• Title/Summary/Keyword: Activity Determination

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Activity Determination, Kinetic Analyses and Isoenzyme Identification of Gamma Glutamyltransferase in Human Neutrophils

  • Sener, Azize;Yardimci, Turay
    • BMB Reports
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    • v.38 no.3
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    • pp.343-349
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    • 2005
  • Gamma-glutamyltransferase (GGT, EC 2.3.2.2) which hydrolyzes glutathione (GSH), is required for the maintenance of normal intracellular GSH concentration. GGT is a membrane enzyme present in leukocytes and platelets. Its activity has also been observed in human neutrophils. In this study, GGT was purified from Triton X-100 solubilized neutrophils and its kinetic parameters were determined. For kinetic analyses of transpeptidation reaction, $\gamma$-glutamyl p-nitroanilide was used as the substrate and glycylglycine as the acceptor. Apparent $K_m$ values were determined as 1.8 mM for $\gamma$-glutamyl p-nitroanilide and 16.9 mM for glycylglycine. The optimum pH of GGT activity was 8.2 and the optimum temperature was $37^{\circ}C$. It had thermal stability with 58% relative activity at $56^{\circ}C$ for 30 min incubation. L-serine, in the presence of borate, was detected as the competetive inhibitor. Bromcresol green inhibited neutrophil GGT activity as a noncompetetive inhibitor. The neutrophils seem to contain only the isoenzyme that is present in platelets. We characterized the kinetic properties and compared the type of the isoenzyme of neutrophil GGT with platelet GGT via polyacrylamide gel electrophoresis (PAGE) under a standart set of conditions.

Analysis of Nitrate Reductase Activity for Dominant Tree Leaves in the Northern Aspect Forest of Changbai Mountain, China (중국 장백산 북사면 산림에서 우세목의 잎 내 질소 환원 효소 활성도 분석)

  • wen, Li-Yu;Kim, Ji-Hong
    • Journal of Korea Foresty Energy
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    • v.22 no.3
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    • pp.29-36
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    • 2003
  • This paper concerned the application of improvement in Vivo of Traditional Method for determination of nitrate reductase (NR) activity of leaves to dominant tree species in five forest communities of northern aspect of Changbai Mountain. The results indicated that the NR activity of tree species was related to shade tolerance, and the intolerant tree species had higher NR activity. The NR of a species was also related to the vertical structure and ecological site condition. The tree species, which have higher NR activities should be selected for fast growing and high yield tree species.

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Antifungal Activity-Guided Analysis of Actinostemma lobatum Extracts through Serial Sub-fractions

  • Seonwoo Choi;Song Hee Lee;Byeong Su Hwang;Young Taek Oh;Junhyun Jeon
    • The Plant Pathology Journal
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    • v.40 no.2
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    • pp.218-224
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    • 2024
  • Plants are treasure trove of novel compounds that have potential for antifungal chemicals and drugs. In our previous study, we had screened plant extracts obtained from more than eight hundred plant materials collected in Korea, and found that butanol fraction of the Actinostemma lobatum were most potent in suppressing growth of diverse fungal pathogens of plants. Here in this study, we describe further analysis of the butanol fraction, and summarize the results of subsequent antifungal activity test for the sub-fractions against a selected set of plant pathogenic fungi. This line of analyses allowed us to identify the sub-fractions that could account for a significant proportion of observed antifungal activity of initial butanol fraction from A. lobatum. Further analysis of these sub-fractions and determination of structure would provide the shortlist for novel compounds that can be a lead to new agrochemicals.

Factors Influencing the Bone Status of Korean Elderly Women (한국 노년기 여성의 골격 상태에 영향을 미치는 요인에 관한 연구)

