• 제목/요약/키워드: quantitative profiles

검색결과 243건 처리시간 0.035초

생쥐 초기배아의 유전자 활성에 미치는 Protein Kinase Inhibitors의 영향 (Effects of Protein Kinase Inhibitors on Gene Activation of Early Embryos in Mouse)

  • 이정은;채영규;배인하;윤용달;김문규
    • Clinical and Experimental Reproductive Medicine
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    • 제22권2호
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    • pp.191-201
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    • 1995
  • Transcriptional activation of the embryonic genome initiates at 2-cell stage in mouse embryo and is characterized by the synthesis of TRC which is restricted to 2-cell stage. To investigate the roles of various protein kinases on the embryonic gene activation, the effects of protein kinase inhibitors on in vitro development and protein synthetic profiles of the early mouse embryos were examinded. None of ${\alpna}-amanitin$ which is a mRNA synthetic inhibitor, H8 which is a PKA inhibitor, and H7 which is a PKC inhibitor, affected on first cleavage of mouse 1-cell embryos in vitro. However, all of these drugs inhibited the second cleavage. When the drugs were removed following treatment for 6 hours, H8 or H7 treatment showed little inhibition on subsequent development of 1-cell embryos to 2-cell stage or further. In contrast, ${\alpna}-amanitin$ irreversibly inhibited the development of 1-cell embryos to 2-cell stage following removal of the drug. Genistein, a TPK inhibitor, inhibited both the first cleavage of 1-cell embryos and the second cleavage of 2-cell embryos, suggesting that TPK activity may be important during the early cleavages. All of the above four drugs inhibited TRC synthesis as shown by the fluorographic analysis of $[^{35}S]-Met$ labeled protein profiles. When late 1-cell embryos were treated with H7 and analyzed synthetic patterns of $[^{35}S]-Met$ labeled protein, the quantitative differences of protein synthesis on SDS-PAGE appeared on 77 kD and 33 kD region at $32{\sim}38$ hours post hCG. From these studies, transcriptional activation of embryonic genome is not essenting to the mouse 1-cell embryos to develop to 2-cell stage. Hawever, TPK activity is reguisite for both the first cleavage and second cleavage. Similarly, both PKC and PKA activities are required for the second cleavage of mouse embryos, but not for the first cleavage.

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Genome-wide identification, organization, and expression profiles of the chicken fibroblast growth factor genes in public databases and Vietnamese indigenous Ri chickens against highly pathogenic avian influenza H5N1 virus infection

  • Anh Duc Truong;Ha Thi Thanh Tran;Nhu Thi Chu;Huyen Thi Nguyen;Thi Hao Vu;Yeojin Hong;Ki-Duk Song;Hoang Vu Dang;Yeong Ho Hong
    • Animal Bioscience
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    • 제36권4호
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    • pp.570-583
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    • 2023
  • Objective: Fibroblast growth factors (FGFs) play critical roles in embryo development, and immune responses to infectious diseases. In this study, to investigate the roles of FGFs, we performed genome-wide identification, expression, and functional analyses of FGF family members in chickens. Methods: Chicken FGFs genes were identified and analyzed by using bioinformatics approach. Expression profiles and Hierarchical cluster analysis of the FGFs genes in different chicken tissues were obtained from the genome-wide RNA-seq. Results: A total of 20 FGF genes were identified in the chicken genome, which were classified into seven distinct groups (A-F) in the phylogenetic tree. Gene structure analysis revealed that members of the same clade had the same or similar exon-intron structure. Chromosome mapping suggested that FGF genes were widely dispersed across the chicken genome and were located on chromosomes 1, 4-6, 9-10, 13, 15, 28, and Z. In addition, the interactions among FGF proteins and between FGFs and mitogen-activated protein kinase (MAPK) proteins are limited, indicating that the remaining functions of FGF proteins should be further investigated in chickens. Kyoto encyclopedia of genes and genomes pathway analysis showed that FGF gene interacts with MAPK genes and are involved in stimulating signaling pathway and regulating immune responses. Furthermore, this study identified 15 differentially expressed genes (DEG) in 21 different growth stages during early chicken embryo development. RNA-sequencing data identified the DEG of FGFs on 1- and 3-days post infection in two indigenous Ri chicken lines infected with the highly pathogenic avian influenza virus H5N1 (HPAIV). Finally, all the genes examined through quantitative real-time polymerase chain reaction and RNA-Seq analyses showed similar responses to HPAIV infection in indigenous Ri chicken lines (R2 = 0.92-0.95, p<0.01). Conclusion: This study provides significant insights into the potential functions of FGFs in chickens, including the regulation of MAPK signaling pathways and the immune response of chickens to HPAIV infections.

