• Title/Summary/Keyword: Diluted factor

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Characterization of Aspergillus niger Mutants Deficient of a Protease

  • Chung, Hea-Jong;Park, Seung-Moon;Kim, Dae-Hyuk
    • Mycobiology
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    • v.30 no.3
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    • pp.160-165
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    • 2002
  • Aspergillus niger has been used as a host to express many heterologous proteins. It has been known that the presence of an- abundant protease is a limiting factor to express a heterologous protein. The protease deficient mutant of A. niger was obtained using UV-irradiation. A total of $1{\times}10^5$ spores were irradiated with $10{\sim}20%$ survival dose of UV, 600 $J/m^2$ at 280 nm, and the resulting spores were screened on the casein-gelatin plates. Ten putative protease deficient mutants showing the reduced halo area around colonies were further analyzed to differentiate the protease deficient mutant from other mutant types. Among ten putative mutants, seven mutants showed significant growth defect on nutrient rich medium and two mutants appeared to be the secretory mutants, which resulted in the impaired secretion of extracellular proteins including proteases. A mutant $pro^--20$ showed reduced halo zone without any notable changes in growth rate. In addition, the starchdegrading and glucose oxidase activities in the culture filtrate of $pro^--20$ mutant showed the similar range as that of the parental strain, which suggested that the $pro^--20$ mutant ought to be the protease deficient mutant rather than a secretory mutant. The reduced proteolytic activity of the $pro^--20$ was demonstrated using SDS-fibrin zymography gel. The reduced extracellular proteolysis was quantified by casein degradation assay and, comparing with the parental strain, less than 30% residual extracellular protease activity was detected in the culture filtrate of the $pro^--20$ mutant. The bio-activity of an exogenously supplemented hGM-CSF(human Granulocyte-Macrophage Colony Stimulating Factor) in the culture filtrate of $pro^--20$ mutant was detected until eight times more diluted preparations than that of the parental strain.

Uncertainty evaluation for the determination of creatinine in urine by LC-MS/MS (LC-MS/MS를 이용한 소변 중 크레아티닌 분석의 측정불확도 평가)

  • Kim, Jin-Young;Kwon, Woon-Yong;Suh, Sung-Ill;In, Moon-Kyo
    • Analytical Science and Technology
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    • v.25 no.1
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    • pp.83-90
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    • 2012
  • The objective of the study was to estimate the measurement uncertainty associated with determination of creatinine (Cr) in urine samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Centrifuged urine samples (10 ${\mu}L$) were diluted with 390 ${\mu}L$ of distilled water. To 20 ${\mu}L$ aliquots of diluted urine samples, 30 ${\mu}L$ of internal standard solution (Cr-$d_3$, 5 ${\mu}g/mL$) and 10 ${\mu}L$ of acetonitrile were added and filtered. The samples (1 ${\mu}L$) were introduced into LC-MS/MS with no further pretreatment. Cr was separated on a multi-mode ODS column (Scherzo SM-C18, 75 ${\times}$ 2.0 mm I.D., 3 ${\mu}m$) and quantified by LC-MS/MS operating in MRM mode (Cr, m/z 114.0${\rightarrow}$ 86.0; Cr-$d_3$, m/z 117.0${\rightarrow}$ 89.1). The four factors that contribute uncertainty to the final result were extracted and evaluated. The principal factors of contribution to combined standard uncertainty were sample dilution, calibration curve and repeatability, while the preparation of standard solution was only a minor factor. Relative extended uncertainty of the measured concentration was 14.2% in a real urine sample.

Pharmacokinetics of Anticancer Agent SB-31 in Rats & Rabbits and the Cardiovascular Effect on the Isolated Perfused Rat Heart & Blood Coagulation (SB-31의 Glycyrrhizin을 지표로 한 Rat과 Rabbit에서의 약물동태 및 심혈관계에 대한 효과 연구)

