• 제목/요약/키워드: semi-quantitative analysis

검색결과 258건 처리시간 0.025초

Apoptosis and Anti-proliferaction by Saussurea lappa and Pharbitis nil in AGS Human Gastric Cancer Cell Line

  • Ko Seong-Gyu;Oh Hee-Rah;Lee Sun-Dong;Hwang Gwi-Seo
    • 대한한방내과학회지
    • /
    • 제24권1호
    • /
    • pp.134-143
    • /
    • 2003
  • Objectives : We performed this study to understand the molecular basis of the antitumor effect of Saussurea lappa, Pharbitis nil, Plantago asiatica and Taraxacum mongolicum, which have been used for cancer treatment in Korean traditional medicine. Design: We analyzed, the effect of these medicinal herbs on proliferation and apoptosis of tumor cells and its association with gene expression, We performed semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) analysis of cell cycle- and apoptosis-related genes using a gastric cancer cell line AGS. Results : Cell counting assay and $[^3H]thymidine$ uptake analysis showed that Saussurea lappa and Pharbitis nil strongly inhibit cell proliferation of AGS in a dose-dependent manner. Interestingly, gene espression assay revealed that mRNA espression levels of c-Jun, c-Fos, c-Myc, and Cyclin D1 were markedly decreased by Saussurea lappa and Pharbitis nil. Furthermore, Saussurea lappa was identified to activate expression of the p53 tumor suppressor and its downstream effector $p21^{Wafl}$, which leads to $G_1$ cell cycle arrest and apoptosis. These observations suggest that the anticancer effect of Saussurea lappa and Pharbitis nil might be associated with their regulatory capability of tumor-related gene expression.

  • PDF

노인시설의 물리적 환경에 대한 거주노인 의견 조사방법의 분석 : 1990년 이후 해외 학술논문자료를 중심으로 (An Analysis on the Research Method of Elderly Residents' Opinion towards the Physical Environments of the Facilities for the Elderly : Focusing on Foreign Academic Journal Articles since 1990)

  • 이민아
    • 가정과삶의질연구
    • /
    • 제33권2호
    • /
    • pp.35-51
    • /
    • 2015
  • The purpose of this study was to provide information for research about residents' opinion toward the physical environments of elderly facilities, through the analysis and investigation on the research methodology of foreign academic journal articles from 1990 to 2014. The study results were as follows: Firstly, purposive sampling was a large majority of both facilities and elderly residents. In quantitative studies, many researchers have conducted simple random, cluster, or stratified sampling. Diverse facilities in area, size, location, and etc. should be considered for participation. The qualifications for residents' participation should be considered as well, so that they all could have autonomy for study participation. Secondly, questionnaire and semi-structured guide were likely to be used in independent and resident care facilities. On the other hand in assisted living and long-term care facilities, open questions and visual material were used as well. A compatible scale should be developed so that elderly having variable functional level could participate independently in the study. Thirdly, in data collection process, compliance with research ethics and well trained interviewer's skill were important for residents' active responses and minimization of response errors. Enough research period of time and mixed study in data collection will decrease the response error.

Differential Expression of Cyclic AMP-Response Element Binding Protein Zhangfei (CREBZF) in the Mouse Testis during Postnatal Development

  • Jang, Hoon
    • 한국발생생물학회지:발생과생식
    • /
    • 제22권1호
    • /
    • pp.65-72
    • /
    • 2018
  • Cyclic AMP-response element binding protein zhangfei (CREBZF), a member of ATF/CREB (activating transcription factor/ cAMP response element binding protein) family, regulates numerous cellular functions and development of cells by interacting transcription factors. This study discovered the expression pattern of CREBZF in seminiferous tubule of testes during the postnatal development of mice. In testis, CREBZF mRNA expression was the highest among other organs. Immunofluorescence analyses showed that the CREBZF was specifically expressed on spermatocyte but not in spermatogonia and Sertoli cells in seminiferous epithelium of mouse testis. Semi-quantitative polymerase chain reaction (PCR) analysis showed that CREBZF transcript level was significantly elevated during postnatal development of mouse testis. Confocal imaging analysis indicated that the protein expression of CREBZF in seminiferous tubule remained low until postnatal day (PD) 14, and was dramatically increased in PD 21. Interestingly, only one type of the spermatocyte expressed CREBZF specifically among SCP3-positive spermatocytes. Taken together, these results suggest that CREBZF may be novel putative marker of the spermatocyte and regulate meiosis during postnatal development of mice.

