Kim, Hak-Jun;Shin, Hee-Jae;Kim, Hyun-Woo;Kang, Sung-Ho;Kim, Young-Tae
Bulletin of the Korean Chemical Society
/
v.28
no.12
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pp.2303-2309
/
2007
Expressed protein ligation (EPL) technique, joining recombinantly expressed proteins to polypeptides, has been widely adopted for addressing various biological questions and for drug discovery. However, joining two recombinant proteins together is sometimes difficult when proteins are expressed insoluble and unrefoldable, because ligation-active proteins via intein-fusion are obtainable when they are folded correctly. We overcame this limitation coexpressing target protein with additional methionine aminopeptidase (MAP) which enhances removal of the initiation methionine of recombinantly expressed protein. Our approach demonstrated that two domains of 46 kDa 5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase, a target of herbicide glyphosate, were successfully joined by native chemical ligation, although its C-terminal domain was expressed as an inclusion body. The intein-fused N-terminal fragment of EPSP synthase (EPSPSN, residues 1-237) was expressed and the ligation-active thioester tagged N-terminal fragment (EPSPSN-thioester) was purified using a chitin affinity chromatography and mercapto-ethanesulphonate (MESNA) as intein thiolysis reagent. Its Cterminal fragment (EPSPSC, residues Met237-238CYS-427), expressed as an inclusion body, was prepared from an additional MAP-expressing strain. Protein ligation was initiated by mixing ~1 mM of EPSPSN-thioester with ~2 mM of EPSPSCCYS (residues 238CYS-427). Also we found that addition of 2% thiophenol increased the ligation efficiency via thiol exchange. The ligation efficiency was ~85%. The ligated full-length EPSP synthase was dissolved in 6 M GdHCl and refolded. Circular dichroism (CD) and enzyme activity assay of the purified protein showed that the ligated enzyme has distinct secondary structure and ~115% specific activity compared to those of wild-type EPSP synthase. This work demonstrates rare example of EPL between two recombinantly expressed proteins and also provides hands-on protein engineering protocol for large proteins.
Kim, Rae-Oh;Cho, Kyoung-Suok;Yoo, Do-Sung;Huh, Pil-Woo;Park, Chun-Kun;Kim, Dal-Soo;Kang, Jun-Ki
Journal of Korean Neurosurgical Society
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v.29
no.4
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pp.555-558
/
2000
Crowth of an angiolipoma of the central nervous system is rare. Only about 60 cases were searched in the literature. We report one case of spinal angiolipoma recently treated in our institution. A 67-year-old man was admitted because of burning sensation on the both lower extremities and paraparesis. He felt burning sensation on the both great toes and it progressively involved to thighs and inguinal area during the past 1 year which gradually worsened. MR image showed a spinal mass lesion at the level of T4 to T6. The lesion was isodense and hyperdense in periphery on T1-weighted image and hyperintense on T2-weighted image. The tumor was located on the posterior area of the spinal cord and markedly enhanced after intravenous Gd-DTPA. At the operation, a soft, dark-red mass was totally removed. On histological examination, the tumor was shown to be mainly composed of mature fatty cells and numerous blood vessels with enlarged lumens. Postoperative course was uneventful and the symptoms improved gradually.
