• BMB Reports
• /
• 제37권5호
• /
• pp.546-551
• /
• 2004

The increasing pace of development in molecular biology during the last decade has had a direct effect on mass testing and diagnostic applications, including blood screening. We report the model Microarray that has been developed for Hepatitis B virus (HBV) and Hepatitis D virus (HDV) detection. The specific primer pairs of PCR were designed using the Primer Premier 5.00 program according to the conserved regions of HBV and HDV. PCR fragments were purified and cloned into pMD18-T vectors. The recombinant plasmids were extracted from positive clones and the target gene fragments were sequenced. The DNA microarray was prepared by robotically spotting PCR products onto the surface of glass slides. Sequences were aligned, and the results obtained showed that the products of PCR amplification were the required specific gene fragments of HBV, and HDV. Samples were labeled by Restriction Display PCR (RD-PCR). Gene chip hybridizing signals showed that the specificity and sensitivity required for HBV and HDV detection were satisfied. Using PCR amplified products to construct gene chips for the simultaneous clinical diagnosis of HBV and HDV resulted in a quick, simple, and effective method. We conclude that the DNA microarray assay system might be useful as a diagnostic technique in the clinical laboratory. Further applications of RD-PCR for the sample labeling could speed up microarray multi-virus detection.

• Molecules and Cells
• /
• 제38권2호
• /
• pp.171-179
• /
• 2015

Prostatic acid phosphatase (PAP) expression increases proportionally with prostate cancer progression, making it useful in prognosticating intermediate to high-risk prostate cancers. A novel ligand that can specifically bind to PAP would be very helpful for guiding prostate cancer therapy. RNA aptamers bind to target molecules with high specificity and have key advantages such as low immunogenicity and easy synthesis. Here, human PAP-specific aptamers were screened from a 2'-fluoropyrimidine (FY)-modified RNA library by SELEX. The candidate aptamer families were identified within six rounds followed by analysis of their sequences and PAP-specific binding. A gel shift assay was used to identify PAP binding aptamers and the 6N aptamer specifically bound to PAP with a Kd value of 118 nM. RT-PCR and fluorescence labeling analyses revealed that the 6N aptamer bound to PAP-positive mammalian cells, such as PC-3 and LNCaP. IMR-90 negative control cells did not bind the 6N aptamer. Systematic minimization analyses revealed that 50 nucleotide sequences and their two hairpin structures in the 6N 2'-FY RNA aptamer were equally important for PAP binding. Renewed interest in PAP combined with the versatility of RNA aptamers, including conjugation of anti-cancer drugs and nano-imaging probes, could open up a new route for early theragnosis of prostate cancer.

• Molecules and Cells
• /
• 제44권11호
• /
• pp.851-860
• /
• 2021

Label-free optical diffraction tomography (ODT), an imaging technology that does not require fluorescent labeling or other pre-processing, can overcome the limitations of conventional cell imaging technologies, such as fluorescence and electron microscopy. In this study, we used ODT to characterize the cellular organelles of three different stem cells-namely, human liver derived stem cell, human umbilical cord matrix derived mesenchymal stem cell, and human induced pluripotent stem cell-based on their refractive index and volume of organelles. The physical property of each stem cell was compared with that of fibroblast. Based on our findings, the characteristic physical properties of specific stem cells can be quantitatively distinguished based on their refractive index and volume of cellular organelles. Altogether, the method employed herein could aid in the distinction of living stem cells from normal cells without the use of fluorescence or specific biomarkers.

• Asian Pacific Journal of Cancer Prevention
• /
• 제17권3호
• /
• pp.1023-1035
• /
• 2016

The purpose of this study was to investigate the role of colon cancer stem cells (CSCs) during chemically-induced rat multi-step colon carcinogenesis with or without the treatment with a specific cyclooxygenase-2 inhibitor drug (celecoxib). Two experiments were performed, the first, a short term 12 week colon carcinogenesis bioassay in which only surrogate markers for colon cancer, aberrant crypt foci (ACF) lesions, were formed. The other experiment was a medium term colon cancer rat assay in which tumors had developed after 32 weeks. Treatment with celecoxib lowered the numbers of ACF, as well as the tumor volumes and multiplicities after 32 weeks. Immunohistochemical proliferating cell nuclear antigen (PCNA) labeling indexes LI (%) were downregulated after treatment by celecoxib. Also different cell surface antigens known to associate with CSCs such as the epithelial cell adhesion molecule (EpCAM), CD44 and CD133 were compared between the two experiments and showed differential expression patterns depending on the stage of carcinogenesis and treatment with celecoxib. Flow cytometric analysis demonstrated that the numbers of CD133 cells were increased in the colonic epithelium after 12 weeks while those of CD44 but not CD133 cells were increased after 32 weeks. Moreover, aldehyde dehydrogenase-1 activity levels in the colonic epithelium (a known CSC marker) detected by ELISA assay were found down-regulated after 12 weeks, but were up-regulated after 32 weeks. The data have also shown that the protective effect of celecoxib on these specific markers and populations of CSCs and on other molecular processes such as apoptosis targeted by this drug may vary depending on the genetic and phenotypic stages of carcinogenesis. Therefore, uncovering these distinction roles of CSCs during different phases of carcinogenesis and during specific treatment could be useful for targeted therapy.

