• 제목/요약/키워드: 2D-gel

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Gel filtration에 의한 콩나물 제조중(製造中) 단백질(蛋白質)의 변화조사(變化調査) (Changes of Protein During Growth of Soybean Sprout)

  • 양차범;박상기;윤석권
    • 한국식품과학회지
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    • 제16권4호
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    • pp.472-474
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    • 1984
  • 대두(大豆)와 대두(大豆)의 발아과정별(發芽過程別) 수추출단백질(水抽出蛋白質)을 Sephadex G-200에 의해서 gel fietration 으로 분획(分劃)한 결과 자엽부(子葉部)에서는 5 개의 분획 (A, B, C, D 및 E) 이 얻어졌고 표품(標品)의 gel filtration과 disc gel electrophoresis 로서 그중(中) A분획이 혼탁물질, B가 11S, C가 7 S, D가 2S로 동정(同定)되었으며 E는 전기영동상에 1 peak를 보였다. 발아(發芽)에 따른 이들 분획(分劃) 변화(變化)는 자명부(子蓂部)에서는 7 S가 먼저 감소되고 2 S가 그 다음이며 11S가 맨나중에 감소되었다. A분획(分劃)은 6 일까지 증가되다가 감소를 보이고 E분획(分劃)은 계속 증가추세를 보이었다. 배축(胚軸)에서는 단지 2 개의 분획(分劃)이 나타났고 그 하나는 B+C(11S+ 7S)의 혼합물이었고 다른 하나는 E분획(分劃)이었다. 발아(發芽)에 따라서 11S+ 7 S 분획(分劃)은 큰 변화(變化)가 없었고 E 분획(分劃)은 약간의 증가를 보였다.

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Isolation and Properties of $\beta$-N-Acetyl-D-glucosaminidase B from Rat Uterus

  • Jung, Jin-Ha;Yang, Chul-Hak
    • Bulletin of the Korean Chemical Society
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    • 제4권3호
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    • pp.139-143
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    • 1983
  • ${\beta}$-N-Acetyl-D-glucosaminidase B was highly purified with the following sequence of steps; DEAE-cellulose, CM-cellulose, and Sephadex G-200 gel filtration chromatograpies. The specific activity of the purified ${\beta}$ -N-acetyl-D-glucosaminidase B was 2.2 units/mg protein with 12.9 % yield and 196.2 fold purity. The purified ${\beta}$-N-acetyl-D-glucosaminidase B showed single band on polyacrylamide gel electrophoresis. The final preparation of ${\beta}$ -N-acetyl-D-glucosaminidase B was completely free friom arylsulfatase and ${\beta}$-glucuronidase. ${\beta}$ -N-Acetyl-D-glucosaminidase B had pH optimum of 4.5 in 0.5 M sodium citrate buffer. The molecular weight of ${\beta}$-N-acetyl-D-glucosaminidase B was 133,000 by Sephadex G-200 gel filtration. The Km value of ${\beta}$-N-acetyl-D-glucosaminidase B using p-nitrophenyl-N-acetyl-${\beta}$-D-glucosaminide as substrate was 1.0 mM and $V_{max}$ was 0.014 ${\mu}$ mole/min. ${\beta}$-N-Acetyl-D-glucosaminidase B was stable at $55^{circ}C$ for 70 minutes. The crude ${\beta}$ -N-acetyl-D-glucosamiinidase in 70 % ammonium sulfate retained 93 % activity after 7 months storage at -$55^{circ}C$. Bovine serum albumin, sodium chloride, and phosphate activated ${\beta}$ -N-Acetyl-D-glucosaminidase B. N-Acetyl-D-glucosamine, ${\alpha}$-methyl-D-mannoside, and acetate inhibited ${\beta}$ -N-acetyl-D-glucosaminidase B.

