• 제목/요약/키워드: linear assay

검색결과 272건 처리시간 0.026초

Streptomycin-anionic linear globular dendrimer G2: Novel antibacterial and anticancer agent

  • Javadi, Sahar;Ardestani, Mehdi Shafiee
    • Advances in nano research
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    • 제7권4호
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    • pp.241-248
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    • 2019
  • Recent researches demonstrated well promising anticancer activities for antibiotics. Such effects would be significantly increased while nanoparticle based delivery systems were applied. In this study, the goal was aim to improve anticancer and antitoxic effects of Streptomycin by loading on special kind of dendrimer (anionic-linear-globular second generation). In the current study, Size and zeta potential as well as AFM techniques have been used to prove the fact that the loading was performed correctly. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of the drug loaded on dendrimer nanoparticle were determined and compared with both of dendrimer alone and free drug with respect to staphylococcus aureus as the test microorganism. The anticancer activity among three groups including Streptomycin, Streptomycin -G2 dendrimer, and control was measured in vitro. In vitro studies showed that G2 anionic linear-globular polyethylene-glycol-based dendrimer, which loaded on Streptomycin was able to significantly improve the treatment efficacy over clinical Streptomycin alone with respect to proliferation assay. Maximal inhibitory concentration (IC50) was calculated to be $257{\mu}g/mL$ for streptomycin alone and $55{\mu}g/mL$ for Streptomycin -G2 dendrimer. In addition, Streptomycin -G2 dendrimer conjugate prevented the growth of MCF-7 cancerous cells in addition to enhance the number of apoptotic and necrotic cells as demonstrated by an annexin V-fluorescein isothiocyanate assay. Streptomycin -G2 dendrimer conjugate was able to increase Bcl-2/Bax ratio in a large scale compared with the control group and Streptomycin alone. Based on results a new drug formulation based nano-particulate was improved against S. aureus with sustained release and enhanced antibacterial activity as well as anticancer activity shown for functional cancer treatment with low side effects.

Metabolic Signatures of Adrenal Steroids in Preeclamptic Serum and Placenta Using Weighting Factor-Dependent Acquisitions

  • Lee, Chaelin;Oh, Min-Jeong;Cho, Geum Joon;Byun, Dong Jun;Seo, Hong Seog;Choi, Man Ho
    • Mass Spectrometry Letters
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    • 제13권1호
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    • pp.11-19
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    • 2022
  • Although translational research is referred to clinical chemistry measures, correct weighting factors for linear and quadratic calibration curves with least-squares regression algorithm have not been carefully considered in bioanalytical assays yet. The objective of this study was to identify steroidogenic roles in preeclampsia and verify accuracy of quantitative results by comparing two different linear regression models with weighting factor of 1 and 1/x2. A liquid chromatography-mass spectrometry (LC-MS)-based adrenal steroid assay was conducted to reveal metabolic signatures of preeclampsia in both serum and placenta samples obtained 15 preeclamptic patients and 17 age-matched control pregnant women (33.9 ± 4.2 vs. 32.8 ± 5.6 yr, respectively) at 34~36 gestational weeks. Percent biases in the unweighted model (wi = 1) were inversely proportional to concentrations (-739.4 ~ 852.9%) while those of weighted regression (wi = 1/x2) were < 18% for all variables. The optimized LC-MS combined with the weighted linear regression resulted in significantly increased maternal serum levels of pregnenolone, 21-deoxycortisol, and tetrahydrocortisone (P < 0.05 for all) in preeclampsia. Serum metabolic ratio of (tetrahydrocortisol + allo-tetrahydrocortisol) / tetrahydrocortisone indicating 11β-hydroxysteroid dehydrogenase type 2 was decreased (P < 0.005) in patients. In placenta, local concentrations of androstenedione were changed while its metabolic ratio to 17α-hydroxyprogesterone responsible for 17,20-lyase activity was significantly decreased in patients (P = 0.002). The current bioanalytical LC-MS assay with corrected weighting factor of 1/x2 may provide reliable and accurate quantitative outcomes, suggesting altered steroidogenesis in preeclampsia patients at late gestational weeks in the third trimester.