Amit Sharma;GD Puri;Rajeev Chauhan;Ankur Luthra;Gauri Khurana;Amarjyoti Hazarika;Shyam Charan Meena
Journal of Dental Anesthesia and Pain Medicine
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v.24
no.1
/
pp.67-73
/
2024
Background: Nitrous oxide has been an integral part of surgical anesthesia for many years in the developed world and is still used in developing countries such as India. The other main concerns in low-resource countries are the lack of an advanced anesthesia gas-scavenging system and modular surgical theatres. As a greenhouse gas that has been present in the atmosphere for more than 100 years and damages the ozone layer, nitrous oxide is three times worse than sevoflurane. Here, we conducted an observational study to quantify the annual nitrous oxide consumption and its environmental impact in terms of carbon dioxide equivalence in one of busiest tertiary health care and research centers in Northern India. Methods: Data related to nitrous oxide expenditure' from the operation theatre and manifold complex of our tertiary care hospital and research center from 2018 to 2021 were collected monthly and analyzed. The outcomes were extracted from our observational study, which was approved by our institutional ethics board (INT/IEC/2017/1372 Dated 25.11.2017) and registered prospectively under the Central Registry (CTRI/2018/07/014745 Dated 05.07.2018). Results: The annual nitrous oxide consumption in our tertiary care hospital was 22,081.00, 22,904.00, 17,456.00, and 18,392.00 m3 (cubic meters) in 2018, 2019, 2020, and 2021, respectively. This indicates that the environmental impact of nitrous oxide (in terms of CO2 equivalents) from our hospital in 2018, 2019, 2020, and 2021 was 13,016.64, 13,287.82, 10,289.94, and 10,841.24 tons, respectively. Conclusion: This huge amount of nitrous oxide splurge is no longer a matter of laughter, and serious efforts should be made at every central and peripheral health center level to reduce it.
Journal of the Korean Society of Food Science and Nutrition
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v.46
no.9
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pp.1071-1080
/
2017
This study examined the adhesiveness of beneficial intestinal bacteria to whole-grains using confocal scanning laser microscopy (CLSM), to demonstrate the prebiotic effects of whole-grains, and to develop prebiotic puffed snacks with these whole-grains. CLSM has been used to observe the adhesiveness of Lactobacillus acidophilus, which belongs to beneficial intestinal bacteria, to whole-grain powders using optical sectioning techniques. The enhanced effects on the growth of beneficial intestinal bacteria with the hot water grain extract were verified using an indirect count method. Finally, a puffed snack was produced with the prebiotic effect and the quality was evaluated by checking the chromaticity and degree of hardness. As a result, L. acidophilus exhibited adhesive ability to whole-grain powders and growth of selected beneficial intestinal bacteria were improved significantly. The Hunter L value of the developed puffed snack increased when seasoning was added. The hardness of the puffed snack with seasoning was higher than that of the control. The results of a sensory evaluation showed that the puffed snack with seasoning was highly rated in the overall preference compared to the control.
Journal of Korean Society of Industrial and Systems Engineering
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v.40
no.3
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pp.1-6
/
2017
Process capability is well known in quality control literatures. Process capability refers to the uniformity of the process. Obviously, the variability in the process is a measure of the uniformity of output. It is customary to take the 6-sigma spread in the distribution of the product quality characteristic as a measure of process capability. However there is no reference of process capability when maximum material condition is applied to datum and position tolerance in GD&T (Geometric Dimensioning and Tolerancing). If there is no material condition in datum and position tolerance, process capability can be calculated as usual. If there is a material condition in a feature control frame, bonus tolerance is permissible. Bonus tolerance is an additional tolerance for a geometric control. Whenever a geometric tolerance is applied to a feature of size, and it contains an maximum material condition (or least material condition) modifier in the tolerance portion of the feature control frame, a bonus tolerance is permissible. When the maximum material condition modifier is used in the tolerance portion of the feature control frame, it means that the stated tolerance applies when the feature of size is at its maximum material condition. When actual mating size of the feature of size departs from maximum material condition (towards least material condition), an increase in the stated tolerance-equal to the amount of the departure-is permitted. This increase, or extra tolerance, is called the bonus tolerance. Another type of bonus tolerance is datum shift. Datum shift is similar to bonus tolerance. Like bonus tolerance, datum shift is an additional tolerance that is available under certain conditions. Therefore we try to propose how to calculate process capability index of position tolerance when maximum material condition is applied to datum and position tolerance.
