• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.15 no.2
• /
• pp.132-144
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• v.8 no.6
• /
• pp.676-684
• /
• 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.

• KSBB Journal
• /
• v.26 no.5
• /
• pp.471-476
• /
• 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.

• Korean Journal of Agricultural Science
• /
• v.46 no.3
• /
• pp.653-660
• /
• 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.

• Journal of Veterinary Science
• /
• v.21 no.5
• /
• pp.79.1-79.12
• /
• 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.

• Applied Chemistry for Engineering
• /
• v.35 no.2
• /
• pp.67-78
• /
• 2024

Light absorption has potential as a signal in biochemical analyses due to its simplicity in measurement and interpretational clarity. Among substances that generate absorption signals, gold nanoparticles possess advantages such as chemical stability, biological compatibility, and unique optical properties from the localized surface plasmon resonance (LSPR) in the visible light range. They also exhibit versatility compared to other colorimetric substances effective only for specific target molecules, as they easily conjugate with various detection active substances like antibodies and aptamers. Particularly due to advantages such as low cost, ease of particle synthesis, and high environmental stability compared to enzyme-based colorimetric methods, gold nanoparticles are extensively researched as signal substances in colorimetric assays. This review summarizes various strategies utilizing gold nanoparticles as absorption signal substances, focusing on recent research. Based on the characteristics of gold nanoparticles, where the optical property is influenced by particle morphology, literature is classified and reviewed based on strategies controlling the shape of gold nanoparticles during signal generation. Through this, it is observed that gold nanoparticles, which have been used as absorption signal substances, continue to be actively researched, affirming their potential for broad and continuous improvement in the future.

• Journal of Biomedical Engineering Research
• /
• v.41 no.6
• /
• pp.228-234
• /
• 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.

• Journal of Korean Medicine Rehabilitation
• /
• v.23 no.3
• /
• pp.15-26
• /
• 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.

• Korean Journal of Clinical Laboratory Science
• /
• v.39 no.3
• /
• pp.196-200
• /
• 2007

In clinical practice, homocysteine has gained popularity because its elevated values are strongly associated with an increased risk of cardiovascular disease. More recently, a new enzymatic colorimetric assay for homocysteine in biological sample, suitable for automated clinical analyzers, has been proposed. To evaluate one of these enzymatic methods and compare the results obtained with this method with those of an immunoenzymatic method, thirty-two samples were analyzed for total homocysteine by HiSens$^{(R)}$ homocysteine reagent on the automated chemistry analyzers TBA 200FR and compared to the widely used immunoenzymatic method ADVIA Centaur. In TBA 200FR, the within-run CVs of two control materials were 3.23% and 0.92%, respectively; the between run CVs were 4.58% and 2.55%, respectively. And in ADVIA 1650, the within-run CVs were 6.81% and 0.99%, respectively; the between run CVs were 9.0% and 3.9%, respectively. The recovery for homocysteine was 100% ($60.8{\mu}mol/L$), 99.1% ($48.64{\mu}mol/L$), 96.3% ($36.48{\mu}mol/L$), 96.1% ($24.32{\mu}mol/L$), and 92.1% ($12.16{\mu}mol/L$). The regression equation of TBA 200FR vs. ADVIA Centaur was y=0.9095x-2.5086 (r=0.9632). And the regression equation for the ADVIA 1650 chemistry vs. Immulite 2000 was y=0.8418x + 0.3207 (r=0.9625). In conclusion, this enzymatic method using automated chemistry analyzer for homocysteine assay shows acceptable analytical performance. I suggest that this assay will be suitable for routine analysis.

• CELLMED
• /
• v.10 no.4
• /
• pp.27.1-27.6
• /
• 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.