• 제목/요약/키워드: Colorimetric assay

검색결과 241건 처리시간 0.027초

神授衛生湯의 抗腫瘍 效果 (Effects of Shinsuwuisaeng-Tang on the Anti-Tumor)

  • 배진석;최정화;김종한
    • 한방안이비인후피부과학회지
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    • 제15권2호
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    • pp.132-144
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    • 2002
  • Shinsuwuisaeng-Tang was a drug that treated carbuncle and cellulitis. So, the purpose of this Study was to investigate effect of Shinsuwuisaeng-Tang on the anti-cancer and nitric oxide(NO) production of peritoneal macrophages. We used Shinsuwuisaeng-Tang extract(SWT) with freeze-dried, 8wks-old male mice. and cancer cell lines(L1210, sarcoma-180) for this Study. The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). The results of this Study were obtained as follow; SWT was showed cytotoxicity on the L1210 and sacoma-180(S-180) cell lines, SWT inhibited significantly proliferation of L1210 cells in L1210 cells transplanted mice, SWT accelerated NO production of peritoneal macrophages in L1210 cells transplanted mice. And SWT inhibited significantly tumor weight, increased significantly body weight and mean survival days in S-180 cells transplanted mice. This results suggest that SWT has anti-cancer by producing NO of peritoneal macrophages.

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Construction of a Baculovirus Expression System Using Hyphantria cunea Nuclear Polyhedrosis Virus for Eukaryotic Cells

  • Lee, Hyung-Hoan;Kang, Bong-Joo;Park, Kap-Ju;Cha, Soung-Chul
    • Journal of Microbiology and Biotechnology
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    • 제8권6호
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    • pp.676-684
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    • 1998
  • Baculovirus transfer and expression vectors with Hyphantria cunea nuclear polyhedrosis virus (HcNPV) were constructed. An initial transfer vector, pHcEV, constructed using HcNPV was previously reported (Park et al. 1993. J. Kor. Soc. Viral. 23: 141-151). Herein, the size of the vector was properly reduced, and a functionally perfect vector was constructed and named pHcEV-IV (6.7 kb). The vector has a 2.2-kb HcNPV DNA sequence in the 5'-flanking region of the vector's polyhedrin gene promoter. The 1.8-kb HcNPV DNA sequence, poly A signal sequence, T3 primer sequence, and 13 multicloning site sequences, in order, were ligated in front of the translation start codon of the polyhedrin gene. The cloning indicating marker lacZ gene was inserted into the pHcEV-IV, named pHcEV-IV-lacZ, and transferred into the wild-type virus. Recombinant expression virus, lacZ-HcNPV, was constructed by replacing the lacZ gene in the pHcEV-IV-lacZ with the polyhedrin gene of the wild-type virus. The recombinant virus was isolated from blue plaques that produce $\beta$-galactosidase without polyhedra. The lacZ gene insertion was confirmed by Southern hybridization analysis. The expression of the lacZ gene in Spodoptera frugiperda cells infected with the lacZ-HcNPV was examined by SDS-PAGE and colorimetric assay. One 116-kDa LacZ protein band appeared on the PAGE. The production rate of the $\beta$-galactosidase was approximately 50 international units (IU) per min per ml between 2 to 5 days postinfection (p.i.). The highest activity occurred at five days p.i. was 170 IU/min/$m\ell$. The enzyme activity first appeared about 20 h p.i. as measured by colorimetric assay.

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말라카이트 그린의 색엷음 현상을 이용한 글리세롤의 정량: 바이오디젤 내 반응물 분석의 적용 가능성 (Quantification of Glycerol by Malachite Green Fading Phenomenon: Application in Reaction By-Product of Biodiesel)

