• Title/Summary/Keyword: protein half life

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Recombinant human BMP-2/-7 heterodimer protein expression for bone tissue engineering using recombinant baculovirus expression system

  • Park, Seung-Won;Goo, Tae-Won;Kim, Seong Ryul;Choi, Kwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.32 no.2
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    • pp.49-53
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    • 2016
  • Bone morphogenetic proteins (BMPs) are essential growth factors for bone formation, skeletal development and bone regeneration. The BMP-2/7 heterodimer is known to have remarkable effects on osteogenic induction that are even stronger than the BMP-2 or BMP-7 homodimers. We designed a recombinant human BMP-2/7 (rhBMP-2/7) heterodimer protein with four glycine residues between BMP-2 and BMP-7 protein to facilitate free bond rotation of domains. The Baculovirus Expression Vector System (BEVS) is routinely used to produce recombinant proteins in the milligram scale. In this study, the BEVS was used to express the rhBMP-2/7 protein whrer the recombinant baculovirus was recovered in the host Sf9 cells. To confirm the biological activity of rhBMP-2/7 protein secreted from the BEVS as an osteogenic differentiation and induction factor, we measured the BMP-induced ALP activity. rhBMP-2/7 could be used as an alternative to BMPs to overcome limitations like short half-life and requirement for high concentrations. Furthermore, rhBMP-2/7 may be an efficient tool for various application studies such as bone regeneration and skeletal development.

Effect of Particle Size on the Physicochemical and Nutritional Properties of Egg Yolk Porridge (입자 크기에 따른 난황죽의 이화학적 및 영양학적 특성)

  • Kim, Hye-Ran;Kim, Mee-Ree
    • Journal of the East Asian Society of Dietary Life
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    • v.20 no.1
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    • pp.77-83
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    • 2010
  • The study was conducted to investigate the effect of particle size on the physicochemical properties of egg yolk-rice porridge. The pH of egg yolk-rice porridge was decreased when compared to that of the control, while the lightness and yellowness was increased as the rice particle size increased. The viscosity of whole particle egg yolk porridge was highest among the three porridges at $40^{\circ}C$. The protein content of the egg yolk-rice porridge was increased three-fold, when compared to that of the rice porridges. The total amino acid content of egg yolk-rice porridge was 1,500.6 mg/100 g, while that of rice was 1,147.5 mg/100 g. The Lys and Thr content of the amino acid content of egg yolk-rice porridge were also increased. Sensory evaluation results revealed that the half particle size rice egg yolk-rice porridge had the highest scores in color, taste and over-all preference. Based on these results, the half particle size egg yolk-porridge had good quality with respect to both the physicochemical and nutritional properties.

Quality Characteristics and Shelf-life of Rice Noodles Prepared with Red Ginseng Powder (홍삼 첨가에 따른 쌀국수의 품질특성과 저장성)

  • Kim, Eun-Mi
    • Culinary science and hospitality research
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    • v.14 no.1
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    • pp.161-169
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    • 2008
  • This study was performed to find out the quality characteristics of rice noodles by addition of red ginseng(0, 2, 6, 10%). The quality characteristics of the sample were estimated in terms of general composition, growth of microorganism and sensory evaluation. The results from this study were as follows. The protein, lipid and ash contents did not show significant difference in any of the groups. In dry rice noodles, moisture content significantly decreased in red ginseng groups but, in half-cooked rice noodles, moisture content significantly increased in 6 % and 10% red ginseng added groups(p<0.05). The microbial count showed less growth in red ginseng added groups after 3 months(p<0.05). According to sensory evaluation, surface color proved very good in the 10% red ginseng added group among the training panel while very good in the 2% red ginseng added group among consumers. Flavor was good in red ginseng added groups(p<0.05). Taste was very good in the 3% red ginseng added group. Appearance and overall quality were highest in the 2% and 6% red ginseng added groups(p<0.05). Therefore, rice noodles containing 2% or 6% red ginseng were most preferable and safe during 5 months and 6 days in dry and half-cooked noodles respectively.

