• Title/Summary/Keyword: 2-DE gel

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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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    • v.47 no.1
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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.

Comparative Study of Protein Profile during Development of Mouse Placenta

  • Han, Rong-Xun;Kim, Hong-Rye;Naruse, Kenji;Choi, Su-Min;Kim, Baek-Chul;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.31 no.4
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    • pp.253-260
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    • 2007
  • To examine the differential protein expression pattern in the 11.5 day post-coitus (dpc) and 18.5 dpc placenta of mouse, we have used the global proteomics approach by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. The differential protein patterns of 3 placentae at the 11.5 dpc and 18.5 dpc from nature mating mice were analyzed. Proteins within isoelectric point range of $3.0{\sim}10.0$, separately were analyzed in 2DE with 3 replications of each sample. A total of approximately 1,600 spots were detected in placental 2-D gel stained with Coomassie-blue. In the comparison of 11.5 dpc and 18.5 dpc placentae, a total of 108 spots were identified as differentially expressed proteins, of which 51 spots were up-regulated proteins such as alpha-fetoprotein, mKIAA0635 protein and transferrin, annexin A5, while 48 spots were down-regulated proteins such as Pre-B-cell colony-enhancing factor l(PBEF), aldolase 1, A isoform, while 4 spots were 11.5 dpc specific proteins such as chaperonin and Acidic ribosomal phosphoprotein P0, while 3 spots were 18.5 dpc specific proteins such as aldo-keto reductase family 1, member B7 and CAST1/ERC2 splicing variant-1. Most identified proteins in this analysis appeared to be related with catabolism, cell growth, metabolism and regulation. Our results revealed composite profiles of key proteins involved in mouse placenta during pregnancy.

Comprehensive proteome analysis using quantitative proteomic technologies

  • Kamal, Abu Hena Mostafa;Choi, Jong-Soon;Cho, Yong-Gu;Kim, Hong-Sig;Song, Beom-Heon;Lee, Chul-Won;Woo, Sun-Hee
    • Journal of Plant Biotechnology
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    • v.37 no.2
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    • pp.196-204
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    • 2010
  • With the completion of genome sequencing of several organisms, attention has been focused to determine the function and functional network of proteins by proteome analysis. The recent techniques of proteomics have been advanced quickly so that the high-throughput and systematic analyses of cellular proteins are enabled in combination with bioinformatics tools. Furthermore, the development of proteomic techniques helps to elucidate the functions of proteins under stress or diseased condition, resulting in the discovery of biomarkers responsible for the biological stimuli. Ultimate goal of proteomics orients toward the entire proteome of life, subcellular localization, biochemical activities, and their regulation. Comprehensive analysis strategies of proteomics can be classified as three categories: (i) protein separation by 2-dimensional gel electrophoresis (2-DE) or liquid chromatography (LC), (ii) protein identification by either Edman sequencing or mass spectrometry (MS), and (iii) quanitation of proteome. Currently MS-based proteomics turns shiftly from qualitative proteome analysis by 2-DE or 2D-LC coupled with off-line matrix assisted laser desorption ionization (MALDI) and on-line electrospray ionization (ESI) MS, respectively, to quantitative proteome analysis. Some new techniques which include top-down mass spectrometry and tandem affinity purification have emerged. The in vitro quantitative proteomic techniques include differential gel electrophoresis with fluorescence dyes, protein-labeling tagging with isotope-coded affinity tag, and peptide-labeling tagging with isobaric tags for relative and absolute quantitation. In addition, stable isotope labeled amino acid can be in vivo labeled into live culture cells through metabolic incorporation. MS-based proteomics extends to detect the phosphopeptide mapping of biologically crucial protein known as one of post-translational modification. These complementary proteomic techniques contribute to not only the understanding of basic biological function but also the application to the applied sciences for industry.

Photoperiodic Proteins in Plant Cells (식물세포의 일주기성 단백질)

  • Hwang, Hee-Youn;Bhoo, Seong-Hee
    • Journal of Applied Biological Chemistry
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    • v.53 no.3
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    • pp.121-125
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    • 2010
  • In the past 10 years, a lot of plant circadian rhythm researches have published in molecular biology and biochemistry. We discussed with published molecular studies of circadian clock and rhythmic genes in Arabidopsis, rice and algae. However past this studies are not sufficient to explain the whole rhythmic metabolism. Recently many researchers have concerned post-transcriptional, translational and post-translational modification of rhythmic proteins. From the view point of the high-throughput study, we could suggest the proteomic analysis with 2-DE gel electrophoresis and MS/MS techniques for the identification of modified proteins.

