• Title/Summary/Keyword: in vitro fruit assay

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Detection and Quantification of Apple Stem Grooving Virus in Micropropagated Apple Plantlets Using Reverse-Transcription Droplet Digital PCR

  • Kim, Sung-Woong;Lee, Hyo-Jeong;Cho, Kang Hee;Jeong, Rae-Dong
    • The Plant Pathology Journal
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    • v.38 no.4
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    • pp.417-422
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    • 2022
  • Apple stem grooving virus (ASGV) is a destructive viral pathogen of pome fruit trees that causes significant losses to fruit production worldwide. Obtaining ASGV-free propagation materials is essential to reduce economic losses, and accurate and sensitive detection methods to screen ASGV-free plantlets during in vitro propagation are urgently necessary. In this study, ASGV was sensitively and accurately quantified from in vitro propagated apple plantlets using a reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) assay. The optimized RT-ddPCR assay was specific to other apple viruses, and was at least 10-times more sensitive than RT-real-time quantitative PCR assay. Furthermore, the optimized RT-ddPCR assay was validated for the detection and quantification of ASGV using micropropagated apple plantlet samples. This RT-ddPCR assay can be utilized for the accurate quantitative detection of ASGV infection in ASGV-free certification programs, and can thus contribute to the production of ASGV-free apple trees.

Comparison of antioxidant activity and prevention of lymphocyte DNA damage by fruit and vegetable juices marketed in Korea (시판 천연 과일주스와 채소주스의 항산화능과 임파구 DNA 손상 방지 효능 비교)

  • Cho, Miran;Lee, Hye-Jin;Kang, Myung-Hee;Min, Hyesun
    • Journal of Nutrition and Health
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    • v.50 no.1
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    • pp.1-9
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    • 2017
  • Purpose: Fruit and vegetable juices are known to be rich sources of antioxidants, which have beneficial effects on diseases caused by oxidative stress. The purpose of this study was to directly compare the antioxidant activities of fruit and vegetable juices marketed in Korea. Methods: We analyzed four fruit juices, two vegetable juices, two yellow-green juices, and six mixed vegetable juices. Antioxidant activities were analyzed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) test, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonate) (ABTS) test, and oxygen radical absorbance capacity (ORAC) assay. Protective effects against DNA damage were determined using an ex vivo comet assay with human lymphocytes. Results: DPPH radical scavenging activities were in the following order: blueberry juice > mixed vegetable C juice > kale juice > mixed vegetable P juice > grape juice. ABTS radical scavenging activities were in the following order: blueberry juice > mixed vegetable C juice > grape juice > mixed vegetable P juice > kale juice. Peroxyl radical scavenging activities as assessed by ORAC assay were in the following order: blueberry juice > kale juice > mixed vegetable C juice > grape juice. Grape or blueberry juice showed strong abilities to prevent DNA damage in lymphocytes, and the difference between them was not significant according to the GSTM1/GSTT1 genotype. Conclusion: Antioxidant activities of fruit and vegetable juices and ex vivo DNA protective activity increased in the order of blueberry juice, grape juice, and kale juice, although the rankings were slightly different. Therefore, these juices rich in polyphenols and flavonoids deserve more attention for their high antioxidant capacity.

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.

Effect of the Cultivated Fruit body of Phellinus linteus on Low Density Lipoprotein Oxidation (배양상황버섯의 저밀도지질단백질 (LDL)산화에 미치는 영향)

  • 정은주;성재모;양기숙
    • YAKHAK HOEJI
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    • v.48 no.2
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    • pp.130-134
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    • 2004
  • Phellinus linteus (Hymenocaetaceae) has been used for the treatment of gastric cancer, noninsulin dependant diabetes, diarrhea, and menstrual irregularity. The antioxidative effect of cultivated fruit body of Phellinus linteus on low density lipoprotein oxidation was investigated. The MeOH and water extracts were examined by TEARS assay on human plasma LDL in vitro system. The MeOH ex. showed antioxidative activity, and then was fractionated into Precipitates, Water Insoluble fr., Water fro and ether fro The results showed that the lipophilic fractions, Water Insoluble fr. and Ether fr. of cu 1-tivated fruit body of Phellinus linteus, inhibited the oxidative modification of LDL.

