• Title/Summary/Keyword: horticultural activity

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A qPCR Method to Assay Endonuclease Activity of Cas9-sgRNA Ribonucleoprotein Complexes

  • Minh Tri Nguyen;Seul-Ah Kim;Ya-Yun Cheng;Sung Hoon Hong;Yong-Su Jin;Nam Soo Han
    • Journal of Microbiology and Biotechnology
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    • v.33 no.9
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    • pp.1228-1237
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    • 2023
  • The CRISPR-Cas system has emerged as the most efficient genome editing technique for a wide range of cells. Delivery of the Cas9-sgRNA ribonucleoprotein complex (Cas9 RNP) has gained popularity. The objective of this study was to develop a quantitative polymerase chain reaction (qPCR)-based assay to quantify the double-strand break reaction mediated by Cas9 RNP. To accomplish this, the dextransucrase gene (dsr) from Leuconostoc citreum was selected as the target DNA. The Cas9 protein was produced using recombinant Escherichia coli BL21, and two sgRNAs were synthesized through in vitro transcription to facilitate binding with the dsr gene. Under optimized in vitro conditions, the 2.6 kb dsr DNA was specifically cleaved into 1.1 and 1.5 kb fragments by both Cas9-sgRNA365 and Cas9-sgRNA433. By monitoring changes in dsr concentration using qPCR, the endonuclease activities of the two Cas9 RNPs were measured, and their efficiencies were compared. Specifically, the specific activities of dsr365RNP and dsr433RNP were 28.74 and 34.48 (unit/㎍ RNP), respectively. The versatility of this method was also verified using different target genes, uracil phosphoribosyl transferase (upp) gene, of Bifidobacterium bifidum and specific sgRNAs. The assay method was also utilized to determine the impact of high electrical field on Cas9 RNP activity during an efficient electroporation process. Overall, the results demonstrated that the qPCR-based method is an effective tool for measuring the endonuclease activity of Cas9 RNP.

The effect of storage temperature on antioxidant capacity and storability of paprika

  • Me-Hea Park;Hyang Lan Eum;Pue Hee Park;Dong Ryeol Baek;Siva Kumar Malka
    • Food Science and Preservation
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    • v.31 no.1
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    • pp.15-23
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    • 2024
  • Storage temperature profoundly influences the storability of paprika (Capsicum annuum L.). However, the impact of storage temperature on storability and its association with the antioxidant activity of paprika are poorly understood. In this study, we evaluated the storage attributes, activity, and gene expression levels of antioxidant enzymes in paprika stored at 4, 10, and 20℃ for 14 d and then at 20℃ for an additional 5 d (14+5 d; retail conditions). Storage at 10℃ effectively mitigated pitting, stalk browning, shriveling, and decay while significantly enhancing the marketability of paprika. The fruits stored at 4℃ were prone to pitting, whereas those stored at 20℃ were sensitive to stalk browning and decay. Moreover, paprika stored at 10℃ exhibited higher 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) activity and total phenolic content than those stored at 4 and 20℃, indicating improved antioxidant activity. Additionally, storage at 10℃ upregulated the expression levels of the antioxidant genes, catalase and peroxidase, suggesting the mechanism underlying the quality enhancement of paprika. Our findings suggest that paprika storage at 10℃ alleviates chilling injuries, preserves the quality and marketability, and enhances the antioxidant potential of paprika. These findings provide insights into how temperature influences the quality and minimizes post-harvest losses during the storage and distribution of paprika.