방사선 감수성 측정법으로서 MTT 법 시행 시의 최적 조건에 대한 연구 (The Optimal Condition of Performing MTT Assay for the Determination of Radiation Sensitivity)

  • 홍세미;김일한
    • Radiation Oncology Journal
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    • 제19권2호
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    • pp.163-170
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    • 2001
  • 목적 : 방사선조사후 세포의 생존 분획은 세포집락 측정기법으로 확인하는 것이 표준이나 많은 비용과 시간이 소요되는 단점을 갖고 있다. 이에 생존 세포의 tetrazolium염의 자색 formazan 침전물로의 환원시키는 능력에 그 기반을 둔 MTT 기법을 사용하여 세포집락 측정기법을 대체하기 위한 기법으로서의 유용성과 그 실행상의 최적조건을 규명하고자 하였다. 방법 : PCI-1, SNU-1066, NCI-H630, RKO등의 세포주에 0, 2, 4, 6, 8, 10 Gy의 방사선을 조사한 후 세포집락 측정 기법과 MTT기법으로 세포 생존 분획을 조사하였다. 세포집락 측정기법은 $25\;cm^2$ 폴리스티렌 배양 플라스크에 방사선량에 따라 다른 수의 세포를 분주한 후 24시간 동안 배양 후 방사선을 조사하였고 이를 $10\~14$일 동안 배양 후 염색하여 생성된 세포집락의 수를 측정하였다. MTT기법은 침전물의 용해과정이 필요없는 Premix WST-1 시약을 이용하여 시행하였다. MTT기법은 각각의 세포주에서 세포수와 흡광도간의 선형관계와 최적 실험조건을 확인한 이후 시행하였다. 이 기법은 방사선을 조사받은 세포에서는 지수적 성장을 회복한 이후와 방사선을 조사받지 않은 세포는 4회 이상의 세포분열을 거친 후에 시행하였다. 세포집락 측정기법 및 MTT기법을 통하여 얻은 세포 생존율을 구한 후 이를 지표로 비교하였다. 결과 : 각 기법으로 얻은 세포 생존율의 표준편차는 $5\%$ 내외였다. 2가지 방법으로 구한 세포 생존율은 t-test로 비교하였을 때 $0\~4\;Gy$에서는 통계적으로 유의한 차이가 없었으며 회귀분석 결과는 선형적 관계가 있었다$(R^2=0.975-0.992)$. MTT 기법의 시행에 최적인 세포수는 배양효율에 따라 다른 것으로 나타났는데, 배양효율이 $30\%$ 이상이면 300개 이하가, $30\%$ 미만인 경우는 $500\~1,000$개가 적당한 것으로 확인되었다. MTT기법은 6 배가시간 경과 이후에 시행하는 것이 세포집락 측정기법과 가장 근접하였으며 적어도 4 배가시간 이후에 시행하는 것이 필요할 것으로 사료되었다. 이에 따르면 배가시간이 3일 이하인 세포주가 세포 민감성 측정방법으로서 MTT 기법이 세포 집락 측정 기법을 대체하여 사용하기에 적합한 것으로 사료되었다. 결론 : 이상에서, MTT기법을 이용하여 방사선조사 후의 세포생존을 측정하기 위해서는 예비실험을 통해 각 세포주에서의 최적의 조건을 찾는 것이 필수적이며 이 조건하에서 MTT기법을 시행해야만 방사선에 의한 세포 민감성 측정에 이용될 수 있음을 확인하였다.

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종이기반 농도 구배 형성 칩을 통한 포도당 발색 반응 검사 (A Colorimetric Glucose Assay via Concentration Gradient Paper Chip)

  • 김태훈;신현영;이윤일;태기식;김민석
    • 대한의용생체공학회:의공학회지
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    • 제38권6호
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    • pp.302-307
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    • 2017
  • This paper presents a paper-based concentration gradient chip to analyze colorimetric glucose assay. The paper-based concentration gradient chip was fabricated through a wax patterning technique that can design the fluidic channel by selectively printing hydrophobic and hydrophilic areas. Afterwards, glucose and dilution solutions were loaded into the inlet of a concentration gradient chip and each solution was then mixed sequentially at mixing channel. Finally, concentration gradient was formed at each outlet of the chip. To measure the glucose concentration of the solution in outlets, we conducted colorimetric glucose assay with fixed concentration of glucose solution (0, 5, 10, 15 and 20 mM) and obtained normalized intensity. Subsequently, glucose concentrations of the outlets were calculated by substituting the normalized intensity to linear regression function based on the normalized intensity of fixed glucose concentration. Finally, the concentration gradient of glucose was formed on the chip with the result of colorimetric assay. The concentration gradient paper chip has the potential to accurately analyze unknown glucose concentration.