  • 이미화;이영철;신현재
    • KSBB Journal
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    • 제26권5호
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    • pp.471-476
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    • 2011
  • Nowadays biodiesel (fatty acid methyl ester, FAME) has been becoming an important issue as a desired alternative of energy products because of non-toxic, biodegradable properties, and lower exhaust emissions. During esterification of fatty acids or transesterification of oils and fats with short chain alcohols by the alkali-catalyzed methanolysis, FAME and unrefined glycerol are generated. Quantification of glycerol as a by-product is important because of a determinant of biodiesel quality. However, the glycerol analysis by gas chromatography (GC) method has laborious works with sample preparation, long time and cost of sample analysis. Thus, there is a need to analyze glycerol more simply. Herein we demonstrate that the colorimetric assay for glycerol analysis conducted by UV-vis spectrophotometer at the wavelength 617 nm whose peak is maximum intensity of malachite green, resulting in the red-shift occurred proportionally as a function of glycerol amount. Thus, it is considered the solvent media for malachite green fading for biodiesel production: (1) water, (2) MeOH, and (3) EtOH. The resulting findings show that the peak intensity at 617 nm in glycerol-malachite green mixture had a relationship between glycerol concentration and degree of peak shift as increase in pure glycerol concentration approximately at pH 7.0. However, when it was measured the unrefined glycerol concentration by diluting and adjusting with water to buffer (pH 7.0), it was not observed the absorption peak at 617 nm because of impurities and OH ions. In case of glycerol from biodiesel production factories, glycerol concentration could be successfully measured.

Anti-inflammatory activity of Ganoderma lucidum by inhibition of NF-κB p65 phosphorylation

  • Kim, Hyung Don;Park, Jeong-Yong;Noh, Hyung-Jun;Lee, Seung Eun;Lee, Jeong Hoon;Seo, Kyung Hye
    • 농업과학연구
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    • 제46권3호
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    • pp.653-660
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    • 2019
  • Ganoderma lucidum, an oriental polypore fungus and medicinal mushroom, has a long history of use for promoting health and longevity in Korea, China, and other Asian countries. This study was aimed at determining the anti-inflammatory activity and mechanism of action of Ganoderma lucidum in murine macrophage RAW 264.7 cells. Ganoderma lucidum was extracted with ethanol and freeze-dried. The anti-inflammatory effect (nitrite production) of Ganoderma lucidum extracts was tested using a nitric oxide (NO) colorimetric assay. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was performed to quantify the mRNA expression of cytokines including tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, and IL-6. Western blotting was performed to measure the expression levels of inflammation-related proteins, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor kappa B ($NF-{\kappa}B$) p65, and phosphorylated $NF-{\kappa}B$ p65. The NO colorimetric assay showed that NO production increased with the treatment of lipopolysaccharide in (LPS)-activated RAW 264.7 macrophages and decreased with the cotreatment of Ganoderma lucidum extracts and LPS. Ganoderma lucidum extracts repressed the mRNA expressions of cytokines, which were increased after the LPS treatment. In addition, Ganoderma lucidum extracts inhibited the LPS-induced expression of iNOS and COX-2 and the LPS-induced phosphorylation of $NF-{\kappa}B$ p65. These results suggest that the Ganoderma lucidum extracts exert an anti-inflammatory activity by inhibiting $NF-{\kappa}B$ related proteins and cytokines.

Development of a biosensor from aptamers for detection of the porcine reproductive and respiratory syndrome virus

  • Kuitio, Chakpetch;Rasri, Natchaya;Kiriwan, Duangnapa;Unajak, Sasimanas;Choowongkomon, Kiattawee
    • Journal of Veterinary Science
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    • 제21권5호
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    • pp.79.1-79.12
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    • 2020
  • Background: Recently, the pork industry of Thailand faced an epidemic of highly virulent strains of porcine reproductive and respiratory syndrome virus (PRRSV), which spread throughout Southeast Asia, including the Lao People's Democratic Republic and Cambodia. Hence, the rapid and on-site screening of infected pigs on a farm is essential. Objectives: To develop the new aptamer as a biosensor for detection PRRSV which are rapid and on-site screening of infected pig. Methods: New aptamers against PRSSV were identified using the combined techniques of capillary electrophoresis, colorimetric assay by gold nanoparticles, and quartz crystal microbalance (QCM). Results: Thirty-six candidate aptamers of the PRRSV were identified from the systematic evolution of ligands by exponential enrichment (SELEX) by capillary electrophoresis. Only 8 out of 36 aptamers could bind to the PRSSV, as shown in a colorimetric assay. Of the 8 aptamers tested, only the 1F aptamer could bind specifically to the PRSSV when presented with the classical swine fever virus and a pseudo rabies virus. The QCM was used to confirm the specificity and sensitivity of the 1F aptamer with a detection limit of 1.87 × 1010 particles. Conclusions: SELEX screening of the aptamer equipped with capillary electrophoresis potentially revealed promising candidates for detecting the PRRSV. The 1F aptamer exhibited the highest specificity and selectivity against the PRRSV. These findings suggest that 1F is a promising aptamer for further developing a novel PRRSV rapid detection kit.