The Effect of Horticultural Therapy on Cognitive Function, Depression, Self-esteem and Activity Daily Living(ADL) in Dementia Care - A Systematic Review and Meta-Analysis - (치매환자의 인지기능, 우울, 자아존중감, 일상생활수행능력에 미치는 원예치유 효과 - 체계적 문헌고찰 및 메타분석 -)

  • Bae, Seung-Jong;Lee, Sun-Hee;Kim, Dae-Sik
    • Journal of Korean Society of Rural Planning
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    • v.27 no.4
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    • pp.97-107
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    • 2021
  • The purpose of this study is to systematically review and to objectively identify the effect size in order to understand the effects of horticultural therapy on cognitive function, depression, self-esteem, and activity daily living(ADL) of dementia patients. Related studies published in Korean and English were selected by searching domestic and foreign academic databases. The risk of bias assessment of individual studies was performed on the included 18 literatures. Horticultural therapy was found to be significantly effective in cognitive function(effect size=1.06), depression(effect size=0.64), self-esteem(effect size=0.99), and ADL(effect size=1.98) in dementia care. It is expected that related experimental studies will be repeatedly conducted to produce a more objective effect size calculation.

Effects of REBT Applied Horticultural Activity Program on Irrational Beliefs, Job-Seeking Stress, and Career Maturity of University Students

  • Jo, Hyun-Soo;Lee, Yu-Bin;Kim, Yong-Hyun;Huh, Moo-Ryong
    • Journal of People, Plants, and Environment
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    • v.22 no.6
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    • pp.601-609
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    • 2019
  • The purpose of this study was to investigate the applicability of rational emotive behavior therapy(REBT) technique to the horticultural activity program as a method of reducing irrational beliefs and job-seeking stress, and improving career maturity through the change of irrational beliefs. The subjects were 30 university students with irrational beliefs, 15 of them in the control group and 15 in the experimental group. The horticultural therapy program was carried out in total 10 sessions once a week for one and a half hours. For irrational beliefs, the experimental group showed a significant decrease (p=.002) in the total score after the program, but the control group showed an insignificant change (p=.529). Total job-seeking stress score was decreased significantly in the experimental group (p=.002) after the program, whereas it was increased in the control group (p=.023). For the career maturity, the experimental group showed a significant increase (p=.008) in the total score, whereas the control group showed a significant decrease (p=.028). Therefore, it can be found that REBT applied horticultural activity programs are helpful for reducing irrational beliefs and job-seeking stress and improving career maturity for young adults.

Antioxidative and Hepatoprotective Effects of Lythrum salicaria (털부처꽃의 항산화 및 간장보호활성)

  • Lee, Seung-Eun;Park, Chun-Geun;Ahn, Young-Sup;Son, Yeong-Deck;Cha, Sun-Woo;Seong, Nak-Sul
    • Korean Journal of Medicinal Crop Science
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    • v.17 no.1
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    • pp.1-7
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    • 2009
  • Several parts of Lythrum salicaria were used for this study. Scavenging activities on radicals, inhibitory activity on linoleic acid peroxidation and total phenol contents of extracts from root, flower, and aerial part were evaluated. Flower and root selected from in vitro assay were subjected to in vivo assay on $CCL_4$-induced liver injury rat model for two weeks. Carbon tetrachloride intoxication on rats produced large amounts of hepatic lipid peroxidation product, thiobarbituric acid reactive substance (TBARS) compared with normal rats. Treatment with root extract of L. salicaria (LSR) showed effective inhibitory activity on lipid peroxidation product. Administration with LSR extract significantly alleviated $CCL_4$-induced increase in GPT activity which were more effective than silymarin. The results of this study suggest that root and flower of L. salicaria have antioxidant and liver protecting activities, and root part is the most effective candidate to develop a new functional material.

Influence of Essential Oil in 'Shiranuhi' Immature Fruit on Antioxidant and Antimicrobial Activities (부지화 미숙과 에센셜 오일의 항산화 및 항균 활성 효과)

