• Title/Summary/Keyword: biological assays

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Silver Materials Induce Differential Cytotoxicity and Pulmonary Toxicity Based on Size and Shape

  • Pak, Pyo June;Kang, Beob Hwa;Chung, Namhyun
    • Journal of Applied Biological Chemistry
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    • v.58 no.2
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    • pp.113-116
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    • 2015
  • Silver materials may be toxic in humans because they can enter the body and accumulate, typically in the lungs. We hypothesized that the cytotoxicity of naive silver materials is affected by their size and shape. Our in vitro assays revealed that the overall toxicity was in the following order: submicro-particles>wires>micro-particles. These results contrast with previous studies, which showed that silver wires are the most toxic among the three tested materials, possibly due to differences in cell lines. Evaluations of in vivo pulmonary toxicity revealed eryptosis in the cavity lining of the lung sections. The observed eryptosis was consistent with the in vitro results. Our results indicate that silver materialinduced cytotoxicity must be measured and compared using various methods.

Antioxidant and Anticancer Activities of Methanol and Water Extracts from Leaves of Cirsium japonicum

  • Yin, Yu;Heo, Seong-Il;Wang, Myeong-Hyeon
    • Journal of Applied Biological Chemistry
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    • v.51 no.4
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    • pp.160-164
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    • 2008
  • Potential antioxidant activities of methanol and water extracts of Cirsium japonicum var. ussuriense (CJ) leaves were examined. The reducing power and hydroxyl raical-scavenging activity assays showed that the methanol extract had a significantly higher activity than the water extract. In addition, the methanol extract showed a concentration-dependent reducing power, ranging from 0.228 to 1.072($0.1{\sim}0.5\;mg/mL$), as well as a high DPPH free radical-scavenging activity($EC_{50}=40.25\;{\mu}/mL$). The total phenolic(as tannic acid) and flavonoid(as quercetin) contents of the extract were 62.41 mg/g and 13.48 mg/g, respectively. The cytotoxic activity indicated that the methanol extract has an inhibition activity in the stomach carcinoma cell (35.40%), suggesting that the methanol extract of CJ leaves could be used as a potential source of pharmaceutical material.

Characterization of F2 Progenies of Wound Minus Arabidopsis Mutant Crossed with Wild Type Plant

  • Park, Sanggyu
    • Journal of Applied Biological Chemistry
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    • v.43 no.1
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    • pp.12-17
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    • 2000
  • To understand the signal transduction pathway that leads to the activation of the wound-inducible proteinase inhibitor II (pin2) promoter. $F_2$ progenies of wound (-) mutant crossed with wild-type Arabidopsis plants were biochemically and genetically characterized. Wound (-) mutant was derived from transgenic Arabidopsis plants containing bacterial cytosine deaminase gene under the control of pin2 promoter. The cytosine deaminase assays indicated that wound (-) mutant is a dominant inhibitor of wound-inducibility as only 3 of the $20F_2$ progenies showed cytosine deaminase (CDase) activity, To construct a structural map of the wound (-) mutant chromosomal regions, cleaved, amplified polymorphic sequences (CAPS) markers that cover all Chromosomes were used. Chromosomal regions covered by three different CAPS markers could be candidates for further fine mapping of the location of the wound (-) mutation. g4026, RGA1 and ASA1 located at 84.9 on recombinant inbred (RI) map of chromosome I, at 1.75 on RI map of chromosome II, and 18.35 on RI map of chromosome V, respectively.

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Anti-oxidant activity of avicularin and isovitexin from Lespedeza cuneata

  • Lee, Ju Sung;Lee, Ah Young;Quilantang, Norman G.;Geraldino, Paul John L.;Cho, Eun Ju;Lee, Sanghyun
    • Journal of Applied Biological Chemistry
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    • v.62 no.2
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    • pp.143-147
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    • 2019
  • This study evaluated the anti-oxidant activity of avicularin and isovitexin from Lespedeza cuneata using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl (OH) scavenging assays. The results showed that among the four fractions, the highest OH radical scavenging activity was exhibited by the n-butanol fraction. The DPPH radical scavenging activities of avicularin and isovitexin were higher than 50% at a concentration of $100{\mu}g/mL$. Moreover, one hundred micrograms per liter of avicularin and isovitexin exhibited effective anti-oxidant activity, with the OH radical scavenging rates being 87.54 and 91.48%, respectively. These results suggest that Lespedeza cuneata would be useful anti-oxidant agents from natural sources.

Comparison of the contents of total polyphenol, total flavonoid, and flavonoid derivatives in unfermented and fermented barley sprouts

  • Neil Patrick Uy;Hak-Dong Lee;Dae Cheol Byun;Sanghyun Lee
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.353-358
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    • 2023
  • Barley (Hordeum vulgare) belongs to the Poaceae family. This study compared the polyphenol and flavonoid levels of unfermented and fermented barley sprouts using spectrophotometric assays. The findings indicated that fermentation greatly boosted the flavonoid content but caused only a slight increase in the polyphenol content. However, this does not imply that fermentation has no effects whatsoever on the polyphenol content of barley sprouts. This was due to the fact that some flavonoids cannot be detected by the wavelength used to calculate the overall polyphenol concentration. Both samples were subjected to high-performance liquid chromatography analysis and detected the flavonoids lutonarin, saponarin, isoorientin, isovitexin, and tricin-all of which have bioactive properties-most notably known for their antioxidant activity. These results augment the ongoing phytochemical profiling research and can possibly valorize the already thriving barley industry.