• Asian-Australasian Journal of Animal Sciences
• /
• 제28권6호
• /
• pp.788-795
• /
• 2015

Two dimensional-fluorescence difference gel electrophoresis (2D DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum.

• 한국콘텐츠학회논문지
• /
• 제10권1호
• /
• pp.275-285
• /
• 2010

지금까지 일반적인 청소년비행을 설명하는 이론들은 많은 발전을 이루었지만, 여성 특히 여자청소년의 비행에 관련되는 요인들을 설명하는 이론들은 극히 소수이고, 대표적인 청소년비행 이론들은 성 중립적인 사회이론으로 소년들의 비행을 설명하는데 적합한 것들이다. 따라서 여성의 특수하고 독특한 경험을 반영하는 여자청소년의 비행에 보다 설득력 있는 요인을 도출해낼 수 있는 성-특성적(gender-specific) 이론이 필요하다고 본다. 이러한 문제에 관심을 가졌던 페미니스트 학자들은 여자청소년의 비행을 설명하는 성 특성적 요인들의 검증을 수행하였고, 소녀들의 비행을 설명하는데 적절한 요인들에 대한 학자들 간에 일치가 이루어지지는 않고 있지만, 일부 요인들은 소년들보다는 소녀들의 비행을 설명하는데 보다 적절하다는데 합의하였다. 이에 이 연구에서는 여자청소년의 비행을 설명하는데 적절한 성 특성적 요인들로 알려진 요인들로서 개인적 상황에 대한 불만, 스스로의 일탈적 낙인과 주위의 일탈적 낙인, 자존감, 부모로부터의 학대나 주변 사람으로부터의 범죄 피해 경험 등의 요인과 비행의 관계를 한국청소년 패널조사(KYPS) 자료를 이용하여 검증하였다. 연구결과 선행연구 결과와 완전히 일치하지는 않지만, 대부분의 요인들이 소년들의 비행 보다는 소녀들의 비행을 설명하는데 설명력이 높은 것으로 나타나 이러한 요인들에 대한 후속연구의 수행을 통해 성-특성적 이론의 가능성을 고려해 볼 수 있다는 점에서 의미 있는 연구라고 생각된다.

• 한국식품위생안전성학회지
• /
• 제33권2호
• /
• pp.146-150
• /
• 2018

고추는 한국에서 매우 중요한 양념 중 하나이다. 하지만 수입 고춧가루와 다진 양념(다대기)에 부과되는 관세율(45%/270%)의 차이로 인해, 다진 양념이 수입된 후, 건조 및 분쇄 과정을 거쳐 고춧가루로 제작되고 있는 실정이다. 본 연구에서는 종 특이 PCR 기술과 whole-genome amplification 방법을 접목하여 고춧가루(N=45) 및 다진 양념(N=5) 제품의 사용원료(고추, 마늘, 양파, 파, 생강)를 분석하였다. 모니터링 결과, 39개 고춧가루 제품은 표시사항을 준수하였으며, 6개 고춧가루 및 5개 다진 양념 제품은 제조 기준을 충족시키지 못했다. 따라서 분석 제품의 22%가 표시사항을 준수하지 못한 것으로 밝혀졌으며, 본 연구에 사용한 분석 방법은 고춧가루 제품에 사용된 원료분석에 적합한 방법임을 입증하였다.

