clinically useful when we evaluate fasting patients in hypoglycemia, classify and predict diabetes, assess the activity of ${\beta}$-cell, research insulin resistance. We are going to increase usability of insulin assay by establishing normal reference value according to statistical analysis. Material & Method: We selected 6,648 patients who visited asan health medical center from May to August in 2008. We set exclusion criteria as family of diabetes, diabetes medication, the past history of blood glucose rise, more than 100 mg/dL in normal fasting blood glucose, outside the scope of BMI 18.5~22.9 $kg/m^2$, and more than HbA1c 6.5%. We determine whether the subgroup is portioned as sex and age or not and establish normal reference value by conducting statistical analysis as Bayesian's method and Hoffman's method. Result: Portioning of subgroup as sex and age is not needed. By statistical analysis of Bayesian method, results 1.5-11.0 uIU/mL. By statistical analysis of Hoffman method, results 1.8~12.8 uIU/mL. Conclusion: We established 1.8~12.8 uIU/mL as Insulin normal reference value by Hoffman method. This is a similar value with reporting reference value 1.7~11.8 uIU/mL in kit. This will enhance the usability of insulin assay by establishing normal reference value.

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Genes of Wild Rice (Oryza grandiglumis) Induced by Wounding and Yeast Extract (상처와 효모추출물 처리조건에서 유발되는 야생벼 유전자 스크린)

  • Shin, Sang-Hyun;Im, Hyun-Hee;Lee, Jai-Heon;Kim, Doh-Hoon;Chung, Won-Bok;Kang, Kyung-Ho;Cho, Sung-Ki;Shin, Jeong-Sheop;Chung, Young-Soo
    • Journal of Life Science
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    • v.14 no.4
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    • pp.650-656
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    • 2004
  • Oryza grandiglumis (CCDD, 2n=48), one of the wild rice species, has been known to possess fungal-,bacterial-, and insect-resistance against sheath blight, rice blast, bacterial leaf blight and brown plant hopper (Nilaparvata lugens). To rapidly isolate differentially expressed genes responding to fungal and wounding stress, wounding and yeast extract were treated to O. grandiglumis for 24 hrs. Suppression subtractive hybridization (SSH) method was used to obtain differentially expressed genes from yeast extract and wounding treated plants. Seven hundreds and seventy six clones were obtained by subcloning PCR product, and colony array and screening were carried out using radio-isotope labeled cDNA probes prepared from the wounding and yeast extract treated plants. One hundred and fifteen colonies were confirmed as true positive ones. Average insert size of the clones were ranged from 400 bp to 700 bp and all the inserts were sequenced. To decide the identity of those clones, sequences were analyzed by sequence homology via GenBank database. The homology search result showed that 68 clones were matched to the genes with known function; 16 were related to primary metabolism, 5 to plant retrotransposons, 5 to defense related metallothionein-like genes. In addition to that, others were matched to various genes with known function in amino acid synthesis and processing, membrane transport, and signal transduction, so on. In northern blot analysis, induced expressions of ogwfi-161, ogwfi-646, ogwfi-663, and ogwfi-695 by wounding and yeast extract treatments were confirmed. The result indicates that SSH method is very efficient for rapid screening of differentially expressed genes.

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.

Hypolipidemic Effects of Peptide Fractions of Casein on Serum Lipids in Rats Fed Normal or High Fat Diet (정상 또는 고지방식을 섭취한 흰쥐에서 Casein 펩타이드 분획물이 혈청 지질농도에 미치는 영향)

  • 오주환;이연숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.2
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    • pp.263-270
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    • 2002
  • In the recent studies, many researchers are interested in foods as functional components rather than nutrient sources. Cow's milk is considered as an excellent food sources because of its many nutrients. Casein is a major milk protein and has been reported to have hyperlipidemic and hypercholesterolemic effects. But several reporters have suggested that peptide fractions and hydrolysate of casein have hypolipidemic effects differing from intact protein, casein. Therefore, the objective of the study was to investigate how the casein peptide fractions affect lipid metabolism in rats fed normal or high fat diets. The peptide fractions and hydrolysate of casein were obtained by casein hydrolysis with trypsin. The male rats (Sprague-Dawley), weighing approximately 150 g, were fed each experimental diet containing casein (CAS), casein hydrolysate (CH), casein hydrolysate precipitate (Cpt) and two kinds of peptide fractions (CL & CB) for three weeks, respectively. In the exit I, the male rats were fed normal fat diets (7% soybean oil & cholesterol-free; Expt. I), and in the expt II, fed high fat diets (18% beef tallow & 1% cholesterol; Expt. II). Crude protein contents were calculated from nitrogen contents. Amino acid composition of each fraction was also analyzed. The concentration of total lipid, total cholesterol and triglyceride in serum, liver and feces were measured. As the results of study, tole rats fed peptide fractions with normal fat diets (Expt. I) had no effects on total lipid, total cholesterol and triglyceride concentration in serum and liver and fecal excretion. However, in the rats fed hydrophobic casein peptide fractions (CB) with high fat diet, fecal lipids excretion were significantly increased and the lipids concentration of serum and those of liver tended to decrease, numerically.