APPLICATION STUDY OF CHEMOINFOMETRICAL NEAR-INFRARED SPECTROSCOPY IN PHARMACEUTICAL INDUSTRY

  • Otsuka, Makoto;Kato, Fumie;Matsuda, Yoshihisa
    • 한국근적외분광분석학회:학술대회논문집
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    • 한국근적외분광분석학회 2001년도 NIR-2001
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    • pp.2111-2111
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    • 2001
  • A chemoinfometrical method for evaluating the quantitative determination of crystallinity one polymorphs based on fourie-transformed near-infrared (FT-NIR) spectroscopy was established. A direct comparison of the data with the ones collected from using the and compared with the conventional powder X-ray diffraction method was performed. [Method] The pPure a and g forms of indomethacin (IMC) were prepared by reportedusing published methods. Six kinds of standard samples obtained by physically mixing of a and g forms. After the powder X-ray diffraction profiles of samples have been measured, the intensity values were normalized to against the intensity of silicon powder as the as an external standard. The calibration curves for quantification of crystal content were based upon the total relative intensity of four diffraction peaks from of the form g crystal. FT-NIR spectra of six calibration sample sets were recorded 5 times with the NIR spectrometer (BRAN+LUEBBE). Chemoinfometric analysis was performed on the NIR spectral data sets by applying the principal component regression (PCR). [Results] The relation between the actual and predicted polymorphic contents of form g IMC measured using by the X-ray diffraction method shows a good straight linen linear relation., and it has slope of 0.023, an intercept of 0.131 and a correlation coefficient of 0.986. PCR analyses wereis was performed based on normalized NIR spectra sets offer standard samples of known content of IMC g form. IMC. A calibration equation was determined to minimize the root mean square error of the predictionthe prediction. Figure 1 shows a plot of the calibration data obtained by NIR method between the actual and predicted contents of form g IMC. The predicted values were reproducible and had a smaller standard deviation. Figure 2 shows that the plot for the predicted transformation rate (%) of form a IMC to form g as measured by X-ray diffractomeoy against to those as measured by NIR method. The plot has a slope of 1.296, an intercept of 1,109, and a correlation coefficient of 0.992. The line represents a satisfactory correlation between the two predicted values of form g IMC content. Thus NIR spectroscopy is an effective method for the evaluation to the pharmaceutical products of quantitative of polymorph.

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Annealing control primer system을 이용한 어류 재조합 myostatin prodomain 단백질에 의해 성장이 증가된 무지개송어의 특이적 발현 유전자 탐색 (Identification of Differentially Expressed Genes in Improved Rainbow Trout Growth by Treatment with a Fish Myostatin Prodomain Using the Annealing Control Primer System)

  • 이상범;진형주
    • 한국어류학회지
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    • 제24권2호
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    • pp.118-124
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    • 2012
  • 이전 연구에서 넙치유래 재조합 마이오스타틴 프로도메인을 무지개송어에 한달간 침지법을 통하여 처리한 결과 대조군에 비하여 무게가 최대 약 42% 증가되었다. 따라서 본 연구는 재조합 마이오스타틴 프로도메인을 침지법에 의해 처리된 무지개송어와 대조군의 근육으로부터 발현되는 cDNA를 제작하여 마이오스타틴 프로도메인에 의해서 유도된 특정유전자를 선발하기 위하여 ACP (annealing control primer)를 이용한 DDRT법을 통하여 분석하였다. 총 20가지의 ACP를 이용한 결과 2개의 특정 유전자를 분석하였으며, NCBI BLAST 분석결과 Cytochrome P450 mono oxygenase와 Profilin으로 판명되었다. 이 중 Cytochrome P450 mono oxygenase는 대조군보다 발현량이 증가하였으며, Profilin는 대조군에 비해서 발현량이 감소하였다. 이러한 결과를 재확인하기 위하여 두 유전자의 primer를 각각 제작하여 semi-quantitative RT-PCR를 시행한 결과 DDRT법에 의한 분석과 동일하였다. 본 결과는 어류의 성장에서 마이오스타틴 프로도메인의 기능 및 메카니즘에 대한 연구에 유용한 자료가 될 것으로 사료된다.

염분과 수온 스트레스에 따른 감성돔의 glucocorticoid receptor mRNA 발현 특징과 생리적 변화에 관한 연구 (Profiles of Glucocorticoid Receptor mRNA Expression and Physiological Changes in Response to Osmotic and Thermal Stress Conditions in Black Porgy (Acanthopagrus schlegeli))