A High-speed Miniature Screening Gaschromatograph with Flame Ionization Detector

  • Banik Rahul;Lee Dong-Yeon;Gweon Dae-Gab
    • Journal of Mechanical Science and Technology
    • /
    • 제19권12호
    • /
    • pp.2197-2204
    • /
    • 2005
  • The combination of Gas chromatography (GC) for separation and Flame Ionization Detection (FID) for detection and identification of the components of a mixture of compounds is a fast and strongly proved method of analytic chemistry. The objective of this research was to design a combined High-speed miniature screening Gas chromatograph along with a Flame Ionization Detector for quick, quantitative and qualitative analysis of gas components. This combined GC-FID system is suitable to detect the volatile and semi-volatile hydrocarbons present in a gas mixture. The construction made it less expensive, easy to use and movable. The complete gas path was developed. On/off valves, temperature and flow sensors and their interface electronics were used for controlling purpose. A Microcontroller was programmed to measure the temperature and gas flow using the sensors and to control and regulate them using the electronics and valves. A pocket PC with its touch screen served as a user interface for the system. Software was developed for the pocket PC, which makes the communication possible with the Microcontroller. The system parameters can be indicated in the Pocket PC as simple text and also the analysis result can be displayed.

Analysis of S-glutathionylated proteins during adipocyte differentiation using eosin-glutathione and glutaredoxin 1

  • Hwang, Sungwon;Iram, Sana;Jin, Juno;Choi, Inho;Kim, Jihoe
    • BMB Reports
    • /
    • 제55권3호
    • /
    • pp.154-159
    • /
    • 2022
  • Protein S-glutathionylation is a reversible post-translational modification on cysteine residues forming a mixed disulfide with glutathione. S-glutathionylation, not only protects proteins from oxidation but also regulates the functions of proteins involved in various cellular signaling pathways. In this study, we developed a method for the detection of S-glutathionylated proteins (ProSSG) using eosin-glutathione (E-GSH) and mouse glutaredoxin 1 (mGrx1). ProSSG was efficiently and specifically labeled with E-GSH to form ProSSG-E via thiol-disulfide exchange. ProSSG-E was readily luminescent allowing the detection of ProSSG with semi-quantitative determination. In addition, a deglutathionylation enzyme mGrx1 specifically released E-GSH from ProSSG-E, which increased fluorescence allowing a sensitive determination of ProSSG levels. Application of the method to the adipocyte differentiation of 3T3-L1 cells showed specific detection of ProSSG and its increase upon differentiation induction, which was consistent with the result obtained by conventional immunoblot analysis, but with greater specificity and sensitivity.

배양세포에서 Semi-quantitative RT-PCR에 의한 조류인플루엔자 H9N2의 전사활성 분석 최적 시기 결정 및 전사체 분석 (Deterimination of an Optimal Time Point for Analyzing Transcriptional Activity and Analysis of Transcripts of Avian Influenza Virus H9N2 in Cultured Cell)

