• Title/Summary/Keyword: Molecular weight determination

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Determination of Genetic Diversity Using 15 Simple Sequence Repeats Markers in Long Term Selected Japanese Quail Lines

  • Karabag, Kemal;Balcioglu, Murat Soner;Karli, Taki;Alkan, Sezai
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.12
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    • pp.1696-1701
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    • 2016
  • Japanese quail is still used as a model for poultry research because of their usefulness as laying, meat, and laboratory animals. Microsatellite markers are the most widely used molecular markers, due to their relative ease of scoring and high levels of polymorphism. The objective of the research was to determine genetic diversity and population genetic structures of selected Japanese quail lines (high body weight 1 [HBW1], HBW2, low body weight [LBW], and layer [L]) throughout 15th generations and an unselected control (C). A total of 69 individuals from five quail lines were genotyped by fifteen microsatellite markers. When analyzed profiles of the markers the observed ($H_o$) and expected ($H_e$) heterozygosity ranged from 0.04 (GUJ0027) to 0.64 (GUJ0087) and 0.21 (GUJ0027) to 0.84 (GUJ0037), respectively. Also, $H_o$ and $H_e$ were separated from 0.30 (L and LBW) to 0.33 (C and HBW2) and from 0.52 (HBW2) to 0.58 (L and LBW), respectively. The mean polymorphic information content (PIC) ranged from 0.46 (HBW2) to 0.52 (L). Approximately half of the markers were informative ($PIC{\geq}0.50$). Genetic distances were calculated from 0.09 (HBW1 and HBW2) to 0.33 (C and L). Phylogenetic dendrogram showed that the quail lines were clearly defined by the microsatellite markers used here. Bayesian model-based clustering supported the results from the phylogenetic tree. These results reflect that the set of studied markers can be used effectively to capture the magnitude of genetic variability in selected Japanese quail lines. Also, to identify markers and alleles which are specific to the divergence lines, further generations of selection are required.

Collaborative Study for the Establishment of KFDA Reference Standard for Somatropin (KS 98/674) (재조합 사람성장호르몬(소마트로핀)의 KFDA 표준품(KS 98/674) 설정 연구)

  • Shin, Won;Joung, Jee-Won;Jin, Jae-Ho;Bristow, Adrian F.;Sohn, Yeo-Won
    • YAKHAK HOEJI
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    • v.45 no.2
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    • pp.227-236
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    • 2001
  • The complexity and variability of both the biologicals and the bioassays used to test them led to the use of the reference standard- a sample of the product of defined purity and potency, against which all preparations of that product must be calibrated. In order to prepare and establish KFDA reference standard for recombinant human growth hormone (somatropin), somatropin substance was filled in ampoules in National Institute for Biological Standards and Control (NIBSC). The candidate KFDA reference standard for somatropin (designated as 98/674) was evaluated to determine the suitability of serving as a KFDA reference standard for somatropin by the collaborative study, in which 10 laboratories participated. Physicochemical analysis and in vivo bioassay were performed by direct comparison with the international somatropin standard 88/624. 98/674 was identified as somatropin by SDS-PAGE, IEF, peptide mapping, and HPLC. Determination of somatropin content by SE-HPLC yielded a mean estimate of 2.01 mg somatropin per ampoule. Data from the study also yielded mean values of 0.39 $\pm$ 0.26% for high molecular weight impurities by SE-HPLC and mean values of 2.13 $\pm$ 1.29% for somatropin related proteins by RP-HPLC. Estimates of relative potency by weight gain bioassay in the hypophysectomised rats showed that relative potency of KS 98/674 was 1.07 aganist IS 88/624. Based on the results of the collaborative study, the candidate reference standard for somatropin is suitable to serve as a KFDA reference standard for somatropin.

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Antioxidation Effect of Leg Bone Extracts and Enzyme Hydrolysates from Jeju Crossbred Horses (Jeju Native Horse×Thoroughbred) (제주 재래마와 더러브렛종과의 교잡종인 한라마 사골추출물과 효소분해물의 항산화 효과)