Anti-Oxidant and Hepatoprotective Activities of Ziziphus mucronata Fruit Extract Against Dimethoate-Induced Toxicity

  • Kwape, Tebogo Elvis;Chaturvedi, Padmaja;Kamau, Macharia;Majinda, Runner
    • Journal of Pharmacopuncture
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    • v.16 no.1
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    • pp.21-29
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    • 2013
  • Objective: The study was carried out to evaluate the hepatoprotective and antioxidant potential of Ziziphus mucronata (ZM) fruit extract. Methods: The different types of fruit extract were prepared by soaking the dry powdered fruit in different solvents followed by rotary evaporation. Each extract was tested for its phenol content and antioxidant activities. An in vivo study was performed in Sprague-Dawley (SD) rats. Thirty adult male SD rats (aged 21 weeks) were divided into six groups of five rats each and treated as follows: The normal control (NC) received distilled water while the dimethoate control (DC) received 6 mg/kg.bw.day-1 dimethoate dissolved in distilled water. The experimental groups E1, E2, E3, and E0 received dimethoate (6 mg/kg.bw) + ZMFM (100 mg/kg.bw-1), dimethoate (6 mg/kg.bw) + ZMFM (200 mg/kg.bw-1), dimethoate (6 mg/kg.bw) + ZMFM (300 mg/kg.bw-1), and ZMFM (300 mg/kg.bw-1) only. Both the normal control and the dimethoate control groups were used to compare the results. After 90 days, rats were sacrificed, blood was collected for biochemical assays, and livers were harvested for histological study. Results: High phenol content was estimated, and 2, 2-diphenyl-1-picryl hydrazyl radical (DPPH) spectrophotometric, thin layer chromatography (TLC) and 2, 2-Azobis-3-ethyl benzothiazoline-6-sulphonic acid (ABTS) assays showed a high antioxidant activity among the extracts. The preventive effects observed in the E1, E2 and E3 groups proved that the extract could prevent dimethoate toxicity by maintaining normal reduced glutathione (GSH), vitamin C and E, superoxide dismutase, catalase, cholineasterase and lipid profiles. The preventive effect was observed to be dose dependent. The EO group showed no extract-induced toxicity. Histological observations agreed with the results obtained in the biochemical studies. Conclusion: The study demonstrated that ZM methanol fruit extract is capable of attenuating dimethoate-induced toxicity because of its high antioxidant activity.

Mechanism of Action of Nigella sativa on Human Colon Cancer Cells: the Suppression of AP-1 and NF-κB Transcription Factors and the Induction of Cytoprotective Genes

  • Elkady, Ayman I;Hussein, Rania A;El-Assouli, Sufian M
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7943-7957
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    • 2015
  • Background and Aims: Colorectal cancer is one of the leading causes of death in the world. The aim of this study was to investigate the growth-suppression potentiality of a crude saponin extract (CSENS) prepared from medicinal herb, Nigella sativa, on human colon cancer cells, HCT116. Materials and Methods: HCT116 cells were subjected to increasing doses of CSENS for 24, 48 and 72 h, and then harvested and assayed for cell viability by WST-1. Flow cytometry analyses, cell death detection ELISA, fluorescent stains (Hoechst 33342 and acridine orange/ethidium bromide), DNA laddering and comet assays were carried out to confirm the apoptogenic effects of CSENS. Luciferase reporter gene assays, quantitative reverse transcription-polymerase chain reaction and Western blot analyses were performed to assess the impact of CAERS and CFEZO on the expression levels of key regulatory proteins in HCT116 cells. Results: The results demonstrated that CSENS inhibited proliferation and induced apoptosis. Apoptosis was confirmed by flow cytometry analyses, while CSENS-treated cells exhibited morphological hallmarks of apoptosis including cell shrinkage, irregularity in cellular shape, cellular detachment and chromatin condensation. Biochemical signs of apoptosis, such as DNA degradation, were observed by comet assay and gel electrophoresis. The pro-apoptotic effect of CSENS was caspase-3-independent and associated with increase of the Bax/Bcl-2 ratio. CSENS treatment down-regulated transcriptional and DNA-binding activities of NF-${\kappa}B$ and AP-1 proteins, associated with down-regulation of their target oncogenes, c-Myc, cyclin D1 and survivin. On the other hand, CSENS up-regulated transcriptional and DNA-binding activities of Nrf2 and expression of cytoprotective genes. In addition, CSENS modulated the expression levels of ERK1/2 MAPK, p53 and p21. Conclusions: These findings suggest that CSENS may be a valuable agent for treatment of colon cancer.

