• Title/Summary/Keyword: Fluorescence loss

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Investigation on the Effects of Three X${\rightarrow}$Histidine Replacements on Thermostability of ${\alpha}$-Amylase from Bacillus amyloliquefaciens

  • Haghani, Karimeh;Khajeh, Khosro;Naderi-Manesh, Hossein;Ranjbar, Bijan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.592-599
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    • 2012
  • Bacillus licheniformis ${\alpha}$-amylase (BLA), a thermophilic counterpart of Bacillus amyloliquefaciens ${\alpha}$-amylase (BAA), is an appropriate model for the design of stabilizing mutations in BAA. BLA has 10 more histidines than BAA. Considering this prominent difference, in the present study, three out of these positions (I34, Q67, and P407; located in the thermostability determinant 1 region and Ca-III binding site of BAA) were replaced with histidine in BAA, using the site-directed mutagenesis technique. The results showed that the thermostability of P407H and Q67H mutants had increased, but no significant changes were observed in their kinetic parameters compared to that of the wild type. I34H replacement resulted in complete loss of enzyme activity. Moreover, fluorescence and circular dichroism data indicated a more rigid structure for the P407H variant compared with that of the wild-type BAA. However, the flexibility of Q67H and I34H mutants increased in comparison with that of wild-type enzyme.

Fast High-throughput Screening of the H1N1 Virus by Parallel Detection with Multi-channel Microchip Electrophoresis

  • Zhang, Peng;Park, Guenyoung;Kang, Seong Ho
    • Bulletin of the Korean Chemical Society
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    • v.35 no.4
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    • pp.1082-1086
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    • 2014
  • A multi-channel microchip electrophoresis (MCME) method with parallel laser-induced fluorescence (LIF) detection was developed for rapid screening of H1N1 virus. The hemagglutinin (HA) and nucleocapsid protein (NP) gene of H1N1 virus were amplified using polymerase chain reaction (PCR). The amplified PCR products of the H1N1 virus DNA (HA, 116 bp and NP, 195 bp) were simultaneously detected within 25 s in three parallel channels using an expanded laser beam and a charge-coupled device camera. The parallel separations were demonstrated using a sieving gel matrix of 0.3% poly(ethylene oxide) ($M_r$ = 8,000,000) in $1{\times}$ TBE buffer (pH 8.4) with a programmed step electric field strength (PSEFS). The method was ~20 times faster than conventional slab gel electrophoresis, without any loss of resolving power or reproducibility. The proposed MCME/PSEFS assay technique provides a simple and accurate method for fast high-throughput screening of infectious virus DNA molecules under 400 bp.

Efficient Complex Surfactants from the Type of Fatty Acids as Corrosion Inhibitors for Mild Steel C1018 in CO2-Environments

  • Abbasov, Vagif M.;El-Lateef, Hany M. Abd;Aliyeva, Leylufer I.;Ismayilov, Ismayil T.;Qasimov, Elmar E.;Narmin, Mamedova M.
    • Journal of the Korean Chemical Society
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    • v.57 no.1
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    • pp.25-34
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    • 2013
  • The efficiency of three complex surfactants based on sunflower oil and nitrogen containing compounds as corrosion inhibitors for mild steel in $CO_2$-saturated 1% NaCl solution, has been determined by weight loss and LPR corrosion rate measurements. These compounds inhibit corrosion even at very low concentrations. The inhibition process was attributed to the formation of an adsorbed film on the metal surface that protects the metal against corrosive media. The inhibition efficiency increases with increasing the concentration of the studied inhibitors. Maximum inhibition efficiency of the surfactants is observed at concentrations around its critical micellar concentration (CMC). Adsorption of complex surfactants on the mild steel surface is in agreement with the Langmuir adsorption isotherm model, and the calculated Gibbs free energy values confirm the chemical nature of the adsorption. Energy dispersive X-ray fluorescence microscopy (EDRF) observations of the electrode surface confirmed the existence of such an adsorbed film.