  • Kang, Won Ku;Park, Yong Soon;Lee, Dong Heum;Kwon, Kwang Il
    • Korean Journal of Clinical Pharmacy
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    • v.8 no.2
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    • pp.122-132
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    • 1998
  • SB-31 which contains Pursatilla, Licoris and Ginseng extracts was recently proved as an anticancer agent. In a preclinical effort to be applied this drug to human, pharmacokinetics of SB-31 was carried out in rats and rabbits. Glycyrrhizin(GZ), a saponin of Licoris was used as a standard ingradient for the pharmacokinetics of SB-31. The rat's blood, bile and urine samples were serially collected in femoral vein, common bile duct and bladder, respectively, after bolus i.v. injection at a dose of 1 or 1/5 ampul/rat and rabbit's blood samples from the marginal ear vein at a dose of 1 or 3 amp./rabbit. GZ and glycyrrhetic acid(GA), a major metabolite of GZ in the physiological samples were analysed by HPLC with UV detection. The decline of GZ in plasma concentration was generally biexponential at each dose. GZ was almost completely recovered in bile within 18 hour. GA wasn't detected in the samples with UV detector. In the rat, Vss and Kel at a dose of 1 and 1/5 ampul of SB-31 were $98.06\pm6.07\;ml,\;0.33\pm0.05\;hr^{-1}\;and\;65.46\pm11.19\;ml,\;0.68\pm0.25\;hr^{-1}$, respectively. Those in rabbits at a dose of 3 and 1 ampul of SB-31 were $235.24\pm30.72\;ml,\;0.13\pm0.36\;hr^{-1}\;and\;341.32\pm28.58\;ml,\;0.27\pm0.04\;hr^{-1}$, respectively. 'WinNonlin' was utilized for the compartmental analysis. A two-compartment model was chosen as the most appropriate pbarmaco-kinetic model. The data were best described by using a weighting factor of $1/y^2$. To evaluate the effect of SB-31 on cardiovascular system, serially diluted SB-31 was directly injected into coronary artery in the isolated perfused rat heart and the effect of PSF, PSH, saponins of Pursatilla, and SB-31 on PT, APTT of healthy human plasma was examined. Except the positive inotropic effect of ten times diluted solution of SB-31, there was no significant effect on LVDP, (- dp/dt)/(+dp/dt), heart rate and coronary flow in comparision with that of vehicle. SB-31 had no effect on PT but slightly delayed APTT about $6.9{\sim}11.5\%$. There was no significant effect of PSF and PSH on PT & APTT. Conclusively, SB-31 did not show any notable toxic effects on cardiovascular system.

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A study on the development of SRM for XRF analysis of PZT[$Pb(ZrTi)O_3$] (PZT[$Pb(ZrTi)O_3$]의 XRF 분석용 SRM 개발에 관한 연구)

  • Kim, Young Man;Jeong, Chan Yee;Lim, Chang Ho;Choi, Beom Suk
    • Analytical Science and Technology
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    • v.10 no.6
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    • pp.439-444
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    • 1997
  • Twelve kinds (1set) standard materials of chemical ingredients of lead zirconate titanate[$Pb(ZrTi)O_3$] have been developed in order to determine fast and accurate measurement of X-ray fluorescence spectrometry. Especially, we used diluted(ahout sixteen times) filling compound($Li_2B_4O_7/LiBO_2=4/1$) to consider removal effect of matrix, storage convenience, and homogenous characteristics. As a result from the four different laboratories, we obtained extremly good agreement about the standard curve on twelve standard materials which containing eleven elements, PbO, $ZrO_2$, $TiO_2$, SrO, $WO_3$, $La_2O_3$, $Cr_2O_3$, MgO, $Nb_2O_5$, and $MnO_2$. The correlation factor of standard curve was over 0.998. However, ZnO has relatively low correlation factor, 0.977, because the concentration was 10ppm lower than other original materials. This analysis reveals that ZnO has shown the poor linearity as well as low fluorescence intensity. In present work, XRF standard materials are useful for determining a rapid and accurate results for major and minor elements concentration among PZT.

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Trophic Transfer of Disinfectant Chemical in Aquatic Ecosystem (소독살균제 기인 화학물질의 수생태계 내 생물 전이 특성 연구)