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In Vitro Ovulation and Prostaglandin Synthesis by Ovarian Follicles of Rana dybowskii

  • Kong, Hye-Young;Chang, Kyung-Ja;Im, Wook-Bin;Kwon, Hyuk-Bang
    • Animal cells and systems
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    • v.3 no.4
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    • pp.385-391
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    • 1999
  • Changes in the levels of prostaglandian F$_{2a}$ (PGF$_{2a}$) and E$_2$ (PGE$_2$) in culture medium during in vitro ovulation of Rana dybowskii follicles were examined. The ovulation was induced by frog pituitary homogenate (FPH) or TPA (12-O-tetradecanoylphorbol-13-acetate, a protein kinase activator) and the levels of PGs were measured by radioimmunoassay. When the ovarian follicles were cultured, only a few oocytes were ovulated by 12 h, but half of them were ovulated by 24 h in response to FPH, whereas around 30% of oocytes were ovulated by 12 h and maximum ovulation (around 50%) occurred by 24 h in response to TPA. Without any stimulation (control), no ovulation occurred. TPA elevated the level of PGF$_{2a}$ to high levels when compared to control (basal levels), but the increase by FPH was less evident. Likewise, the levels of PGE$_2$ increased markedly in response to TPA, but rather decreased by FPH treatment. Interestingly, PGF$_{2a}$ induced ovulation but PGE$_2$ suppressed FPH- or PGF$_{2a}$-induced oocyte ovulation. Basal levels of PGs Increased steadily during culture. When theca/epithelium (THEP) layer and granulosa cell-enclosed oocytes (GCEOs) were separated by microdissection and cultured independently, higher levels of both PGs were secreted by THEP than by GCEOS. Synthesis of PGs by follicle or follicular components was strongly suppressed by exogenous cAMP or indomethacin. These results suggest that: 1) PGF$_{2a}$ plays an important role in Rana ovulation, 2) protein kinase C is involved in PGs production, and 3) thecal epithelium layer is responsible for the PGs production in Rana.

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Pharmacokinetic profile and tissue distribution of sulfamethazine in pigs and rats (돼지와 랫트에서 sulfamethazine의 약물동태학 및 조직분포)

  • Yun, Hyo-in;Park, Seung-chun;Oh, Tae-kwang;Cho, Joon-hyoung;Park, Jong-myeong
    • Korean Journal of Veterinary Research
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    • v.37 no.2
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    • pp.291-299
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    • 1997
  • In order to establish optimal dosage schedules and withdrawal times for sulfamethazine(SMZ) in pigs, pharmacokinetic and tissue distribution experiments were conducted in pigs. For comparative purposes, tissue depletion kinetics are also studied in rats. From three pigs administered with SMZ i.v., the pharmacokinetic profile of SMZ in two pigs was adequately described by a one-compartment open model whereas that in one pig was patterned after a two-compartment open model. Volume of distribution(Vd) was 0.48~0.57 L/kg and biological half-life($t_{1/2}$) was 11.8-16.8 h. From three pigs dosed with SMZ p.o., pharmacokinetic profile was explainable with a one-compartment open model. Time to reach maximum SMZ concentration in serum (Tmax) was 2.8 h, 3.2 h and 7.5 h. Elimination half-life was 2.8-7.5 h. The descending order in concentration of SMZ was plsama > kidney > liver > lung > heart > pancreas > spleen > duodenum > ileum > brain > adipsoe tissue from three pigs sacrificed at 5h, 29h and 54h after the administration of SMZ, p.o.. The protein binding of SMZ in pigs was 55.2%($2.5{\mu}g/ml$), 71.5% ($5{\mu}g/kg$) and 71.5%($10{\mu}g/ml$). The mean systemic bioavailability (F) of SMZ p.o. was 49.1 %. Meanwhile the pharmacokinetic profile of SMZ in rats was adequately described by a one-compartment open model. Absorption of SMZ p.o. in the rat was very rapid. In conclusion, the oral optimal dosage regimen of SMZ for pigs was the initial dose of 45.7 mg/kg followed by the maintenance dose of 30.2 mg/kg for high specific pathogens to SMZ. The time to reach below the stipulated residual allowable concentration (0.1 ppm) was calculated 93 h after oral administration of 200 mg/kg recommended by manufactureres.