The reactive intermediates formed during the metabolism of therapeutic agents, toxicants and carcinogens by cytochromes P450 are frequently capable of covalently binding to tissue macromolecules and causing tissue damage. It has been shown that YH439, a congener of malotilate, is effective in suppressing hepatic P450 2E1 expression. The present study was designed to further establish the mechanistic basis of YH439 protection against toxicant by assessing its effects against chemical-mediated potentiated hepatotoxicity. Retinoyl palmitate (Vit-A) pretreatment of rats for 7 days substantially enhanced carbon tetrachloride hepatotoxicity, as supported by an ${\sim}5-fold$ increase in serum alanine aminotransferase (ALT) activity, as compared to $CCl_4$ treatment alone. The elevation of ALT activity due to Vit-A was completely blocked by the treatment of $GdCl_3$ a selective inhibitor of Kupffer cell activity. Concomitant pretreatment of rats with both YH439 and Vit-A resulted in a 94% decrease in Vit-A-potentiated $CCl_4$ hepatotoxicity. YH439 was also effective against propyl sulfide-potentiated $CCl_4-induced$ hepatotoxicity. Whereas propyl sulfide (50 mg/kg, 7d) enhanced $CCl_4-induced$ hepatotoxicity by >5-fold, relative to $CCl_4$ treatment alone, concomitant treatment of animals with both propyl sulfide and YH439 at the doses of 100 and 200 mg/kg prevented propyl sulfide-potentiated $CCl_4$ hepatotoxicity by 35% and 90%, respectively. Allyl sulfide, a suppressant of hepatic P450 2E1 expression, completely blocked the propyl sulfide-enhanced hepatotoxicity, indicating that propyl sulfide potentiation of $CCl_4$ hepatotoxicity was highly associated with the expression of P450 2E1 and that YH439 blocked the propyl sulfide-enhanced hepatotoxicity through modulation of P450 2E1 levels. Propyl sulfide- and $CCl_4-induced$ stimulation of lipid peroxidation was also suppressed by YH439 in a dose-related manner, as supported by decreases in malonedialdehyde production. The role of P450 2E1 induction in the potentiation of $CCl_4$ toxicity and the effects of YH439 were further evaluated using pyridine as a P450 2E1 inducer. Pyridine pretreatment substantially enhanced the $CCl_4$ hepatotoicity by 23-fold, relative to $CCl_4$ alone. YH439, however, failed to reduce the pyridine-potentiated toxicity, suggesting that the other form(s) of cytochroms P450 inducible by pyridine, but not suppressible by YH439 treatment, may play a role in potentiating $CCl_4-induced$ hepatotoxicity. YH439 was capable of blocking cadmium chloride-induced liver toxicity in mice. These results demonstrated that YH439 efficiently blocks Vit-A-enhanced hepatotoxiciy through Kupffer cell inactivation and that the suppression of P450 2E1 expression by YH439 is highly associated with blocking of propyl sulfide-mediated hepatotoxicity.
Jeong, Young Ae;Jang, Dong Cheol;Kwon, Jin Kyung;Kim, Dae Hyun;Choi, Eun Young
Journal of Bio-Environment Control
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v.31
no.3
/
pp.221-229
/
2022
This study was aimed to investigate spatial and vertical characteristics of greenhouse environments according to the location of the environmental sensors, and to investigate the correlations between temperature, light intensity, and carbon dioxide (CO2) concentration according to the type of greenhouse. Temperature, relative humidity (RH), CO2, and light sensors were installed in the four-different vertical positions of the whole canopy as well as ground and roof space at the five spatial locations of the Venlo greenhouse. Also, correlations between temperature, light intensity, and CO2 concentration in Venlo and semi-closed greenhouses were analyzed using the Curve Expert Professional program. The deviations among the spatial locations were larger in the CO2 concentration than other environmental factors in the Venlo greenhouse. The average CO2 concentration ranged from 465 to 761 µmol·mol-1 with the highest value (646 µmol·mol-1) at the Middle End (4ME) close to the main pipe (50Ø) of the liquefied CO2 gas supply and lowest (436 µmol·mol-1) at the Left Middle (5LM). The deviation among the vertical positions was greater in temperature and relative humidity than other environments. The time zone with the largest deviation in average temperature was 2 p.m. with the highest temperature (26.51℃) at the Upper Air (UA) and the lowest temperature (25.62℃) at the Lower Canopy (LC). The time zone with the largest deviation in average RH was 1 p.m. with the highest RH (76.90%) at the LC and the lowest RH (71.74%) at the UA. The highest average CO2 concentration at each hour was Roof Air (RF) and Ground (GD). The coefficient of correlations between temperature, light intensity, and CO2 concentration were 0.07 for semi-closed greenhouse and 0.66 for Venlo greenhouse. All the results indicate that while the CO2 concentration in the greenhouse needs to be analyzed in the spatial locations, temperature and humidity needs to be analyzed in the vertical positions of canopy. The target CO2 fertilization concentration for the semi-closed greenhouse with low ventilation rate should be different from that of general greenhouses.