In-office dental bleaching with violet light emitting diode: bleaching efficacy and pulpal temperature rise

  • Brunna Katyuscia de Almeida Guanaes;Talyta Neves Duarte;Gisele Maria Correr;Marina da Rosa Kaizer;Carla Castiglia Gonzaga
    • Restorative Dentistry and Endodontics
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    • 제47권1호
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    • pp.7.1-7.14
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    • 2022
  • Objectives: This study evaluated the bleaching efficacy of different in-office protocols associated with violet light emitting diode (V-LED), and measured the pulpal temperature rise caused by V-LED with or without gel application. Materials and Methods: Bovine incisors were distributed in 4 groups (n = 10): VL - V-LED; HP - 35% hydrogen peroxide (control); HYB - hybrid protocol, V-LED applied without gel for 10 irradiation cycles followed by V-LED applied with gel for another 10 irradiation cycles; and HPVL - gel and V-LED applied for 20 irradiation cycles. Three bleaching sessions were performed with 7-day intervals. Bleaching efficacy was evaluated with ΔEab*, ΔE00 and ΔWID. Data were recorded at baseline, 7, 14, 21 and 70 days. For pulpal temperature rise, thermocouples were placed inside the pulp chamber of human incisors. To determine intrapulpal temperature, the teeth were irradiated with V-LED with or without application of bleaching gel. Color difference data were analyzed by 2-way repeated measures ANOVA and Tukey's test. Pulpal temperature was analyzed by t-test (α = 5%). Results: VL exhibited lower color (ΔEab* and ΔE00) and whiteness changes (ΔWID) than the other groups. HPVL presented higher color change values than HYB. HYB and HPVL showed not different ΔWID values; and HP showed the highest whiteness changes at all times. There were significant differences comparing ΔT with gel (8.9℃) and without gel application (7.2℃). Conclusions: HPLV was more efficient than HYB. The 2 protocols with VL showed similar results to control. Gel application combined with VL promoted higher pulpal temperature than to the no gel group.

Study on the Evaluation of Stability of Gel Structured Cosmetics

  • Park, Chan-Ik;Kim, Ki-Sun;Lee, Sung-Jun;Yoon, Myeong-Suk;Kang, Seh-Hoon
    • 대한화장품학회지
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    • 제22권2호
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    • pp.167-173
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    • 1996
  • The stability of gel structured emulsion and the effect of polyols on it have been studied by rheological property and interfacial tension. In this paper, three types of gel structured emulsions were prepared by using three polyols respectively(glycerine for sample 1, 1.3 BG for sample 2, PG for sample 3). And both complex modulus($G^*$) and loss angle[$\delta$ = tan-1(G"/G')] of samples were investigated against oscillating shear stress and frequency($\omega$). The results show sample 1 is most highly consistent with oscillating shear stress. And the results were compared with those of accelerated tests concerning storage stability of gel structured emulsion. To correlate consistency of rheological property with storage stability, interfacial tension from which adsorption efficiency of surfactant(Octyldodecyl Ether) could be known was measured. Sample 1 showed the largest value of [$d{\gamma}/dIn_{Cconc. of surfactant}$] in Gibbs equation. In summary, the prediction of stability could be correctly made by the consistency of rheological property(G*,$\gamma$) of gel structured emulsion against oscillating shear stress and it could be supported by measuring interfacial tension. And polyol affected the value of [$d{\gamma}/dIn_{Cconc. of surfactant}$], consequently affected the stability.lity.

