In order to screen interactor(s) of the Aspergillus nidulans ${\alpha}$-COP of COPI vesicle, we performed the yeast two hybrid screening by using the gene for A. nidulans ${\alpha}$-COP as a bait and identified ${\varepsilon}$-COP of the COPI vesicle as an interacting protein. The A. nidualns gene for the ${\varepsilon}$-COP was designated $aneA^+$ ($\underline{A.}$$\underline{n}$idulans $\underline{e}$psi-lone-COP), which encoded 296 amino acid residues with high level of identity with orthologs from other fungi. Domain analyses with yeast two-hybrid system suggested that the interaction between ${\alpha}$-COP and ${\varepsilon}$-COP relied on the C-terminus of both proteins, and that the N-terminal WD domian of ${\alpha}$-COP and the TPR region of ${\varepsilon}$-COP were not essential but required for the enhancement of the interaction. These results indicate that the interaction mode between ${\alpha}$-COP and ${\varepsilon}$-COP of COPI vesicle is evolutionarily well conserved in eukaryotes.
Proceedings of the Korean Society of Toxicology Conference
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2006.11a
/
pp.65-74
/
2006
Modem drug discovery requires rapid pharmacokinetic evaluation of chemically diverse compounds for early candidate selection. This demands the development of analytical methods that offer high-throughput of samples. Naturally, liquid chromatography / tandem mass spectrometry (LC-MS/MS) is choice of the analytical method because of its superior sensitivity and selectivity. As a result of the short analysis time(typically 3-5min) by LC-MS/MS, sample preparation has become the rate- determining step in the whole analytical cycle. Consequently tremendous efforts are being made to speed up and automate this step. In a typical automated 96-well SPE(solid-phase extraction) procedure, plasma samples are transferred to the 96-well SPE plate, internal standard and aqueous buffer solutions are added and then vacuum is applied using the robotic liquid handling system. It takes only 20-90 min to process 96 samples by automated SPE and the analyst is physically occupied for only approximately 10 min. Recently, the ultra-high flow rate liquid chromatography (turbulent-flow chromatography)has sparked a huge interest for rapid and direct quantitation of drugs in plasma. There is no sample preparation except for sample aliquotting, internal standard addition and centrifugation. This type of analysis is achieved by using a small diameter column with a large particle size(30-5O ${\mu}$m) and a high flow rate, typically between 3-5 ml/min. Silica-based monolithic HPLC columns contain a novel chromatographic support in which the traditional particulate packing has been replaced with a single, continuous network (monolith) of pcrous silica. The main advantage of such a network is decreased backpressure due to macropores (2 ${\mu}$m) throughout the network. This allows high flow rates, and hence fast analyses that are unattainable with traditional particulate columns. The reduction of particle diameter in HPLC results in increased column efficiency. use of small particles (<2 urn), however, requires p.essu.es beyond the traditional 6,000 psi of conventional pumping devices. Instrumental development in recent years has resulted in pumping devices capable of handling the requirements of columns packed with small particles. The staggered parallel HPLC system consists of four fully independent binary HPLC pumps, a modified auto sampler, and a series of switching and selector valves all controlled by a single computer program. The system improves sample throughput without sacrificing chromatographic separation or data quality. Sample throughput can be increased nearly four-fold without requiring significant changes in current analytical procedures. The process of Bioanalytical Method Validation is required by the FDA to assess and verify the performance of a chronlatographic method prior to its application in sample analysis. The validation should address the selectivity, linearity, accuracy, precision and stability of the method. This presentation will provide all overview of the work required to accomplish a full validation and show how a chromatographic method is suitable for toxirokinetic sample analysis. A liquid chromatography/tandem mass spectrometry (LC-MS/MS) method developed to quantitate drug levels in dog plasma will be used as an example of tile process.
