• Title/Summary/Keyword: MALDI-TOF-TOF

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MALDI-TOF Analysis of Polyhexamethylene Guanidine (PHMG) Oligomers Used as a Commercial Antibacterial Humidifier Disinfectant

  • Hwang, Hyo Jin;Nam, Jungjoo;Yang, Sung Ik;Kwon, Jung-Hwan;Oh, Han Bin
    • Bulletin of the Korean Chemical Society
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    • v.34 no.6
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    • pp.1708-1714
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    • 2013
  • Polyhexamethylene guanidine (PHMG) polymers used as an active ingredient in an antibacterial humidifier disinfectant were reported to cause harm to the human health when inhaled, although physical contact with this material was known to present low toxicity to humans. It is therefore necessary to develop an optimal analysis method which enables detection and analysis of PHMG polymers. MALDI-TOF investigations of PHMG are performed with a variety of matrices, and it is found that CHCA and 2,5-DHB are excellent matrices which well reflects the polymer population even at high mass. For the provided PHMG sample, the number-average ($M_n$) and weight-average ($M_w$) molecular masses were determined to be 744.8 and 810.7, respectively, when the CHCA was used as a matrix. The rank of the matrices in terms of averaged molecular weight was CHCA ~2,5-DHB > 5-NSA > DHAP, THAP > ATT > IAA ~ super-DHB ~ HABA. In addition, PSD of the PHMG oligomer ions exhibited a few unique fragmenation characteristics. The formation of a- and c-type fragments was the major fragmentation pathway, and the 25-Da loss peaks generally accompanied a- and c-type fragments.

Discrimination of Bacillus anthracis Spores by Direct in-situ Analysis of Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry

  • Jeong, Young-Su;Lee, Jonghee;Kim, Seong-Joo
    • Bulletin of the Korean Chemical Society
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    • v.34 no.9
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    • pp.2635-2639
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    • 2013
  • The rapid and accurate identification of biological agents is a critical step in the case of bio-terror and biological warfare attacks. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been widely used for the identification of microorganisms. In this study, we describe a method for the rapid and accurate discrimination of Bacillus anthracis spores using MALDI-TOF MS. Our direct in-situ analysis of MALDI-TOF MS does not involve subsequent high-resolution mass analyses and sample preparation steps. This method allowed the detection of species-specific biomarkers from each Bacillus spores. Especially, B. anthracis spores had specific biomarker peaks at 2503, 3089, 3376, 6684, 6698, 6753, and 6840 m/z. Cluster and PCA analyses of the mass spectra of Bacillus spores revealed distinctively separated clusters and within-groups similarity. Therefore, we believe that this method is effective in the real-time identification of biological warfare agents such as B. anthracis as well as other microorganisms in the field.

Isolation and Purification of Antimicrobial Substance from the Giant Snail, Achatina fulica (식용 달팽이 [Achatina fulica]로부터 항균성 물질의 분리 및 정제)

  • Kim In-Hae;Hyun Jin-Won;Lee Jae-Hwa
    • Environmental Mutagens and Carcinogens
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    • v.26 no.1
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    • pp.20-24
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    • 2006
  • An antimicrobial substance was purified from the giant snail body extract (Achatina fulica) using solid-phase extraction and separation on HPLC reversed-phase chromatography. The primary structure were determined by a combination of an automated amino acid sequence and MALDI-TOF Mass. Its molecular mass was found to be 1392.64 Da. This result was in excellent agrement with the theoretical molecular mass calculated from the amino acid sequence. purified peptide showed antimicrobial activity in vitro against Escherichia coli D31. This result indicate that giant snail whole body was potentially antimicrobial.