  • 나기윤;이영민;변승준;전익수;박종현;조인수;주이석;이윤정;권준헌;구용범
    • 미생물학회지
    • /
    • 제45권3호
    • /
    • pp.286-290
    • /
    • 2009
  • 조류인플루엔자 바이러스 mRNA의 전사는 감염 동안에 시간적으로 조절되어진다. 감염 후 세포 내에서 증식된 바이러스가 방출되어 세포를 다시 감염하게 되면 전사 수준 분석에 오차가 발생할 수 있으므로 바이러스가 방출 되기 이전에 전사 수준의 측정이 이루어져야 한다. 본 연구에서는 조류인플루엔자 H9N2를 감염시킨 닭의 섬유아세포주 UMNSAH/DF-1에서 바이러스 감염 후 증식된 바이러스의 방출까지의 시간을 측정하기 위하여 배양액 중에 방출되는 바이러스 RNA 게놈 수준을 semi-quantitative RT-PCR에 의해 측정하였다. 배양액 중의 바이러스 RNA 게놈의 분리 과정에서 발생할 수 있는 RNA 회수율의 오차를 보정하기 위해 mouse의 전체 RNA를 배양액 시료에 넣어서 바이러스 RNA 분리에 carrier로 사용하고 그 속에 포함된 mouse의 GAPDH를 RNA를 역전사 반응 및 PCR의 내부 대조 RNA로 사용하였다. 그 결과 감염 후 방출까지 16~20시간이 소요됨을 알 수 있었다. 따라서 감염 후 12시간 후에 세포 내에서 합성된 8종의 전사체의 수준을 측정하였다. 8종의 mRNA 중 PA를 제외한 7종의 mRNA (HA, NA, PB1, PB2, NP, M, NS를 암호화하는 mRNA)는 뚜렷한 증폭 band를 보였으며, 이것은 이들이 전사활성 측정에 사용될 수 있는 것으로 나타났다. 이상과 같은 전사수준의 측정 방법은 조류인플루엔자 바이러스의 RNA polymerase를 표적으로 한 항바이러스제의 검색에 사용할 수 있을 것으로 보인다.

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
    • /
    • 제24권2호
    • /
    • pp.118-124
    • /
    • 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.

Accuracy and reproducibility of 3D digital tooth preparations made by gypsum materials of various colors

  • Tan, Fa-Bing;Wang, Chao;Dai, Hong-Wei;Fan, Yu-Bo;Song, Jin-Lin
    • The Journal of Advanced Prosthodontics
    • /
    • 제10권1호
    • /
    • pp.8-17
    • /
    • 2018
  • PURPOSE. The study aimed to identify the accuracy and reproducibility of preparations made by gypsum materials of various colors using quantitative and semi-quantitative three-dimensional (3D) approach. MATERIALS AND METHODS. A titanium maxillary first molar preparation was created as reference dataset (REF). Silicone impressions were duplicated from REF and randomized into 6 groups (n=8). Gypsum preparations were formed and grouped according to the color of gypsum materials, and light-scanned to obtain prepared datasets (PRE). Then, in terms of accuracy, PRE were superimposed on REF using the best-fit-algorithm and PRE underwent intragroup pairwise best-fit alignment for assessing reproducibility. Root mean square deviation (RMSD) and degrees of similarity (DS) were computed and analyzed with SPSS 20.0 statistical software (${\alpha}=.05$). RESULTS. In terms of accuracy, PREs in 3D directions were increased in the 6 color groups (from 19.38 to $20.88{\mu}m$), of which the marginal and internal variations ranged $51.36-58.26{\mu}m$ and $18.33-20.04{\mu}m$, respectively. On the other hand, RMSD value and DS-scores did not show significant differences among groups. Regarding reproducibility, both RMSD and DS-scores showed statistically significant differences among groups, while RMSD values of the 6 color groups were less than $5{\mu}m$, of which blue color group was the smallest ($3.27{\pm}0.24{\mu}m$) and white color group was the largest ($4.24{\pm}0.36{\mu}m$). These results were consistent with the DS data. CONCLUSION. The 3D volume of the PREs was predisposed towards an increase during digitalization, which was unaffected by gypsum color. Furthermore, the reproducibility of digitalizing scanning differed negligibly among different gypsum colors, especially in comparison to clinically observed discrepancies.