  • Ha-Eun Cho;Dokyun Kim;Eun-Ji Won;Hyeong-Wook Jo;Kwang-Guk An;Kyung-Hoon Shin
    • Korean Journal of Ecology and Environment
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    • v.55 no.4
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    • pp.376-389
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    • 2022
  • Despite the consumption of disinfectants have been increased by COVID-19 pandemic, the fate of the chemicals in aquatic food webs are still unclear. In order to understand the trophic transfer of the chemicals, the concentration of disinfectants including six benzalkonium chloride (BACs) and five didecyldimethylammonium chlorides(DDACs) were measured at the Geum (2020), Han (2021), and Yeongsan River (2021), before and after rainfall. The highest concentration of ∑BACs (mainly C12 and C14) and ∑DDACs (mainly C10 and C14) were observed in the Han River, followed by Yeongsan River, Geum River Estuary, and Gapcheon. After rainfalls, both concentration and detection frequency were decreased in all sites. Although the BAC and DDAC seems to be accumulated in organisms, they were bio-diluted rather than magnified in the aquatic food web with the biomagnification factor(BMF) of less than 1, trophic magnification slope (TMS) from -0.236 to 0.001, and trophic magnification factor(TMF) from 0.85 to 1.01.

Preconcentration of Ultra Trace Amounts Bismuth in Water Samples Using Cloud Point Extraction with Na-DDTC and Determination by Electrothermal Atomic Absorption Spectrometry (ET-AAS) (Na-DDTC로 흐림점 추출을 사용한 물시료에서 초미량 비스머스의 예비농축)

  • Moghimi, Ali
    • Journal of the Korean Chemical Society
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    • v.52 no.2
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    • pp.140-147
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    • 2008
  • .A new approach for a cloud point extraction-electrothermal atomic absorption spectrometric method was used for determining bismuth. The aqueous analyte was acidified with sulfuric acid (pH 3.0-3.5). Triton X-114 was added as a surfactant and natriumdiethyldithiocarbaminat (Na-DDTC) was used as a complexing agent. After phase separation at 50oC based on the cloud point separation of the mixture, the surfactant-rich phasen was diluted using tetrahydrofuran (THF). Twenty microliters (20 L) of the enriched solution and 10 l of 0.1% (w/v) Pd(NO3)2 as chemical modifier were dispersed into the graphite tube and the analyte determined by electrothermal atomic absorption spectrometry. After optimizing extraction conditions and instrumental parameters, a preconcentration factor of 195 was obtained for a sample of only 10 mL. The detection limit was 0.04 ng ml-1 and the analytical curve was linear for the concentration range of 0.04-0.70 ng mL-1. Relative standard deviations were <5%. The method was successfully applied for the extraction and determination of bismuth in water samples.

Characterization of carrier transport and trapping in semiconductor films during plasma processing

  • Nunomura, Shota;Sakata, Isao;Matsubara, Koji
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.391-391
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    • 2016
  • The carrier transport is a key factor that determines the device performances of semiconductor devices such as solar cells and transistors [1]. Particularly, devices composed of in amorphous semiconductors, the transport is often restricted by carrier trapping, associated with various defects. So far, the trapping has been studied for as-grown films at room temperature; however it has not been studied during growth under plasma processing. Here, we demonstrate the detection of trapped carriers in hydrogenated amorphous silicon (a-Si:H) films during plasma processing, and discuss the carrier trapping and defect kinetics. Using an optically pump-probe technique, we detected the trapped carriers (electrons) in an a-Si:H films during growth by a hydrogen diluted silane discharge [2]. A device-grade intrinsic a-Si:H film growing on a glass substrate was illuminated with pump and probe light. The pump induced the photocurrent, whereas the pulsed probe induced an increment in the photocurrent. The photocurrent and its increment were separately measured using a lock-in technique. Because the increment in the photocurrent originates from emission of trapped carriers, and therefore the trapped carrier density was determined from this increment under the assumption of carrier generation and recombination dynamics [2]. We found that the trapped carrier density in device grade intrinsic a-Si:H was the order of 1e17 to 1e18 cm-3. It was highly dependent on the growth conditions, particularly on the growth temperature. At 473K, the trapped carrier density was minimized. Interestingly, the detected trapped carriers were homogeneously distributed in the direction of film growth, and they were decreased once the film growth was terminated by turning off the discharge.

