• 제목/요약/키워드: Molecular weight determination

검색결과 105건 처리시간 0.026초

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
    • /
    • 제29권12호
    • /
    • pp.1696-1701
    • /
    • 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.

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

  • 신원;정지원;진재호;;손여원
    • 약학회지
    • /
    • 제45권2호
    • /
    • pp.227-236
    • /
    • 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.

  • PDF

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

  • 김동욱;박재인;채현석;김영붕;장애라
    • 생명과학회지
    • /
    • 제23권9호
    • /
    • pp.1147-1154
    • /
    • 2013
  • 본 연구는 제주 재래마와 더러브렛(Thoroughbred)종 교잡종인 제주산 마(한라마) 사골 추출분말(HLBE)과 효소분해물의 항산화 효과를 DPPH, ABTS 라디칼 소거능, FRAP, ORAC 방법을 이용하여 확인하기 위해 실시하였다. 말 사골 추출물은 열수로 8시간씩 3회 추출하여 혼합한 후 동결 건조하여 분말로 이용하였다. 동결 건조된 말 사골 추출물은 식품효소인 multifect PR 6L (MP), pepsin (PS), pepsin+ pancreatin 혼합효소(1:1, PSPC)를 첨가하고 4시간 동안 각 효소의 최적활성 온도인 60, 50, $50^{\circ}C$에서 분해하였다. 분해 후 3 kDa 이상의 분획과 이하의 분획으로 분리하였다. 말 사골추출물의 수율을 100%으로 고정하였을 때 각 효소에 의해 분해되어 분리한 3 kDa 이상의 크기의 분획의 수율은 각각 10.86%, 3.26%, 8.00%이었다. 단백질 함량은 말 사골 추출물보다 3 kDa 이하의 저분자 분획에서 유의적으로 감소하였다. MP와 PSPC처리군의 3 kDa 이하의 저분자 분획은 말 사골추출물과 3 kDa 이상의 효소분해물에 비해 유의적으로 높은 DPPH와 ABTS 라디칼 소거능과 ORAC 활성을 나타내었다. 그러나 PS처리군 내 3 kDa 이하의 저분자 분획은 MP처리군 보다 유의적으로 높은 FRAP 활성을 나타내었다. 이러한 결과는 말 사골 추출물 자체 보다는 효소를 이용하여 분리한 3kDa 이하의 저분자 펩타이드가 더욱 강한 항산화 효과를 갖는 것을 제시하는 것으로 특히 pepsin에 의해 생성된 저분자펩타이드 보다는 multifect와 pepsin+pancreatin 혼합효소(1:1)에 의해 생성된 분해물이 더욱 높은 항산화 효과를 나타내었다. 그러므로 말 사골추출물을 multifect와 pepsin+pancreatin 혼합효소(1:1)를 이용하여 효소 분해하여 얻은 3 kDa 이하의 저분자 펩타이드 분획은 항산화제로서 식품산업계에서 이용될 수 있을 것으로 예상되며 이를 뒷받침하기 위해 in vivo 테스트가 추후 진행되어야 할 것으로 판단된다.

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

  • 정해강;심형섭;전승호;김지흥;남성우;전붕수;김영준
    • 폴리머
    • /
    • 제39권3호
    • /
    • pp.487-492
    • /
    • 2015
  • Palladium(II) 촉매를 이용한 norobornene carboxylic acid estsers의 중합 시 단량체의 알킬 작용기의 종류, endo/exo 비율이 촉매의 활성도 및 중합 특성에 미치는 영향을 조사하였다. Norbornene esters 단량체는 5-norborene-2-carboxylic acid와 다양한 알코올을 반응시켜 합성하였다. 중합 촉매로는 di-$\mu$-chloro-bis(-methoxybicyclo[2,2,1]-hept-2-ene)palladium(II)(DCBMP)를 합성하여 사용하였고, 짝음이온으로 실버 헥사플루오로안티모네이트($AgSbF_6$)를 이용하였다. 고분자의 분석을 위해 젤 투과 크로마토그래피(gel permeation chromatography, GPC), 열 중량 분석법(thermogravimetric analysis, TGA), 시차주사 열량측정법(differential scanning calorimetry, DSC), 화학구조 분석을 위해 $^1H$ NMR spectroscopy를 이용하였다. 분자량 분석 결과 모든 작용기의 경우 endo-이성체의 비율이 exo-이성체의 비율보다 높을 경우 촉매의 구조적 방해로 인하여 반응성이 감소됨을 보였다. 또한 단량체와 촉매의 비율이 중합 거동에 미치는 영향을 조사하기 위해 단량체와 촉매의 몰비율을 100:1, 200:1, 300:1로 변화시켜 실험을 진행하였으며, 이 때 exo-norbornene carboxylic acid octyl ester의 경우 300:1 촉매비에서 필름형성이 가능한 높은 분자량($M_n=27500g/mol$)의 고분자를 합성할 수 있었다.

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
    • 한국축산식품학회지
    • /
    • 제35권1호
    • /
    • pp.35-40
    • /
    • 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.

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

  • 조성윤;고정민;이경아
    • 대한유전성대사질환학회지
    • /
    • 제16권2호
    • /
    • pp.70-78
    • /
    • 2016
  • 선천성 부신 과형성증 중 가장 흔한 21-hydroxy-lase deficiency (21-OHD)는 신생아 선별검사에서 17-hydroxyprogesterone (17-OHP)의 증가로 조기 진단이 가능하다. 17-OHP가 애매하게 증가되는 경우에는 ACTH 자극 검사가 필요하며, 이 검사는 nonclassical (NC형) 21-OHD 진단의 gold standard이다. 전형적인 임상 증상이 없는 경우, 예를 들어 남성화가 심하지 않은 여아, 경한 simple virilizing (SV)형 남아나 신생아 선별 검사에서 발견되지 않을 수 있는 NC형 환자의 경우, 분자유전학적 검사가 진단에 도움을 줄 수 있으며, 이는 예후 에측 및 유전 상담에도 도움이 된다. 미숙아와 저체중 출생아의 경우는 17-OHP가 위양성을 보이기 쉬우므로 출생 주수나 출생 체중에 따른 cutoff 값 설정이 필요하다. 높은 위양성률을 극복하기 위해 기존 RIA방법에 비해 최근 LC-MS/MS가 민감도와 특이도를 높이는 검사로 주목 받고 있다. 21-OHD 신생아 선별 검사의 효율성을 높이기 위해서는 SW형 남아를 조기에 발견하고, 여아에서 성별 결정을 조기에 올바르게 하고, NC형 환자를 찾아내고, 미숙아/저체중 출생아/아픈 신생아에서 위양성률을 낮추어서 불필요한 재검 및 경제적/심리적 부담을 최소화 하기 위한 노력이 필요하다. 무엇보다 21-OHD가 임상적으로 확실하게 의심되는 경우에는 확진 검사에 앞서 적절한 치료가 조기에 시작되어야 한다. 저자들은 본 종설에서 21-OHD의 신생아 선별 검사 후 진단 알고리즘에 대해 최신 문헌들에 근거하여 가이드라인을 제시하는 바이다.

  • PDF

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
    • /
    • 제6권1호
    • /
    • pp.33-40
    • /
    • 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
    • /
    • 제4권4호
    • /
    • pp.283-289
    • /
    • 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.

  • PDF

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
    • /
    • 제8권4호
    • /
    • pp.341-346
    • /
    • 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.

  • PDF

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

  • 조미라;김풍호;이태식;오은경;유홍식;이희정
    • 한국수산과학회지
    • /
    • 제39권6호
    • /
    • pp.454-459
    • /
    • 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.