• Title/Summary/Keyword: esterase

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Intergeneric Transfer of Isolated Nuclei from Lentinus edodes into Protoplasts of Pleurotus florida (사철느타리버섯 원형질체내에 표고 핵의 전이)

  • Yoo, Young-Bok;Shin, Pyung-Gyun
    • The Korean Journal of Mycology
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    • v.24 no.2 s.77
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    • pp.111-126
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    • 1996
  • Transfer of the isolated nuclei from Lentinus edodes into protoplasts of Pleurotus florida was induced with polyethlene glycol (PEG) and $CaCl_2$. The intergeneric transfer products were classified into nuclear hybrid, heterokaryon or synkaryon, and reconstituted cell. These progenies except nuclear hybrids formed mature fruiting bodies on sawdust rice bran medium. Formation of fruit bodies was influenced by several factors such as light, temperature, nutrition and physic state of the culture media. Most of fruiting body characters were similar to those of P. florida in synkaryon and L. edodes in reconstituted cell, respectively. All these basidiocarps had clamp connections though initial heterokaryon colonies were lacking. Isozyme patterns of intergeneric progenies were quite different from those of parents. DNA polymorphisms of transfer products were also compared by random amplified polymorphic DNAs (RAPD) analysis based on polymerase chain reaction. The RAPD patterns were different from those of donor and recipient. DNA fingerprints ranged in size from 0.25 to 4.0 Kb. On the basis of RAPD, the transfer products were classified into five groups. Two synkaryon were analysed with distribution of progenies and segregation of genetic markers by random spore analyses. The genetic markers were segregated into wild type and riboflavine requiring auxotrophs.

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The Changes in Firmness, Ca Content and Polygalacturonase and Pectinesterase Activities during Oyijangachi Preparation (오이장아찌 제조중 경도, 칼슘 함량, Polygalacturonase 및 Pectinesterase 활성 변화에 관한 연구)

  • 정순택;이홍열;박현진
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.5
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    • pp.796-802
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    • 1995
  • Oyijangachi, a traditional Korean brinded cucumber, was prepared by brinning the cucumbers in five different solutions for 48 hrs and then, was dipped into dipping sources(Kochujang, Doenjang and Ganjang) for 30 days of aging. Firmness, calcium content and enzyme activities(pectinesterase and polygalacturonase) changes were measured among the cucumbers which were treated by five different solutions during aging. The firmness of Kochujang Oyijangachi were the lowest after 10 days of aging for all from the five brining solutions because of "hollow phenomena" of cucumbers. Calcium contents of cucumbers after dipping into the five solutiosn increased as calcium content of the solutions increased and also increased when the cucumbers dipped into the dipping bases(Kochujang, Doenjang and Ganjang) because of calcium migration from the dipping sources into the cucumbers during aging. The calcium contents of the three dipping bases were ranged from 70mg% to 120mg% of Ca. The activity of polygalacturonase in the Oyijangachi decreased generally during aging and decreased rapidly during initial 5 days of aging. The activity of pectinesterase of cucumbers treated with 12% salts solutions(treatment 3, 4 and 5) were higher than those of cucumbers treated with 6% salts solutions(treatment 1 and 2).

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Comparison of the Results of Multistix$^{(R)}$SG and Comber-9-Test$^{(R)}$ RL Urine Dipstick Assay (Multistix$^{(R)}$-SG와 Comgur-9-test$^{(R)}$RL에 의한 요시험지봉검사 성적의 비교)

  • Kim, Dae-Chul;Kim, Kyung-Dong;Jung, Bo-Chan;Kim, Chung-Sook;Cho, Kil-Ho
    • Journal of Yeungnam Medical Science
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    • v.8 no.1
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    • pp.42-52
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    • 1991
  • Two types of urine dipstick assays, Multistix-SG and Combur-9-test RL, were compared for compatibility, accuracy, specificity and predictive values of a positive and negative test in 501 patients urine and artificially prepared specimen. We found that the results of semiquantitative tests of Multistix-SG and Combur-9-Test RL performed were statistically similar in patients specimen. The urinary leukocyte estrase tests of Combur-9-Test RL assays compared with urine sediment microscopy in regard to compatibility, sensitivity, specificity, and predictive values of a positive and negative test 83.7%, 48.1%, 90.3%, 47.4% and 90.1%, respectively. The urinar nitrite tests of Comber-9-Test RL assays compared with urine culture tests, in regard to compatibility, sensitivity, specificity, and predictive values of a positive and negative tests were 90.3%, 19.4%, 84.7%, 53.8% and 94.1%, respectively. For the urinary protein, the sulfosalicylic acid method was the most sensitive test for any kinds of protein, and Multistix-SG appeared more sensitive than Compur-9-Test RL for the albuminuria. For the urinary bilirubin and glusose, two dipstick assays were similar in their diganostic efficiency. Finally in the urinary occult blood tests, Combur-9-Test RL assays was more sensitive than Multistix-SG.

