• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1229-1235
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• 2006

A lab-on-a-chip (LoC) was designed for simultaneous monitoring of microorganisms, antibiotic residues, somatic cells, and pH in raw milk. The LoC was fabricated from polydimethylsiloxane (PDMS) using microelectromechanical system (MEMS) technology, which consisted of two parts; a protein array and microchannel. The protein array was fabricated by immobilizing five types of antibodies corresponding to two microorganisms, two antibiotic residues, and somatic cells. A sol-gel film was deposited on a glass substrate to immobilize the antibodies. The target analytes in raw milk could be bound with the corresponding antibody by an immunoreaction, and the antigen-antibody complex was detected using fluorescence microscopy. SNARF-dextran was used as a pH indicator, and the SNARF-entrapped hydrogel was attached to the microchannel in the chip. After injecting the milk sample into the channel, the pH was measured by monitoring the change in fluorescence intensity by fluorescence microscopy. The on-chip simultaneous assay of two microorganisms (E. coli O157:H7 and Streptococcus agalactiae), two antibiotic residues (penicillin G and dihydrostreptomycin), and neutrophils was successfully accomplished using the proposed LoC system.

• Asian Pacific Journal of Cancer Prevention
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• 제14권1호
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• pp.507-510
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• 2013

Background: More and more research indicate that the immediately early response gene 3 (IER3) is involved inmany biological provesses, such as apoptosis and immunoreaction, as well as viral infection, tumorigenesis and tumour progression. Methods: Here we describe the construction of an eukaryotic expression vector containing IER3 gene and its expression in A549 cells as assessed through fluorescence microscopyand Western-blotting. Results: Fluorescence detection displayed that GFP in cytoplasm was high during 48 and 72 hours post-transfection. In addition, Western blotting showed significant increase in IER3 gene expression in the transfected cells compared with controls. Conclusion: The recombinate plasmid expression vector was constructed successfully, which may provide a basis for further exploration of function of IER3 in lung cancer.

• 센서학회지
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• 제20권1호
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• pp.35-39
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• 2011

A disposable electrochemical immunosensor system has been developed for the detection of herbicide in aqueous samples. Disposable screen printed carbon electrodes(SPCE) were used as basic electrodes and an enzyme, horseradish peroxidase (HRP), and anti-herbicide antibodies was immobilised on to the working electrode of SPCE by using avidin-biotin coupling reactions. An herbicide-glucose oxidase conjugates have been used for the competitive immunoreaction with sample herbicides. The enzymatic reaction between the conjugated glucose oxidase and glucose added generates hydrogen peroxide, which was reduced by the peroxidase immobilised. The latter process caused an electrical current change, due to direct re-reduction of peroxidase by a direct electron transfer mechanism, which was measured to determine the herbicides in the sample. The optimal operational condition was found to be: $20\;{\mu}gl-1$ deglycosylated avidin loading to the working electrode and working potential +50 mV vs. Ag/AgCl. The total assay time was 15 min after sample addition. The detection limits for herbicides, atrazine and simazine, were found to be 3 ppb and 10 ppb, respectively.

• Bulletin of the Korean Chemical Society
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• 제31권11호
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• pp.3259-3264
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• 2010

A label-free amperometric immunosensor has been proposed for the detection of myeloperoxidase (MPO) in human serum. To fabricate such an immunosensor, a composite film consisting of N,N-dimethylformamide (DMF), multiwall carbon nanotubes (MWCNTs) and 1-ethyl-3-methyl imidazolium tetrafluoroborate ($EMIMBF_4$) suspension was initially formed on a glassy carbon electrode (GCE). Then cerium dioxide ($CeO_2$) dispersed by chitosan was coated on the GCE. After that, MPO antibodies (anti-MPO) were attached onto the nano$CeO_2$ surface. With a noncompetitive immunoassay format, the antibody-antigen complex formed between the immobilized anti-MPO and MPO in sample solution. The immunosensor was characterized by cyclic voltammetry, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The factors influencing the performance of the immunosensor were studied in detail. Under optimal conditions, the current change before and after the immunoreaction was proportional to MPO concentration in the range of 5 to $300\;ng\;mL^{-1}$ with a detection limit of $0.2\;ng\;mL^{-1}$.

