• Title/Summary/Keyword: IGF-Ⅰ

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Expression of Porcine Acid-labile Subunit (pALS) of the 150-kilodalton Ternary Insulin-like Growth Factor Complex and Initial Characterization of Recombinant pALS Protein

  • Lee, Dong-Hee;Chun, Choa;Kim, Sang-Hoon;Lee, C.-Young
    • BMB Reports
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    • v.38 no.2
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    • pp.225-231
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    • 2005
  • Acid-labile subunit (ALS) is a component of the 150-kDa insulin-like growth factor-binding protein-3 (IGFBP-3) complex, which, by sequestering the majority of IGFs-I and -II and thereby prolonging the half-life of them in plasma, serves as a circulating reservoir of IGFs in mammalian species. A pGEX-2T plasmid and a baculovirus expression constructs harboring a coding sequence for glutathione-S transferase (GST)-porcine ALS (pALS) fusion protein were expressed in BL21(DE3) E. coli and Sf9 insect cells, respectively. The expressed protein was purified by glutathione or Ni-NTN affinity chromatography, followed by cleavage of the fusion protein using Factor Xa. In addition, pALS and hIGFBP-3 were also produced in small amounts in the Xenopus oocyte expression system which does not require any purification procedure. A 65-kDa pALS polypeptide was obtained following the prokaryotic expression and the enzymatic digestion, but biochemical characterization of this polypeptide was precluded because of an extremely low expression efficiency. The baculovirus-as well as Xenopus-expressed pALS exhibited the expected molecular mass of 85 kDa which was reduced into 75 and 65 kDa following deglycosylation of Asn-linked carbohydrates by Endo-F glycosidase, indicating that the expressed pALS was properly glycosylated. Moreover, irrespective of the source of pALS, the recombinant pALS and hIGFBP-3 formed a 130-kDa binary complex which could be immunoprecipitated by anti-hIGFBP-3 antibodies. Collectively, results indicate that an authentic pALS protein can be produced by the current expression systems.

Effect of LED Lighting Time on Productivity, Blood Parameters and Immune Responses of Dairy Cows (LED 점등시간이 젖소의 생산성, 혈액 매개변수 및 면역 반응에 미치는 영향)

  • Park, Jin-Ryong;Yoon, Nam-Jin;Belal, Shah-Ahmed;Shim, Kwan-Seob
    • Korean Journal of Organic Agriculture
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    • v.26 no.3
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    • pp.515-532
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    • 2018
  • Light is an essential and powerful element to animals. A light-emitting diode (LED) is most efficient in terms of economic benefits. The aim of the present study was to evaluate the effects of LED lighting time on milk production, milk composition, and the immune response of Holstein cows. Forty lactating cows were assigned to four experimental groups: control; natural daylight, treatment; am3-6, pm6-12 and pm6-am6. We found that there was no significant effect on the decrease ratio in milk production among the groups. Milk urea nitrogen (MUN) was significantly decreased in pm6-am6 and pm6-12 than the control. With regard to the hemolytic biochemical analysis, GLU was significantly increased and CRE, T-BIL were significantly decreased in the pm6-12 than the control. IGF-1 levels were significantly increased in pm6-12 compared to other groups. Besides, cortisol was significantly lowered in the pm6-12 than the control, while prolactin, IgA and IgG were not significant among the groups. In addition, catalase and glutathione peroxidase were also significantly increased in pm6-12 than the control. However, antioxidant enzyme activity and superoxide dismutase were not significant among the experimental groups. Therefore, it was concluded that LED lighting time had some impact on blood parameters and immune responses in dairy cows without any changes in milk production.

