• Title/Summary/Keyword: Specific reactivity

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Molecular Orbital Theory on Cellulolytic Reactivity Between pNP-Cellooligosccharides and ${\beta}$-Glucosidase from Cellulomonas uda CS1-1

  • Yoon, Min-Ho;Nam, Yun-Kyu;Choi, Woo-Young;Sung, Nack-Do
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1789-1796
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    • 2007
  • A ${\beta}$-glucosidase with the molecular mass of 160,000 Da was purified to homogeneity from cell extract of a cellulolytic bacterium, Cellulomonas uda CS1-1. The kinetic parameters ($K_m$ and $V_{max}$) of the enzyme were determined with pNP-cellooligosccharides (DP 1-5) and cellobiose. The molecular orbital theoretical studies on the cellulolytic reactivity between the pNP-cellooligosaccharides as substrate (S) molecules and the purified ${\beta}$-glucosidase (E) were conducted by applying the frontier molecular orbital (FMO) interaction theory. The results of the FMO interaction between E and S molecules verified that the first stage of the reaction was induced by exocyclic cleavage, which occurred in an electrophilic reaction based on a strong charge-controlled reaction between the highest occupied molecular orbital (HOMO) energy of the S molecule and the lowest occupied molecular orbital (LUMO) energy of the hydronium ion ($H_3O^+$), more than endocyclic cleavage, whereas a nucleophilic substitution reaction was induced by an orbital-controlled reaction between the LUMO energy of the oxonium ion ($SH^+$) protonated to the S molecule and the HOMO energy of the $H_2O_2$ molecule. A hypothetic reaction route was proposed with the experimental results in which the enzymatic acid-catalyst hydrolysis reaction of E and S molecules would be progressed via $SN_1$ and $SN_2$ reactions. In addition, the quantitative structure-activity relationships (QSARs) between these kinetic parameters showed that $K_m$ has a significant correlation with hydrophobicity (logP), and specific activity has with dipole moment, respectively.

IgE Binding Reactivity of Peptide Fragments of Bla g 4, a Major German Cockroach Allergen

  • Shin, Kwang-Hyun;Jeong, Kyoung-Yong;Hong, Chein-Soo;Yong, Tai-Soon
    • Parasites, Hosts and Diseases
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    • v.47 no.1
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    • pp.31-36
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    • 2009
  • Cockroaches have been recognized as a major cause of asthma. Bla g 4 is one of the most important German cockroach allergens. The aim of this study is to investigate IgE reactivity to the recombinant Bla g 4 (rBla g 4) in the sera of allergic patients and identify linear IgE binding epitope. For protein expression, full-length Bla g 4 (EF202172) was divided into 5 overlapping peptide fragments (E1: aa 1-100, E2: aa 34-77, E3: aa 74-117, E4: aa 114-156, and E5: aa 153-182). The full-length and 5 peptide fragments of Bla g 4 was generated by PCR and over-expressed in E. coli BL21 (DE3). The IgE binding reactivities of the full-length and peptide fragments were measured by ELISA using 32 serum samples of cockroach allergy. The sera of 8 patients (25%) reacted with rBla g 4. Four sera (100%) showed IgE-binding reactivity to full-length and peptide fragment 4, and 2 sera (50%) reacted with peptide fragment 2. One (20%) serum reacted with peptide fragment 3. The results of ELISA using overlapping recombinant fragments indicated that the epitope region was located at amino acid sequences 34-73 and 78-113, and major IgE epitope of Bla g 4 was located at amino acid sequences 118-152 of C-terminal. B-cell epitope analysis of German cockroach allergen Bla g 4 could contribute to the strategic development of more specific and potentially efficacious immunotherapy.

Psychophysiological Reactivity to Affective Visual Stimulation of Negative Emotional Valence: Comparative Analysis of Autonomic and Frontal EEG Responses to the IAPS and the KAPS