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The Effects of Exercise of Diverse Intensities on the Expression of TNF-α in the Spinal Cord in Osteoarthritic Rats (다양한 운동 강도가 골관절염 흰쥐의 척수 내 TNF-α 발현에 미치는 영향)

  • Park, Soo-Jin
    • Journal of the Korean Society of Physical Medicine
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    • v.8 no.4
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    • pp.539-547
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    • 2013
  • PURPOSE: This study examined the effects of exercise of diverse intensities on the expression of TNF-${\alpha}$(tumor necrosis factor-${\alpha}$) in the spinal cord in osteoarthritic rats. METHODS: Over a period of four weeks, the authors applied treadmill exercise programs of diverse intensities to Sprague-Dawley rats, to which intra-articular injection of monosodium iodoacetate(MIA, $3mg/50{\mu}l$, diluted in saline) was applied to the right knee joint to induce osteoarthritis. The four-week exercise program was not carried out with the control group(CG, n=10). Exercise programs of applicable intensities were applied to the low-intensity exercise group(LEG, n=10), moderate-intensity exercise group (MEG, n=10), and high-intensity exercise group(HEG, n=10) over the four weeks. Observations were made of expression of TNF-${\alpha}$ in the spinal cord in osteoarthritic rats using western blot analysis. RESULT: there were significant differences(p<.05) in the comparison of expression of TNF-${\alpha}$ between the four groups involved. The expression of TNF-${\alpha}$ in the spinal cord, the LEG and HEG had more elevated expression significantly than the CG(p<.05). But the MEG had reduced expression significantly than the CG(p<.05). CONCLUSION: These study results suggest that moderate-intensity exercise is effective in inhibition TNF-${\alpha}$ in the spinal cord. They also indicate that in prescribing exercise to treat osteoarthritic patients, exercise of moderate intensity is most suitable to patients' chronic pain, rather than low or high intensity.

Response to Bee Venom Acupuncture and Polymorphism of Interleukin 4 Receptor and Interleukin 10 in Rheumatoid Arthritis

  • Kim, Yung-Tae;Lee, Jae-Dong;Lee, Yun-Ho
    • The Journal of Korean Medicine
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    • v.25 no.2
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    • pp.207-219
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    • 2004
  • Objectives : We investigated to find the relationship between single-nucleotide polymorphism (SNP) of IL4R, IL-10 and bee venom therapy efficacy in patients with RA treated with bee venom for 8 weeks. Methods : Korean RA patients (n=114) and healthy subjects (n=109) were included in this prospective study. Korean bee venom was dissolved in saline (diluted 1:3000) and administrated into acupuncture points. Bee venom therapy was applied twice a week and continued for 8 weeks. The clinical response was evaluated using various assessments before and after treatment. Disease severity was measured by determining the number of tender joints and swollen joints. Laboratory studies included ESR, CRP, and rheumatoid factor. Genotyping for IL-4R and IL-10 polymorphism was done by pyrosequencing analysis. Results : 1. In IL4R genotypes, there was significant difference between RA ptitients tind controls group. 2. In IL4R genotypes, there was significant difference among Good, Mild and Bad responders to in RA patients, but in the frequency of alleles and carriers, there were no significant difference. 3. There was no significant difference between RA patients and controls group in IL-10 gene genotypes. 4. In IL-10 genotypes, there was no significant difference among Good, Mild and Bad responders to in RA patients. 5. There was no significant difference in the improvement of ESR, CRP and KHAQ scores after bee venom therapy in RA patients among the IL4R or IL-10 genotypes. Conclusions : In IL-4R genotypes, there was significant difference between RA patients and control group, and among Good, Mild and Bad responders in RA patients. However, in IL-10 genotypes, there was no significant difference between RA patients and controls group and among Good, Mild and Bad responders in RA patients.

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Development of an Agrobacterium-mediated Transient Expression System for Intact Leaves of Chili Pepper (Agrobacterium을 이용한 고추의 Transient Expression 시스템)

  • Seong, Eun-Soo;Joung, Young-Hee;Choi, Doil
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.185-190
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    • 2004
  • We established a transient gene expression system in chili pepper leaves based on Agrobacterium-mediated transformation of GUS gene. For the best GUS transient expression, two step culture system was adopted. When the Agrobacterium tumefaciens cell density of pre-culture was $A_{600nm}$ 0.3, the cells were harvested and diluted to $A_{600nm}$ 0.8 with virulence induction medium after cell harvested. The addition of acetosyringone (200 $\mu$M) in virulence induction step was a key factor for successful transient expression. Additionally, Younger leaves showed more effective transient expression than older leaves. Temporally, the strongest intensity of GUS expression was detected at 2 days after infiltration. These results demonstrate that Agrobacterium-mediated transient expression can be used for a simple in vivo assays of plant promoters, transcription factors and furthermore provide efficient protocol for chili pepper transformation.