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DentalVibe reduces pain during the administration of local anesthetic injection in comparison to 2% lignocaine gel: results from a clinical study

  • Joshi, Sagar;Bhate, Kalyani;Kshirsagar, Kapil;Pawar, Vivek;Kakodkar, Pradnya
    • Journal of Dental Anesthesia and Pain Medicine
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    • 제21권1호
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    • pp.41-47
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    • 2021
  • Background: This study was designed to compare the efficacy of DentalVibe against 2% lidocaine gel in reducing pain during the administration of local anesthetic injection in the adult population. Methods: This was a split-mouth open-label, randomized, controlled clinical study conducted in the Department of Oral and Maxillofacial Surgery of a dental institute. Fifty patients who were scheduled for bilateral dental extractions requiring an inferior alveolar nerve block were enrolled in the study. Site A (n = 50) was coated with 2% lidocaine gel followed by a local anesthetic injection, and DentalVibe with local anesthetic injection was used for Site B (n = 50). The primary outcome was pain, which was recorded immediately after the administration of anesthetic injection using the Visual Analogue Scale [VAS 0 - 10]. Results: The VAS pain scores ranged from 4 to 10 for site A and 0 to 6 for site B. Comparison between the two sites showed a statistically significant difference [Mann-Whitney U test value = 51.50, P < 0.001] favoring site B. Conclusion: This study showed that DentalVibe reduces pain during injection of local anesthesia compared to topical anesthetic gel.

Neurospora crassa의 L-Ascorbic Acid 생산효소의 순수 분리 및 이의 특성에 관한 연구 (Isolation and Characterization of L-Ascorbic Acid-Producing Enzyme in Neurospora crassa)

  • 김인실;이연희
    • 미생물학회지
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    • 제32권2호
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    • pp.132-138
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    • 1994
  • Neurospora crass에서 L-ascorbic acid 합성 효소는 mitochondria에 위치하며 배지에 D- glucono-${\gamma}$-lactone과 L-gluno-${\gamma}$-lactone을 첨가하였을 때 각각의 기질에 대한 효소 활성도가 증가함을 볼 수 있었다. 이 효소의 순수 분리에는 ammonium sulfate 분획, DEAE-Sepharose CL-6B 에 의한 이온 교환 크로마토그래피, Sephacryk S-200에 의한 gel filtrarion 크로마토그래피, Reactive yellow 3-agarose dye column 크로마토그래피 등의 방법들이 이용되었다. 그 결과 2.1%의 수율에 specific activity가 239.6배 증가되었다. Sephacryl S-200 gel filtration을 통해 본 이 효소의 분자량은 약 150,000 dalton 이었다. 그리고 SDS-polyacrylamide gel electrophoresis를 실시하였을 때는 분자량이 약 75,000 dalton이었으므로 이 효소는 동일한 단위체로 구성된 이합체로 생각되었다. 이 효소의 최적 반은 pH는 9.0으로 나타났으며 D-glucono-${\gamma}$-lactone을 기질로 하였을때의 $K_m$값은 0.073이었다.

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한외여과 공정에 의한 알로에 베라 겔 농축 (Concentration of Fresh Gel from Aloe vera L. by Using Ultrafiltration Process)

  • 백진홍;김성아;이신영
    • KSBB Journal
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    • 제23권2호
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    • pp.169-176
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    • 2008
  • The concentration of fresh gel from Aloe vera L. by using ulfrafiltration (UF) process was investigated and analyzed. The two membranes (organic and ceramic) with different molecular weight cut-off (MWCO) and modules (flat sheet and tubular) was used. Under optimum operation conditions, ceramic (zirconium dioxide) tubular membrane with MWCO of 50 kDa resulted in higher flux, less fouling, more turbid, higher total solid, higher polysaccharide and less aloin content. Optimum operation conditions were transmembrane pressure of 1.0 bar, feed velocity of 240 L/hr and temperature of $23^{\circ}C$. Volume concentration factor of aloe gel was 3.13 at permeate flux of $51.1\;L/m^2{\cdot}hr$ after processing time of 1.66 hr. Aloin in fresh aloe gel by UF process was effectively removed as permeate and bioactive polysaccharide content was 2.1 times higher than that of fresh aloe gel. These results allowed a very good level of concentration degree and polysaccharide content. Thus, ultrafiltration process of this study was suitable for the concentration of fresh aloe gel though the aloe concentrate showed both the viscosity decrease and partially separation of liquid layer during storage at $4^{\circ}C$.