Sheikh, M. Mominul Islam;Kim, Chul-Hwan;Park, Hyun-Jin;Kim, Sung-Ho;Kim, Gyeong-Chul;Lee, Ji-Yong;Kim, Jae-Won
Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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2011.10a
/
pp.167-177
/
2011
Renewable energy resources and technologies have the potential to provide long-lasting solutions of the global energy-requirements faced by the economic and environmental sectors of a nation. Therefore, waste money bills were used as renewable energy source for the production of bio-ethanol. In this study, different concentrated NaOH 0.5%. 1.0%, 2.0%, 3.0% and 0.0% (as a control) were used for 10, 20 and 30 mins at $121^{\circ}C$/15 psi in an autoclave. Saccharification and fermentation (aerobic and anaerobic) were carried out through commercial enzyme Celluclast 1.5 L, Novozymes 188 and Saccharomyces cerevisiae KCCM 11304 respectively. The results of pretreatment showed that the NaOH pre-treated substrate enhanced enzyme action and released more amount of glucose. The amount of glucose was found with the increasing concentration of NaOH and time $44996.95{\pm}6.30$, $46763.10{\pm}3.56$, $53421.32{\pm}4.72$, $63431.25{\pm}6.95$ and $56850.98{\pm}6.75\;ng/{\mu}l$ for 30 min respectively. As for bioethanol, the conversion rate of NaOH resulted $1010.08{\pm}4.71$, $1050.25{\pm}4.37$, $1109.49{\pm}4.39$, $1139.25{\pm}3.26$ and $1020.77{\pm}3.89$ ppm for aerobic; $16730.54{\pm}6.67$, $17076.45{\pm}6.25$, $17516.17{\pm}4.49$, $19782.68{\pm}6.19$ and $17973.39{\pm}7.50$ ppm for anaerobic and $18935.02{\pm}4.59$, $19895.45{\pm}5.39$, $21912.95{\pm}4.83$, $24895.21{\pm}6.72$ and $18961.21{\pm}4.90$ ppm for anaerobic condition with benzoic acid for respective condition. Thus, the results of the present work clearly revealed that with the increasing of alkali concentration might be more effective for bio-ethanol production from waste money bill, which is economic and environmental friendly.
Park, Hong-Hyun;Kim, Sung-Kon;Pyun, Yu-Ryang;Lee, Shin-Young
Korean Journal of Food Science and Technology
/
v.21
no.6
/
pp.815-819
/
1989
Rheological properties of chestnut starch suspensions (3 and 4%, db) and gelatinized starch (4%, db) were investigated with a capillary and rotational viscometer, respectively. Starch suspensions had no yield stress and showed dilatant flow behavior in the temperature ranges of $30-65^{\circ}C$. However, starch suspension showed pseudoplastic flow behavior at $70^{\circ}C\; and\;above\; 65^{\circ}C$ for 3 and 4% concentration, respectively Flow activation energy below $50^{\circ}C$ was 0.56 kcal/mole but increased to 51.9-80.8 kcal/mole at $60-70^{\circ}C$. The behavior of gelatinized starch (4%) was pseudoplastic regardless of heating temperature $(65-80^{\circ}C)$ and time (15-60 min). The apparent viscosity of the starch remained constant after heating at $80^{\circ}C$ for 45 min. The swelling power and log apparent viscosity showed similar pattern. The activation energy of the apparent viscosity of the geletinized starch at $70-80^{\circ}C$ was 13.09kcal/mole. The apparent viscosity of thermal-gelatinized $(90^{\circ}C)$ starch was lower than that of 15 psi-gelatinized starch.
Journal of the Korean Society for Aeronautical & Space Sciences
/
v.43
no.9
/
pp.822-829
/
2015
In this paper, the piezoelectric-hydraulic pump with a piezostack actuator as a driving source has been designed, fabricated, and evaluated for its application to UAV's brake system. The performance requirements of the piezoelectric-hydraulic pump were decided based on the requirements analysis of the target aircraft brake system. The geometric design of the piezoelectric-hydraulic pump to meet the performance requirements of the pump was conducted, and all components of the pump including the spring sheet type check valves were machined with close tolerance. By constructing a test apparatus for the performance check of the piezoelectric-hydraulic pump, the performance characteristics of the pump, such as the outlet flow rate for load-free condition and the outlet oil pressure for closed loop condition, have been evaluated. It has been found by the performance test result that the developed piezoelectric-hydraulic pump satisfies the design requirements effectively.
