• 제목/요약/키워드: non-specific binding

검색결과 149건 처리시간 0.024초

Wnt/$\beta$-catenin/Tcf Signaling Induces the Transcription of a Tumor Suppressor Axin2, a Negative Regulator of the Signaling Pathway

  • Jho, Eek-hoon;Tong Zhang;Claire Domon;Joo, Choun-Ki;Freund, Jean-Noel;Frank Costantini
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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    • pp.108-108
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    • 2001
  • Axin2/Conductin/Axil and its ortholog Axin are negative regulators of the Wnt signaling pathway, which promote the phosphorylation and degradation of ${\beta}$-catenin. While Axin is expressed ubiquitously, Axin2 mRNA was seen in a restricted pattern during mouse embryogenesis and organogenesis. Because many sites of Axin2 expression overlapped with those of several Wnt genes, we tested whether Axin2 was induced by Wnt signaling. Endogenous Axin2 mRNA and protein expression could be rapidly induced by activation of the Wnt pathway, and Axin2 reporter constructs, containing a 5.6 kb DNA fragment including the promoter and first intron, were also induced. This genomic region contains eight Tcf/LEF consensus binding sites, five of which are located within longer, highly conserved non-coding sequences. The mutation or deletion of these Tcf/LEF sites greatly diminished induction by ${\beta}$-catenin, and mutation of the Tcf/LEF site T2 abolished protein binding in an electrophoretic mobility-shift assay. These results strongly suggest that Axin2 is a direct target of the Wnt pathway, mediated through Tcf/LEF factors. The 5.6 kb genomic sequence was sufficient to direct the tissue specific expression of d2EGFP in transgenic embryos, consistent with a role for the Tcf/LEF sites and surrounding conserved sequences in the in vivo expression pattern of Axin2. Our results suggest that Axin2 participates in a negative feedback loop, which could serve to limit the duration or intensity of a Wnt-initiated signal.

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Association of Two Polymorphisms of DNA Polymerase Beta in Exon-9 and Exon-11 with Ovarian Carcinoma in India

  • Khanra, Kalyani;Panda, Kakali;Bhattacharya, Chandan;Mitra, A.K.;Sarkar, Ranu;Bhattacharyya, Nandan
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권4호
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    • pp.1321-1324
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    • 2012
  • Background: DNA polymerase beta ($pol{\beta}$) is a key enzyme in the base excision repair pathway. It is 39kDa protein, with two subunits, one large subunit of 31 kDa having catalytic activity between exon V to exon XIV, and an 8 kDa smaller subunit having single strand DNA binding activity. Exons V to VII have double strand DNA binding activity, whereas exons VIII to XI account for the nucleotidyl transferase activity and exons XII to XIV the dNTP selection activity. Aim: To examine the association between $pol{\beta}$ polymorphisms and the risk of ovarian cancer, the present case control study was performed using 152 cancer samples and non-metastatic normal samples from the same patients. In this study, mutational analysis of $pol{\beta}$ genomic DNA was undertaken using primers from exons IX to XIV - the portion having catalytic activity. Results: We detected alteration in DNA polymerase beta by SSCP. Two specific heterozygous point mutations of $pol{\beta}$ were identified in Exon 9:486, A->C (polymorphism 1; 11.18%) and in Exon 11:676, A->C (polymorphism 2; 9.86%). The correlation study involving polymorphism 1 and 4 types of tissue showed a significant correlation between mucinous type with a Pearson correlation value of 4.03 (p=0.04). The association among polymorphism 2 with serous type and stage IV together have shown Pearson ${\chi}^2$ value of 3.28 with likelihood ratio of 4.4 (p=0.07) with OR =2.08 (0.3-14.55). This indicates that there is a tendency of correlation among polymorphism 2, serous type and stage IV, indicating a risk factor for ovarian cancer. Conclusion: Hence, the results indicate that there is a tendency for $pol{\beta}$ polymorphisms being a risk factor for ovarian carcinogenesis in India.