Jet Entrainment Effect in Buoyant Jet and Iso-Thermal Fire Modeling (부력제트의 주위공기 유입효과 및 등온기체 모델링)

  • Lee, Eui-Ju
    • Fire Science and Engineering
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    • v.23 no.6
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    • pp.111-115
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    • 2009
  • Acetone LIF and Rayleigh scattering measurements were performed to identify the entrainment of ambient air in the buoyant jet qualitatively. The air entrainment near nozzle exit was enhanced with increasing both an axial distance and Reynolds number. The results supported that the air entrainment had to be considered in isothermal model for the development of its accuracy. Also, this paper provided an isothermal model based on the ideal plume, of which radiative heat loss fraction was assumed to 0.35 and the entainment of isothermal jet was considered. This simple model could be used in compartment or semi-enclosure fires such as tunnel, and it is more reliable because of introducing entrainment effect in isothermal jet.

Processing Characteristics of Freeze-Dried Pork Powder for Meat Emulsion Gel

  • Lee, Seonmin;Choi, Yun-Sang;Jo, Kyung;Jeong, Hyun Gyung;Yong, Hae In;Kim, Tae-Kyung;Jung, Samooel
    • Food Science of Animal Resources
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    • v.41 no.6
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    • pp.997-1011
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    • 2021
  • The processing characteristics of freeze-dried pork powder as raw meat for comminuted meat products were compared with those of freeze-thawed pork. The tertiary structural properties, oxidation, and solubility of proteins in the freeze-dried pork powder were investigated. In addition, the properties of the emulsion gels manufactured with freeze-dried pork powder (GFD) and freeze-thawed pork (GFT) at 1.5% and 2.0% NaCl were evaluated. The surface hydrophobicity and intrinsic tryptophan fluorescence intensity of myofibrillar proteins between the freeze-dried pork powder and freeze-thawed pork were similar. However, freeze-dried pork powder had higher carbonyl compounds and lower solubility of sarcoplasmic and myofibrillar proteins than freeze-thawed pork (p<0.05). GFD had higher cooking loss than GFT in 2.0% NaCl, and lower hardness and a* value of GFD were observed regardless of NaCl level (p<0.05). Moreover, GFD had higher malondialdehyde content than GFT at the two NaCl concentrations (p<0.05). Therefore, our study demonstrated that freeze-dried pork powder has lower functional properties than freeze-thawed pork as raw meat for comminuted meat products.

REMINERALIZATION EFFECT OF INTERPROXIMAL CARIES ADJACENT TO GLASS IONOMER RESTORATIONS: IN VITRO STUDY USING QLF (QLF(Quantitative light-induced fluorescence)를 이용한 글라스 아이오노머 수복물의 인접면 우식 재광화 효과에 관한 연구)

  • Lee, Hyeok-Sang;Hyun, Hong-Keun;Jang, Ki-Taeg
    • Journal of the korean academy of Pediatric Dentistry
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    • v.38 no.3
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    • pp.244-249
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    • 2011
  • This in vitro study compared the remineralization of incipient interproximal caries in the presence of three glass ionomer cements(highly-filled glass ionomer cement, resin-modified glass ionomer cement, compomer) and a resin composite(control). Thirty-two extracted premolars were selected based upon the lack of any visible demineralization. The teeth were coated in a transparent acid resistant nail varnish leaving $3{\times}3$ mm square. The teeth were subjected to the demineralizing buffer for 3 days and quantitative light-induced fluorescence(QLF) images of the subjects were taken. Proximal restoration was simulated by placing tooth specimens and the various glass ionomer cements in closed containers with artificial saliva at $37^{\circ}C$ and pH 7.0 with constant circulation. Further QLF images were subsequently taken at 30, 60, and 90 days. The changes of mineral loss(${\Delta}Q$) were evaluated by QLF and the change of ${\Delta}Q$(${\Delta}{\Delta}Q$) were compared between groups in order to evaluate the effects of remineralization. All data were analyzed using ANOVA and the post-HOC Dunnett C multiple comparison test at p<0.05. While ${\Delta}Q$(changes of mineral loss) increased for all treatments, the increases for three glass ionomer groups were significantly higher than that for the resin group at first month period. As time went on, the amount of ${\Delta}{\Delta}Q$ decreased.

