• Title/Summary/Keyword: Enzyme assays

Search Result 308, Processing Time 0.032 seconds

Trends in Rapid Detection Methods for Marine Organism-derived Toxins (해양 생물 유래 독소의 나노 기술 기반 신속 진단법 개발 동향)

  • Park, Chan Yeong;Kweon, So Yeon;Moon, Sunhee;Kim, Min Woo;Ha, Sang-Do;Park, Jong Pil;Park, Tae Jung
    • Journal of Food Hygiene and Safety
    • /
    • v.35 no.4
    • /
    • pp.291-303
    • /
    • 2020
  • Marine organism-derived toxins have negative effects not only on human health but also in aquaculture, fisheries, and marine ecosystems. However, traditional analytical methods are insufficient in preventing this threat. In this paper, we reviewed new rapid methods of toxin detection, which have been improved by adopting diverse types of nanomaterials and technologies. Moreover, we herein describe the main strategies for toxin detection and their related sensing performance. Notably, to popularize and commercialize these newly developed technologies, simplifying the process of pre-treating real samples real samples is very important. As part of these efforts, numerous studies have reported pretreatment methods based on the antibody-immobilized magnetic nanoparticles, and some cases have applied nanoparticles to enhance the sensing performance by utilizing the intrinsic catalytic activity. Furthermore, some reports have introduced fluorescent nanoparticles, such as quantum dots, to represent the lower detection limits of conventional enzyme-based colorimetric methods and lateral flow assays. Some studies using electrochemical measurements based on aptamer-nanoparticle complexes have also been announced. In addition, as the response to new toxins generated by changes in the marine environment is still lacking, further research on diagnostic and detection is also greatly needed for these kinds of marine toxins and their derivatives.

Physiological Activity of Sarcodon aspratus Extracts (능이버섯(Sarcodon aspratus) 추출물의 생리활성)

  • 송재환;이현숙;황진국;한정환;노정근;금동혁;박기문
    • Food Science of Animal Resources
    • /
    • v.23 no.2
    • /
    • pp.172-179
    • /
    • 2003
  • This study was carried out to find the preventive medical and therapeutic effects of Sarcodon asparatus on adult disease by employing several biological and biochemical assays. Nitrate scavenging ability(NSA) of Sarcodan asparatus extracts was displayed up to 99.9% at pH 1.2 in a dose-dependent manner. They also had 90.4% electron donating ability(EDA) at the concentration of 0.1 mg/mL. Extracts of Sarcodon asparatus were also able to function as a powerful antioxidant at all concentrations(0.01∼l.0 mg/mL). Furthermore, we observed that 1 mg/mL concentration of the extracts was more powerful than BHT, With respect to fibrolytic activity, Sarcodon asparatus showed 1,843.8 unit/g, which was higher than streptokinase(1,189 unit/g). The inhibitory effects of the extracts on angiotensin converting enzyme, measured by the normal and pretreatment methods, were 53 and 58%, respectively. We also performed cytotoxicity effect of Sarcodon asparatus extracts on a various cancer cell lines. The growth inhibitory effects of the extracts(5.0 mg/mL) on A549, HeLa, AGS, and SK-Hep-1 cells were 78.9, 55.3, 69.0, and 42.5 %, respectively. Interestingly, Sarcodon asparatusextracts induced mutation on Salmonella typhimurium TA98 and TA100 when Ames test was done.