플라즈모닉 금 나노입자의 흡광 특성을 활용한 생화학적 비색 분석법 연구 동향 (Recent Progress in Colorimetric Assays Using the Absorption of Plasmonic Gold Nanoparticles)

  • 김봉근;윤상빈;황수경;나현빈
    • 공업화학
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    • 제35권2호
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    • pp.67-78
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    • 2024
  • 흡광은 측정이 간편하고 해석의 직관성이 높다는 점에서 생화학 기반 분석법의 신호로서 강점을 가진다. 흡광을 가지는 물질 중에서 금 나노입자는 화학적 안정성, 생물학적 친화성, 가시광선 범위에서 야기되는 국소 표면 플라즈몬 공명(localized surface plasmon resonance, LSPR)에 의한 독특한 광학적 특성 등의 유용한 성질을 지니며, 특정 표적 물질에만 유효한 다른 발색물질과 비교해 항체나 압타머 등 다양한 검출 활성물질과 접합이 용이하여 확장성을 가진다. 특히, 기질의 산화로 발색을 야기하는 효소 기반 발색법에 비해 낮은 가격, 쉬운 입자 합성, 높은 환경안정성 등의 장점으로 인해 비색화 분석법의 신호물질로서 광범위하게 연구되고 있다. 본 총설에서는 이와 같은 금 나노입자를 신호물질로 활용하는 다양한 전략을 최근의 연구들을 중심으로 요약 정리하였으며, 입자의 형태에 광학 특성이 영향을 받는 금 나노입자의 특징에 착안하여 신호생성 시에 활용한 금 나노입자의 형태 제어 전략을 기준으로 문헌들을 분류하고 검토하였다. 이를 통해 이미 오랜 기간 활용되어온 금 나노입자가 현재에도 흡광 신호물질로서 여전히 활발하게 연구되고 있다는 사실을 고찰하고, 향후에도 광범위하고 지속적으로 개선될 여지를 가진다는 점을 확인하였다.

클러스터 초상자성체 산화철 나노입자를 이용한 색채학적 해석 기반 당 측정 (Colorimetric Based Analysis Using Clustered Superparamagnetic Iron Oxide Nanoparticles for Glucose Detection)

  • 최원석;기재홍
    • 대한의용생체공학회:의공학회지
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    • 제41권6호
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    • pp.228-234
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    • 2020
  • Superparamagnetic iron oxide nanoparticles (SPIONs) are approved by the Food and Drug Administration (FDA) in the United States. SPIONs are used in magnetic resonance imaging (MRI) as contrast agents and targeted delivery in nanomedicine using external magnet sources. SPIONs act as an artificial peroxidase (i.e., nanozyme), and these reactions were highly stable in various pH conditions and temperatures. In this study, we report a nanozyme ability of the clustered SPIONs (CSPIONs) synthesized by the oil-in-water (O/W) method and coated with biocompatible poly(lactic-co-glycolic acid) (PLGA). We hypothesize that the CSPIONs can have high sensitivity toward H2O2 derived from the reaction between a fixed amount of glucose and glucose oxidase (GOX). As a result, CSPIONs oxidized a 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) commonly used as a substrate for hydrogen peroxidase in the presence of H2O2, leading to a change in the color of the substrate. We also utilized a colorimetric assay at 417 nm using various glucose concentrations from 5 mM to 1.25 μM to validate β-D-glucose detection. This study demonstrated that the absorbance value increases along with increasing the glucose level. The results were highly repeated at concentrations below 5 mM (all standard deviations < 0.03). Moreover, the sensitivity and limit of detection were 1.50 and 5.44 μM, respectively, in which CSPIONs are more responsive to glucose than SPIONs. In conclusion, this study suggests that CSPIONs have the potential to be used for glucose detection in diabetic patients using a physiological fluid such as ocular, saliva, and urine.