소형 표면 플라즈몬 공명 센서와 이합체 구조를 가진 SAM을 이용한 CRP 검출 (A portable surface plasmon resonance sensor system for detection of C-reactive protein using SAM with dimer structure)

  • 신은정;정은정;조진희;황동환;손영수
    • 센서학회지
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    • 제19권6호
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    • pp.456-461
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    • 2010
  • The detection of C-reactive protein(CRP) using self-assembled monolayer(SAM) was investigated by a portable surface plasmon resonance(SPR) sensor system. The CRP is a biomarker for the possible cardiovascular disease. The SAM was formed on gold(Au) surface to anchor the monoclonal antibody of CRP(anti-CRP) for detection of CRP. Sequence injection of the anti-CRP and bovine serum albumin(BSA) into the sensor system has been carried out immobilize the antibody and to prevent non-specific binding. The portable SPR system has two flow channels: one for the sample measurements and the other for the reference. The output SPR signal was increased with the injection of the anti-CRP, BSA and CRP due to binding of the proteins on the sensor chip. The valid output SPR signals was linearly related to the critical range of the CRP concentration. The experimental results showed the feasibility of the portable SPR system with newly developed SAM to diagnose a risk of the future cardiovascular events.

용존성 방류수 유기물질이 비스페놀 A의 흡착 및 에스트로겐 활성에 미치는 영향 (Effect of Dissolved Effluent Organic Matter on Adsorption and Estrogenic Activity of Bisphenol A)

  • 유지수;나주림;정진호
    • Ecology and Resilient Infrastructure
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    • 제6권2호
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    • pp.128-135
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    • 2019
  • 본 연구는 하수종말처리장의 용존성 방류수 유기물질이 비스페놀 A (BPA)의 흡착과 에스트로겐 활성에 미치는 영향을 조사하였다. 특이적 자외선 흡수 (SUVA)와 형광 지수 (FI) 분석 결과, 방류수 유기물질은 참조물질로 사용한 스와니강 자연 유기물질과는 달리 미생물 기원의 비휴믹성 유기물질 특성을 나타내었다. Langmuir와 Freundlich 모델 모두 방류수 및 자연 유기물질에 대한 BPA의 흡착을 잘 설명하였다. 방류수 유기물질은 자연 유기물질 보다 더 높은 BPA 흡착능을 보여주었고, 이에 따라 에스트로겐 활성 역시 더 많이 감소시켰다. 이러한 결과들은 유기물질의 기원에 따라 BPA 흡착능과 에스트로겐 활성 저감이 크게 영향을 받는 다는 것을 제시한다.

Evaluation of Radiological Effects on the Aptamers to Remove Ionic Radionuclides in the Liquid Radioactive Waste

  • Minhye Lee;Gilyong Cha;Dongki Kim;Miyong Yun;Daehyuk Jang;Sunyoung Lee;Song Hyun Kim;Hyuncheol Kim;Soonyoung Kim
    • Journal of Radiation Protection and Research
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    • 제48권1호
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    • pp.44-51
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    • 2023
  • Background: Aptamers are currently being used in various fields including medical treatments due to their characteristics of selectively binding to specific molecules. Due to their special characteristics, the aptamers are expected to be used to remove radionuclides from a large amount of liquid radioactive waste generated during the decommissioning of nuclear power plants. The radiological effects on the aptamers should be evaluated to ensure their integrity for the application of a radionuclide removal technique. Materials and Methods: In this study, Monte Carlo N-Particle transport code version 6 (MCNP6) and Monte Carlo damage simulation (MCDS) codes were employed to evaluate the radiological effects on the aptamers. MCNP6 was used to evaluate the secondary electron spectrum and the absorbed dose in a medium. MCDS was used to calculate the DNA damage by using the secondary electron spectrum and the absorbed dose. Binding experiments were conducted to indirectly verify the results derived by MCNP6 and MCDS calculations. Results and Discussion: Damage yields of about 5.00×10-4 were calculated for 100 bp aptamer due to the radiation dose of 1 Gy. In experiments with radioactive materials, the results that the removal rate of the radioactive 60Co by the aptamer is the same with the non-radioactive 59Co prove the accuracy of the previous DNA damage calculation. Conclusion: The evaluation results suggest that only very small fraction of significant number of the aptamers will be damaged by the radioactive materials in the liquid radioactive waste.