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Comparison of toxicity and detoxifying enzyme activity in carp (Cyprinus carpio) treated with some synergistic pesticides (농약 상호간의 협력작용에 의한 잉어의 독성과 해독효소 활성의 비교)

  • Yang, Kwang-Rok;Shim, Jae-Han;Suh, Yong-Tack
    • Applied Biological Chemistry
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    • v.35 no.5
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    • pp.367-374
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    • 1992
  • This study was performed to investigate effects of probable detoxifying enzyme activity and toxicity by pesticides and their combinations in the fresh water fish. Seven pesticides including IBP, isoprothiolane, cartap, ridomil, chlorothalonil, captafol and endosulfan were subjected to investigate for their acute toxicites and synergism possibilities. The $LC_{50}$ value of endosulfan was the lowest at showing 0.0079 ppm and that of metalaxyl was the highest as showing 40 ppm over. The synergism effects of relative pesticides were observed in the combinations of isoprothiolane+IBP and isoprothiolane+cartap. The changes of glycogen contents in fish liver were assayed for 5 pesticides and its highest inhibition effect of glycogen showed in IBP treated fish. The activity of probable detoxifying enzymes including carboxylesterase (CE), glutathion S-transferase (GST) and lactate dehydrogenase (LDH) were assayed in carp liver at dose of sublethal concentrations. Effects of pesticides on changes in each enzyme activities were as follows: carboxylesterase (CE) activities were the highest in IBP and gtutathion S-transferase (GST) activities were the highest in iosoprothiolane+IBP. Both activities of carboxylesterase (CE) and glutahtion S-transferase (GST) were increased by 5 chemicals. The highest LDH activity showed in isoprothiolane treated fish, while the lowest activity was observed in isoprothiolane+cartap. Sublethal exposure to cartap and isoprothiolane+cartap in carp exerted various effects on LDH activity.

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A staining method to determine marine microplanktonic organism viability and investigate the efficacy of a ship's ballast water treatment system (선박평형수 처리장치 효율 검증을 위한 해양미소부유생물 생사판별기법)

  • Baek, Seung Ho;Shin, kyoungsoon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.6
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    • pp.4328-4334
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    • 2015
  • We determined a method to determine marine planktonic organism viability using Evan's blue, Aniline blue, and 5-choromethyfluorescein diacetate (CMFDA). The Evan's blue and Aniline blue methods produced bright blue light for dead phytoplankton and zooplankton and were the best dyes to detect dead cells. The staining efficiency of Evan's blue and Aniline blue were ${\geq}90%$ of the original field sample. However, it was difficult to test the efficiency of a ship's ballast water treatment system because detection of living cells. In contrast, the CMFDA method, which is based on measuring cell esterase activity using a fluorimetric stain, was the best dye to detect live cells of almost all phytoplankton species, and staining efficiency was 70%. The CMFDA method is similar to the fluorescein diacetate (FDA) staining method. Therefore, we estimated viability of phytoplankton species using a double-staining method by combining CMFDA and FDA to determine optimum staining efficiency. As a result, the frequency of dying cells based on the double-staining method was 95%, which was significantly higher than that of single CMDFA staining. Our results suggest that a CMDFA + FDA assay is more effective to determine survival of marine plankton and that this method was applicable to investigate the efficacy of a ship's ballast water treatment system.