  • Kim, Dongwook;Pak, Jae-In;Chae, Hyun-Seok;Kim, Young-Boong;Jang, Aera
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1147-1154
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    • 2013
  • This study was conducted to evaluate the antioxidation activity of Jeju crossbred horse (Jeju native horse ${\times}$ thoroughbred) leg bone extracts (HLBE) and its enzyme hydrolysates by determination of 1,1-diphenyl-2picryl-hydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzo thiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity, ferric reducing/antioxidant power (FRAP), and oxygen radical absorbance capacity (ORAC). HLBE was extracted with hot water for 24 hr and lyophilized. The lyophilized HLBE was hydrolyzed using multifect PR6L (MP), pepsin (PS), and a pepsin and pancreatin mixture (PSPC) for 4 hr at 60, 50, and $50^{\circ}C$, respectively. The hydrolysates were separated by a molecular weight of 3 kDa more or less. When the yield of HLBE was 100%, the yield of hydrolysates less than 3 kDa of MP, PS, and PSPC was 10.86, 3.26, and 8.00%, respectively. Enzyme hydrolysates with low molecular weight less than 3 kDa in MP and PSPC showed significantly higher DPPH, ABTS radical scavenging activity, and ORAC compared to HLBE and its hydrolysates with more than 3 kDa. However, the FRAP of the hydrolysates less than 3 kDa in PS was significantly higher than in MP. These results suggest that low molecular weight enzyme hydrolysates less than 3 kDa have more powerful antioxidation activity, especially when they are hydrolyzed by MP and PSPC rather than PS. Therefore, low molecular weight enzyme hydrolysates of HLBE hydrolyzed with MP and PSPC have significant potential as antioxidants in the food industry. Further in vivo studies are required to support the antioxidation activities of the hydrolysates in vitro.

Addition Polymerization of 5-Norbornene-2-carboxylic Acid Esters Using Palladium Catalyst System: Synthesis of Monomers, Effect of Their Stereochemistry on Polymerization Behavior (Palladium 촉매를 이용한 5-Norbornene-2-carboxylic Acid Esters의 부가 중합: 단량체의 합성, 단량체의 Stereochemistry(Endo-, Exo-이성질체)가 고분자의 중합 거동에 미치는 영향)

  • Chung, Hae-Kang;Shim, Hyoug-Sub;Jeon, Seung-Ho;Kim, Ji-Heung;Nam, Sung Woo;Jeon, Boong Soo;Kim, Young Jun
    • Polymer(Korea)
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    • v.39 no.3
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    • pp.487-492
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    • 2015
  • The effects of chemical structure of alkyl groups of norbornene carboxylic alkyl esters(methyl, octyl, 4-chlorobenzyl) and endo/exo ratios of norbornene monomers on activity of palladium catalyst and polymerization behavior were investigated. Norbornene ester monomers were synthesized from the reaction of 5-norborene-2-carboxylic acid and various alcohols. Polymerization catalyst, di-${\mu}$-chloro-bis(-methoxybicyclo[2,2,1]-hept-2-ene)palladium(II) (DCBMP), was synthesized according to the literature procedure and silver hexafluoroantimonate ($AgSbF_6$) was used as a conjugate anion source. Gel permeation chromatography (GPC), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) were the principal techniques for polymer characterization and $^1H$ NMR spectroscopy was used for chemical structures determination of monomers and polymers. For all of the norbonene alkyl esters GPC data showed that when the amounts of endo isomers exceeded those of exo isomers decreased molecular weight polymers were obtained probably due to the decreased catalyst activity. Polymerizations were conducted by varying the monomer/catalyst mole ratios (100:1, 200:1, 300:1). When 300:1 monomer/catalyst ratio was employed it was possible to synthesize high molecular weight ($M_n=27500g/mol$), film forming polymer from exo-norbornene carboxylic acid octyl ester.

Effect of Sub- and Super-critical Water Treatment on Physicochemical Properties of Porcine Skin

  • Jo, Yeon-Ji;Kim, Jae-Hyeong;Jung, Kyung-Hun;Min, Sang-Gi;Chun, Ji-Yeon
    • Food Science of Animal Resources
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    • v.35 no.1
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    • pp.35-40
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    • 2015
  • Super- and sub-critical water treatments have been of interest as novel methods for protein hydrolysis. In the present study, we studied the effect of sub-critical water (Sub-$H_2O$, $300^{\circ}C$, 80 bar) treatment as well as super-critical water (Super-$H_2O$, $400^{\circ}C$, 280 bar) treatment on the physicochemical properties of porcine skin (PS), which has abundant collagen. Porcine skin was subjected to pre-thermal treatment by immersion in water at $70^{\circ}C$, and then treated with sub- or super-critical water. Physicochemical properties of the hydrolysates, such as molecular weight distribution, free amino acid content, amino acid profile, pH, color, and water content were determined. For the molecular weight distribution analysis, 1 kDa hydrolyzed porcine skin (H-PS) was produced by Super-$H_2O$ or Sub-$H_2O$ treatment. The free amino acid content was 57.18 mM and 30.13 mM after Sub-$H_2O$ and Super-$H_2O$ treatment, respectively. Determination of amino acid profile revealed that the content of Glu (22.5%) and Pro (30%) was higher after Super-$H_2O$ treatment than after Sub-$H_2O$ treatment, whereas the content of Gly (28%) and Ala (13.1%) was higher after Sub-$H_2O$ treatment. Super-$H_2O$ or Sub-$H_2O$ treatment affected the pH of PS, which changed from 7.29 (Raw) to 9.22 (after Sub-$H_2O$ treatment) and 9.49 (after Super-$H_2O$ treatment). Taken together, these results showed that Sub-$H_2O$ treatment was slightly more effective for hydrolysis than Super-$H_2O$ was. However, both Sub-$H_2O$ and Super-$H_2O$ treatments were effective processing methods for hydrolysis of PS collagen in a short time and can be regarded as a green chemistry technology.