항산화 활성의 평가를 위한 연구법: 타우린의 적용 (Methods for Evaluation of Antioxidant Activity: Application to Taurine)

  • 김봉희;오정민;윤강욱;김충현;김상겸
    • Toxicological Research
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    • 제23권3호
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    • pp.263-269
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    • 2007
  • Although taurine (2-aminoethanesulfonic acid) can inhibit oxidative stress in both animal and epidemiological studies, it is obscure whether taurine directly scavenges oxy-radicals or indirectly regulates oxidant production and/or antioxidant defense system. The reason for this discrepancy remains unknown but may be due, in part, to the lack of a validated assay system for evaluating oxy-radical scavenging capacity. The antioxidant activities of taurine and hypotaurine (2-aminoethanesulfinic acid), a precursor of taurine, against peroxyl radicals, hydroxyl radicals and peroxynitrites were determined by the total oxy-radical scavenging capacity (TOSC) assay and cell-based assay using H4IIE cells. tert-Butylhydroperoxide or hydrogen peroxide-induced cell toxicity determined by MTT assay was markedly inhibited by 10mM taurine or hypotaurine. The tert-butylhydroperoxide- or hydrogen peroxide-induced changes in oxidative stress markers, such as cellular glutathione and malondialdehyde, were ameliorated by 10mM taurine or hypotaurine. However, specific TOSC values calculated from the slope of the linear regression for taurine against peroxyl radicals, hydroxyl radicals or peroxynitrites were all less than 1 TOSC/mM. On the other hand specific TOSC values for hypotaurine against peroxyl radicals, hydroxyl radicals or peroxynitrites were 48, 2096, or 69 TOSC/mM, respectively. These results suggest that taurine protects cells against oxidative insults, which is not ascribed to directly scavenging activity of taurine against oxy-radicals. These results support the idea that the oxidation state of sulfur in antioxidants may be a determinant of oxy-radical scavenging capacity.

DEVELOPMENT OF LEAD SLOWING DOWN SPECTROMETER FOR ISOTOPIC FISSILE ASSAY

  • Lee, YongDeok;Park, Chang Je;Ahn, Sang Joon;Kim, Ho-Dong
    • Nuclear Engineering and Technology
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    • 제46권6호
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    • pp.837-846
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    • 2014
  • A lead slowing down spectrometer (LSDS) is under development for analysis of isotopic fissile material contents in pyro-processed material, or spent fuel. Many current commercial fissile assay technologies have a limitation in accurate and direct assay of fissile content. However, LSDS is very sensitive in distinguishing fissile fission signals from each isotope. A neutron spectrum analysis was conducted in the spectrometer and the energy resolution was investigated from 0.1eV to 100keV. The spectrum was well shaped in the slowing down energy. The resolution was enough to obtain each fissile from 0.2eV to 1keV. The detector existence in the lead will disturb the source neutron spectrum. It causes a change in resolution and peak amplitude. The intense source neutron production was designed for ~E12 n's/sec to overcome spent fuel background. The detection sensitivity of U238 and Th232 fission chamber was investigated. The first and second layer detectors increase detection efficiency. Thorium also has a threshold property to detect the fast fission neutrons from fissile fission. However, the detection of Th232 is about 76% of that of U238. A linear detection model was set up over the slowing down neutron energy to obtain each fissile material content. The isotopic fissile assay using LSDS is applicable for the optimum design of spent fuel storage to maximize burnup credit and quality assurance of the recycled nuclear material for safety and economics. LSDS technology will contribute to the transparency and credibility of pyro-process using spent fuel, as internationally demanded.

Preliminary Application of Molecular Monitoring of the Pacific Herring (Clupea pallasii) Based on Real-time PCR Assay Utilization on Environmental Water Samples