The experiment was designed to compare the char combustion kinetics of pulverized Indonesia coals commonly utilized in Korea power plants. The reaction rate of coal char has been formulated using the external and internal effectiveness factors to describe the diffusion effect quantitatively. The Random Pore Model (RPM) was used for applying internal specific surface area as a function of carbon conversion ratio. Reaction rate was obtained from reaction time using the Wire Heating Reactor (WHR) which can heat and measure the char particle temperature at the same time. BET and TGA were used to obtain physical properties such as internal specific surface area and structural parameter. Three kinds of Indonesia Sub-bituminous coals "BARAMULTI, ENERGYMAN, AGM" were used in order to derive the activation energy and pre-exponential factor. The results of this study showed that the effect of internal diffusion than that of external diffusion is the dominant as comparison of kinetics was reflected in external and internal effectiveness factors. For three kinds of coal char, finally, activation energy of intrinsic kinetics indicates 110~118 kJ/mol.
Fasanmade, Adedigbo A.;Owuor, Edward D.;Ee, Rachel P.L.;Qato, Dima;Heller, Mark;Kong, Ah Ng Tony
Archives of Pharmacal Research
/
v.24
no.2
/
pp.126-135
/
2001
Quinacrine (QU), a phospholipase-A2 (PLA-2) inhibitor has been used clinically as a chemotherapeutic adjuvant. To understand the mechanisms leading to its chemotherapeutic effect, we have investigated QU-induced apoptotic signaling pathways in human cervical squamous carcinoma HeLa cells. In this study, we found that QU induced cytochrome c-dependent apoptotic signaling. The release of pro-apoptotic cytochrome c was QU concentration- and time-dependent, and preceded activation of caspase-9 and -3. Flow cytometric FACScan analysis using fluorescence intensities of $DiOC_6$/ demonstrated that QU-induced cytochrome c release was independent of mitochondrial permeability transition (MPT), since the concentrations of QU that induced cytochrome c release did not alter mitochondrial membrane potential (${\blacktriangle}{\Psi}_m$). Moreover, kinetic analysis of caspase activities showed that cytochrome c release led to the activation of caspase-9 and downstream death effector caspase-3, Caspase-3 inhibitor (Ac-DEVD-CHO) partially blocked QU-induced apoptosis, suggesting the importance of caspase-3 in this apoptotic signaling mechanism. Supplementation with arachidonic acid (AA) sustained caspase-3 activation induced by QU. Using inhibitors against cellular arachidonate metabolism of lipooxygenase (Nordihydroxyguaiaretic Acid, NDGA) and cyclooxygenase (5,8,11,14-Eicosatetraynoic Acid, ETYA) demonstrated that QU-induced apoptotic signaling may be dependent on its role as a PLA-2 inhibitor. Interestingly, NDCA attenuated QU-induced cytochrome c release, caspase activity as well as apoptotic cell death. The blockade of cytochrome c release by NDCA was much more effective than that attained with cyclosporin A (CsA), a MPT inhibitor. ETYA was not effective in blocking cytochrome c release, except under very high concentrations. Caspase inhibitor z-VAD blocked the release of cytochrome c suggesting that this signaling event is caspase dependent, and caspase-8 activation may be upstream of the mitochondrial events. In summary, we report that QU induced cytochrome c-dependent apoptotic signaling cascade, which may be dependent on its role as a PLA-2 inhibitor. This apoptotic mechanism induced by QU may contribute to its known chemotherapeutic effects.
