• Food Science and Biotechnology
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• 제17권3호
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• pp.651-654
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• 2008

In order to enhance the efficacy of norovirus detection by reverse transcriptase-polymerase chain reaction (RT-PCR) and nested PCR, this study developed a norovirus mRNA concentration method using poly oligo dT-conjugated magnetic beads. An efficient norovirus detection protocol was performed on commercial ham using 2 viral elution buffers (glycine buffer and Tris beef extract buffer) and 2 concentration solutions [polyethylene glycol (PEG) and zirconium hydroxide]. The different approaches were verified by RT-PCR and nested PCR. This method was performed on ham in less than 8 hr by artificial inoculation of serial dilutions of the virus ranging from 1,000 to 1 RT-PCR unit/mL. The viral extraction and concentration method had 10-fold higher sensitivity using the combination of Tris beef extract buffer and PEG as compared to glycine buffer and zirconium hydroxide. This method proved that RT-PCR and nested PCR have the sensitive ability to detect norovirus in commercial ham, in that norovirus was successfully detected in artificially contaminated samples at a detection level as low as 1-10 RT-PCR unit/mL. Overall, such a detection limit suggests this protocol is both quick and efficient in terms of its potential use for detecting norovirus in meat products.

• Journal of Microbiology and Biotechnology
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• 제31권9호
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• pp.1323-1329
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• 2021

Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a double-stranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.

• 한국자기학회지
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• 제24권4호
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• pp.101-106
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• 2014

Dc 마그네트론 스퍼터링법으로 자기저항비 5.0 %와 자장감응도 1.5 %/Oe를 갖는 glass/Ta(5.8 nm)/NiFe(5 nm)/Cu(2.3 nm)/NiFe(3 nm)/IrMn(12 nm)/Ta(5.8 nm) 다층구조 GMR-SV 박막을 증착하였다. 광 리소그래피 공정으로 적혈구 직경 크기인 $7{\mu}m{\sim}8{\mu}m$ 이내의 선폭인 GMR-SV 소자를 제작하였다. 직경 $1{\mu}m$ 크기인 여러 개의 자성비드가 결합된 적혈구를 자기저항비 1.06 % 자장감응도 0.3 %/Oe를 갖는 GMR-SV 소자에 떨어뜨려 -0.6 Oe 부근에서 약 $0.4{\Omega}$과 약 0.15 %의 변화를 관찰하였다. 본 연구로부터 미크론 크기의 선폭을 갖는 GMR-SV 소자가 자성비드를 결합한 적혈구내 헤모글로빈의 새로운 자성 특성을 분석하는 바이오센서로 활용할 수 있음을 보여주었다.

• 대한의용생체공학회:의공학회지
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• 제28권1호
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• pp.102-107
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• 2007

In molecular imaging studies via magnetic resonance imaging, in vivo cell tracking is an important issue for the observation of cell therapy or disease behavior. High resolution imaging and longitudinal study are necessary to track the cell movement. Since the field inhomogeneity extends over several voxels, we have performed the numerical analysis using the sub-voxel method dividing a voxel of MR image into several elements and the information about the field inhomogeneity distribution around the micro-beads. We imbedded ferrite-composite micro-beads with the size of $20-150{\mu}m$ in the subject substituted for cells to induce local field distortion. In the phantom imaging with the isotropic voxel size of $200{\mu}m^3$, we could confirm the feasibility of sub-voxel tracking in a 3.0 T MRI.

• 동의생리병리학회지
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• 제17권5호
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• pp.1330-1334
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• 2003

Astragali Radix(AR), one of the strong tonic herbs, is known to improve immunological responses in mice and human. In this study, AR's ai-reinforcing effect was examined in the context of CD4/sup +/ T cells' TCR/CD3 induced activation responses. In order to evaluate the direct effect of AR on helper T cells, CD4/sup +/ T cells are isolated using magnetic bead and their proliferation and CD69 expression in AR treated medium were assessed with anti-CD3/anti-CD28 activation for 48h. CD4 T cells' proliferation was slightly increased but there was little effect on CD69 expression. RT PCR and ELISA equally demonstrated that IL-2 and IL-4 production was increased but IFN-ν was down-regulated. This shows AR ethanol extract favors Th2 cytokine profile under neutral conditions.

• 한국어병학회지
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• 제7권2호
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• pp.95-103
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• 1994

미꾸라지의 성장호르몬 유전자를 분리하기 위하여 미꾸라지의 cDNA library를 준비하였다. total RNA는 미꾸라지의 뇌하수체로부터 얻었으며 oligo (dT)-coupled magnetic bead를 이용하여 total RNA로부터 mRNA를 순수분리하였다. 정제된 mRNA는 cDNA를 합성하기 위한 기질로 사용하였으며, 합성된 cDNA는 EcoRV/Smal으로 절단된 pBlueKS+ plasmid vector에 삽입하였다. 모든 ligation 반응용액을 E. coli, JM109 균주에 형질전환을 유도하였으며 형질전환 효율을 최대화시키기 위하여 전기천공법을 이용하였다. 얻어진 모든 형질전환주들을 DIG로 표지된 Tilapia의 성장호르몬 유전자를 이용하여 고밀도 colony hybridization 에 의하여 검색하였다. 양성반응을 나타내는 10개의 형질전환주를 분리하여 2차 colony hybridization 및 southern hybridization에 의하여 성장호르몬 유전자가 cloning 되었음을 확인하였다. 10 개의 형질전환주 중 하나인 pCGH1을 probe로 사용한 Tilapia 성장 호르몬 유전자의 염기서열과 비교분석하였으며 53.2%의 유사성을 나타냄을 확인하였다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권10호
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• pp.6069-6075
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• 2013