Reevaluation of the Change of Leuconostoc Species and Lactobacillus plantarum by PCR During Kimchi Fermentation

  • Choi, Jae-Yeon;Kim, Min-Kyun;Lee, Jong-Hoon
    • Journal of Microbiology and Biotechnology
    • /
    • 제12권1호
    • /
    • pp.166-171
    • /
    • 2002
  • The genus Leuconostoc is generally recognized as a favorable microorganism associated with a good taste of Kimchi and Lactobacillus plantarum is responsible for the overripening and acidification of Kimchi. A rapid and reliable PCR-based method to monitor the change of these lactic acid bacterial populations during Kimchi fermentation was attempted. A Leuconostoc-specific primer set was chosen from the conserved sequences of 16S rRNA genes among Leuconostoc species. The Lb. plantarum-specific primer set was the internal segments of a Lb. plantarum-specific probe which was isolated after randomly amplified polymorphic DNA (RAPD) analysis and tested for identification. The specificity of this protocol was examined in DNA samples isolated from a single strain. In agarose gel, as little as 10 pg of template DNA could be used to visualize the PCR products, and quantitative determination was possible at the levels of 10 pg to 100 ng template DNA. For the semi-quantitative determination of microbial changes during Kimchi fermentation, total DNAs from the 2 h-cultured microflora of Kimchi were extracted for 16 days and equal amounts of DNA templates were used for PCR. The intensities of DNA bands obtained from PCR using Leuconostoc-specific and Lb. plantarum-specific primer sets marked a dramatic contrast at the 1 ng and 100 ng template DNA levels during Kimchi fermentation, respectively. As the fermentation proceeded, the intensity of the band for Leuconostoc species increased sharply until the 5th day and the levels was maintained until the 11 th day. The sharp increase for Lb. plantarum occurred after 11 days with the decrease of Leuconostoc species. The results of this study indicate that Leuconostoc species were the major microorganisms at the beginning of Kimchi fermentation and reach their highest population during the optimum ripening period of Kimchi.

열량 및 열량영양소 섭취량과 관련된 유전자 변이에 대한 전장유전체 연관성 분석연구 (Genetic Variants Associated with Calorie and Macronutrient Intake in a Genome-Wide Association Study)

  • 백인경;안윤진;이승구;김소리울;한복기;신철
    • Journal of Nutrition and Health
    • /
    • 제43권4호
    • /
    • pp.357-366
    • /
    • 2010
  • There has been no genome-wide association study (GWAS) for macronutrient intake as a quantitative trait. To explore genetic loci associated with total calorie and macronutrient intake, genome-wide association data of autosomal single nucleotide polymorphisms (SNPs) from Korean adults were analyzed. We conducted a GWAS in 3,690 men and women aged 40 to 60 years from an urban population-based cohort. At the baseline examination (June 18, 2001 through January 29, 2003), DNA samples of the study subjects were collected and analyzed for genotyping. The information of average daily consumption of total calorie, carbohydrate, protein, and fat was obtained from a semi-quantitative food frequency questionnaire and transformed by natural logarithm for analyses after adjustment of calorie intake. Using multivariate linear regression analysis adjusted for age, sex, and height, we tested for 352,021 SNPs and found weak associations, which do not reach genome-wide association significance, with calorie and macronutrient intake. However, a number of SNPs were found to have potential associations with macronutrient intake; in particular, signals in SORBS1 and those in PRKCB1 were likely associated with carbohydrate and fat intake, respectively. We observed an inverse association between the minor allele of the SNPs in these genes and the amount of consumption of carbohydrate or fat. Our GWAS identified loci and minor alleles weakly associated with macronutrient intake. Because SORBS1 and PRKCB1 are reportedly associated with the metabolism of glucose and lipid as well as with obesity-related diseases, further investigations on biological and functional roles of polymorphism of these genes in the relation to macronutrient intake are warranted.