• 한국동물생명공학회지
• /
• 제34권3호
• /
• pp.190-196
• /
• 2019

Tight junctions are constituents of the blood-epididymis barrier that play roles in regulating the unidirectional transcellular transport of ions, water, and solutes to maintain optimal conditions for sperm maturation and storage. Claudin 1 (Cldn1) and 4 (Cldn4) are known as tight junction proteins and are expressed in the basolateral membranes as well as tight junctions in the epididymis of rodents. Here, we examined the expression and localization of Cldn1 and 4 to determine the function of these proteins in the pig epididymis. Cldn1 was highly expressed in the basolateral membrane of epithelial cells in the caput and corpus regions of the epididymis. In the cauda region, however, Cldn1 labeling was significantly decreased in the basolateral membrane of epithelial cells. In contrast, labeling indicated that Cldn4 was expressed in the basolateral membrane in the cauda region of the epididymis and was present at punctate reactive sites in the caput and corpus regions. However, in no region of the epididymis did we detect colocalization of Cldn1 and 4 with labeled ZO-1, the distribution of which is restricted to the tight junctions. Our results indicate that Cldn1 and 4 were region-specifically expressed in the pig epididymis but not present in the tight junctions of epididymal epithelium. In addition, reciprocal regulation in specific regions of the epididymis between Cldn1 and 4 may play an important role in generating an optimal luminal environment for sperm maturation and storage in the pig epididymis.

• Toxicological Research
• /
• 제34권1호
• /
• pp.1-6
• /
• 2018

The purpose of this study was to detect cell death in the liver of mice treated with thioacetamide (TAA) using fluorescence bioimaging and compare this outcome with that using conventional histopathological examination. At 6 weeks of age, 24 mice were randomly divided into three groups: group 1 (G1), control group; group 2 (G2), fluorescence probe control group; group 3 (G3), TAA-treated group. G3 mice were treated with TAA. Twenty-two hours after TAA treatment, G2 and G3 mice were treated with Annexin-Vivo 750. Fluorescence in vivo bioimaging was performed by fluorescence molecular tomography at two hours after Annexin-Vivo 750 treatment, and fluorescence ex vivo bioimaging of the liver was performed. Liver damage was validated by histopathological examination. In vivo bioimaging showed that the fluorescence intensity was increased in the right upper part of G3 mice compared with that in G2 mice, whereas G1 mice showed no signal. Additionally ex vivo bioimaging showed that the fluorescence intensity was significantly increased in the livers of G3 mice compared with those in G1 or G2 mice (p < 0.05). Histopathological examination of the liver showed no cell death in G1 and G2 mice. However, in G3 mice, there was destruction of hepatocytes and increased cell death. Terminal deoxynucleotidyl transferase dUTP nick end labeling staining confirmed many cell death features in the liver of G3 mice, whereas no pathological findings were observed in the liver of G1 and G2 mice. Taken together, fluorescence bioimaging in this study showed the detection of cell death and made it possible to quantify the level of cell death in male mice. The outcome was correlated with conventional biomedical examination. As it was difficult to differentiate histological location by fluorescent bioimaging, it is necessary to develop specific fluorescent dyes for monitoring hepatic disease progression and to exploit new bioimaging techniques without dye-labeling.

• 한국생활과학회지
• /
• 제13권6호
• /
• pp.943-957
• /
• 2004

The purpose of this study is to 1) understand the importance of brands, which are known as one of the key purchasing points to consumers today; 2) 10 clearly clarify the various attributes of goods well-known to consumers; 3) to research the effect of the goods attributes on brand attitude. We examined 465 consumers to understand a purchase or a shopping habit with national and non-brand. The study used SPSS 11.0 package for a data analysis. Then using varimax rotation, we employed a factor analysis to analyze the data and obtain Cronbach ${\alpha}$ value. For more specific data analysis, we conducted t-test, regression analysis, and x2 analysis. The results are as followings: 1. The goods attribute is classified into product attribute, shop attribute, and price attribute. The product attribute is divided into wearability, labeling of size and quality, aesthetic expression, brand expression-harmony, textile-suitability, manageability and product assortment. The shop attribute is into shop environment, shopping convenience, promotion, salesperson service, convenience of location, shop's reputation. The price attribute is into price reasonability, price value, price economy, and price information. 2. The product attribute is different, by national and non-brands, in wearability, labeling of size and quality, brand expression-harmony, textile-suitability, manageability. The shop attribute had a significant difference in environment of shop and its reputation, and the price attribute had one only in price economy. 3. The brand attitude was affected by wearability, aesthetic expression, shop environment, price value, and price information. 4. Also, there exists a difference by demographic variables(age, job, academic background, marriage, income level) in brand attitude. For example, national brands have its consumers with the following variables: higher education, higher income, professional job, over 20s, and married.