  • Sohn, Jin-Hun;Estate M. Sokhadze;Lee, Kyung-Hwa
    • Science of Emotion and Sensibility
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    • v.3 no.2
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    • pp.29-40
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    • 2000
  • Autonomic and EEG responses were analyzed in 32 college students exposed to visual stimulation with Korean Affective Picture System (KAPS) and 36 students exposed to the International Affective Picture System (IAPS). Cardiac, electrodermal, and electrocortical measures were recorded during 30 sec of viewing affective pictures. The slides intended to elicit basic emotions (fear, anger, surprise, disgust, and sadness) were presented to subjects via Kodak slide-projector. The aim of the study was to differentiate autonomic and EEG responses associated with the same negative valence emotions elicited by KAPS and IAPS stimulation and to identify the influence of cultural relevance on physiological reactivity. The analysis of obtained results revealed significant differences in physiological responsiveness to emotionally negative valence slides from KAPS and IAPS. The typical response profile for all emotions elicited by the KAPS included HR acceleration (except surprise), and increase of electrodermal activity, slow and fast alpha blocking and fast beta power increase in EEG, which was not associated with significant asymmetry (except fast alpha in sadness). Stimulation with the IAPS evoked HR deceleration, specific electrodermal responses with relatively high tonic electrodermal activation, alpha-blocking and fast beta increase, and was accompanied also by theta power increase and marked frontal asymmetry (e.g., fast beta, theta asymmetries in sadness, fast alpha in fear). Physiological responses to fear and anger-eliciting slides from the IAPS were significantly less profound and were accompanied by autonomic and EEG changes more typical for attention rather than negative affect. Higher cardiovascular and electrodermal reactivity to fear emotion observed in the KAPS, e.g., as compared to data with the IAPS as stimuli, can be explained by cultural relevance and higher effectiveness of the KAPS in producing certain emotions such as fear in Koreans.

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Variability of laying hen behaviour depending on the breed

  • Kozak, Agnieszka;Kasperek, Kornel;Zieba, Grzegorz;Rozempolska-Rucinska, Iwona
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.1062-1068
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    • 2019
  • Objective: For many generations, most species of farm animals have been subjected to intense and strictly targeted selection for improvement of their performance traits. This has led to substantial changes in animal anatomy and physiology, which resulted in considerable differences between the current animal breeds and their wild ancestors. The aim of the study was to determine whether there is breed-specific variability in behaviour as well as differences in emotional reactivity and preferences of laying hens. Methods: The investigations involved 50 Green-legged Partridge, 50 Polbar, and 50 Leghorn hens. All birds were kept in the same conditions, and the behavioural tests were carried out at 30 weeks of age. We used the tonic immobility test and a modified open-field test including such objects as water, commercial feed, feed enriched with cereal grains, finely cut straw, and insect larvae, a sandpit, a mirror, and a shelter imitating a hen nest. Results: The research results demonstrate that the birds of the analysed breeds differ not only in the excitability and emotional reactivity but, importantly, also in the preferences for environment-enriching elements. Ensuring hens' well-being should therefore be based on environmental modifications that will facilitate acquisition of essential elements of chickens' behaviour. The greatest emotional reactivity was found in the Leghorn breed, which may be a result of correlated selection aimed at an increase in chicken productivity. Conclusion: The differences in the behaviour of the birds from the analysed breeds indicate that laying hens cannot be regarded as one group of animals with the same environmental requirements.

Oxytocin-induced endothelial nitric oxide dependent vasorelaxation and ERK1/2-mediated vasoconstriction in the rat aorta

  • Xu, Qian;Zhuo, Kunping;Zhang, Xiaotian;Zhang, Yaoxia;Xue, Jiaojiao;Zhou, Ming-Sheng
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.4
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    • pp.255-262
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    • 2022
  • Oxytocin is a neuropeptide produced primarily in the hypothalamus and plays an important role in the regulation of mammalian birth and lactation. It has been shown that oxytocin has important cardiovascular protective effects. Here we investigated the effects of oxytocin on vascular reactivity and underlying the mechanisms in human umbilical vein endothelial cells (HUVECs) in vitro and in rat aorta ex vivo. Oxytocin increased phospho-eNOS (Ser 1177) and phospho-Akt (Ser 473) expression in HUVECs in vitro and the aorta of rat ex vivo. Wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase (PI3K), inhibited oxytocin-induced Akt and eNOS phosphorylation. In the rat aortic rings, oxytocin induced a biphasic vascular reactivity: oxytocin at low dose (10-9-10-8 M) initiated a vasorelaxation followed by a vasoconstriction at high dose (10-7 M). L-NAME (a nitric oxide synthase inhibitor), endothelium removal or wortmannin abolished oxytocin-induced vasorelaxation, and slightly enhanced oxytocin-induced vasoconstriction. Atosiban, an oxytocin/vasopressin 1a receptor inhibitor, totally blocked oxytocin-induced relaxation and vasoconstriction. PD98059 (ERK1/2 inhibitor) partially inhibited oxytocin-induced vasoconstriction. Oxytocin also increased aortic phospho-ERK1/2 expression, which was reduced by either atosiban or PD98059, suggesting that oxytocin-induced vasoconstriction was partially mediated by oxytocin/V1aR activation of ERK1/2. The present study demonstrates that oxytocin can activate different signaling pathways to cause vasorelaxation or vasoconstriction. Oxytocin stimulation of PI3K/eNOS-derived nitric oxide may participate in maintenance of cardiovascular homeostasis, and different vascular reactivities to low or high dose of oxytocin suggest that oxytocin may have different regulatory effects on vascular tone under physiological or pathophysiological conditions.