Background: At present, the diagnosis of colorectal cancer (CRC) requires a colorectal biopsy which is an invasive procedure. We undertook this pilot study to develop an alternative method and potential new biomarkers for diagnosis, and validated a set of well-integrated tools called ClinProt to investigate the serum peptidome in CRC patients. Methods: Fasting blood samples from 67 patients diagnosed with CRC by histological diagnosis, 55 patients diagnosed with colorectal adenoma by biopsy, and 65 healthy volunteers were collected. Division was into a model construction group and an external validation group randomly. The present work focused on serum proteomic analysis of model construction group by ClinProt Kit combined with mass spectrometry. This approach allowed construction of a peptide pattern able to differentiate the studied populations. An external validation group was used to verify the diagnostic capability of the peptidome pattern blindly. An immunoassay method was used to determine serum CEA of CRC and controls. Results: The results showed 59 differential peptide peaks in CRC, colorectal adenoma and health volunteers. A genetic algorithm was used to set up the classification models. Four of the identified peaks at m/z 797, 810, 4078 and 5343 were used to construct peptidome patterns, achieving an accuracy of 100% (> CEA, P<0.05). Furthermore, the peptidome patterns could differentiate the validation group with high accuracy close to 100%. Conclusions: Our results showed that proteomic analysis of serum with MALDI-TOF MS is a fast and reproducible approach, which may provide a novel approach to screening for CRC.

• 대한의생명과학회지
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• 제29권4호
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• pp.363-370
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• 2023

Human respiratory viral infections such as COVID-19 are highly contagious, so continuous management of airborne viruses is essential. In particular, indoor air monitoring is necessary because the risk of infection increases in poorly ventilated indoors. However, the current method of detecting airborne viruses requires a lot of time from sample collection to confirmation of results. In this study, we proposed a system that can monitor airborne viruses in real time to solve the deficiency of the present method. Air samples were collected in liquid form through a bio sampler, in which case the virus is present in low concentrations. To detect viruses from low-concentration samples, viral RNA was concentrated and extracted using silica-magnetic beads. RNA binds to silica under certain conditions, and by repeating this binding reaction, bulk samples collected from the air can be concentrated. After concentration and extraction, viral RNA is specifically detected through real-time qPCR (quantitative polymerase chain reaction). In addition, based on liquid handling technology, we have developed an automatic machine that automatically performs the entire testing process and can be easily used even by non-experts. To evaluate the system, we performed air sample collection and automated testing using bacteriophage MS2 as a model virus. As a result, the air-collected samples concentrated by 45 times then initial volume, and the detection sensitivity of PCR also confirmed a corresponding improvement.

• 대한용접접합학회:학술대회논문집
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• 대한용접접합학회 2002년도 Proceedings of the International Welding/Joining Conference-Korea
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• pp.361-365
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• 2002

Advancements in silicon phase control (SCR) technologies provide an arc welding power supply that has the capability to allow the alteration of the Alternating Current (AC) welding output. These technologies provide a square wave output involving sixteen frequency selections and multiple balance selections. While an AC out put is known to minimize magnetic disturbances associate with Direct Current (DC), the potentials of a non-sinusoidal waveform have not been explored. The focus of the paper is to determine the effects that the frequency and balance of an AC wave form output will have upon a high speed Submerge Arc (SAW) application. The test matrix of the project includes welding .250" steel plate. Joint type is square groove with a travel speed of 65 IPM. Each of the weld parameters was held constant, only the frequency and/or balance were altered between welds. Each frequency/balance combination involved three-gap spacing. Upon completion of the welds the bead profiles were measured and recorded. A relationships/trends were observed with various frequency and balance values. Optimum frequency and balance values were found for the .250" square groove application which permit consistent weld sizing, ease of slag removal, and minimal plate distortion.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2002년도 제9차 국제심포지움 및 추계정기학술발표회
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• pp.52-52
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• 2002

Chitosan is a key compound of shrimp waste. It is a biopolymer, which is widely used in the field of medical Sciences. Chitosan-TPP (Tripolyphosphate) complex has more or less similar physical properties as Ca-alginate which can be used for the production of synthetic seeds. Possibility of the use of Chitosan-TPP complex as a matrix for encapsulation of somatic embryos was tested against the Ca-alginate complex (2.5w/v Na-alginate, 100mM CaCl2 at pH 5.5). Somatic embryos grown in the induction medium (IM) were drawn into the viscous chitosan solution (1%) and mixed well by inverting the tube carefully. Then the mixture was dropped at regular intervals into the tripolyphosphate (TPP) solution kept on a magnetic stirrer for bead formation. Synthetic seeds formed were washed and transferred into the incubation medium, then allowed either to air-dry or freeze-dry.(중략)