ATF3 Mediates Anti-Cancer Activity of Trans-10, cis-12-Conjugated Linoleic Acid in Human Colon Cancer Cells

  • Kim, Kui-Jin;Lee, Jihye;Park, Yeonhwa;Lee, Seong-Ho
    • Biomolecules & Therapeutics
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    • v.23 no.2
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    • pp.134-140
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    • 2015
  • Conjugated linoleic acids (CLA) are a family of isomers of linoleic acid. CLA increases growth arrest and apoptosis of human colorectal cancer cells through an isomer-specific manner. ATF3 belongs to the ATF/CREB family of transcription factors and is associated with apoptosis in colorectal cancer. The present study was performed to investigate the molecular mechanism by which t10, c12-CLA stimulates ATF3 expression and apoptosis in human colorectal cancer cells. t10, c12-CLA increased an apoptosis in human colorectal cancer cells in dose dependent manner. t10, c12-CLA induced ATF3 mRNA and luciferase activity of ATF3 promoter in a dose-dependent manner. The responsible region for ATF3 transcriptional activation by t10, c12-CLA is located between -147 and -1850 of ATF3 promoter. mRNA stability of ATF3 was not affected by t10, c12-CLA treatment. t10, c12-CLA increases $GSK3{\beta}$ expression and suppresses IGF-1-stimulated phosphorylation of Akt. The knockdown of ATF3 suppressed expression of $GSK3{\beta}$ and NAG-1 and PARP cleavage. The results suggest that t10, c12-CLA induces apoptosis through ATF3-mediated pathway in human colorectal cancer cells.

Study on the Effect of Gamissanghwa-tang and each Medicinal Plant Extract for the Hair Growth of the Mice using In vivo and In vitro Test (In vivo와 In vitro 실험에서 가미쌍화탕 및 구성한약재가 마우스의 모발 성장에 미치는 실험적 연구)

  • Yun Jeong Hun;Kim Nam Kwen;Lim Kyu Sang;Roh Seok Seon;Hwang Chung Yeon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.2
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    • pp.561-570
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    • 2004
  • To screen the effective materials for hair loss treatment, the Gamissanghwa-tang extracts were tested. As a result we found that the Gamissanghwa-tang extracts have the hair growth promoting effect. After topical application of each test materials to the back of CS7BL/6 mice, the earlier conversion of telogen-to-anagen phase was induced. In the experiments of 5α-reductase type II inhibition assay, Radix Paeoniae Alba, Semen Cuscutae showed effective potential to inhibit the activity of 5α-reductase type II. And hair growth index of the Gamissanghwa-tang extracts ranked as 1.2, especially the hair growth index of Fructus Rubi is highest as 1.8. But there were no plant extracts which have effect on the DNA proliferation of hair dermal papilla cell measured by [³H]thymidine incorporation, the expression of growth factors such as IGF-I, KGF, HGF estimated by RT-PCR and protein synthesis of vibrissae hair follicle measured by [/sup 35/S] cysteine incorporation. Cortex Cinnamomi showed anti-bacterial effect on P. ovale, Radix Paeoniae Alba has the highest radical scavening activity and Radix Glycyrrhizae has the highest effects of NO synthesis. These results suggest that Gamissanghwa-tang can be used as a potent treatment agent for helping hair growth stimulation.

Modification of Hydrogen Determinator for Total Hydrogen Analysis in Irradiated Zircaloy Cladding Tube (수소분석기 개조 및 조사후 지르칼로이 피복관의 총수소분석)

  • Park, Soon-Dal;Choi, Kwang-Soon;Kim, Jong-Goo;Joe, Kih-Soo;Kim, Won-Ho
    • Analytical Science and Technology
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    • v.12 no.6
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    • pp.490-497
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    • 1999
  • A hydrogen determinator was modified and installed in the glove box to analyse total hydrogen content in irradiated zircaloy tube. The analysis method of hydrogen is Inert Gas Fusion(IGF)-Thermal Conductivity Detection(TCD). The hydrogen recoveries of no tin method using Ti and Zr matrix standards, respectively, were available within $3{\mu}g$ of hydrogen. Also the smaller size of sample showed the better hydrogen recovery. It was found that the hydrogen standard of Ti matrix is avaliable to hydrogen analysis in zircaloy sample. The mean radioactivity of irradiated zircaloy sample was 10 mR/hr and hydrogen concentration was 130 ppm.