Anti-Glyaction and Radical Scavenging Activities of Ligustri Fructus by Extraction Method (추출 방법에 따른 여정실의 최종당화산물 생성 저해 및 라디칼 소거 활성)

  • Jeong, Yun Hee;Kim, Seo Yoon;Jeong, Gyeong Han;Kim, Tae Hoon
    • The Korea Journal of Herbology
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    • v.33 no.4
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    • pp.95-100
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    • 2018
  • Objectives : Ligustri Fructus has been used since ancient times as a medicinal usages in folk medicines against antitumor purpose. Many biological active constituents have been identified from this biomass such as several terpenoids and lignans. In current study, the properties of antioxidant and anti-diabetic complications using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)($ABTS^+$) radicals scavenging, and advanced glycation end products (AGEs) inhibition assays were evaluated by different extraction methods of Ligustri Fructus. Methods : In present continuing research for development of bioactive natural ingredients, antioxidant and AGEs formation inhibitory capacities of Ligustri Fructus extracts using different organic solvents were prepared and the biological potentials were investigated using in vitro bioassays. Antioxidant properties were evaluated employing radical scavenging assays using DPPH and $ABTS^+$ radicals. In addition, the anti-diabetic complications effects of Ligustri Fructus extracts were tested via AGEs formation inhibitory assay. The total phenolic contents were determined using a spectrophotometric method. Results : All the tested extracts exhibited dose-dependent radical scavenging and AGEs formation inhibitory activities. Among the tested samples, hot water extract of Ligustri Fructus was showed the most potent activity with $IC_{50}$ value of $494.8{\pm}6.7{\mu}g/m{\ell}$ against DPPH radical scavenging assay. Also, $ABTS^+$ radical scavenging activity of hot water extract was higher than those of other extracts. In addition, AGEs formation inhibitory effects of each extacts and total phenolic contents were evaluated. Conclusions : These results suggested that Ligustri Fructus can be considered as a new effective source of natural antioxidant and anti-diabetic complications resources.

Feature Analysis of Different In Vitro Antioxidant Capacity Assays and Their Application to Fruit and Vegetable Samples (In Vitro 항산화능 측정법에 대한 특징 분석과 채소.과일 시료에 대한 적용 사례 고찰)

  • Kim, Min-Jung;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.7
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    • pp.1053-1062
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    • 2011
  • Reactive oxygen species (ROS), including singlet oxygen (${O_2}^1$), superoxide anion radical ($O_2{\cdot}^-$), hydroxyl radical ($HO{\cdot}$), peroxyl radical ($ROO{\cdot}$), hydrogen peroxide ($H_2O_2$), and hypochlorous (HOCl), are generated as byproducts of normal cellular metabolism. ROS induce damage to many biological molecules, such as lipids, proteins, carbohydrates, and DNA. It is widely believed that some degenerative diseases caused by ROS can be prevented by the high intake of fruits and vegetables due to their antioxidant activities. Recently, research on natural antioxidants has become increasingly active in various fields. Several assays have been developed to measure the total antioxidant capacity of antioxidants in fruits and vegetables in vitro. These assays include those for DPPH radical scavenging activity, SOD-like activity, total polyphenol content, oxygen radical absorbance capacity, reducing power, trolox equivalent antioxidant capacity (ABTS assay), single-cell gel electrophoresis (comet assay), and a cellular antioxidant activity assay. Because different antioxidant compounds may act through different mechanisms in vitro, no single assay can fully evaluate the total antioxidant capacity of foods. Due to the complexity of the composition of foods, it is important to be able to measure antioxidant activity using biologically relevant assays. In this review, recently used assays were selected for extended discussion, including a comparison of the advantages and disadvantages of each assay and their application to fruits and vegetables.

High Level Production of Glycoprotein H of HSV-1 (F) Using HcNPV Vector System

  • Kang, Hyun;Cha, Soung-Chul;Han, You-Jin;Park, In-Ho;Lee, Min-Jung;Byun, Si-Myung;Lee, Hyung-Hoan
    • BMB Reports
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    • v.33 no.6
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    • pp.483-492
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    • 2000
  • The Herpes simplex virus type 1 (HSV-1) strain F glycoprotein H (gH) gene in the pHLB-4 plasmid was recombinated into a baculovirus expression vector (lacZ-HcNPV) to construct a recombinant virus GH-HcNPV expressing gH. The sequences of gH and its expression were analyzed. The gH gene was located in the 6.41 kb BglII fragment. The open reading frame (ORF) of the gH gene was 2,517 bp and codes 838 amino acid residues. Insect cells infected with this recombinant virus synthesized a high level of the matured and gX-gH fusion protein with approximately 112 kDa. The fusion gH protein was localized on the membrane of the insect cells as seen by using immunofluorescence assay and accumulated in the cultured media by the SDS-PAGE and immunoprecipitation assays. The amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. Antibodies raised in mice to this recombinant protein recognized viral gH and neutralized the infectivity of HSV-1 in vitro. These results demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV; accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

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