Remifentanil Protects Human Keratinocyte Through Autophagic Expression

  • Kim, Eok Nyun;Park, Chang Hoon;Woo, Mi Na;Yoon, Ji Young;Park, Bong Soo;Kim, Yong Ho;Kim, Cheul Hong
    • Journal of The Korean Dental Society of Anesthesiology
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    • v.14 no.2
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    • pp.101-106
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    • 2014
  • Background: Remifentanil, an ultra-short-acting mu-opioid receptor agonist, is unique from other opioids because of its esterase-based metabolism, minimal accumulation, and very rapid onset and offset of clinical action. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. However, the effects of remifentanil on human keratinocyte and autophagy have yet to be fully elucidated during hypoxia-reoxygenation. Here we investigated whether remifentanil confers protective effect against hypoxia-reoxygenation in human keratinocyte and, if so, whether autophagy mediates this effect. Methods: The human keratinocytes were cultured under 1% oxygen tension. The cells were gassed with 94% $N_2$, and 5% $CO_2$ and incubated for 24 h at $37^{\circ}C$. To determine whether the administration of affects human keratinocytes hypoxia-reoxygenation injury, cells were then exposed to various concentrations of remifentanil (0.01, 0.1, 0.5 and 1 ng/ml) for 2 h. After remifentanil treatment, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at $37^{\circ}C$. Control group did not receive remifentanil treatment. Normoxia group did not receive hypoxia and remifentanil treatment for 36 h. 3-MA group was treated 3-methyladenine (3-MA) for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with MTT, showing the mitochondrial activity of living cells. Cells were stained with fluorescence and analyzed with Western blot analysis to find out any relations with activation of autophagy. Results: Prominent accumulation of autophagic specific staining MDC was observed around the nuclei in RPT group HaCaT cells. Similarly, AO staining, red fluorescent spots appeared in RPT group HaCaT cells, while the Normoxia, control and 3-MA groups showed mainly green cytoplasmic fluorescence. We here examined activation of autophagy related protein under H/R-induced cells by Western blotting analysis. Atg5, Beclin-1, LC3-II (microtubule-associated protein 1 light chain 3 form II) and p62 was elevated in RPT group cells. But they were decreased when autophagy was suppressed by 3-MA (Fig. 5). Conclusions: Although the findings of this study are limited to an in vitro interpretation, we suggest that remifentanil may have a beneficial effect in the recovery of wound from hypoxia-reoxygenation injury.

Characteristics of Fibrinolytic Enzymes of Bacillus licheniformis CY-24 Isolated from Button Mushroom Compost (양송이 배지로부터 분리한 Bacillus licheniformis CY-24의 섬유소분해 효소의 특성)

  • Min, Gyeong-Jin;Park, Hea-sung;Lee, Een-ji;Lee, Chan-Jung
    • The Korean Journal of Mycology
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    • v.49 no.2
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    • pp.199-209
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    • 2021
  • The present study was performed to improve the technique used for fermenting the mushroom growth medium. Taxonomic analysis of 16S rDNA sequence from the predominant Bacillus strain CY-24 isolated during the fermentation phase of the rice straw medium identified it as Bacillus licheniformis. In addition, the growth environment of B. licheniformis was also examined in this study, which revealed the optimal growth temperature and pH to be 30 ℃ and 6.0, respectively. This study also revealed that carboxymethyl cellulase (CMCase) and polygalacturonase (PGase) enzymes isolated from B. licheniformis achieved their maximal activities at 50 ℃ and 60 ℃ respectively. Furthermore, the study confirmed that the two enzymes, i.e., CMCase and PGase in B. licheniformis are stable at temperatures above 60 ℃. The present study thus demonstrates that B. licheniformis CY-24 possesses excellent enzymatic properties. It also reveals that the action of enzymes during the production of growth mediums used for the cultivation of mushrooms is closely associated with the promotion of fermentation and softening of the rice straw. Overall, this study provides elementary information regarding the role of B. licheniformis enzymes during growth medium fermentation for Agaricus bisporus cultivation.

Efficacy of combination of endo-xylanase and xylan-debranching enzymes in improving cereal bran utilization in piglet diet

  • Wang, Weiwei;Zheng, Dawen;Zhang, Zhenzhen;Ye, Hui;Cao, Qingyun;Zhang, Changming;Dong, Zemin;Feng, Dingyuan;Zuo, Jianjun
    • Animal Bioscience
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    • v.35 no.11
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    • pp.1733-1743
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    • 2022
  • Objective: This study was aimed to explore the efficacy of combination of endo-xylanase (Xyn) and xylan-debranching enzymes (arabinofuranosidase, Afd and feruloyl esterase, FE) in improving utilization of bran in piglet diet. Methods: In vitro experiments were firstly conducted to examine the enzymological properties of Xyn, Afd, and FE, concurrent with their effect on degradation of arabinoxylan (Abx) in bran. In vivo experiment was then implemented by allocating two hundred and seventy 35-d-old postweaning piglets into 3 groups (6 replicates/group), which received bran-containing diet supplemented with Xyn (1,600 U/kg) or its combination with Afd (0.8 U/kg) and FE (4 U/kg) or without enzyme. Results: Both Xyn, Afd, and FE are relatively stable against the changes in temperature and pH value. Combining Xyn with Afd and FE had a superiority (p<0.05) over Xyn alone and its combination with Afd or FE in promoting (p<0.05) degradation of Abx in different brans. Combined treatment with Xyn, Afd, and FE was more beneficial than Xyn alone to induce increasing trends (p<0.10) of average daily gain, final body weight and feed efficiency of piglets fed bran-containing diet. Moreover, combination of Xyn, Afd, and FE showed advantages (p<0.05) over Xyn alone in causing reductions (p<0.05) in diarrhea rate and cecal pH value, concurrent with increases (p<0.05) in cecal and colonic acetic acid and total volatile fatty acid concentrations, as well as cecal butyric acid concentration of piglets fed bran-containing diet. Conclusion: Combining Xyn with Afd and FE was more beneficial than Xyn alone in promoting degradation of Abx in bran, along with growth performance and intestinal volatile fatty acid profile of piglets received bran-containing diet. Thereby, combination of Xyn, Afd, and FE had a superior efficacy relative to Xyn alone in improving application of cereal bran in piglet diet.