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Polymorphism of Insulin-like Growth Factor Binding Protein-4 Gene in 17 Pig Breeds and Its Relationship with Growth Traits

  • Wang, Wenjun;Hu, Xiaoxiang;Fei, Jin;Meng, Qinyong;Li, Ning
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.10
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    • pp.1490-1495
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    • 2007
  • Insulin-like growth factor binding protein-4 (IGFBP-4) is a member of the IGF super family, and regulates the action of IGFs. The polymorphism of porcine IGFBP-4 gene in 17 pig breeds (total n = 570) was detected by PCR-SSCP, and alleles A and B were detected. In these pig breeds, it was found that exotic pig breeds carried high frequencies of allele A, while Chinese native pig breeds carried high frequencies of allele B. The role of porcine IGFBP-4 was investigated in 172 F2 offspring of a $Lantang{\times}Lantang $ population. Forty eight growth traits were recorded for analyzing the association between IGFBP-4 gene polymorphism and quantitative performance traits. In this resource family, pigs with AA genotype had higher fore-body weight, bone weight of mid-body, bone weight of rear-body, fore-leg weight and rear-leg weight than those pigs with BB genotype (p<0.05); while pigs which carried BB genotype had higher back-fat thickness at C point and lard weight than those pigs with AA genotype (p<0.05); pigs with AA genotype had higher body weight than those with BB genotype; for meat quality traits, pigs with AA genotype had higher meat color than those of BB genotype (p<0.01), and pigs with BB genotype had higher marbling than those of AA and AB genotypes (p<0.01 and p<0.05, respectively).Based on these results, it is necessary to do more studies on IGFBP-4 before using the IGFBP-4 locus for the application of marker-assisted selection programs.

Effects of dietary omega-3 polyunsaturated fatty acids on growth and immune response of weanling pigs

  • Li, Qizhang;Brendemuhl, Joel H.;Jeong, Kwang C.;Badinga, Lokenga
    • Journal of Animal Science and Technology
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    • v.56 no.3
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    • pp.7.1-7.7
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    • 2014
  • The recognition that omega-3 polyunsaturated fatty acids (n-3 PUFA) possess potent anti-inflammatory properties in human models has prompted studies investigating their efficacy for animal growth and immunity. This study examined the effect of feeding an n-3 PUFA-enriched diet on growth and immune response of weanling piglets. Newly weaned pigs (averaging $27{\pm}2$ days of age and $8.1{\pm}0.7kg$ of body weight) were assigned randomly to receive a control (3% vegetable oil, n = 20) or n-3 PUFA-supplemented (3% marine n-3 PUFA, n = 20) diet for 28 day after weaning. Female pigs consuming the n-3 PUFA-enriched diet were lighter at week 4 post-weaning than those fed the vegetable oil supplement. Weanling pigs gained more weight, consumed more feed and had better growth to feed ratios between days 14 and 28 than between days 0 and 14 post-weaning. Plasma insulin-like growth factor I (IGF-I) decreased between days 0 ($87.2{\pm}17.0ng/mL$) and 14 ($68.3{\pm}21.1ng/mL$) after weaning and then increased again by day 28 ($155.2{\pm}20.9ng/mL$). In piglets consuming the vegetable oil-enriched diet, plasma tumor necrosis factor alpha (TNF-${\alpha}$) increased from $37.6{\pm}14.5$ to $102.9{\pm}16.6pg/mL$ between days 0 and 14 post-weaning and remained high through day 28 ($99.0{\pm}17.2pg/mL$). The TNF-${\alpha}$ increase detected in the piglets fed vegetable oil was not observed in the piglets fed n-3 PUFA. Results indicate that weaning induces considerable immune stress in piglets and that this stress can be mitigated by dietary supplementation of n-3 PUFA.

Insulin Resistance in Obese and Non-obese Patients with Polycystic Ovarian Syndrome (다낭성 난소 증후군 환자에서 비만군과 비비만군에서의 인슐린 저항성)

  • Lim, Y.K.;Lee, B.S.;Cho, E.J.;Cha, D.H.;Park, W.I.;Park, K.H.;Cho, D.J.;Lee, K.;Song, C.H.;Lee, E.J.
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.1
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    • pp.83-88
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    • 1994
  • This study was designed to investigate the relationship between insulin resistance and obesity in the pathogenesis of polycystic ovarian syndrome(PCO). Twenty-two women with PCO, of whom thirteen were non-obese with body mass index(BMI, kg/$m^2$) of <25 and nine were obese with BMI${\geq}$25 were studied. Eight non-obese control women and seven obese control women were studied. Serum concentrations of testosterone, lutenizing hormone(LH)/follicle-stimulating hormone(FSH) ratio, and insulin-like growth factor I (IGF-I) were found to be significantly higher(P<0.05) in PCO women compared with control women, which clearly is not related to obesity. Serum glucose, insulin, and C-peptide levels were measured during a 2-hour oral glucose tolerance test(OGTT). Non-obese and obese women with PCO both(P<0.05) compared with control women demonstrated significant hyperinsulinemia after OGTT. The degree of hyperinsulinemia was found to be significantly higher in the obese women with PCO compared with the non-obese women with PCO. We concluded that obesity may contribute to hyperinsulinemia, however may not playa central role in the pathogenesis of PCO.