  • 김혜경;윤진숙
    • Journal of Nutrition and Health
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    • v.24 no.1
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    • pp.30-39
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    • 1991
  • This study was designed to investigate the effects of dietary calcium. serum estrogen level and physical activity on the bone status of 116 healthy elderly women living in urban area. Current calcium intake was assessed by convenient method(refered to as Ca intake) and calcium containing food frequency method(refered to as Ca index) Daily activity record was used for the estimation of physical activity level, and serum estrogen level was measured from fasting blood of subjects. The rate of bone resorption was evaluated by the determination of hydroxyproline(Hpr) in fasting urine with correction for creatinine excretion. The results of this study are summarized as follows : 1) Average daily Ca intake of subjects was 621.4$\pm$155.8mg, which is above the Korean recommended dietary allowances. However 44.8% of the subjects consumed Ca below RDA level. Ca index score was significantly correlated with the bone status(P<0.05), Ca intake did not show significant correlation with the bone status although a positive trend of influence was evident. 2) Average serum estrogen level of subjects was 18.7$\pm$9.8pg Contrary to our anticipation. estrogen level did not show any significant relation to age and bone status. 3) Daily physical activity was classified into four categories by activity intensity : sedentary. moderate, active and severe. The average physical activity of subjects belong to moderate level. and the bone status was significantly related to the physical activity(P<0.01) 4) Among other influential factors such as age, pocket-money. family type. drinking, smoking and BMI, there was a significant difference between bone status and BMI(P<0.05). 5) Multiple regression analysis of variables showed that physical activity has greater effect than other variables when the entire subjects were taken into account. However. eliminating the subjects whose bone status rated as excellent(Hpr/cr<0.009), Ca index showed higher correlation than physical activity. These results have demonstrated that dietary calcium intake is the primary important factor for keeping good bone health and that bone status of subjects with a sufficient calcium intake is affected by various factors such as physical activity, age, smoking. BMI and others.

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The effects of an increase in self - determination experience on the behavior of young children with autism spectrum disorder by telepresence robot. (텔레프레젠스 로봇을 이용한 자기결정 경험의 증대가 자폐범주성 장애유아의 행동에 미치는 효과 (자기결정 활동 멀티미디어 콘텐츠의 적용을 통하여))

  • Kim, Su-Jin
    • Journal of rehabilitation welfare engineering & assistive technology
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    • v.12 no.1
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    • pp.38-45
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    • 2018
  • The purpose of this study was to investigate the effects of an increase in self - determination experience on the behavior of young children with autism spectrum disorder by telepresence robot. As for research method, the study used AB design, two selected children engaged in activities with a telepresence robot in free play time in the morning. The activities were conducted in 19 sessions, twice a week, 15 to 40 minutes each. To investigate the effect of the activity on the child's behavior was observed using the behaviors of free play time and work time in the afternoon. All the process was recorded by a camera and then analyzed by frequency recording. The results of the study are as follows. First, the participation of young children with autism spectrum disorder in free play time increased. Second, choice-making or preference behavior of young children with autistic spectrum disorder were increased. This study suggests that increasing the self-determination experience of young children with autism spectrum disorders using telepresence robots increases their participation and increases their choice-making or preference behavior.

Factors Indicating Culture Status During Cultivation of Spirulina (Arthrospira) platensis

  • Kim, Choong-Jae;Jung, Yun-Ho;Oh, Hee-Mock
    • Journal of Microbiology
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    • v.45 no.2
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    • pp.122-127
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    • 2007
  • Factors indicating culture status of two Spirulina platensis strains were monitored in a batch mode cultivation for 36 days. Changing mode in all factors showed a common turning point, indicating shift of cell or culture status. Mean biomass productivity was highly sustained until day 22, chlorophyll a concentration peaked on day 22, pH value was > 12 on day 22, coil number was abruptly shortened on day 22, and floating activity was sustained at greater than 79% after day 22, indicating that day 22 is a criterion reflecting phase-transfer in cell physiology in a batch culture system. Many of these changes may have been caused by increased pH, suggesting that pH control is essential for mass production of S. platensis. Fluctuations in floating activity were likely induced by the number of cellular gas vacuoles. Consequently, coil number per trichome and floating activity of S. platensis could readily act as simple indicators for determination of culture status or harvesting time of cells.

Characterization of Bacteriocin Produced by Lactobacillus bulgaricus

  • Kim, Hyun-Jin;Kim, Ji-Hyun;Son, Jeong-Hwa;Seo, Hyo-Jin;Park, So-Jin;Paek, Nam-Soo;Kim, Sung-Koo
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.503-508
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    • 2004
  • The antimicrobial substance produced by Lactobacillus bulgaricus was inactivated by protease. It showed inhibitory activity against Staphylococcus aureus ATCC6538, Streptococcus agalactiae ATCC14364, some Gram-positive and Gram-negative bacteria, and characteristics of a bacteriocin. The optimal temperature and culture time for the production of bacteriocin were $30^{\circ}C$ and 10 h, respectively, in the culture of L. bulgaricus. The bacteriocin production started in the exponential phase and reached a maximum at the early stationary phase. Using Staph. aureus ATCC6538 and Strep. agalactiae ATCC14364, known as common bovine mastitis pathogens, as indicator strains for determination of the bacteriocin activity, the antimicrobial activity of the bacteriocin was found to be stable in acidic and neutral pH's (2- 7) even at lOOT, whereas it was lost at high pH (10- 11) and $100^{\circ}C$. The mode of action for the antimicrobial activity was bacteriocidal, and the molecular weight determined by SDS-PAGE and overlay method was 14 kDa.