• 대한한의학회지
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• 제26권1호
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• pp.18-25
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• 2005

Objective & Methods: We examined the effects of Platycodi radix on the process of lipopolysaccharide (LPS)-induced nuclear factor $NF-{\kappa}Bp65$ and inhibitory $(I)-{\kappa}B{\alpha}$ alteration in RAW 264.7 cells and acute lung injury in rats. Results: Immunoblot analysis showed that LPS-induced degradation of $I-{\kappa}B{\alpha}$ in RAW 264.7 was inhibited by pretreatment of Platycodi radix. The total cells of bronchoalveolar lavage fluid by LPS challenge markedly decreased in the Platycodi radix pretreatment rats. Platycodi radix pretreatment also caused a decline in neutrophils infiltration into interstitium of the lung. In the alveolar macrophages and neutrophils, decreased $NF-{\kappa}Bp65$ and inducible nitric oxide synthase and increased $I-{\kappa}B{\alpha}$ immunoreaction were detected in Platycodi radix pretreated rats compared with LPS alone treated ones. Conclusion : It may be concluded that Platycodi radix attenuates the development of LPS-induced inflammation by reduction of $NF-{\kappa}Bp65$ activation and neutrophil-mediated acute lung injury. Platycodi radix would be useful as a therapeutic agent for endotoxin-induced lung disease.

• Journal of Chest Surgery
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• 제31권4호
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• pp.422-426
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• 1998

53 세 남자가 하부 식도에서 발생한 거대한 용종성 종괴에 의한 연하곤란을 주소로 내원하여 식도 절제술과 식도-위 문합술을 시행하였다. 현미경소견상 기저세포양 편평세포암과 연골 분화가 여러 곳에서 관찰되는 연골육종이 혼재하는 진성암육종이었다. 위장주위 임파절과 기도분기부하 임파절에 편평세포암이 전이되어 있었다. 면역조직화학적 검사상 편평세포 지역은 cytokeratin 에 강한 양성이었고 기저세포양 지역은 고분자량 cytokeratin 에 양성이었다. 방추세포육종은 vimentin 과 smooth muscle actin 에 양성이었고 연골육종 지역은 vimentin 과 S-100 단백질에 양성이었다. 술후 항암제 치료와 방사선 치료의 보조병용요법을 시행하였으며 술후 11 개월인 현재까지 재발의 소견 없이 정상 식사를 하고 있다.

• Journal of Life Science
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• 제10권2호
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• pp.55-60
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• 2000

We examined the effects of Kamgil-Tang on the process of lipopolysaccharide (LPS)-induced unclear factor (NF)-${\kappa}$ Bp65 and inhibitory (I)-${\kappa}$ B${\alpha}$ alteration in RAW 264.7 cell and acute lung injury in rats. Immunoblot analysis showed that LPS-induced degradation of I-${\kappa}$ B${\alpha}$ in RAW 264.7 was inhibited by pretreatment of Kamgil-Tang. The total cells of bronchoalveolar lavage fluid by LPS challenge markedly decreased in the Kamgil-Tang pretreatment rats. Kamgil-Tang pretreatment caused also a decline in neutrophils infiltration into interstitium of the lung. In the alveolar macrophages and neutrophils, decreased NF-${\kappa}$ Bp65 and inducible nitric oxide synthase and increased I-${\kappa}$ B${\alpha}$ immunoreaction were detected in Kamgil-Tang pretreated rats compared with LPS alone treated ones. It may be concluded that Kamgil-Tang attenuates the development of LPS-induced inflammation by reduction of NF-${\kappa}$ Bp65 activation and neutrophil-mediated acute lung injury. Kamgil-Tang would be useful as a therapeutic agent for endotoxin-induced lung disease.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권4호
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• pp.309-314
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• 2005