Perilla Frutescens Extract Protects against Scopolamine-Induced Memory Deficits in Mice (스코폴라민으로 유도한 기억력 손상 모델에서 소엽 추출물의 보호 효과)

  • Lee, Jihye;Lee, Eunhong;Jung, Eun Mi;Kim, Dong Hyun;Kim, Sung-kyu;Park, Mi Hee;Jung, Ji Wook
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.35 no.3
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    • pp.97-103
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    • 2021
  • Perilla frutescens (P. frutescens) is an important herb used for many purposes such as medicinal, aromatic, and functional food in Asian countries and has beneficial effects such as antioxidant activity, anti-inflammation activity, anti-depression activity, and anxiolytic activity. However, there have been no studies on the protective effect of P. frutescens extract (PFE) on amnesia in vivo. The present study aimed to investigate whether PFE protects memory deficit using a scopolamine-induced mice model and elucidate the underlying mechanisms involved. The protective effect of PFE against scopolamine-induced memory deficits was investigated using Y-maze, passive avoidance, and Morris water maze tests. Furthermore, the potential mechanisms of PFE in improving memory capabilities related to the cholinergic system and antioxidant activity were examined. PFE significantly increased spontaneous alternation in the Y-maze test, step-through latency in the passive avoidance test, and swimming time in the target quadrant in the probe test when compared to the scopolamine-treated group. Likewise, PFE significantly decreased escapes latency in the Morris water maze test. PFE could not regulate cholinergic function in acetylcholine level and acetylcholine esterase activity. However, PFE increased DPPH radical scavenging activity dose-dependently and total polyphenol content was 127.7±1.2 ㎍ GAE/mg. The results showed that the PFE could be a preventive and/or therapeutic candidate for memory and cognitive dysfunction in Alzheimer's disease.

Mountain-cultivated ginseng protects against cognitive impairments in aged GPx-1 knockout mice via activation of Nrf2/ChAT/ERK signaling pathway

  • Bao Trong Nguyen;Eun-Joo Shin;Ji Hoon Jeong;Naveen Sharma;Ngoc Kim Cuong Tran;Yen Nhi Doan Nguyen;Dae-Joong Kim;Myung Bok Wie;Yi Lee;Jae Kyung Byun;Sung Kwon Ko;Seung-Yeol Nah;Hyoung-Chun Kim
    • Journal of Ginseng Research
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    • v.47 no.4
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    • pp.561-571
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    • 2023
  • Background: Escalating evidence shows that ginseng possesses an antiaging potential with cognitive enhancing activity. As mountain cultivated ginseng (MCG) is cultivated without agricultural chemicals, MCG has emerged as a popular herb medicine. However, little is known about the MCG-mediated pharmacological mechanism on brain aging. Methods: As we demonstrated that glutathione peroxidase (GPx) is important for enhancing memory function in the animal model of aging, we investigated the role of MCG as a GPx inducer using GPx-1 (a major type of GPx) knockout (KO) mice. We assessed whether MCG modulates redox and cholinergic parameters, and memory function in aged GPx-1 knockout KOmice. Results: Redox burden of aged GPx-1 KO mice was more evident than that of aged wild-type (WT) mice. Alteration of Nrf2 DNA binding activity appeared to be more evident than that of NFκB DNA binding activity in aged GPx-1 KO mice. Alteration in choline acetyltransferase (ChAT) activity was more evident than that in acetylcholine esterase activity. MCG significantly attenuated reductions in Nrf2 system and ChAT level. MCG significantly enhanced the co-localization of Nrf2-immunoreactivity and ChAT-immunoreactivity in the same cell population. Nrf2 inhibitor brusatol significantly counteracted MCG-mediated up-regulation in ChAT level and ChAT inhibition (by k252a) significantly reduced ERK phosphorylation by MCG, suggesting that MCG might require signal cascade of Nrf2/ChAT/ERK to enhance cognition. Conclusion: GPx-1 depletion might be a prerequisite for cognitive impairment in aged animals. MCG-mediated cognition enhancement might be associated with the activations of Nrf2, ChAT, and ERK signaling cascade.