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Molecular Characterization and Expression Pattern of Gene IGFBP-5 in the Cashmere Goat (Capra hircus)

  • Wang, X.J.;Shi, J.J.;Yang, J.F.;Liang, Y.;Wang, Y.F.;Wu, M.L.;Li, S.Y.;Guo, X.D.;Wang, Z.G.;Liu, D.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.606-612
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    • 2012
  • Insulin-like growth factor-binding protein-5 (IGFBP-5) is one of the six members of IGFBP family, important for cell growth, apoptosis and other IGF-stimulated signaling pathways. In order to explore the significance of IGFBP-5 in cells of the Inner Mongolian Cashmere goat (Capra hircus), IGFBP-5 gene complementary DNA (cDNA) was amplified by reverse transcription polymerase chain reaction (RT-PCR) from the animal's fetal fibroblasts and tissue-specific expression analysis was performed by semi-quantitative RT-PCR. The gene is 816 base pairs (bp) in length and includes the complete open reading frame, encoding 271 amino acids (GenBank accession number JF720883). The full cDNA nucleotide sequence has a 99% identity with sheep, 98% with cattle and 95% with human. The amino acids sequence shares identity with 99%, 99% and 99%, respectively. The bioinformatics analysis showed that IGFBP-5 has an insulin growth factor-binding protein homologues (IB) domain and a thyroglobulin type-1 (TY) domain, four protein kinase C phosphorylation sites, five casein kinase II phosphorylation sites, three prenyl group binding sites (CaaX box). The IGFBP-5 gene was expressed in all the tested tissues including testis, brain, liver, lung, mammary gland, spleen, and kidney, suggesting that IGFBP-5 plays an important role in goat cells.

A study on the effect on obesity and lipid metabolism in liver hypofunction animal-experimental model induced by Acetaminophen(AAP) injection (Acetaminophen(AAP)으로 유발한 간기능 저하 동물 모델에서의 비만 및 지질대사에 대한 영향 평가)

  • Park, Junghwan;Kim, Yoonha;Kwak, Jinyoung;Hong, Seojin;Park, Jungmi;Ahn, Taekwon
    • The Journal of Korean Medicine
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    • v.37 no.3
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    • pp.47-61
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    • 2016
  • Objectives: The objective of this research is to develop new animal-experimental model for Sasang Constitutional Medicine, especially for partial Taeyangin(one of four constitution which has good pulmonary function and poor hepatic function) by AAP intraperitoneal injection, and to estimate from the viewpoint of obesity and lipid metabolism. Methods: The C57bl/6J mice was divided into 4 groups ; Normal group, AAP group, High-Fat-Diet(HFD) group, and HFD+AAP group. 200mg AAP was injected intraperitoneally to the AAP group twice a week for six weeks, and HFD group was fed with 60%-High-fat Diet for six weeks. HFD+AAP group got both AAP injection and 60%-High-fat Diet at the same time for the same period. In this period, We measured the weight and Food Efficiency Ratio(FER, %) once a week. After six weeks, We conducted the blood chemical test from the groups, and extracted the fat tissue to measure weight. Results & conclusion: In the liver function test, two AAP groups had higher AST and ALP, and normal LDH. The blood level of creatinine from all groups were normal. The rate in weight was lesser by 7.8% in HFD+AAP group, and had lesser FER than HFD group. Also They had lesser Total cholesterol and LDL cholesterol, and had more HDL cholesterol than HFD group. HFD+AAP group hadmore glucose in serum and lesser Insulin-like Growth Factor 1(IGF-1) than HFD group.