LncRNA CRNDE Promotes the Progression of B-cell Precursor Acute Lymphoblastic Leukemia by Targeting the miR-345-5p/CREB Axis

  • Wang, Weimin;Wu, Feifei;Ma, Ping;Gan, Silin;Li, Xue;Chen, Li;Sun, Ling;Sun, Hui;Jiang, Zhongxing;Guo, Feng
    • Molecules and Cells
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    • 제43권8호
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    • pp.718-727
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    • 2020
  • The imbalance between the proliferation and apoptosis of B-cell precursors is an important contributor to the pathogenesis of B-cell precursor acute lymphoblastic leukemia (BCP-ALL), while its specific regulatory mechanism remains perplexing. This study aimed to expound the underlying mechanism of the proliferation and apoptosis of BCP-ALL cells from the perspective of non-coding RNA. In this study, long non-coding RNA colorectal neoplasia differentially expressed (LncRNA CRNDE) was upregulated in the bone marrow of BCP-ALL patients and BCP-ALL cell lines (NALM-6 and RS4;11). Functionally, LncRNA CRNDE knockdown restrained cell proliferation and boosted cell apoptosis in NALM-6 and RS4;11 cells. The subsequent investigation confirmed that LncRNA CRNDE bound to miR-345-5p and negatively regulated miR-345-5p expression. The overexpression of miR-345-5p suppressed cell proliferation and boosted cell apoptosis in NALM-6 and RS4;11 cells. Further experiments revealed that miR-345-5p downregulated cyclic AMP response element-binding protein (CREB) expression by targeting its mRNA directly. CREB overexpression reversed the effect of miR-345-5p mimic on cell proliferation and apoptosis in NALM-6 and RS4;11 cells. Finally, in vivo experiments showed that LncRNA CRNDE knockdown prolonged the survival of mice xenotransplanted with NALM-6 cells. In conclusion, LncRNA CRNDE upregulated CREB expression by suppressing miR-345-5p, thus promoting cell proliferation and reducing cell apoptosis in BCP-ALL.

Gene Expression Profiling of Non-Hodgkin Lymphomas

  • Zekri, Abdel-Rahman Nabawy;Hassan, Zeinab Korany;Bahnassy, Abeer Ahmed;Eldahshan, Dina Hassan;El-Rouby, Mahmoud Nour Eldin;Kamel, Mahmoud Mohamed;Hafez, Mohamed Mahmoud
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권7호
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    • pp.4393-4398
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    • 2013
  • Background: Chromosomal translocations are genetic aberrations associated with specific non-Hodgkin lymphoma (NHL) subtypes. This study investigated the differential gene expression profile of Egyptian NHL cases based on a microarray approach. Materials and Methods: The study included tissue samples from 40 NHL patients and 20 normal lymph nodes used as controls. Total RNA was extracted and used for cDNA microarray assays. The quantitative real time polymerase chain reaction was used to identify the aberrantly expressed genes in cancer. Results: Significant associations of 8 up-regulated and 4 down-regulated genes with NHL were observed. Aberrant expression of a new group of genes not reported previously was apparent, including down-regulated NAG14 protein, 3 beta hydroxy-delta 5-c27 steroid oxi-reductase, oxi-glutarate dehydrogenase (lipo-amide), immunoglobulin lambda like polypeptide 3, protein kinase x linked, Hmt1, and caveolin 2 Tetra protein. The up-regulated genes were Rb binding protein 5, DKFZP586J1624 protein, protein kinase inhibitor gamma, zinc finger protein 3, choline ethanolamine phospho-transferase CEPT1, protein phosphatase, and histone deacetylase-3. Conclusions: This study revealed that new differentially expressed genes that may be markers for NHL patients and individuals who are at high risk for cancer development.