  • 안광운;신현숙;민병화;길경석;최철영
    • 한국어류학회지
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    • 제22권1호
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    • pp.17-24
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    • 2010
  • 본 연구에서는 감성돔의 염분과 수온 변화에 따른 스트레스 반응을 알아보기 위하여 glucocorticoid receptor (GR) mRNA 발현을 조사하였다. 감성돔 신장으로부터 전장의 GR cDNA를 클로닝하였고, 염분과 수온이 변화하는 동안 아가미, 신장 및 장에서 GR mRNA 발현 변화를 quantitative real-time PCR (QPCR)을 이용하여 조사하였다. 염분 변화시, 아가미, 신장 및 장에서 GR mRNA 발현은 0 psu에서 가장 높게 나타났으며, 혈장 cortisol과 glucose 농도도 증가한 반면, triiodothyronine ($T_3$) 농도는 감소하였다. 수온 변화시, 아가미, 신장 및 장에서 GR mRNA 발현은 $30^{\circ}C$에서 가장 높게 관찰되었다. 혈장 cortisol, glucose 및 $T_3$ 농도 또한 고수온 ($30^{\circ}C$)에서 증가하였다. GR mRNA 발현의 증가는 염분과 수온 변화와 같은 환경 요인에 대한 좋은 스트레스 지표로 여겨진다.

녹차씨 주정추출물의 카페인 함량 평가와 고지방식이 비만유도 C57BL/6 마우스와 3T3-L1 지방전구세포의 항비만효과 (Quantitative Determination of Caffeine of Green Tea Seed Ethanol Extract on Anti-obesity in C57BL/6 Mice Fed a High-fat Diet and 3T3-L1 Cells)

  • 정명아;이현미;오둘리;김유진;배동혁;오교녀;신희찬;김선오
    • 동의생리병리학회지
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    • 제30권2호
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    • pp.88-94
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    • 2016
  • The study was carried out to examine the anti-obesity effects of 40% ethanol extract from green tea seed (GS) and quantitative determination of caffeine as its major compound. The specificity was satisfied with retention time and UV spectrum by analysis of caffeine using HPLC and comparison with standard compound. It showed a high linearity in the calibration curve with a coefficient of correlation (R2) of 0.9974. The amount of caffeine in GS was about 4.649 mg/g (0.465%) in the three times analysis, and relative standard deviation (RSD) was less than 0.452% by the validated method. The anti-obesity effects of GS were evaluated by using Oil Red O staining in 3T3-L1 adipocytes and body weight, visceral fat and lipid profiles in high fat diet (HFD)-induced C57BL/6 obese mice. Our results indicated that treatment with GS dose-dependently decreased lipid accumulation contents (p<0.001). Moreover, after oral administration for 30 days feeding with HFD-induced obses mice, GS (100 and 300 mg/kg/day) produced a significant decrease in serum total cholesterol (TC), glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and visceral fat. Thus, the result of this study indicate that the GS may be a useful resource for the management of obesity.

Tc-99m-MIBI 심근 SPECT 극성지도 분석에 의한 심근 기능의 정량적 평가 (A Quantitative Method for the Assessment of Myocardial Function using the Polar Analysis of Tc-99m-MIBI Myocardial SPECT)

  • 곽철은;이동수;여정석;이경한;정준기;이명철;서정돈;고창순
    • 대한핵의학회지
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    • 제28권2호
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    • pp.172-176
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    • 1994
  • 심근 SPECT에 의한 심근질환의 심도 및 허혈부위를 평가하는 방법으로 정상인의 극성지도와의 비교에 의한 결손지도 평가방법이 상용화되었으나 본 연구에서는 결손부위의 범위만을 산출할 수 있었던 종래의 방법을 개선할 수 있는 관류회복정도 및 심도성적 산출 방법을 시도하여 보았다. 허혈 및 심도성적의 산출에서 부하 및 휴식기 극성지도의 차이를 3등급으로 분할하여 각 등급에 해당되는 화소의 수를 가중하므로서 관류결손부위의 면적이 반영된 허혈 및 심도성적 산출방법을 제안하고자 하였으며, 등급 및 가중치의 할당은 정상인의 잔상동맥영역별 관류분포를 반영하여 고른 등급 할당방법보다 상관도가 개선된 결과를 얻을 수 있었다.

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Discrimination of Panax ginseng Roots Cultivated in Different Areas in Korea Using HPLC-ELSD and Principal Component Analysis