A Diagnostic Algorithm after Newborn Screening for 21-hydroxylase Deficiency (선천성 부신 과형성증(21-hydroxylase 결핍)의 신생아 선별 검사 후 진단 알고리즘)

  • Cho, Sung Yoon;Ko, Jung Min;Lee, Kyung-A
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.16 no.2
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    • pp.70-78
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    • 2016
  • 21-hydroxylase deficiency (21-OHD), most common form of congenial adrenal hyperplasia, is categorized into classical forms, including the salt-wasting (SW) and the simple virilizing (SV) types, and nonclassical (NC) forms based on the severity of the disease. Newborn screening for 21-OHD has been performed in Korea since 2006. $17{\alpha}$-hydroxyprogesterone (17-OHP) is a marker for 21-OHD and is measured using a radioimmunoassay or a fluoroimmunoassay. Premature and low birth weight infants are likely to give false positive 17-OHP findings, therefore, cutoff values for these infants should be determined based on gestational weeks or birth weight. ACTH simulation test is helpful when the 17-OHP shows equivocal increase, and it is gold standard for diagnosis of NC type. Recently, liquid chromatography linked with tandem mass spectrometry was developed for rapid, highly specific, and sensitive analysis of multiple analytes. Molecular analysis of CYP21A2 is useful for confirming diagnosis of mild SV or NC type, predicting prognoses, and genetic counseling. In order to make newborn screening for 21-OHD more efficient, early detection of boy with SW type, early determination of girl with ambiguous genitalia, detection of NC type, and overcoming of false positive in premature and low birth weight infants should be considered. Above all, early treatment should be started when the patient is suspected as having 21- OHD clinically before confirming the diagnosis to prevent adrenal crisis. Here, author reviewed recent articles of guideline and proposed guideline for 21-OHD.

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Characterization of the Physical Form of Allergenic Cry j 1 in the Urban Atmosphere and Determination of Cry j 1 Denaturation by Air Pollutants

  • Wang, Qingyue;Morita, Jun;Gong, Xiumin;Nakamura, Shinichi;Suzuki, Miho;Lu, Senlin;Sekiguchi, Kazuhiko;Nakajima, Takuya;Nakajima, Daisuke;Miwa, Makoto
    • Asian Journal of Atmospheric Environment
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    • v.6 no.1
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    • pp.33-40
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    • 2012
  • In this study, we characterized the physical form of allergenic Cry j 1 in the urban atmosphere. Through an immunofluorescence antibody method, we showed that allergenic Cry j 1 exists as fine particles (${\leq}1.1{\mu}m$). To determine Cry j 1 concentrations and its particle size distribution, we used the ELISA method to confirm that most Cry j 1 exists as fine particles in the urban atmosphere and is found at high concentrations on fine day next to rainy day. Furthermore, we evaluated Cry j 1 denaturation by using the Biacore J system based on the surface plasmon resonence (SPR) principle and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). We showed that the dissociation constant ($K_D$) of Cry j 1 that has been exposed to urban polluted air is lower ($1.76{\times}10^{-14}$ M) than that of Cry j 1 ($1.32{\times}10^{-9}-3.37{\times}10^{-9}$ M) of original pollen grains that has not been exposed to air pollutants. Cry j 1 turns into low molecular weight proteins by reacting with various acidic solutions. In sum, we showed that allergenic Cry j 1 exists as fine particles that can deposit in the lower respiratory tract. This finding clarifies the relationship between Japanese cedar pollinosis and air pollutants.