We performed a basic experiment for the rapid, on-line, real-time measurement of hepatitis B surface antigen using a surface plasmon resonance biosensor. We immobilized anti­HBsAg (hepatitis B surface antigen) polyclonal antibody, as a ligand, to the dextran layer on a CM5 chip surface that had previously been activated by N-hydroxysuccinimide. A sample solution containing HBsAg was fed through a microfluidic channel, and the reflecting angle change due to the mass increase from the binding was detected. The binding characteristics between HBsAg and its polyclonal antibody followed the typical monolayer adsorption isotherm. When the entire immobilized antibody had interacted, no additional, non-specific binding occurred, suggesting the immunoreaction was very specific. The bound antigen per unit mass of the antibody was independent of the immobilized ligand density. No significant steric hindrance was observed at an immobilization density of approximately $17.6 ng/mm^2$. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the ligand was linear up to ca. $40{\mu}g$/mL. This linearity was much higher than that of the ELISA method. It appeared the anti­gen-antibody binding increased as the immobilized ligand density increased. In summary, this study showed the potential of this SPR biosensor-based method as a rapid, simple and multi­sample on-line assay. Once properly validated, it may serve as a more efficient method for HBsAg quantification for replacing the ELISA.

• Restorative Dentistry and Endodontics
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• 제21권1호
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• pp.35-53
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• 1996

The development and repair requires the formation of new tissues comprised of various extracellular matrix components. The present study investigated the formation and distribution of the major ECM components such as type I collagen, type III collagen, fibronection, bone sialoprotein, and osteonection during development and repair. For developing observation. Sprague-Dawley rats weighing $27{\pm}1gm$ were sacrificed. For repair observation, Sprague-Dawley rats weighing $110{\pm}5gm$ were used. The pulp perforation were prepared on mesial surface of the maxillary first molar by using 1/2round bur. At 5 days after perforation, rats were sacrificed by perfusion with 3 % paroformaldehyde. The maxillary first molar region were cut, demineralized, dehydrated and embedded in paraffin. Immunostaining the ECM components was achieved by the avidin-biotin complex method. The results as follows : 1. Bright immunoreaction for fibronectin was present in the basement membrane at the inner epithelial-mesenchymal interface, especially concentrated in the blood vessel walls, cell membrane of odontoblasts, and initial predentin. 2. Type I and III collagen was observed in the newly formed pulp tissue, predentin, and its intensity increased as more of these components during repair. 3. Strong immunostaining for bone sialoprotein and osteonectin was found in dentin while no or weaker staining was observed loose connective tissue of the pulp. 4. These results suggest that develpment and repair is achieved through a series of cell differentiation and attachment by the specific ECM components.

• 식물병연구
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• 제18권3호
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• pp.231-235
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• 2012

본 연구는 citrus에 심각한 피해를 초래하는 바이러스인 ICRSV가 국내로 유입되는 것을 차단하여 그로 인한 피해를 방지하기 위해 이를 진단하는 시스템을 구축하고자 하였다. ICRSV가 감염된 시료를 구할 수 없어 외피단백질 유전자를 E. coli의 codon usage를 고려하여 optimization한 뒤 E. coli에서 수용성 단백질로 과발현된 재조합 ICRSV 외피단백질을 정제하였다. 정제한 재조합 단백질을 이용해 제작한 복클론 항체는 $1{\times}10^{-4}$으로 희석하였을 때 western blot과 ELISA를 통해서 각각 10 ng, 5 ng의 재조합 ICRSV 외피단백질을 검출할 수 있었다. 이로써 제작된 항체를 이용하여 소량의 바이러스 입자만으로도 ICRSV를 검출할 수 있을 것이다.