Purpose : This study was conducted to investigate the extent to which the professional self concept between the psychiatric mental health nurse practitioner (PMHNP) and general nurse in psychiatric ward was comparable. The results were expected to provide basic data for developing the construct of professional self concept and making awareness of the importance of the program increasing professional self concept. Method : The subjects of this study were 227 PMHNP and 436 general nurse in psychiatric ward. The instruments used for this study were PSCNI by Arthur (1990), PSI by Heppner and Peterson(1982) and the index of work satisfaction by Slavitt et al.(1978). With the aid of the SAS, t-test. two-way ANOVA and stepwise multiple regression were conducted. Result : 1. The average item score of PSCNI of PMHNP was $2.82\pm0.27$. and that of general nurse was $2.66\pm0.27$. Statistically significant difference between two groups was found(p=0.0000) 2. There was statistically significant difference between two groups in the score of professional practice(p=0.0000), satisfaction(p=0.0024), leadersbip(p=0.0000) , flexibility(p=0.0000) and skill (p=0.0000). 3. Statistically significant differences between the two groups were observed in terms of age(p=0.0003), marital status(p=0.0001). education(p=0.0005), religion(p=-.0144), motive (P=-.0001), length of service as a nurse(p=0.0121), the length of service in psychiatric unit(p=0.0143). However there were no significant interaction effect with group and age, marital state. education, religion, motive, length of service as a nurse, length of service in psychiatric unit. 4. Job satisfaction (JS) and problem solving inventory score(PS) were found to be the highest factor predicting the professional self concept between the PMHNP and general nurse. JS and PS accounted for $43.4\%$ in the professional self concept of PMHNP, whereas PS, JS, age and religion accounted for $53\%$ in the professional self concept of general nurse in psychiatric ward. In conclusion, this study suggested that we need to develop programs and polices to increase the professional self concept of nurse, particularly of psychiatric mental health nurse practitioners.
Kim Jin-Seog;Lee Byung-Hoi;Kwon Suk-Yoon;Kim Yun-Hee;Kim So-Hee;Cho Kwang-Yun
Journal of Plant Biotechnology
/
v.32
no.2
/
pp.97-103
/
2005
Antioxidative responses of transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts was investigated with several herbicides. In greenhouse test, tolerance of SOD/APX-overexpressed tobacco (CA) to photosystem (PS) I inhibitor paraquat was increased by about 40%. However, any response differences between CA and wild type (WT) tobacco was not observed in a treatment with PS II inhibitors (bromoxynil, diuron and bromacil), chlorophyll biosynthesis inhibitor(oxyfluorfen), carotenoid biosynthesis inhibitor (fluridone) and 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase inhibitor (glyphosate). This tendency was also similar in the growth chamber test of low light intensity, using paraquat and diuron. That is, increased antioxidant activity of CA was shown only in paraquat treatment. When paraquat was foliar-treated to 6 to 9-leaf stage plant, the third to fourth placed leaf from shoot tip showed relatively higher antioxidant activity. Ascorbate supplemented to paraquat solution alleviated the phytotoxicity with a similar range in both CA and WT. In conclusion, CA specifically responded to oxidative stress induced by paraquat among tested herbicides in a whole plant assay.
Journal of the Korean Society of Food Science and Nutrition
/
v.40
no.5
/
pp.649-659
/
2011
To examine the anti-cancer effects of Lepidium virginicum L., the anti-proliferative and pro-apoptotic effects of a water extract of L. virginicum leaves (WELVL) and of L. virginicum roots (WELVR) were investigated in HCT116 human colon carcinoma cells. The treatment of HCT116 cells with WELVL and WELVR resulted in the inhibition of growth and morphological changes in a concentration-dependent manner by inducing apoptosis. The growth inhibition and apoptosis induction by WELVR was stronger than that of WELVL thus, we determined that WELVR was the more optimal extract for this study. The increased apoptotic events in HCT116 cells caused by WELVR were associated with an up-regulation of Fas ligand, Bax, and Bad expression, a down-regulation of Bcl-2, Bcl-$_XL$, and Bid expression, and a decrease in the mitochondrial membrane potential (MMP, ${\Delta}{\psi}m$). WELVR treatment induced the proteolytic activation of caspase-3, -8, and -9, and the degradation of caspase-3 substrate proteins, such as poly (ADP-ribose) polymerase (PARP), ${\beta}$-catenin, and phospholipase C-${\gamma}1$ (PLC-${\gamma}1$). In addition, apoptotic cell death induced by WELVR was correlated with a down-regulation of inhibitors of the apoptosis protein (IAP) family, such as the X-linked inhibitor of apoptosis protein (XIAP), cIAP-1, and cIAP-2. These findings suggest that the WELVR-induced inhibition of cell proliferation is associated with the induction of apoptotic cell death. WELVR may be a potential chemotherapeutic agent for the control of HCT116 human colon carcinoma cells.
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