Anti-aging Effects of L-Carnitine on Human Skin (L-카르니틴의 사람피부에 대한 항노화 효과)

  • Lee Bum-Chun;Choe Tae-Boo;Sim Gwan-Sub;Lee Geun-Soo;Park Sung-Min;Lee Chun-Il;Pyo Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.30 no.3 s.47
    • /
    • pp.393-397
    • /
    • 2004
  • L-Carnitine $({\beta}-hydroxy-{\gamma}-trimethyl-ammoniumbutyric{\;}acid)$ is a small water-soluble molecule important in mammalian fat metabolism. It is essential for the normal oxidation of fatty acids by the mitochondria, and is involved in the trans-esterification and excretion of acyl-CoA esters. In this paper, to investigate the relationship between aging and L-carnitine, we investigated the effects of in vitro matrix-metalloproteinase (MMP) inhibition and activity and expression of UYA-induced MMPs in human skin fibroblasts. Also, we studied to develop as anti-aging cosmetics with L-carnitine. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. ELISA (enzyme linked immune sorbent assay), gelatin-substrate zymography, RT-PCR ELISA techniques were used for the effects of L-carnitine on MMP expression, activity, and MMP mRNA expression in UVA irradiated fibroblast $(5\;J/cm^2)$, respectively. In addition, we performed clinical study with L-carnitine cream. L-carnitine inhibited the activities of MMP-1 in a dose-dependent manner and the $IC_{50}$ values calculated from semi-log plots were 2.45 mM, and L-carnitine showed strong inhibition on MMP-2 (gelatinase) activity in UVA irradiated fibroblast by zymography. Also, UVA induced MMP-1, 2 expression was reduced $43\%,\;53\%$ by treated with L-carnitine at 1.25 mM, and MMP-1 mRNA expression was reduced dose-dependent manner. Therefore L-carnitine was able to significantly inhibit the MMP activity, and regulate MMP expression in protein and mRNA level. The results of clinical study showed that $1.0\%$ L-carnitine treated group reduced wrinkle significantly compared with placebo treated group (P<0.05). All these results suggest that L-carnitine may be useful as new anti-aging cosmetics for protection against UVA induced Mm expression and activity.

Extraction of Polyphenols from Apple Peel Using Cellulase and Pectinase and Estimation of Antioxidant Activity (Cellulase와 Pectinase를 이용한 사과껍질 폴리페놀 추출 및 항산화 활성 평가)

  • Park, Min-Kyung;Kim, Cherl-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.38 no.5
    • /
    • pp.535-540
    • /
    • 2009
  • The effect of cellulolytic (Viscozyme) and pectolytic (Pectinex) enzyme treatments on extraction of total polyphenol and antioxidant activity of extract from apple peel have been examined. Extraction was carried out with a dosage of 0.1, 0.25, 0.5, 1 and 2% (v/v) of Viscozyme, Pectinex and Viscozyme+Pectinex at $30{\sim}50^{\circ}C$ for $12{\sim}24$ hours. Total polyphenol contents (mg/mL) of extracts obtained with 2% of Viscozyme, Pectinex or Viscozyme+Pectinex treatment for 12 hours were $0.30{\pm}0.02$, $0.16{\pm}0.01$, and $0.33{\pm}0.02$ at $30^{\circ}C$, $0.34{\pm}0.01$, $0.19{\pm}0.01$, and $0.35{\pm}0.02$ at $40^{\circ}C$ and $0.34{\pm}0.01$, $0.22{\pm}0.01$, and $0.38{\pm}0.02$ at $50^{\circ}C$ respectively. The result shows that Viscozyme was more effective than Pectinex at all experimental temperatures, and Viscozyme+Pectinex resulted in the highest phenolic content at $50^{\circ}C$. Antioxidant activities determined by DPPH, ABTS and FRAP assays were increased with concentrations of extracts produced by 2% of Viscozyme+Pectinex treatment, which ranged from 0.10 to 0.40 vit. C eq mM for $5{\sim}25\;mg$ of dried matters, from 0.09 to 0.28 vit. C eq mM for $1{\sim}5\;mg$ of dried matters, and from 0.06 to 1.85 $FeSO_4$ eq mM for $1{\sim}5\;mg$ of dried matters, respectively.