The Dynamics of Protein Decomposition in Lakes of Different Trophic Status - Reflections on the Assessment of the Real Proteolytic Activity In Situ

  • Siuda, Waldemar;Kiersztyn, Bartosz;Chrost, Ryszard J.
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.897-904
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    • 2007
  • The aim of this paper is to discuss the methodology of our investigation of the dynamics of protein degradation and the total in situ protealytic activity in meso/eutrophic, eutrophic, and hypereutrophic freshwater environments. Analysis of the kinetics and rates of enzymatic release of amino acids in water samples preserved with sodium azide allows determination of the concentrations of labile proteins $(C_{LAB})$, and their half-life time $(T_{1/2})$. Moreover, it gives more realistic information on resultant activity in situ $(V_{T1/2})$ of ecto- and extracellular proteases that are responsible for the biological degradation of these compounds. Although the results provided by the proposed method are general y well correlated with those obtained by classical procedures, they better characterize the dynamics of protein degradation processes, especially in eutrophic or hypereutrophic lakes. In these environments, processes of protein decomposition occur mainly on the particles and depend primarily on a metabolic activity of seston-attached bacteria. The method was tested in three lakes. The different degree of eutrophication of these lakes was clearly demonstrated by the measured real proteolytic pattern and confirmed by conventional trophic state determinants.

Establishment of in vitro 3-Dimensional Tumor Model for Evaluation of Anticancer Activity Against Human Solid Tumors (항고형암제의 활성평가를 위한 in vitro 삼차원 암세포 배양계의 확립)

  • Lee, Sang-Hak;Lee, Joo-Ho;Kuh, Hyo-Jeong
    • Journal of Pharmaceutical Investigation
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    • v.34 no.5
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    • pp.393-399
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    • 2004
  • For the efficient determination of activity against solid tumors, an in vitro tumor model that resembles the condition of in vivo solid tumors, is required. The purpose of this study was to establish a rapid culture method and viability assay for an in vitro 3-dimensional tumor model, multicellular spheroid (MCS). Among 12 human cancer cell lines, a few cell lines including DLD-1 (human colorectal carcinoma cells) formed fully compact MCS which was adequate for in vitro viability assay. DLD-1 MCS showed steady growth reaching $700\;{\mu}m$ diameter after 11 day culture. DLD-1 cells grown as MCS showed significant increase in $G_0/G_1$ phase compared to the monolayer cells (73.9% vs 45.7%), but necrotic regions or apoptotic cells were not observed. The cells cultured as MCS showed resistance to 5-FU (10.3 fold higher $IC_{50}$) compared to monolayers, however, tirapazamine (a hypotoxin) showed similar activity in both culture systems. In summary, MCS may be a valid in vitro model for activity screening of anticancer agents against human solid tumors and also exploitable for studying molecular markers of drug resistance in human solid tumors.

Anti-oxidant Activity of Phenolic Compounds and Simultaneous Contents Determination using HPLC-UV from the Akebiae caulis (목통의 페놀성 화합물의 항산화 활성 및 HPLC-UV 동시 함량분석)

  • Kyung, Min-Ji;Jeon, Hye-Jin;Whang, Wan-Kyunn
    • YAKHAK HOEJI
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    • v.59 no.3
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    • pp.120-126
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    • 2015
  • Akebiae caulis have been used in folk medicines for diuretic, menstrual pain, and diuretic pain. It has been also resolved nephritis and cystitis. In this study, we isolated three phenolic compounds from 70% methanol extract using the open column chromatography. These isolated compounds which were 2-(3,4-dihydroxyphenyl)-ethyl-${\beta}$-D-glucopyranoside, 5-O-caffeoylquinic acid, and syringoylglycerol 2-O-${\beta}$-D-glucoside were determined by physico-chemical apparatus. Furthermore, we conducted DPPH and ABTS assay in order to screen the antioxidant activity of isolated three compounds. Also, we developed a rapidly HPLC-UV analysis method of two phenolic compounds (2-(3,4-dihydroxyphenyl)-ethyl-${\beta}$-D-glucopyranoside, 5-O-caffeoylquinic acid) for evaluating the Akebiae Caulis collected 30 samples from different regions. From the experiments, all three isolated compounds showed the significant antioxidant activity. We suggested that the content criteria of marker compounds were shown by a simple and rapid HPLC-UV method. The contents respectively were 0.009% (2-(3,4-dihydroxyphenyl)-ethyl-${\beta}$-D-glucopyranoside) and 0.036% (5-O-caffeoylquinic acid).