  • Lee, Dae-Young;Cho, Jin-Gyeong;Lee, Min-Kyung;Lee, Jae-Woong;Lee, Youn-Hyung;Yang, Deok-Chun;Baek, Nam-In
    • Journal of Ginseng Research
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    • 제35권1호
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    • pp.31-38
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    • 2011
  • In order to distinguish the cultivation area of Panax ginseng, principal component analysis (PCA) using quantitative and qualitative data acquired from HPLC was carried out. A new HPLC method coupled with evaporative light scattering detection (HPLC-ELSD) was developed for the simultaneous quantification of ten major ginsenosides, namely $Rh_1$, $Rg_2$, $Rg_3$, $Rg_1$, Rf, Re, Rd, $Rb_2$, Rc, and $Rb_1$ in the root of P. ginseng C. A. Meyer. Simultaneous separations of these ten ginsenosides were achieved on a carbohydrate analytical column. The mobile phase consisted of acetonitrile-water-isopropanol, and acetonitrile-water-isopropanol using a gradient elution. Distinct differences in qualitative and quantitative characteristics for ginsenosides were found between the ginseng roots produced in two different Korean cultivation areas, Ganghwa and Punggi. The ginsenoside profiles obtained via HPLC analysis were subjected to PCA. PCA score plots using two principal components (PCs) showed good separation for the ginseng roots cultivated in Ganghwa and Punggi. PC1 influenced the separation, capturing 43.6% of the variance, while PC2 affected differentiation, explaining 18.0% of the variance. The highest contribution components were ginsenoside $Rg_3$ for PC1 and ginsenoside Rf for PC2. Particularly, the PCA score plot for the small ginseng roots of six-year old, each of which was light than 147 g fresh weight, showed more distinct discrimination. PC1 influenced the separation between different sample sets, capturing 51.8% of the variance, while PC2 affected differentiation, also explaining 28.0% of the variance. The highest contribution component was ginsenoside Rf for PC1 and ginsenoside $Rg_2$ for PC2. In conclusion, the HPLC-ELSD method using a carbohydrate column allowed for the simultaneous quantification of ten major ginsenosides, and PCA analysis of the ginsenoside peaks shown on the HPLC chromatogram would be a very acceptable strategy for discrimination of the cultivation area of ginseng roots.

Ginsenoside Content of North American Ginseng (Panax quinquefolius L. Araliaceae) in Relation to Plant Development and Growing Locations

  • Jackson, Chung Ja C.;Dini, Jean-Paul;Lavandier, Clara;Faulkner, Harold;Rupasinghe, H.P. vasantha;Proctor, John T.A.
    • Journal of Ginseng Research
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    • 제27권3호
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    • pp.135-140
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    • 2003
  • North American ginseng (Panax quinquefolius L.) was analysed for total ginsenosides and ten major ginsenosides (R$_{0}$ , Rb$_1$, Rb$_2$, Rc, Rd, Re, Rf, Rg$_1$, pseudoginsenoside F$_{11}$ and gypenoside XVII), and variations in ginsenoside content with age of plant (over a four-year-period) and geographic location (Ontario versus British Columbia) were investigated. In the roots the total ginsenoside content increased with age up to 58-100 mgㆍg$^{-1}$ dry weights in the fourth year, but in leaves it remained constant over time. Roots and leaves, moreover, had different proportions of individual ginsenosides. The most abundant ginsenosides were Rb$_1$ (56mgㆍg$^{-1}$ for Ontario; 37mgㆍg$^{-1}$ for British Columbia) and Re (21mgㆍg$^{-1}$ for Ontario; 15 mgㆍg$^{-1}$ for British Columbia) in roots, and Rd (28-38 mgㆍg$^{-1}$ ), Re (20-25 mgㆍg$^{-1}$ ), and Rb$_2$ (13-19 mgㆍg$^{-1}$ ) in leaves. Measurable quantities of Rf were found in leaves (0.4-1.8 mgㆍg$^{-1}$ ) but not in roots or stems. Our results show that ginsenoside profiles in general, and Rf in particular, could be used for chemical fingerprinting to distinguish the different parts of the ginseng plant, and that ginseng leaves could be valuable sources of the ginsenosides Rd, Re, and Rb$_2$.

Mixture에서 봉우리 면적을 활용한 유전자 증거의 해석 (Interpreting Mixtures Using Allele Peak Areas)

  • 홍유림;이효정;이재원
    • 응용통계연구
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    • 제23권1호
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    • pp.113-121
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    • 2010
  • 형사사건에서 STR(short tandem repeat)을 이용한 개인 식별은 범죄현장에서 발견된 유전자 증거와 용의자의 유전자 검사 결과를 이용하여 평가하게 된다. 특히 범죄현장에서 발견된 유전자 증거 표본에 기여자가 두 명 이상인 경우를 Mixture라고 하며, 이는 강간 등과 같은 사건에서 흔히 볼 수 있다. 이러한 상황에서 법의학적 추론을 위해 유전자 증거 표본을 해석하고자 하는 연구들이 계속되어 왔으며, 최근에는 Mixture에서 발생한 대립유전자의 봉우리 면적을 활용하여 유전자 증거를 해석하기 위한 노력들이 계속되고 있다. 따라서 본 연구에서는 봉우리 변적을 이용하여 유전자 증거 표본을 해석하기 위한 연구 방법들에 대해 살펴볼 것이며, 또한 유전자 증거 표본에 기여한 사람의 수가 세 명 이상안 표본으로 확장시킨 연구를 실시해 볼 것이다. 마지막으로 사례틀 통해 유전자 증거를 해석하는 방법을 살펴보고자 한다.