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Interaction Between Acid-Labile Subunit and Insulin-like Growth Factor Binding Protein 3 Expressed in Xenopus oocytes

  • Park, Kyung-Yi;Lee, Dong-Hee
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.99-99
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    • 2001
  • The acid-labile subunit (ALS) associates with insulin-like growth factor (IGF)-I or -II and IGF binding protein-3 (IGFBP-3) to form a 150-kD complex in the circulation. This complex is thought to regulate the serum IGFs by restricting them in the vascular system and promotes their endocrine actions. Little is known about how ALS binds to IGFBP3, which connects the IGFs to ALS. Xenopus oocyte was utilized to study the function of ALS in assembling IGFs into the ternary complexes. Xenopus oocyte was shown to correctly translate in vitro transcribed mRNAs of ALS and IGFBP3. IGFBP3 and ALS mRNAs were injected in mixture and their products were immunoprecipitated by antisera against ALS and IGFBP3. Contrary to the traditional reports that ALS interacts only with IGF-bound IGFBP3, this study shows that ALS is capable of forming a binary complex with IGFBP3 in the absence of IGF. When cross-linked by disuccinimidyl substrate, band representing ALS-IGFBP3 complex was evident on the PAGE. IGFBP3 movement was monitored according to the distribution between the hemispheres. Following a localized translation in the vegetal hemisphere, IGFBP3 was shown to remain in the vegetal half in the presence of ALS. Different from wild type IGFBP3, however, mutant IGFBP3 freely diffused into the animal half despite the presence of ALS. Taken together, this study suggests that ALS may play an important role in sequestering IGFBP3 polypeptides via the intermolecular aggregation. Studies using this heterologous model will lead to a better understanding of the IGFBP3 and ALS assembling into the ternary structure and circulating IGF system.

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Pharmacokinetics of LB20304, a New Fluoroquinolone, in Rats and Dogs

  • Seo, Mi-Kyeong;Lee, Sun-Hwa;Choi, Yun-Jeong;Jeong, Yi-Na;Lee, Sung-Hack;Kim, In-Chull;Lee, Yong-Hee
    • Archives of Pharmacal Research
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    • v.19 no.5
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    • pp.359-367
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    • 1996
  • The pharmacokinetics of LB20304 was investigated following intravenous (IV) and oral administration to rats and dogs. Additionally, in vitro metabolism and serum protein binding studies were also conducted. The total body clearance, apparent volume of distribution, terminal half-life, and extent of bioavailability were 21.8 ml/min/kg, 2265 ml/kg, 93.6 min, and 30.8% for rats; and 7.95 ml/min/kg, 4144 ml/kg, 363 min, and 81.1% for dogs, respectively. LB20304 was stable in the liver microsome containing NADPH generating system and its serum protein binding was 58.5-65.8% for rats, 19.1-29.6% for dogs, and 56.9-59.6% for humans. Its tissue concentration levels in lever, stomach, small intestine, and kidney were 9.5 to 26.1 times greater than plasma level, but the concentration in testis was quite low and that in brain was negligible in rats. The 48 hr urinary recovery of the dose was 44% for IV dosing and 14% for oral dosing, shereas the 48 hr biliary recovery of the dose was 6.4% for IV dosing and 4.5% for oral dosing in rats. In summary, the pharmacokinetic properties of LB20304 were characterized by its good oral absorption, long plasma half-life, and good tissue distribution.

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Effects of Cholic Acid/CDCA and FGF-19 on the Protein Levels of the Endogenous Small Heterodimer Partner (SHP) in the Mouse Liver and HepG2 Cells (생쥐의 간과 HepG2 세포에 있어서 내인성 small heterodimer partner (SHP)의 단백질 수준에 미치는 cholic acid/CDCA 및 FGF-19의 효과)