Effect of Atrial Natriuretic Peptide on the Proliferation and Activity of Osteoblastic Cells

  • Lee, Jong-Ryeul;Ko, Seon-Yle;Kim, Jung-Keun;Kim, Se-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.4
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    • pp.283-289
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    • 2000
  • Natriuretic peptides comprise a family of three structurally related peptides; atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP). The present study was performed to investigate the effect of ANP on the proliferation and activity of ROS17/2.8 and HOS cells which are well-characterized osteoblastic cell lines. ANP dose-dependently decreased the number of ROS17/2.8 and HOS cells after 48-hour treatment. ANP generally increased the alkaline phosphatase activity of ROS17/2.8 and HOS cells after 48 hr treatment, regardless of the fact that basal activity of alkaline phosphatase was much lower in HOS cells compared to that of ROS17/1.8 cells. ANP increased the NBT reduction by ROS17/2.8 and HOS cells. ANP showed the variable but no significant effect on the nitric oxide production by ROS17/2.8 and HOS cells. ROS17/2.8 and HOS cells produced and secreted gelatinase into culture medium, and this enzyme was thought to be the gelatinase A type with the molecular weight determination. The gelatinase activity produced by ROS17/2.8 cells was increased by the treatment of ANP. However, the enzyme activity was not affected by ANP treatment in the HOS cell culture. In summary, ANP decreased the proliferation and increased the alkaline phosphatase activity and NBT reduction of osteoblasts. These results indicate that ANP is one of the important regulators of bone metabolism.

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Determination of Medium Components in the Flocculating Activity and Production of Pestan Produced by Pestalotiopsis sp. by Using the Plackett-Burman Design

  • Moon, Seong-Hoon;Hong, Soon-Duck;Kwon, Gi-Seok;Suh, Hyun-Hyo;Kim, Hee-Sik;An, Keug-Hyun;Oh, Hee-Mock;Mheen, Tae-Ick;Yoon, Byung-Dae
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.341-346
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    • 1998
  • Optimization for the production of Pest an was followed by the Plackett-Burman Design, using modified Czapek-dox medium as the starting point. At the flask level, $K_2HPO_4$, $MgSO_4{\cdot}7H_2O$, and aeration variables positively affected the Pestan production, DCW (dry cell weight), apparent viscosity, and flocculating activity response. KCI and $FeSO_4{\cdot}7H_2O$ negatively affected the Pestan production, DCW, apparent viscosity, and flocculating activity response. Aeration variable was shown to have a positive effect on only the flocculating activity response among Pestan production, DCW, and apparent viscosity responses. In comparison of the positive and negative variables media conditions, Pestan production and flocculating activity differed by about 9 and 125 times, respectively. In particular, at the jar fermentor level, the aeration variable was the most important factor of the all responses (pestan production, DCW, apparent viscosity, flocculating activity, and anionic charge density). The flocculating activity and apparent viscosity of Pestan were closely related to the molecular chain length and charge density.

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Simultaneous Determination of Amoxicillin and Ampicillin in Fish Meat Using High-Performance Liquid Chromatography (HPLC를 이용한 어류 중의 Amoxicillin과 Ampicillin 항생제 동시 분석법)

  • Jo, Mi-Ra;Kim, Poog-Ho;Lee, Tae-Seek;Oh, Eun-Gyoung;Yu, Hong-Sik;Lee, Hee-Jung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.39 no.6
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    • pp.454-459
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    • 2006
  • A simultaneous high-performance liquid chromatography assay method for amoxicillin and ampicillin in fish products was developed, evaluated, and validated by monitoring these antibiotics in fish samples obtained from aquaculture and distribution. The recovery rate of this method was higher than those of conventional methods and was 95.3-106.6% for amoxicillin and 81.4-92.4% for ampicillin. Our pretreatment procedure sufficiently removed or reduced materials affecting HPLC analysis, such as low-molecular-weight substances. The performance limit of this method was evaluated as 0.01 ppm of amoxicillin and ampicillin in fish muscle. Finally, 171 fish samples, including olive flounder (Paralichthys olivaceus), common sea bass (Lateolabrax japonicus), and black rock fish (Sebastes schlegeli) collected from fish farms in the coastal area between April and September 2005 were analyzed to evaluate the overall efficiency of the method and to monitor the actual of amoxicillin and ampicillin usage in fish farms. The results indicated that the developed method was suitable for analyzing amoxicillin and ampicillin in fish muscle, and determined that those antibiotics were being used for fish farming but were not detected in fish samples during the shipping and distribution stages.