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Identification of antigenic proteins of lymphocystis disease virus (LCDV) by MALDI-TOF mass spectrometry

  • Chung, Chang-Kyun;Kim, Byung-Gwan;Jung, Myung-Hwa;Jung, Sung-Ju
    • Journal of fish pathology
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    • v.28 no.3
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    • pp.133-143
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    • 2015
  • The antigenic proteins of Lymphocystis disease virus (LCDV) from tumors of olive flounder, Paralichthys olivaceus, are described following characterization by mass spectrometry. In SDS-PAGE, predominant protein bands were observed at 114, 88, 70, 54, 52, 47, 42 and 24 kDa. Western blot analysis showed that antisera reacted strongly at molecular weights of 114, 67 and 54 kDa, and reacted weakly at molecular weights of 74, 70, 36, 24 and 22 kDa. In the identification of LCDV antigenic proteins by matrix-assisted laser desorption ionization (MALDI) TOF mass spectrometry, 10 of 14 excised bands consisted mostly of proteins with amino acid sequences that matched LCDV-C (lymphocystis disease virus isolate China) ORFs. Strong antigens with molecular weights of 114, 67 and 54 kDa were identified as LDVICp236 (chromosome segregation ATPase), LDVICp033 (membrane bound metallopeptidase) and LDVICp157 (hypothetical protein), respectively. Minor antigens with molecular weights of 70, 36, 24 and 22 kDa proteins were identified as LDVICp160 (acetyl-coA hydrolase), LDVICp213 (hypothetical protein), LDVICp039 (hypothetical protein) and LDVICp213 (hypothetical protein). However, the major capsid protein (LDVICp043) did not react with the polyclonal antibody.

Highly Dispersed CuO Nanoparticles on SBA-16 Type Mesoporous Silica with Cyclam SBA-16 as a Precursor

  • Prasetyanto, Eko Adi;Sujandi, Sujandi;Lee, Seung-Cheol;Park, Sang-Eon
    • Bulletin of the Korean Chemical Society
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    • v.28 no.12
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    • pp.2359-2362
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    • 2007
  • MALDI-TOF-MS technique was applied to obtain structural and compositional information of synthetic polyamides, Nylon6 and Nylon66. Mass spectra of both the original and the hydrolyzed polyamide samples were analyzed using the self calibration method as well as the internal calibration method with the standard materials of known masses. The MALDI-TOF mass spectra of Nylon6 samples showed the presence of protonated, sodiated, and potassiated ions that were assigned to cyclic and NH2/COOH terminated linear oligomers. From the MALDI-TOF mass spectra of Nylon66 samples, the potassiated linear oligomers with three different end groups are identified as well as the cyclic oligomers, i.e., NH2/COOH terminated oligomers, NH2/NH2 terminated oligomers, and COOH/COOH terminated oligomers. Full characterization of the molecular species and end groups present in the polyamide samples has been achieved, and also the changes in mass spectral patterns after the hydrolysis of the samples are presented.

THE SHEAR STRESS PROTEOME OF CHONDROCYTES (전단력이 연골세포에 미치는 영향에 관한 연구)

  • Kim, Shin-Yeop;Kim, Seong-Gon;Choi, Je-Yong;Nahm, Dong-Seok
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.1
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    • pp.9-15
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    • 2005
  • The objective of this study is screening the shear stress related proteins in chondrocytes using twodimensional electrophresis and MALDI-TOF. C-28/I2 cell line were grown. The fluid-induced shear stress(FISS) was applied using a cone viscometer at a rotational velocity of 80rpm for periods of 12 hours. Control cultures were tested under identical conditions without mechanical load application. Collected samples were used for the two-dimensional electrophoresis and MALDI-TOF. The identified proteins were calcyclin, RPE-spondin, interleukin-2, extracellular signal regulated kinase (ERK), lamin B2, porA protein, and RET-ELE1 protein. All of them showed a decreased expression. In conclusion, seven proteins were identified as a shear stress related proteins in chondrocytes. As the destruction of articular cartilage is one of main pathogenesis of TMJ internal derangement, this study will give useful information for the understanding of the molecular aspect of TMJ disease.