Anti-Oxidative and Anti-Inflammatory Activities of Euptelea Pleiosperma Ethanol Extract (Euptelea pleiosperma 에탄올 추출물의 항산화 및 항염증 활성)

  • Jin, Kyong-Suk;Park, Jung Ae;Lee, Ji Young;Kang, Ji Sook;Kwon, Hyun Ju;Kim, Byung Woo
    • Microbiology and Biotechnology Letters
    • /
    • v.42 no.2
    • /
    • pp.170-176
    • /
    • 2014
  • In this study, the anti-oxidative and anti-inflammatory activities of Euptelea pleiosperma ethanol extract (EPEE) were evaluated using in vitro assays and cell culture model systems. EPEE possessed a more potent scavenging activity against 1,1-diphenyl-2-picryl hydrazyl than the ascorbic acid used as a positive control. EPEE effectively suppressed lipopolysaccharide (LPS), in addition to hydrogen peroxide induced reactive oxygen species on RAW 264.7 cells. Furthermore, EPEE induced the expression of the anti-oxidative enzyme heme oxygenase 1 (HO-1) and its upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2), dose and time dependently. The modulation of HO-1 and Nrf2 expression might be regulated by mitogen-activated protein kinases and phosphatidyl inositol 3 kinase/Akt as their upstream signaling pathways. On the other hand, EPEE inhibited LPS induced nitric oxide (NO) formation without cytotoxicity. Suppression of NO formation was the result of the down regulation of inducible NO synthase (iNOS) by EPEE. Suppression of NO and iNOS by EPEE may be modulated by their upstream transcription factor, nuclear factor ${\kappa}B$, and AP-1 pathways. Taken together, these results provide important new insights into E. pleiosperma, namely that it possesses anti-oxidative and anti-inflammatory activities, indicating that it could be utilized as a promising material in the field of nutraceuticals.

Physico-chemical, Nutritional, and Enzymatic Characteristics of Shiitake Spent Mushroom Substrate (SMS) (표고버섯 수확 후 배지의 이화학적, 영양적, 효소적 특성)

  • Sung, Hwa-Jung;Pyo, Su-Jin;Kim, Jong-Sik;Park, Jong-Yi;Sohn, Ho-Yong
    • Journal of Life Science
    • /
    • v.28 no.11
    • /
    • pp.1339-1346
    • /
    • 2018
  • In Korea, edible mushrooms are produced largely on commercial artificial media, so the annual production of spent mushroom substrate (SMS), as a by-product of the mushroom industry, is estimated at over 200 million tons. This SMS is assumed to contain abundant fungal mycelia and pre-fruiting bodies, as well as various nutritive and bioactive compounds that are presently discarded. This study examined the physico-chemical, nutritional, and enzymatic characteristics of uninoculated sterilized medium (USM) and SMS of shiitake mushrooms with the aim of developing a high-value added product from SMS. The contents of crude protein, crude lipid, and ash were higher after the third SMS harvest ($SMS-A-3^{rd}$) than in USM or $SMS-A-1^{st}$. The contents of Ca, Mg, and P in $SMS-A-3^{rd}$ were 2.95, 2.35, and 2.1-fold higher compared than in USM. No As or Cd was detected in USM or SMS. The pH, Brix, and acidity were 4.6, 20.0, and 1.4, respectively in $SMS-A-3^{rd}$, but 5.6, 6.0, and 0.0, respectively, in USM. These results suggest a highly active production of soluble components and organic acids in $SMS-A-3^{rd}$. The distinct color differences noted for USM, $SMS-A-1^{st}$, and $SMS-A-3^{rd}$ could be used as a mycelial growth indicator. Enzyme activity assays using the APIZYM system showed that SMS is a potent source of hydrolysis-related enzymes, especially esterase (C4) and ${\beta}$-glucuronidase. Our results suggested that the SMS of shiitake has a high potential for use in environmental, agricultural, and stock-breeding industries, for example, as active ingredients for sewage treatment, waste-polymer degradation, and feed additives.