Study on Genotoxicity of Crocin, a Component of Gardenia Fruit, in Bacterial and Mammalian Cell Systems

  • Choi, Hae-Yeon;Kim, Youn-Jung;Jeon, Hee-Kyung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.4 no.4
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    • pp.285-292
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    • 2008
  • Crocin is one of the major components of gardenia fruit and saffron which are widely used as natural food colorants and as traditional Chinese medicines. However, the genotoxicity data on crocin are not sufficient for safety evaluation. The purpose of this study was the examination of the genotoxicity on crocin from gardenia yellow in bacterial and mammalian cells, using various genotoxic battery testing assays and the influence of crocin on methyl methanesulfonate (MMS) and ${H_2}{O_2}$-induced DNA damage in vitro, using single cell gel electrophoresis (comet) assay. From results, no considerable mutagenicity and clastogenicity were seen in bacteria and mammalian cells treated with crocin, by Ames test, chromosomal aberration assay, ${tk}^{+/-}$ gene forward mutation assay and comet assay. And, post-treatment with crocin significantly suppressed ${H_2}{O_2}$-induced DNA damage in a dose-dependent manner. In conclusion, the findings of the present study and other previous observations indicate that crocin has no genotoxic potential. And it showed that crocin clearly repressed the genotoxic potency of ${H_2}{O_2}$. These results suggest that anti-oxidative effects of crocin may be involved in the protective effects of DNA damage.

IN-VITRO STUDY OF CO2 EXTRACT OF TERMINALIA CHEBULA IN BREAST CANCER CELL LINE MD-MBA-231

  • Chandil, Shachi;Bamoriya, Harikishan;More, D.B.
    • CELLMED
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    • v.11 no.3
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    • pp.16.1-16.7
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    • 2021
  • Cancer is an abnormal growth of cells in body which leads to death. These cells are born due to imbalance in cell proliferation mechanism. In 2018, WHO released new statistics on cancer incidence, mortality, and prevalence worldwide i.e., GLOBOCAN 2018 estimates for 28 types of cancer in which more prevalence of cervix and breast cancer. According to survey, in India about 7.8 million cancer deaths and 11.5 million new cases arise in 2018, which will increase to 19.3 million new cases per year by 2025. Though breast cancer as such is not explained anywhere in Ayurvedic compendia, correlations can be done with the Stana Arbuda. Ayurveda, the ancient system of medicine came into existence 1000's of years ago with an objective of maintaining the health of people and treating diseases. Many herbs used in Ayurveda have been screened for activity against cancer and in-vitro and in-vivo studies have given promising leads. The plant, called as "Mother of Medicine", Haritaki has been extensively studied for its various ailments because of its extraordinary healing potency. Haritaki (Terminalia chebula Retz.), Family: Combretaceae have a great therapeutic value and is widely distributed in India. Dried fruit of Terminalia chebula contains high quantities phenolic compounds consist of ellagic acid, gallic acid and chebulic acid. The fruit extract of T. chebula is having different biological properties like anticancer, antioxidant, hepatic and renal protective activities etc. In this study, we focus on the use of CO2 extract of Terminalia chebula, on the breast cancer cell line MDA-MB-231. All tests proved that CO2 extract of Terminalia chebula containing active chemical component, therefore our experiment showed the positive results for CO2 extract of Terminalia chebula against breast cancer cell line cancer MDA-MB-231. The MTT assay results were used to evaluate the anti-cancer activity of the extract. The percentage of cell growth and cell viability were calculated from tabulated result values of MTT assay. Cell viability MTT assay also showed significant growth inhibition, at the same time statistical analysis of MTT assay also proved significant results.

Effect of Fruit Extract of Prunus mume on the Scavenging Activity of Reactive Oxygen Species and Melanin Production in B16F1 Cells (매실추출물이 활성산소종 소거효과와 B16F1 세포에서 멜라닌 생성에 미치는 영향)

  • Park, Hyeong-Joon;Kim, Moon-Moo;Oh, Yung-Hee
    • Journal of Life Science
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    • v.22 no.7
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    • pp.936-942
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    • 2012
  • Prunus mume has been traditionally used as a medicinal food in Korea, Japan, and China. In particular, this fruit has been reported to have beneficial biological effects on gastritis and gastric ulcers. However, its action in relation to skin whitening has remained unclear. Accordingly, the effects of fruit extract of P. mume related to antioxidation and skin whitening were examined in this study. First, using the MTT assay, it was observed that fruit extract of P. mume below 0.1% has no cytotoxicity in B16-F1 cells as a result of cell viability. Second, the direct scavenging effects and the reducing power of the fruit extract of P. mume were evaluated in vitro on DPPH radicals, hydrogen peroxide, and superoxide. It exhibited high reducing power and scavenging activity on the aforementioned reactive oxygen species. Furthermore, we found that its protective effect against genomic DNA damage related to oxidative stress was increased in a dose-dependent manner. In addition, the fruit extract of P. mume had an inhibitory effect on melanin production induced by L-dopa. In addition, it reduced the expression level of NRF-2, SOD-1, and SOD-2 related to antioxidation in western blot analysis. These results suggest that fruit extract of P. mume could exert a whitening effect through inhibition of melanin production by its antioxidant effect.