Galactomannan 이용에 관한 연구 : Affinity Chromatography에 의한 지렁이 유래 /alpha-Galactosidase의 정제 및 응용법 (Purification and Application of Earthworm /alpha-Galactosidase by Affinity Chromatography)

  • 박귀근;정규훈;소림수행
    • 한국미생물·생명공학회지
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    • 제27권4호
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    • pp.298-303
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    • 1999
  • An $\alpha$-D-galactosidase ($\alpha$-D-galactoside galactohydrolase, EC 3. 2. 1. 22) from earthworm was purified by affinity chromatography using N-$\varepsilon$-aminocaproyl-$\alpha$-D-galactopyranosylamine coupled to sepharose and its properties were examined. The specific activity of the purified enzyme, tested with p-nitrophenyl-$\alpha$-D-galactopyranoside as substrate, was 314 units/mg protein, representing an 122-fold purification of the original crude extract. The final preparation obtained from by Sephadex G-25 chromatography showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight was determined to be 48,000 by SDS-polyacrylamide gel electrophoresis. The purified galactosidase was showed maximum activity at pH 4.5 and 4$0^{\circ}C$, and was stable in the pH and temperature ranges from 4.0 to 5.5 and 30 to 5$0^{\circ}C$, respectively. The enzyme activity was inhibited by Zn2+, Hg2+ and Co2+. When the purified $\alpha$-galactosidase treated to guar gum for 6 hour, gel-promoting property was increased. It was clear that enzymatic elimination of galactose from guar gum by purified $\alpha$-galactosidase would lead to a significant increase in gelation ability.

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Proteomic Analysis of Bovine Pregnancy-specific Serum Proteins by 2D Fluorescence Difference Gel Electrophoresis

  • Lee, Jae Eun;Lee, Jae Young;Kim, Hong Rye;Shin, Hyun Young;Lin, Tao;Jin, Dong Il
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권6호
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    • pp.788-795
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    • 2015
  • Two dimensional-fluorescence difference gel electrophoresis (2D DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum.

Trichoderma koningii에서 분비되는 .$\beta$-D-glucosidase의 반응산물에 대한 핵자기공명분석 ($^{1}$H-NMR spectroscopic evidence on the glycosidic linkages of the transglycosylated products of low-molecular-weight $\beta$-D-glucosidase from trichoderma koningii)

  • 이헌주;정춘수;강사욱;하영칠
    • 미생물학회지
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    • 제27권1호
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    • pp.35-42
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    • 1989
  • The mode of transglycosylation reaction observed during the action of low-molecular-weigh $\beta$-D-glucosidase ($\beta$-D-glucoside glucohydrolase, EC3.2.1.21) purified from Trichoderma koningii ATCC 26113 was investigated using $^{1}H$-NMR spectroscopy. The enzyme was purified by the series of procedures including ammonium sulfate precipitation, and fractionations by column chromatographies on Bio-Gel P-150, DEAE-Sephadex A-50, and SP-Sephadex C-50. The final purification was performed by the band eluation after preparative polyacrylamide gel electrophoresis. The enzyme showed its molecular size of 78,000 through the analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and its isoelectric point of 5.80 through the analysis of analytical isoelectric focusing. The H-1 proton resonances were analyzed. After the reaction of the enzyme with cellobiose, the reaction products were separated by high performance liquid chromatography using refractive index detector. H-1 resonances of the products were consisted with those of gentiobiose [$\beta$-D-glucopyranosyl--(1,6)-D-glucopyranose], and cellotriose [$\beta$-D glucopyranosyl-(1,4)-$\beta$-D-glucopyranosyl]-(1,4)-D-glucopyranose] with minor resonances of sophorose [$\beta$-D-glucopyranosyl-(1,2)-D-glucopyranose], respectively.

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