Optimal Fuzzy Sliding-Mode Control for Microcontroller-based Microfluidic Manipulation in Biochip System

  • Chung, Yung-Chiang;Wen, Bor-Jiunn
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 2004년도 ICCAS
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    • pp.196-201
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    • 2004
  • In biometric and biomedical applications, a special transporting mechanism must be designed for the ${\mu}$TAS (micro total analysis system) to move samples and reagents through the microchannels that connect the unit procedure components in the system. An important issue for this miniaturization and integration is microfluid management technique, i.e., microfluid transportation, metering, and mixing. In view of this, this study presents an optimal fuzzy sliding-mode control (OFSMC) design based on the 8051 microprocessor and implementation of a complete microfluidic manipulated system implementation of biochip system with a pneumatic pumping actuator, a feedback-signal photodiodes and flowmeter. The new microfluid management technique successfully improved the efficiency of molecular biology reaction by increasing the velocity of the target nucleic acid molecules, which increases the effective collision into the probe molecules as the target molecules flow back and forth. Therefore, this hybridization chip was able to increase hybridization signal 6-fold and reduce non-specific target-probe binding and background noises within 30 minutes, as compared to conventional hybridization methods, which may take from 4 hours to overnight. In addition, the new technique was also used in DNA extraction. When serum existed in the fluid, the extraction efficiency of immobilized beads with solution flowing back and forth was 88-fold higher than that of free-beads.

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대두 사포닌의 활성에 관한 기전 연구

  • 성미경
    • Journal of Nutrition and Health
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    • 제28권10호
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    • pp.1022-1030
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    • 1995
  • Saponins are glycosidic compounds present in many plant foods. They are characterized by their ability to lyse cell membranes due to their surface-active properties. Saponins are believed to interact primarily with cholesterol in the cell membrane. In this study, the interaction of soybean(SS) with cell membrane was investigated using erythrocytes as a model. Mechanisms of interaction was also investigated by measuring their binding capacity with different membrane lipid fractions. Throughout the study, gypsophilla saponin(GS) and quillaja saponin(QS) were used to evaluate the membranolytic activity of soybean saponins. All saponins released hemoglobin in a concentration-dependent manner. SS induced 40% hemolysis at the concentration of 400 ppm, however there was no increase in hemoglobin release above 400ppm concentration. 5ppm of GS and 8 ppm of QS hemolyzed 100% of erythrocytes. Isolation of SS fractions by thin layer chromatography revealed that only one non-polar saponin possesses strong hemolytic activity. When saponins were incubated decreased the release of cholesterol. When the hemolytic activity of saponins was measured in the presence of other major membrane lipid components, sphingomyelin significantly reduced the hemolytic activity of SS, while cholesterol reduced the activity of QS. GS showed high affinity to other component(s) in the incubation media as well as lipids. These results suggest that the membranolytic activity of saponins are related to their specific chemical structure, which determines the interaction behavior between saponins and different membrane components, and thereby influence the biological activity.

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Tau mis-splicing in the pathogenesis of neurodegenerative disorders

  • Park, Sun Ah;Ahn, Sang Il;Gallo, Jean-Marc
    • BMB Reports
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    • 제49권8호
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    • pp.405-413
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    • 2016
  • Tau proteins, which stabilize the structure and regulate the dynamics of microtubules, also play important roles in axonal transport and signal transduction. Tau proteins are missorted, aggregated, and found as tau inclusions under many pathological conditions associated with neurodegenerative disorders, which are collectively known as tauopathies. In the adult human brain, tau protein can be expressed in six isoforms due to alternative splicing. The aberrant splicing of tau pre-mRNA has been consistently identified in a variety of tauopathies but is not restricted to these types of disorders as it is also present in patients with non-tau proteinopathies and RNAopathies. Tau mis-splicing results in isoform-specific impairments in normal physiological function and enhanced recruitment of excessive tau isoforms into the pathological process. A variety of factors are involved in the complex set of mechanisms underlying tau mis-splicing, but variation in the cis-element, methylation of the MAPT gene, genetic polymorphisms, the quantity and activity of spliceosomal proteins, and the patency of other RNA-binding proteins, are related to aberrant splicing. Currently, there is a lack of appropriate therapeutic strategies aimed at correcting the tau mis-splicing process in patients with neurodegenerative disorders. Thus, a more comprehensive understanding of the relationship between tau mis-splicing and neurodegenerative disorders will aid in the development of efficient therapeutic strategies for patients with a tauopathy or other, related neurodegenerative disorders.