  • Kim, Keun-Yong;Heo, Jung Soo;Moon, Seong Yong;Kim, Keun-Sik;Choi, Jung-Hwa;Yoo, Joon-Taek
    • 생태와환경
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    • 제54권3호
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    • pp.209-220
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    • 2021
  • Pacific herring, Clupea pallasii, a keystone species with significant ecological and commercial importance, is declining globally throughout much of its range. While traditional fishing equipment methods remain limited, new sensitive and rapid detection methods should be developed to monitor fisheries resources. To monitor the presence and quantity of C. pallasii from environmental DNA (eDNA) extracted from seawater samples, a pair of primers and a TaqMan® probe specific to this fish based on mitochondrial cytochrome b (COB) sequences were designed for the real-time PCR (qPCR) assay. The combination of our molecular markers showed high specificity in the qPCR assay, which affirmed the success of presenting a positive signal only in the C. pallasii specimens. The markers also showed a high sensitivity for detecting C. pallasii genomic DNA in the range of 1 pg~100 ng rxn-1 and its DNA plasmid containing COB amplicon in the range of 1~100,000copies rxn-1, which produced linear standard calibration curves (r2=0.99). We performed a qPCR assay for environmental water samples obtained from 29 sampling stations in the southeastern coastal regions of South Korea using molecular markers. The assay successfully detected the C. pallasii eDNA from 14 stations (48.2%), with the highest mean concentration in Jinhae Bay with a value of 76.09±18.39 pg L-1 (246.20±58.58 copies L-1). Our preliminary application of molecular monitoring of C. pallasii will provide essential information for efficient ecological control and management of this valuable fisheries resource.

경사비율 방법에 의한 단백질의 영양가 평가 (Evauation of the Nutritive Quality of Proteins by Using Slope-Ratio Assay)

  • 오훈일
    • 한국식품영양과학회지
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    • 제20권3호
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    • pp.197-202
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    • 1991
  • 서로 다른 3가지 단백질인 락타 알부민, 대두 및 땅콩 단백질의 영양가를 평가하기 위하여 각 단백질의 함량(0-35%)을 달리한 사료를 어린쥐에게 2주간에 걸쳐 먹었다. 선량으로 전체 질소 소비량을 측정하여 이에 대한 체중 증가로 반응을 측정하였다. 직선 부위에 있는 단백질의 회귀분석에 대한 경사를 표준단백질인 락타 알부민의 경사와 비교한 결과 대두 및 땅콩 단백질의 상대적 성장지수는 각각 78.4 및 55.7로 나타났다. 본 실험의 결과 경사 비율방법은 잘 계획된 실험 조건하에서 단백질의 영양가를 평가하는데 매우 유용한 방법이라고 사료된다.

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Progesterone의 단크론성 항체에 관한 특성 및 활용에 관한 연구 II. ELISA 기법의 개발 (Characteristics and application of monoclonal antibody to progesterone II. Development of progesterone enzyme-linked immunosorbent assay(ELISA))

  • 강정부;김종수
    • 대한수의학회지
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    • 제31권4호
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    • pp.403-409
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    • 1991
  • Progesterone의 단크론성 항체를 생산, 이용하여 감도가 높으면서도 신속히 측정할 수 있는 ELISA 기법을 처음으로 개발코져 실시하였다. 단크론성 항체는 종래의 면역방법에 의해 획득한 항혈청에 비해 약 10배의 결합율을 보였고 titer 역시 높았다. Dot-blot 분석 결과 단크론성 항체는 IgM이었다. 경합반응은 2시간으로 충분하였고, progesterone 표준용액을 이용한 표준 곡선은 0~1000pg/well에서 거의 직선적이었다. Progesterone의 단크론성 항체를 이용한 ELISA는 임상적으로는 물론 연구용으로도 신속한 항체의 기능 측정에는 물론 각종 번식 관련의 지표로 충분히 활용될 수 있을 것으로 판단된다.

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Fissile Measurement in Various Types Using Nuclear Resonances

  • YongDeok Lee;Seong-Kyu Ahn
    • 방사성폐기물학회지
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    • 제21권2호
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    • pp.235-246
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    • 2023
  • Neutron resonance transmission technique was applied for assaying isotopic fissile materials produced in the pyro-process. In each process of the pyro-process, a different composition of the fissile material is produced. Simulation was basically performed on 235U and 239Pu assay for TRU-RE product, hull waste, and uranium addition. The resonance energies were evaluated for uranium and plutonium in the simulation, and the linearity in the detection response was examined on the fissile content variation. The linear resonance energies were determined for the analysis of 235U and 239Pu on the different fissile materials. For enriched TRU-RE assay, the sample condition was suggested; The sample density, content, and thickness are the key factors to obtain accurate fissile content. The detection signal is discriminated for uranium and plutonium in neutron resonance technique. The transmitted signal for fissile resonance has a direct relation with the content of fissile. The simulation results indicated that the neutron resonance technique is promising to analyze 235U and 239Pu for various types of the pyro-process material. An accurate fissile assay will contribute toward safeguarding the pyro-processing system.