Comparative Proteomics Analysis of Colorectal Cancer

  • Wang, Jun-Jiang;Liu, Ying;Zheng, Yang;Lin, Feng;Cai, Guan-Fu;Yao, Xue-Qing
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1663-1666
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    • 2012
  • Background and Objective: Protein expression in colon and rectal cancer (CRC) and paired normal tissues was examined by two-dimensional gel electrophoresis (2-DE) to identify differentially expressed proteins. Materials and Methods: Five fresh colorectal cancer and paired adjacent normal tissues were obtained and differentially expressed protein spots were determined using PDQuest software, with identification on the basis of MALDI-TOF mass spectra. Results: Compared with normal colorectal mucosa, protein abnormal expression of 65 spots varying more than 1.5 times were found in 2-DE gels from colorectal cancer samples (P<0.05); forty-two proteins were up-regulated and 23 were down-regulated; twelve protein spots were identified using mass spectrometry, of which 8 were up-regulated, includimng HSPB1and Annexin A4, while 4 were down-regulated, the results being consistent with Western blot findings. Conclusions: Two-dimensional electrophoresis reference maps for CRC tissues and adjacent normal mucosa (NMC) were established and 12 differentially expressed proteins were identified. Up-regulated HSPB1 and Annexin A4 may play many important roles in the pathogenesis of colorectal cancer.

Structural and gelling properties of very low methoxyl pectin produced by an alkali-treatment

  • Lee, Byung-Hoo;Jung, Ho-Tak;Kim, Hyun-Seok;Yoo, Sang-Ho
    • Korean Journal of Food Science and Technology
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    • v.53 no.2
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    • pp.121-125
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    • 2021
  • Very low methoxyl pectin (VLMP) has different physical and rheological properties compared to high and low methoxyl pectins (HMP and LMP). In this study, we produced LMP and VLMP by alkaline de-esterification, and investigated the structural and textural properties. Apple peel pectin was kept at pH 12 using 5.0 M NaOH solution for 3 and 24 h to produce LMP and VLMP, respectively. The molecular weight was decreased due to the removal of an esterified group in the pectin backbones by the alkali treatment, and the VLMP showed a higher calcium ion sensitivity which leads to the production of the gel with increased hardness. The result clearly showed that VLMP has the potential to improve the texture and stability in food products depending on their degree of esterification, and this result can be applied as a functional ingredient in food industrial area application to enhance the current commercial pectins.

Shaping ability and apical debris extrusion after root canal preparation with rotary or reciprocating instruments: a micro-CT study

  • Emmanuel Joao Nogueira Leal da Silva;Sara Gomes de Moura;Carolina Oliveira de Lima;Ana Flavia Almeida Barbosa;Waleska Florentino Misael;Mariane Floriano Lopes Santos Lacerda;Luciana Moura Sassone
    • Restorative Dentistry and Endodontics
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    • v.46 no.2
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    • pp.16.1-16.11
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    • 2021
  • Objectives: The aim of this study was to evaluate the shaping ability of the TruShape and Reciproc Blue systems and the apical extrusion of debris after root canal instrumentation. The ProTaper Universal system was used as a reference for comparison. Materials and Methods: Thirty-three mandibular premolars with a single canal were scanned using micro-computed tomography and were matched into 3 groups (n = 11) according to the instrumentation system: TruShape, Reciproc Blue and ProTaper Universal. The teeth were accessed and mounted in an apparatus with agarose gel, which simulated apical resistance provided by the periapical tissue and enabled the collection of apically extruded debris. During root canal preparation, 2.5% sodium hypochlorite was used as an irrigant. The samples were scanned again after instrumentation. The percentage of unprepared area, removed dentin, and volume of apically extruded debris were analyzed. The data were analyzed using 1-way analysis of variance and the Tukey test for multiple comparisons at a 5% significance level. Results: No significant differences in the percentage of unprepared area were observed among the systems (p > 0.05). ProTaper Universal presented a higher percentage of dentin removal than the TruShape and Reciproc Blue systems (p < 0.05). The systems produced similar volumes of apically extruded debris (p > 0.05). Conclusions: All systems caused apically extruded debris, without any significant differences among them. TruShape, Reciproc Blue, and ProTaper Universal presented similar percentages of unprepared area after root canal instrumentation; however, ProTaper Universal was associated with higher dentin removal than the other systems.

A Comprehensive Identification of Synaptic Vesicle Proteins in Rat Brains by cRPLC/MS-MS and 2DE/MALDI-TOF-MS