  • Min, Gye-Sik
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1731-1736
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    • 2009
  • Recent studies determined that a chronic western-style diet increased the endogenous small heterodimer partner (SHP) protein levels in mice. In experiments with cell cultures, chenodeoxy cholic acid (CDCA) treatment increased endogenous SHP protein levels and reduced the degradation rate of exogenously expressed flag-SHP levels in the human hepatoma cell line, HepG2 cells. In addition, bile acid-induced intestinal fibroblast growth factor-19 (FGF-19) increased the half-life of the exogenously expressed SHP when HepG2 cells were transfected with ad-flag-SHP. However, both the expression level and the degradation rate of the endogenous SHP in response to cholic acid and FGF-19 have not been well understood, either in mice or in cultured HepG2 cells. This study examined the effects of cholic acid treatment on the endogenous SHP protein levels in mice and the effects of FGF-19 on the degradation rate of the endogenous SHP protein in HepG2 cells. Mice fed 0.5% cholic acid in normal chow showed an increase in endogenous SHP protein levels during both 12 hr and 24 hr treatment periods as compared to control mice fed only normal chow. In cultured HepG2 cells, treatment with CDCA did not noticeably change the rate of degradation in the endogenous SHP protein from cells not treated with CDCA. Although consistent with the previous studies on the exogenous ad-flag-SHP protein, treatment with FGF-19 significantly decreased the degradation rate of the endogenous SHP protein when HepG2 cells were treated with cyclohexamide. These results suggest that both bile acids and FGF-19 increase the endogenous SHP protein levels in mouse liver and HepG2 cells.

The Dynamics of Protein Decomposition in Lakes of Different Trophic Status - Reflections on the Assessment of the Real Proteolytic Activity In Situ

  • Siuda, Waldemar;Kiersztyn, Bartosz;Chrost, Ryszard J.
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.897-904
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    • 2007
  • The aim of this paper is to discuss the methodology of our investigation of the dynamics of protein degradation and the total in situ protealytic activity in meso/eutrophic, eutrophic, and hypereutrophic freshwater environments. Analysis of the kinetics and rates of enzymatic release of amino acids in water samples preserved with sodium azide allows determination of the concentrations of labile proteins $(C_{LAB})$, and their half-life time $(T_{1/2})$. Moreover, it gives more realistic information on resultant activity in situ $(V_{T1/2})$ of ecto- and extracellular proteases that are responsible for the biological degradation of these compounds. Although the results provided by the proposed method are general y well correlated with those obtained by classical procedures, they better characterize the dynamics of protein degradation processes, especially in eutrophic or hypereutrophic lakes. In these environments, processes of protein decomposition occur mainly on the particles and depend primarily on a metabolic activity of seston-attached bacteria. The method was tested in three lakes. The different degree of eutrophication of these lakes was clearly demonstrated by the measured real proteolytic pattern and confirmed by conventional trophic state determinants.

Ig G fusion 단백질을 사용한 리간드-수용체의 상호작용

  • 천혜경
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.11a
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    • pp.143-145
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    • 1994
  • Chimeric fusion proteins involving IgG have proven valuable in studying protein-protein interactions and may possess therapeutic applications as well. For example, three receptor subtypes for the natriuretic peptides, when fused to the Fc portion of human IgG ${\gamma}$ chain, were quantitatively and qualitatively indistinguishable from the native receptor, thus allowing detailed structure-function studies of the receptor. In an attempt to block human immunodeficiency virus infectivity with soluble derivatives of CD4, a CD4/IgG Fc chimeric molecule was shown to increase the plasma half life of soluble CD4 and possessed the added advantage of IgG Fc-mediated placental transfer. In the case of the KGFR, this approach provided a framework for dissection of its ligand binding domains and made it possible to demonstrate that high affinity binding sites for two ligands, aFGF and KGF, reside within different receptor Ig-like domains. Chimeric molecules fused to immunoglobulins would have the advantages of secretion from transfected cells as well as detection and purification from medium utilizing Staphylococcus aureus Protein A. In addition, where highly related receptors make their discrimination very hard due to the difficulties in generating specific immunochemical probes, IgG fusion protein with tailor-made specificities confers particular advantages to elucidate patterns of receptor distribution and expression. The approach described here may have general applications in defining ligand-receptor interactions as well as searching for specific agonists and antagonists of receptor function.

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