THE PREVENTIVE EFFECT OF COMMERCIALLY AVAILABLE ANTICARIOGENIC PRODUCTS ON ENAMEL EROSION INDUCED BY CARBONATED BEVERAGE IN VITRO (탄산음료 유발 법랑질 침식에 대한 항우식 제품의 예방 효과)

  • Song, Ju-Hyun;Kim, Young-Jae;Kim, Jung-Wook;Jang, Ki-Taeg;Kim, Chong-Chul;Hahn, Se-Hyun;Lee, Sang-Hoon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.37 no.1
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    • pp.1-12
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    • 2010
  • The aim of this study was to evaluate the preventive effect of commercially available anticariogenic products, specifically, the tooth cream containing Casein phosphopeptide-amorphous calcium phosphate(CPP-ACP), fluoride varnish and low-level fluoride mouthrinse on enamel erosion induced by carbonated beverage in a short period of time. Enamel specimens were treated as follows and were then kept in artificial saliva for 24 hours followed by further processing by alternately soaking them in Cola beverage and in distilled water for 1 minute each five times. Group 1: control group (no treatment) Group 2: tooth cream with CPP-ACP Group 3: fluoride varnish (1,000 ppm F) Group 4: low-level fluoride mouthrinse (227 ppm F) Group 5: fluoride varnish + tooth cream with CPP-ACP Group 6: low-level fluoride mouthrinse + tooth cream with CPP-ACP Microhardness and erosion depth were measured and the mineral loss of each specimen was evaluated by measuring the volumetric fluorescence change(${\Delta}Q$) against the stable fluorescent grid using quantitative light-induced fluorescence(QLF). The experiment lasted for 6 days repeated each day. The results were as follows: 1. The microhardness was increased as follows: Group $1{\leq}2{\leq}4$<6<$3{\fallingdotseq}5$. 2. The mean erosion depth was increased as follows: Group $5{\fallingdotseq}3$<6<$4{\fallingdotseq}2{\fallingdotseq}1$. 3. The ${\Delta}Q$ was increased as follows: Group $1{\fallingdotseq}2{\leq}4{\leq}6{\leq}3{\fallingdotseq}5$. The decrement of ${\Delta}Q$ was similar between group 1 and 2, group 4 and 6 and group 3 and 5. 4. The ${\Delta}Q$ showed positive correlation with microhardness (r=0.96, p<0.05), while it was negatively correlated to erosion depth (r=-0.96, p<0.05).

Effect of Black Rice (Oryza sativa L. indica) Ethanolic Extract on Tyrosinase Activity and Antioxidant Activity Related to Melanin Production (흑미의 주정 추출물이 melanin 생성과 관련된 tyrosinase 및 항산화 활성에 미치는 영향)

  • Lee, Eunbeen;Kim, Moon-Moo
    • Journal of Life Science
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    • v.28 no.5
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    • pp.532-539
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    • 2018
  • Hair graying is processed by loss of melanin production caused by the decrease of activity and number of melanocyte and the accumulation of hydrogen peroxide ($H_2O_2$) in the hair follicle with increase of age. The purpose of this study was to investigate the effect the Black oryzasativa ethanolic extract (BLEE) on the melanin production. In this study BLEE at $8{\mu}g/ml$ or more showed a significant 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity and reduction power. BLEE at $16{\mu}g/ml$ or more showed promoted tyrosinase activity and melanin production. In addition BLEE scavenged intracellular $H_2O_2$ in 2',7'-dichlorodihydrofluorescein (DCF) fluorescence assay in B16F1 cells. However, Western blot analyses displayed that BLEE decreased the expression level of catalase, but no effect on the expression level of tyrosinase, tyrosinase associated protein-1 (TRP-1), tyrosinase associated protein-2 (TRP-2) and microphthalmia-associated transcription factor (MITF) transcription factor involved in melanogenesis. Thus, the promotive effect of BLEE on melanin production is attributed to the increase of tyrosinase activity and the reduction of intracellular $H_2O_2$ level. In conclusion, BLEE played a key role in in promoting melanin production, which suggests that the BLEE could be applied as a potential functional material in the development of hair care cosmetics related to the promotion of melanin production for the growth of black hair.