황금작약탕(黃芩芍藥湯)의 MRSA에 대한 항균활성에 관한 연구 (Antibacterial Effect of Huanggeumjakyak-tang against Methicillin-resistant Staphylococcus aureus)

  • 김에스더;최종환;김일현;이하일;송용선
    • 한방재활의학과학회지
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    • 제23권3호
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    • pp.15-26
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    • 2013
  • Objectives The aim of this study was to investigate antimicrobial activity of Huanggeumjakyak- tang water extract against MRSA. Methods The antibacterial activities of Huanggeumjakyak-tang were evaluated against 3 strains of Methicillin-resistant Staphylococcus aureus (MRSA) and 1 standard Methicillinsusceptible S. aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MICs) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay was performed under dark. Results The MIC (minimum inhibitory concentration) of Huanggeumjakyak-tang water extract against S. aureus strains ranged from 1,000 to $2,000{\mu}g/ml$. So we confirmed that it has a strong antibacterial effect. Also the combinations of Huanggeumjakyak-tang water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. Conclusions The results obtained in this study suggest that Huanggeumjakyak-tang water extract showed antibacterial effect against MRSA, and it also showed reducing effect on the side-effect problems that are the major weak points of traditional antibiotics.

Evaluation of Enzymatic Method using an Automated Chemistry Analyzers for Homocysteine Measurement

  • Shim, Moon-Jung
    • 대한임상검사과학회지
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    • 제39권3호
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    • pp.196-200
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    • 2007
  • 호모시스테인은 관상동맥질환, 뇌혈관 질환의 위험인자로 알려지고 있다. 초기의 호모시스테인 측정은 HPLC를 이용하여 일반적인 검사실에서 이용하기에는 어려움이 많았으며 현재 여러 가지 면역분석기가 도입되어 시행되고 있다. 하지만 최근 자동화학분석기를 이용한 새로운 효소법의 시약이 개발되어 이 시약의 유용성 평가를 하기위해 본 연구에서는 기존의 면역분석기와 비교분석하였다. 그 결과 정밀도 평가에서 변이계수는 0.98 - 3.23%, 2.55~4.58%를 나타내었고, 자동화학분석기(TBA 200FR와 Advia 1650)를 이용한 새로운 효소법(HBI)과 기존의 면역분석기(Advia Centaur와 Immulite 2000)간의 상관관계 평가에서 상관계수는 0.9632와 0.9625로 우수한 상관관계를 보였다. 이 결과를 토대로 호모시스테인 측정에 있어 새로운 효소법의 자동화학분석기를 이용한 방법이 제시되어 보다 간편하고 쉬운 routine analysis에 적합하리라 사료된다.

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In-Vitro Anticancer and Free Radical Scavenging Potential of Compound Formulation Used in Unani System of Medicine

  • Mannan, Mohd Nazir;Kazmi, Munawwar Husain;Chakraborty, Alokananda;Zakir, Mohammad;Ahmad, Tasleem;Lahari, K.
    • 셀메드
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    • 제10권4호
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    • pp.27.1-27.6
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    • 2020
  • Cancer is one of the leading cause of mortality in India as well as worldwide. The management of cancer by conventional therapy has shown life threatening adverse effects. The researchers are now exploring the natural way of treatment. Unani system of medicine have rich literature for cancer and many compound formulations have been described in this system. Unani system of medicine is based on holistic approach and treat human being as a unit with natural herbs, mineral and animal origin drugs. An important compound Unani formulation (CUF) from the literature has been chosen to explore the Unani claim of its anticancer activity. The phytochemical constituents were assessed using standard phytochemical screening method. Antioxidant property of this formulation was assessed by DPPH assay. The DPPH free radical scavenging assay was carried out by colorimetric method and ascorbic acid was taken as a positive control. Three different extracts of CUF on different concentrations were used to screening on human breast cancer (BCC) MCF-7 cell line. For the estimation of in-vitro cytotoxic potency of the investigated extracts was assessed on MTT assay by using trypan blue method and paclitaxel was used as the standard. Hydro-ethanolic (HE) extract showed highest free radical scavenging activity among all extracts. DPPH Assay showed substantial antioxidant activity of these extracts in hydro-ethanol extract at 1㎍ concentration of CUF. The CUF showed antioxidant and anticancer activity. The claim made by Unani physician has been proved.