Kim, Myung Dong;Wahid, Muhamad FR;Raju, Kati;Kim, Seyoung;Yu, Ji Haeng;Park, Chun Dong;Yoon, Dang-Hyok
Journal of the Korean Ceramic Society
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v.55
no.5
/
pp.492-497
/
2018
This paper reports the efficacy of tape casting using an Ag-10 wt% CuO filler for the successful joining of a sintered $Ce_{0.9}Gd_{0.1}O_{2-{\delta}}-La_{0.7}Sr_{0.3}MnO_{3{\pm}{\delta}}$ (GDC-LSM) ceramic with a SUS 460 FC metal alloy by reactive air brazing. The as-prepared green tape was highly flexible without drying cracks, and the handling was easy when used as a filler material for reactive air brazing. Heat treatment for the GDC-LSM/SUS 460 FC joint was performed at $1050^{\circ}C$ for 30 min in air. Microstructural observations indicated a reliable and compact joining. The room temperature mechanical shear strength of the as-brazed joints was $60{\pm}8MPa$ with a cohesive failure. The flexural strength of joints was measured from room temperature up to $850^{\circ}C$, where the strength retention revealed to be almost 100% at $500^{\circ}C$. However, the joints showed a degradation in strengths at 800 and $850^{\circ}C$, exhibiting strength retentions of 57% and 37%, respectively.
The analytical performance of glow discharge mass spectrometer (GD-MS), using a flat type ion source is discussed. The efficiency of ion extraction was maximized at the distance between anode and cathode of 6 mm. At the operation condition of 4 mA, -1000 volt, and 1 mbar for the source, the optimum voltages for sampler and skimmer were40 volt and -280 volt, respectively. The intensities of Cu, Zn, and Mn were increased as a function of square root of current approximately. Korea standard reference materials (KSRM) were tested for an application study. The detection limits of most elements were obtained in the range of several ppm at the optimized operating condition. The peaks of aluminum and chromium were interfered by those of residual gases. The depth profile of nickel coated copper specimens (3, 5, 10 ${\mu}m$ thickness) were obtained by plotting time versus intensities of Ni and Cr after checking the thickness of nickel coated using a scanning electron microscope (SEM). At this moment, the sputtering rate of 0.2 ${\mu}m/min$ at the optimum operating condition was determined from the slope of the plot of time to the coating thickness. The roughness spectra of specimen's crater after 16 min, discharge were obtained using a Talysuf5m-120 roughness tester as well.
For the precise measurements of special nuclear materials (SNM) including Pu and Am isotopes, we have used phoswich detector combination of two single crystal scintillators of Gd3Ga3Al2O12:Ce and CsI:Tl. High detection efficiency and sensitivity along with high figure of merit for the discrimination of these phoswich detectors ensures the detection and discrimination of thermal neutrons and gammas from spontaneous fission of Pu and other isotopes in presence of high gamma background. Using this detector, the low energy gammas, which is stopped completely in 1mm thick disc of GGAG, can be also discriminated from high energies gamma and shows linearity in wide range of sample quantities. By changing only the appropriate shielding, the similar setup was used for thermal neutron detection and shows a very good linearity over wide range. The quantity of a test sample was also calculated accurately by using the measured calibrated plot.
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