Development of a Novel Immunochromatographic Assay for Rapid Detection of OXA-23 β-lactamase-producing Acinetobacter baumannii

  • Ji, Gil Young;Song, Hyung Geun;Jo, Mi Young;Hong, Seung Bok;Shin, Kyeong Seob
    • Biomedical Science Letters
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    • v.22 no.2
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    • pp.29-36
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    • 2016
  • Among the several agents causing carbapenem resistance of Acinetobacter baumannii, the most common cause is OXA-23 ${\beta}$-lactamase, which is known to hydrolyze carbapenem. To effectively control dissemination of carbapenem-resistant Acinetobacter baumannii (CRAB), development of both rapid and easy-to-use detection methods are required. The aim of this study is to develop a novel immunochromatographic assay (ICA) for rapid detection of OXA-23 ${\beta}$-lactamase. Of the seven monoclonal antibodies (mAbs) screened by ELISA, four mAbs (4G6, 4H6, 6G4, 9A4) exhibited high reactivity. Of these four specific antibodies, the combination of 6G4/4G6 showed the greatest reactivity and this combination of mAbs (6G4/4G6 mAbs) was used to develop the OXA-23 ${\beta}$-lactamase ICA. Of 102 A. baumannii isolates tested, the OXA-23 ${\beta}$-lactamase ICA results were consistent with PCR analysis except one false positive and one false negative isolate. The overall sensitivity and specificity were 98.36% and 97.56%, respectively. In conclusion, to the best of our knowledge, we have developed the first specific antibody set to detect OXA-23 ${\beta}$-lactamase using an ICA kit. This novel ICA can be used as a reliable and easy-to-use immunological assay for detection of OXA-23 ${\beta}$-lactamase producing CRAB in clinical laboratories.

RADIOLOGICAL CHARACTERISTICS OF DECOMMISSIONING WASTE FROM A CANDU REACTOR

  • Cho, Dong-Keun;Choi, Heui-Joo;Ahmed, Rizwan;Heo, Gyun-Young
    • Nuclear Engineering and Technology
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    • v.43 no.6
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    • pp.583-592
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    • 2011
  • The radiological characteristics for waste classification were assessed for neutron-activated decommissioning wastes from a CANDU reactor. The MCNP/ORIGEN2 code system was used for the source term analysis. The neutron flux and activation cross-section library for each structural component generated by MCNP simulation were used in the radionuclide buildup calculation in ORIGEN2. The specific activities of the relevant radionuclides in the activated metal waste were compared with the specified limits of the specific activities listed in the Korean standard and 10 CFR 61. The time-average full-core model of Wolsong Unit 1 was used as the neutron source for activation of in-core and ex-core structural components. The approximated levels of the neutron flux and cross-section, irradiated fuel composition, and a geometry simplification revealing good reliability in a previous study were used in the source term calculation as well. The results revealed the radioactivity, decay heat, hazard index, mass, and solid volume for the activated decommissioning waste to be $1.04{\times}10^{16}$ Bq, $2.09{\times}10^3$ W, $5.31{\times}10^{14}\;m^3$-water, $4.69{\times}10^5$ kg, and $7.38{\times}10^1\;m^3$, respectively. According to both Korean and US standards, the activated waste of the pressure tubes, calandria tubes, reactivity devices, and reactivity device supporters was greater than Class C, which should be disposed of in a deep geological disposal repository, whereas the side structural components were classified as low- and intermediate-level waste, which can be disposed of in a land disposal repository. Finally, this study confirmed that, regardless of the cooling time of the waste, 15% of the decommissioning waste cannot be disposed of in a land disposal repository. It is expected that the source terms and waste classification evaluated through this study can be widely used to establish a decommissioning/disposal strategy and fuel cycle analysis for CANDU reactors.