  • Lee, Won-Kyu;Kim, Hye-Jung;Min, Hye-Ki;Kang, Un-Beom;Lee, Cheol-Ju;Lee, Sang-Won;Kim, Ick-Young;Lee, Seung-Taek;Kwon, Oh-Seung;Yu, Yeon-Gyu
    • Bulletin of the Korean Chemical Society
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    • v.28 no.9
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    • pp.1499-1509
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    • 2007
  • Proteomic analyses of synaptic vesicle fraction from rat brain have been performed for the better understanding of vesicle regulation and signal transmission. Two different approaches were applied to identify proteins in synaptic vesicle fraction. First, the isolated synaptic vesicle proteins were treated with trypsin, and the resulting peptides were analyzed using a high-pressure capillary reversed phase liquid chromatography/tandem mass spectrometry (cRPLC/MS/MS). Alternatively, proteins were separated by two-dimensional gel electrophoresis (2DE) and identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF/MS). Total 18 and 52 proteins were identified from cRPLC/MS-MS and 2DE-MALDI-TOF-MS analysis, respectively. Among them only 2 proteins were identified by both methods. Of the proteins identified, 70% were soluble proteins and 30% were membrane proteins. They were categorized by their functions in vesicle trafficking and biogenesis, energy metabolism, signal transduction, transport and unknown functions. Among them, 27 proteins were not previously reported as synaptic proteins. The cellular functions of unknown proteins were estimated from the analysis of domain structure, expression profile and predicted interaction partners.

Purification, Characterization, and Partial Primary Sequence of a Major-Maltotriose-producing $\alpha$-Amylase, ScAmy43, from Sclerotinia sclerotiorum

  • Ben Abdelmalek-Khedher, Imen;Urdad, Maria Camino;Limam, Ferid;Schmitter, Jean Marie;Marzouki, M. Nejib;Bressollier, Philippe
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1555-1563
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    • 2008
  • A novel $\alpha$-amylase ($\alpha$-1,4-$\alpha$-D-glucan glucanohydrolase, E.C. 3.2.1.1), ScAmy43, was found in the culture medium of the phytopathogenic fungus Sclerotinia sclerotiorum grown on oats flour. Purified to homogeneity, ScAmy43 appeared as a 43 kDa monomeric enzyme, as estimated by SDS-PAGE and Superdex 75 gel filtration. The MALDI peptide mass fingerprint of ScAmy43 tryptic digest as well as internal sequence analyses indicate that the enzyme has an original primary structure when compared with other fungal a-amylases. However, the sequence of the 12 N-terminal residues is homologous with those of Aspergillus awamori and Aspergillus kawachii amylases, suggesting that the new enzyme belongs to the same GH13 glycosyl hydrolase family. Assayed with soluble starch as substrate, this enzyme displayed optimal activity at pH 4 and $55^{\circ}C$ with an apparent $K_m$ value of 1.66 mg/ml and $V_{max}$ of 0.1${\mu}mol$glucose $min^{-1}$ $ml^{-1}$. ScAmy43 activity was strongly inhibited by $Cu^{2+}$, $Mn^{2+}$, and $Ba^{2+}$, moderately by $Fe^{2+}$, and was only weakly affected by $Ca^{2+}$ addition. However, since EDTA and EGTA did not inhibit ScAmy43 activity, this enzyme is probably not a metalloprotein. DTT and $\beta$-mercaptoethanol strongly increased the enzyme activity. Starting with soluble starch as substrate, the end products were mainly maltotriose, suggesting for this enzyme an endo action.

Adjuvant therapy with 1% alendronate gel for experimental periodontitis treatment in rats

  • de Campos Kajimoto, Natalia;de Paiva Buischi, Yvonne;Loomer, Peter Michael;Bromage, Timothy G.;Ervolino, Edilson;Fucini, Stephen Enrico;Pola, Natalia Marcumini;Pirovani, Beatriz Ommati;Morabito, Maria Juliana Sismeiro;de Almeida, Juliano Milanezi;Furlaneto, Flavia Aparecida Chaves;Nagata, Maria Jose Hitomi
    • Journal of Periodontal and Implant Science
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    • v.51 no.6
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    • pp.374-385
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    • 2021
  • Purpose: The aim of this study was to evaluate the effects of locally delivered 1% alendronate (ALN) gel used as an adjunct to non-invasive periodontal therapy. Methods: Ligature-induced periodontitis was performed in 96 rats. The ligature was tied in the cervical area of the mandibular left first molar. The animals were randomly divided into 4 groups: 1) NT, no treatment; 2) SRP, scaling and root planning; 3) SRP/PLA, SRP followed by filling the periodontal pocket with placebo gel (PLA); and 4) SRP/ALN, SRP followed by filling the periodontal pockets with 1% ALN gel. Histomorphometric (percentage of bone in the furcation region [PBF]) and immunohistochemical (receptor activator of nuclear factor-κB ligand, osteoprotegerin, and tartrate-resistant acid phosphatase) analyses were performed. Data were statistically analyzed, with the threshold of statistical significance set at P≤0.05. Results: The SRP, SRP/PLA, and SRP/ALN groups presented a higher PBF than the NT group (P≤0.01) at 7, 15, and 30 days. The SRP/ALN group presented a higher PBF than the SRP/PLA group in all experimental periods, as well as a higher PBF than the SRP group at 15 and 30 days. No differences were observed in the immunohistochemical analyses (P>0.05 for all). Conclusions: Locally delivered 1% ALN gel used as an adjunct to SRP enhanced bone regeneration in the furcation region in a rat model of experimental periodontitis.