Preparation of Fullerene Oxides by Fullerenes[$C_{60},C_{70}$] with Several Oxidants under Ultrasonic Condition (초음파 조건에서 산화제를 이용한 풀러렌 산화물의 제조)

  • Kown, Sock-Chan;Jeong, Hong-Seok;Ko, Weon-Bae
    • Elastomers and Composites
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    • v.38 no.2
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    • pp.147-156
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    • 2003
  • Synthesis of fullerene oxides by fullerenes [$C_{60},\;C_{70}$] and several oxidants such as benzoylperoxide, trichloroisocyanuric acid, methyltrioxorhenium(VII), iodosobenzene, phosphorous pentoxide take place under ultrasonic condition at room temperature. The MALDI-TOF MS,UV-visible spectra and HPLC analysis confirmed that the products of fullerenes oxidation are [$C_{60}(O)_n$], ($n=1{\sim}3$ or n=1) and [$C_{70}(O)_n$], ($n=1{\sim}2$ or n=1). As compared with the reactivity of epoxidation of fullerenes [$C_{60},\;C_{70}$], the reaction rate of $C_{70}$ was lower than that of $C_{60}$ under same reaction condition.

Characterization of Yeast and Bacterial Type Strains with Food and Agricultural Applications by MALDI-TOF Mass Spectrometry Biotyping

  • Harnpicharnchai, Piyanun;Jaresitthikunchai, Janthima;Seesang, Mintra;Jindamorakot, Sasitorn;Tanapongpipat, Sutipa;Ingsriswang, Supawadee
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.138-147
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    • 2020
  • Various microorganisms play important roles in food fermentation, food spoilage, and agriculture. In this study, the biotype of 54 yeast and bacterial strains having high potential for utilization in food and agriculture, including Candida spp., Lactobacillus spp., and Acetobacter spp., were characterized by matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS). This characterization using a fast and robust method provides much-needed information on the selected microorganisms and will facilitate effective usage of these strains in various applications. Importantly, the unique protein profile of each microbial species obtained from this study was used to create a database of fingerprints from these species. The database was validated using microbial strains of the same species by comparing the mass spectra with the created database through pattern matching. The created reference database provides crucial information and is useful for further utilization of a large number of valuable microorganisms relevant to food and agriculture.

Identification and Molecular Characterization of Three Isoforms of Iturin Produced by Endophytic Bacillus sp. CY22 (식물 내생균 Bacillus sp. CY22가 생성하는 iturin isoform의 분리 및 특성)

  • Cho, Soo-Jeong;Yun-Han-Dae
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.1005-1012
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    • 2005
  • Endophytic Bacillus sp. CY22 was previously isolated from the interior of balloon flower root and showed strong antifungal activity against phytopathogenic fungi such as Rhizoctonia solnni, Fusarium oxysporum, and Phythium ultimum. Many Bacillus strains produce antifungal compound such as iturin, fengycin, and mycosubtilin. We isolated and identified antifungal compound from cell supernatant of the endophytic strain. By the MALDI-TOF mass result, the antifungal compound was similar to the known antifungal lipopeptide iturin. It was found that the purified iturin had three isoforms with protonated masses of m/z 1,043.39, 1,057.42, and 1,071.42 and different structures in combination with $Na^{+}$ ion using MALDI-TOF MS. The ita22 gene, which transacylase gene is associated with production of antifungal iturin, had an open reading frame (ORF) of 1,200 bp encoding 400 amino acids. Results of deduced amino acids sequence homology search, Ita22 was homologous with FenF (BAB69697) of Bacillus subtilis 168.

Multidrug-resistant Staphylococci Found on Book Surfaces in East London Libraries

  • Idris, Adi;Cutler, Ron R
    • Microbiology and Biotechnology Letters
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    • v.45 no.4
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    • pp.354-357
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    • 2017
  • There is an increase in the presence of drug-resistant staphylococci outside of the nosocomial and healthcare setting. Although the presence of staphylococci has been studied in several public spaces, nothing is known on the presence of staphylococci in public libraries. Book surfaces from public libraries in the East London area, United Kingdom were swabbed and cultured and identity of the isolates determined by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS). Seven different staphylococcal species were identified by MALDI-TOF-MS analysis. This short study provides evidence of the presence of multidrug-resistant staphylococci in public libraries in the East London area.