  • Kim, Sang Suk;Hyun, Ju Mi;Kim, Kwang Sik;Park, Kyung Jin;Park, Suk Man;Choi, Young Hun
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.6
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    • pp.493-497
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    • 2013
  • This study was designed to analyze the chemical composition of essential oil in 'Shiranuhi' immature fruit and to test their biological activities. 'Shiranuhi' immature essential oils (SIEO) were obtained by steam distillation from fruits collected from Jeju Island and were analyzed using gas chromatograph (GC)-flame ionization detectors (FID) and GC-MS. Fourteen components were identified in the essential oil. Limonene (75.21%) and terpineol (8.68%) were the major components in SIEO. Since acne vulgaris is the combined result of a bacterial infection and the inflammatory response to that infection, we examined whether SIEO possessed antibacterial against skin pathogens. As a result, SIEO showed excellent antibacterial activities against drug-susceptible and -resistant Propionibacterium acnes and Staphylococcus epidermidis, which are acne-causing bacteria. In this study, SIEO was examined on DPPH radical scavenging activities, which showed moderate antioxidant activity ($SC_{50}$, $15.36{\mu}L/mL$). In order to determined whether SIEO can be safely applied to human skin, the cytotoxicity effects of SIEO were determined by colorimetric MTT assays in normal human fibroblasts and keratinocyte HaCaT cells. They exhibited low cytotoxicity at $0.5{\mu}L/mL$ in both celllines. Based on these results, we suggest the possibility that essential oil of 'Shiranuhi' maybe considered as an antibacterial and antioxidant agent.

Purification and Characterization of Chitinase from Streptomyces sp. M-20

  • Kim, Kyoung-Ja;Yang, Yong-Joon;Kim, Jong-Gi
    • BMB Reports
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    • v.36 no.2
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    • pp.185-189
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    • 2003
  • Chitinase (EC 3.2.1.14) was isolated from the culture filtrate of Streptomyces sp. M-20 and purified by ammonium sulfate precipitation, DEAE-cellulose ion-exchange chromatography, and Sephadex G-100 gel filtration. No exochitinase activity was found in the culture filtrate. The molecular mass of the purified chitinase was 20 kDa, estimated by a sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and was confirmed by activity staining with Calcofluor White M2R. Chitinase was optimally active at pH of 5.0 and at $30^{\circ}C$. The enzyme was stable from pH 4 to 8, and up to $40^{\circ}C$. Among the metals and inhibitors that were tested, the $Hg^+$, $Hg^{2+}$, and p-chloromercuribenzoic acid completely inhibited the enzyme activity. The chitinase activity was high on colloidal chitin, chitotriose, and chitooligosaccharide. The purified chitinase showed antifungal activity against Botrytis cinerea, and lysozyme activity against the cell wall of Botrytis cinerea.

Comparison of the biological activity of extracts from the mycelium, sclerotium, and fruiting body of Wolfiporia cocos (F.A. Wolf) Ryvarden & Gilb using different extraction solvents (복령균핵, 균사체 및 자실체의 추출용매별 생리활성 성분 비교)

  • An, Gi-Hong;Cho, Jae-Han;Kim, Ok-Tae;Lee, Chan-Jung;Han, Jae-Gu
    • Journal of Mushroom
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    • v.18 no.3
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    • pp.244-253
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    • 2020
  • The aim of this study was to investigate the biological activity of extracts obtained from the mycelium, sclerotium, and fruiting body of Wolfiporia cocos using different extraction solvents (hot water, 70% ethanol, and 70% methanol). Among the three developmental stages, the mycelium extracts showed the highest DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity, ferric reducing antioxidant power (FRAP), nitrite scavenging activity, and total polyphenolic content. Among the extraction solvents in the context of the W. cocos mycelium, the DPPH radical scavenging activity, FRAP, and total polyphenol content in the hot-water extracts were significantly higher than those in the other extracts. In the case of the sclerotium, the reducing power, nitrite scavenging activity, and total polyphenol content were significantly higher in 70% ethanolic extracts. The fruiting body showed the highest DPPH radical scavenging activity, reducing power, nitrite scavenging activity, and total polyphenol content in the context of hot-water extraction. Moreover, the β-glucan content was significantly higher in the sclerotium versus the mycelium or fruiting body. The total amino acid and total essential amino acid contents were remarkably higher in the mycelium and fruiting body than in the sclerotium; of note, and arginine (Arg) and phenylalanine (Phe) were highly detected among the amino acid components.