Quantum Mechanical Simulation for the Analysis, Optimization and Accelerated Development of Precursors and Processes for Atomic Layer Deposition (ALD)

  • Mustard, Thomas Jeffrey Lomax;Kwak, Hyunwook Shaun;Goldberg, Alexander;Gavartin, Jacob;Morisato, Tsuguo;Yoshidome, Daisuke;Halls, Mathew David
    • Journal of the Korean Ceramic Society
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    • v.53 no.3
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    • pp.317-324
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    • 2016
  • Continued miniaturization and increasingly exact requirements for thin film deposition in the semiconductor industry is driving the search for new effective, efficient, selective precursors and processes. The requirements of defect-free, conformal films, and precise thickness control have focused attention on atomic layer deposition (ALD). ALD precursors so far have been developed through a trial-and-error experimental approach, leveraging the expertise and tribal knowledge of individual research groups. Precursors can show significant variation in performance, depending on specific choice of co-reactant, deposition stage, and processing conditions. The chemical design space for reactive thin film precursors is enormous and there is urgent need for the development of computational approaches to help identify new ligand-metal architectures and functional co-reactants that deliver the required surface activity for next-generation thin-film deposition processes. In this paper we discuss quantum mechanical simulation (e.g. density functional theory, DFT) applied to ALD precursor reactivity and state-of-the-art automated screening approaches to assist experimental efforts leading toward optimized precursors for next-generation ALD processes.

Development of Immunological Methods for Analysis of 5' -deoxy-5' -methylthioadenosine

  • Lee, Sung-Ho;Cho, Young-Dong
    • BMB Reports
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    • v.30 no.6
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    • pp.403-409
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    • 1997
  • Studies were undertaken to develop a competitive radioimmunoassay (RIA) and indirect antigen capture enzyme-linked immunosorbent assay (ELISA) for the determination of 5'-deoxy-5'-methylthioadenosine (MTA), which is formed from decarboxylated S-adenosylmethionine by spermidine and spermine synthase. Specific antiserum against MTA was raised in rabbits by immunization with MTA-BSA which was prepared by coupling BSA to oxidized MTA with periodate. Since MTA is oxidized easily to the sulfoxide, the sulfhydryl reagent, DTT. was added to the immunogen. For RIA, immunocomplexes were separated from free MTA by using ammonium sulfate precipitation. The antiserum showed almost no cross-reactivity with a variety of other nucleotides and riboses. But, the level of cross-reactivity of 5'-isobutylthioadenosine (SIBA) was high. These results showed the importance of hydrophobicity adjacent to the 5'-OH for determining antigenicity. The lower limit of detection by this assay was 100 fmol of MTA per tube. Using this assay. MTA levels were more easily and precisely determined in biological samples when compared with HPLC analysis. The RIA procedure is less time consuming. More than 24 analyses can be carried out in 2 h and required only a very small amount of sample ($20{\mu}l$ serum). In ELISA, biotin conjugated MTA-BSA was used as the labelled MTA. The sensitivity limit of this assay was lower than 100 pmol.

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Passive Immunization Approach to Reduce Body Fat in Pigs Using Fat-specific Polyclonal Antiserum

  • Baek, Kyung Hoon;Choi, Chang Weon;Choi, Chang Bon;Lee, Byong Seak
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.10
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    • pp.1594-1599
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    • 2007
  • Plasma membrane proteins from pig adipocytes, brain, heart, kidney, liver and spleen were isolated using a 32% sucrose gradient. An adult male sheep was immunized three times at 3-wk intervals with the purified pig adipocyte plasma membrane (APM) proteins. Blood samples were taken from the immunized sheep 12 d after the third immunization. Antiserum showed strong reactivity with APM proteins determined by ELISA, and the reactivity could be detected at dilutions in excess of 1:128,000. Antiserum showed very low binding affinity with proteins isolated from brain, heart, kidney, liver or spleen. Ninety weanling pigs were allocated randomly to three treatment groups and were injected i.p. with 40 ml of antiserum (n = 30) or 20 ml of lyophilized antiserum (21.5 mg/ml; n = 30). A control group (n = 30) received 40 ml of saline, and all pigs were slaughtered at 24 wk of age. The polyclonal antiserum did not change BW or ADG. Carcass percentage of pigs was numerically increased by the antiserum treatment compared with control. Both antiserum treatments did not significantly (p>0.05) affect body composition, including body fat content, relative to the control group.