Conditioned Media of RAW 264.7 Cells Stimulated with Phellinus linteus Extract Regulates the Epithelial-mesenchymal Transition in Prostate Cancer Cells (상황버섯에 의해 활성화된 RAW 264.7 대식세포주 배양액의 인간 전립선암 세포주의 epithelial-mesenchymal transition 조절)

  • Kang, Taewoo;An, Hyun-Hee;Park, Sul-Gi;Yu, Sun-Nyoung;Hwang, You-Lim;Kim, Ji-Won;Ahn, Soon-Cheol
    • Journal of Life Science
    • /
    • v.29 no.8
    • /
    • pp.904-915
    • /
    • 2019
  • Prostate cancer (PCa) is one of the most metastatic tumor. Although hormone therapy or surgical castration is mostly conducted to treat PCa, it has a lot of side effects. Recently, many researchers have been exploring the tumor microenvironment to remedy these circumstances. Immune cells, especially macrophages, are an important composition of the tumor microenvironment. Under normal conditions, macrophages exhibit mild tumoricidal activity against tumors. However, once activated by interferon gamma or lipopolysaccharides, macrophages can kill cancer cells directly or indirectly by secreting cytokines and chemokines. In this study, murine macrophage RAW 264.7 cells were treated with Phellinus linteus extract. To analyze their pro-inflammatory phenotype, we were used several assays such as a real-time polymerase chain reaction, an enzyme-linked immunosorbent and nitric oxide assay. Prostate cancer cells were treated with the RAW 264.7-conditioned media, which was identified as a pro-inflammatory nature, for 48 h, and the expression of epithelial-mesenchymal transition (EMT)-related genes was determined. Not only N-cadherin, Snail, Twist, Slug, and Cadherin 11, which are mechenchymal-related proteins, were decrease, but epithelial marker of E-cadherin was increased. In addition, the mRNA level of vimentin, ccl2, and vegfa were decreased, as the EMT is closely related to the migration and invasion of cancer cells. In conclusion, the RAW 264.7-conditioned media stimulated with P. linteus extract inhibited migration and invasion and regulated the EMT pathway in human prostate cancer cells.

Antioxidant activity of ethanol extract of Lycium barbarum's leaf with removal of chlorophyll (클로로필을 제거한 영하구기엽 에탄올 추출물의 항산화 활성)

  • Kim, Ji Eun;Bae, Su Mi;Nam, You Ree;Bae, Eun Young;Ly, Sun Yung
    • Journal of Nutrition and Health
    • /
    • v.52 no.1
    • /
    • pp.26-35
    • /
    • 2019
  • Purpose: The aim of this study was to estimate the antioxidant activities of 50%, 70%, and 100% ethanol extracts of Lycium barbarum leaf and chlorophyll removal extract. Methods: The antioxidant activities were estimated by measuring total polyphenol content and by assays of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfate) (ABTS) radical scavenging activities and ferric reducing antioxidant power (FRAP). In addition, reactive oxygen species (ROS) production, DNA fragmentation, and antioxidant enzyme (superoxide dismutase and catalase) activities of the extracts were measured in hydrogen peroxide ($H_2O_2$)-stressed HepG2 cells. Results: The total polyphenol content, DPPH and ABTS radical scavenging activities, and FRAP value of the extracts increased in an ethanol concentration-dependent manner. The antioxidant activities of the chlorophyll-removal extracts were much higher than those of the chlorophyll-containing extracts. Cytotoxicity was not observed in HepG2 cells with extracts up to $1,000{\mu}g/mL$. All extracts inhibited ROS production in a concentration-dependent manner from $31.3{\mu}g/mL$ and inhibited DNA damage at $250{\mu}g/mL$. The SOD and catalase activities of cell lines treated with the extracts and $H_2O_2$ were similar to those of normal cells, indicating a strong protective effect. Conclusion: Lycium barbarum leaf extracts had high antioxidant activities and protected $H_2O_2$-stressed HepG2 cells. Since the chlorophyll-removal extract exhibited higher antioxidant activities than the chlorophyll-containing ones and the cytoprotective effect was similar, chlorophyll removal extract of Lycium barbarum leaf could be developed as ingredients of functional food and cosmetics.