Augmentation of Thermotolerance in Primary Skin Fibroblasts from a Transgenic Pig Overexpressing the Porcine HSP70.2

  • Chen, Ming-Yu;Tu, Ching-Fu;Huang, San-Yuan;Lin, Jyh-Hung;Tzang, Bor-Show;Hseu, Tzong-Hsiung;Lee, Wen-Chuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.1
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    • pp.107-112
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    • 2005
  • A high environmental temperature affects the economic performance of pigs. Heat shock protein 70 (HSP70) has been reported to participate importantly in thermotolerance. This study aims to produce transgenic pigs overexpressing porcine HSP70.2, the highly inducible one of HSP70 members, and to prove the cellular thermotolerance in the primary fibroblasts from the transgenics. A recombinant plasmid in which the sequence that encodes the porcine HSP70.2 gene is fused to green fluorescence protein (GFP) was constructed under the control of cytomegalovirus (CMV) enhancer and promoter. Two transgenic pigs were produced by microinjecting pCMV-HSP70-GFP DNA into the pronucleus of fertilized eggs. Immunoblot assay revealed the varied overexpression level (6.4% and 1.4%) of HSP70-GFP in transgenic pigs. After heating at $45^{\circ}C$ for 3 h, the survival rate (78.1%) of the primary fibroblast cells from the highly expressing transgenic pig exceeded that from the non-transgenic pig (62.9%). This result showed that primary fibroblasts overexpressing HSP70-GFP confer cell thermotolerance. We suggest that transgenic pigs overexpressing HSP70 might improve their thermotolerance in summer and therefore reduce the economic loss in animal production.

Synthesis and Evaluation of Variable Temperature-Electrical Resistance Materials Coated on Metallic Bipolar Plates (온도 의존성 가변 저항 발열체로 표면 처리된 금속 분리판 제조 및 평가)

  • Jung, Hye-Mi;Noh, Jung-Hun;Im, Se-Joon;Lee, Jong Hyun;Ahn, Byung Ki;Um, Sukkee
    • 한국신재생에너지학회:학술대회논문집
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    • 2010.11a
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    • pp.73.1-73.1
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    • 2010
  • For the successful cold starting of a fuel cell engine, either internal of external heat supply must be made to overcome the formation of ice from water below the freezing point of water. In the present study, switchable vanadium oxide compounds as variable temperature-electrical resistance materials onto the surface of flat metallic bipolar plates have been prepared by a dip-coating technique via an aqueous sol-gel method. Subsequently, the chemical composition and micro-structure of the polycrystalline solid thin films were analyzed by X-ray diffraction, X-ray fluorescence spectroscopy, and field emission scanning electron microscopy. In addition, it was carefully measured electrical resistance hysteresis loop over a temperature range from $-20^{\circ}C$ to $80^{\circ}C$ using the four-point probe method. The experimental results revealed that the thin films was mainly composed of Karelianite $V_2O_3$ which acts as negative temperature coefficient materials. Also, it was found that thermal dissipation rate of the vanadium oxide thin films partially satisfy about 50% saving of the substantial amount of energy required for ice melting at $-20^{\circ}C$. Moreover, electrical resistances of the vanadium-based materials converge on an extremely small value similar to that of pure flat metallic bipolar plates at higher temperature, i.e. $T{\geq}40^{\circ}C$. As a consequence, experimental studies proved that it is possible to apply the variable temperature-electrical resistance material based on vanadium oxides for the cold starting enhancement of a fuel cell vehicle and minimize parasitic power loss and eliminate any necessity for external equipment for heat supply in freezing conditions.

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