Genotoxicity Assessment of Gardenia Yellow using Short-term Assays

  • Chung, Young-Shin;Eum, Ki-Hwan;Ahn, Jun-Ho;Choi, Seon-A;Noh, Hong-June;Seo, Young-R.;Oh, Se-Wook;Lee, Michael
    • Molecular & Cellular Toxicology
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    • v.5 no.3
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    • pp.257-264
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    • 2009
  • Gardenia yellow, extracted from gardenia fruit, has been widely used as a coloring agent for foods, and thus, safety of its usage is of prime importance. In the current study, short-term genotoxicity assays were conducted to evaluate the potential genotoxic effects of gardenia yellow. The gardenia yellow used was found to contain 0.057 mg/g of genipin, a known biologically active compound of the gardenia fruit extract. Ames test did not reveal any positive results. No clastogenicity was detected by a chromosomal aberration test, even on evaluation at the highest feasible concentration of gardenia yellow. Gardenia yellow was also shown to be non-genotoxic using an in vitro comet assay and a micronucleus test with L5178Y cells, although a marginal increase in DNA damage and micronuclei frequency was reported in the respective assays. Additionally, in vivo micronucleus test results clearly demonstrated that oral administration of gardenia yellow did not induce micronuclei formation in the bone marrow cells of male ICR mice. Taken together, our results indicate that gardenia yellow is not mutagenic to bacterial cells, and that it does not cause chromosomal damage in mammalian cells, either in vitro or in vivo.

Comparative Studies of Adriamycin and 28-Deacetyl Sendanin on In Vitro Growth Inhibition of Human Cancer Cell Lines

  • Kim, Hwan-Mook;Oh, Goo-Taeg;Han, Sang-Bae;Hong, Dong-Ho;Hwang, Bang-Yeon;Kim, Young-Ho;Lee, Jung-Joon
    • Archives of Pharmacal Research
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    • v.17 no.2
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    • pp.100-103
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    • 1994
  • The limonoid compound (28-deacetyl sendanin0 isolated from the fruit of Melia toosendan SIEB. et ZUCC. was evaluated on anticancer activity. According to a standard in vitro cytotoxicity assy, eight human cancer cell lines and SRB assay were introduced for present evaluation. As a positive standard, adriamycin was tested in parallel. The cell lines were originated from six different organs. In view of dose-response profiles to 28-deacetyl sendanin, the most sensitive cells were SF-539 and PC-3 which were derived from CNS and prostate, respecitively. In contrast, all the cell lines responded similarly to adriamycin to give rise to nearly indentical six cell lines were more sensitive to 28-deacetyl sendanin and two were more resistant. As a result, 28-deacetyl sendanin had more senstive and selective inhibitory effects on in vitro growth of human cancer cell lines in a comparison with adriamycin.

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Antioxidative and Inhibitory Activities of Extract and Juice Powder from Hippophae rhamnoides L. against Nitric Oxide and Elastase Production (비타민나무 추출물 및 착즙 분말의 항산화 활성 및 Nitric Oxide 생성과 Elastase에 대한 저해활성)

  • Park, Ju Hee;Lee, Chan Ok;Yoo, Ji Hye;An, Nguyen Thi Lan;Yoo, Nam Ho;Kim, Myong Jo
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.2
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    • pp.119-126
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    • 2018
  • Background: The flowering plant Hippophae rhamnoides L. has been used for many studies on fruit or leaf extracts. This study was conducted to investigate the development of a new cosmetic material from H. rhamnoides fruits and leaves that have by antioxidant, anti-inflammatory and wrinkle improvement activities. Methods and Results: The antioxidant abilities of H. rhamnoides extracts, including of a water-soluble fruit powder (FW), a fatsoluble fruit powder (FF), a supercritical extract of fruit by-product (BS), and a mixture of leaf and fruit (MIX), were investigated in vitro. A DPPH radical assay for antioxidant activity was performed for these fractions alongside assay to evaluate the total phenolic and flavonoid content (TPC and TFC). As expected, the MIX had the highest DPPH radical scavenging activity ($RC_{50}=10.27{\mu}g/m{\ell}$), and the TPC and TFC also were highest in MIX ($225.7mg{\cdot}GAE/g$, and $25.18mg{\cdot}QE/g$, respectively). Nitric oxide (NO) production in LPS-induced RAW264.7 cells was estimated and the results indicated an over 75% decrease of NO production in FF and MIX. In other assays, the highest elastase inhibitory activity was found in FW. Conclusions: These results revealed that H. rhamnoides extracts have a high potential for antioxidant, anti-inflammatory and antiwrinkle activities. H. rhamnoides products are suggested to be applied as the functional materials of cosmetic ingredients.