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

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Characteristics of Coagulase-negative Staphylococci Isolates from Dental Clinic Environments in Busan, Korea (부산지역 치과환경에서 분리된 coagulase-negative staphylococci의 특성)

  • Jung, Hye-In;Jung, So Young;Park, Indal;Bae, Il Kwon
    • Journal of Life Science
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    • v.26 no.2
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    • pp.220-225
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    • 2016
  • Coagulase-negative staphylococci (CNS) have recently become the bacteria most frequently found in clinical infections. The aim of this study was to investigate the prevalence, antimicrobial susceptibilities, and molecular characteristics of CNS isolates from dental clinic environments in Busan, Korea. One hundred and fifty-four samples were collected from 10 dental clinics and dental hospitals in Busan from December 2014 to January 2015. Species were identified by matrix-assisted laser desorption/ionization–time-of-flight. Antimicrobial susceptibility was determined by disk diffusion methods. A polymerase chain reaction was performed to detect mecA, mupA gene, and SCCmec types. Of the 154 samples, 10(6.5%) isolates were identified as CNS (5 Staphylococcus epidermidis, 2 Staphylococcus capitis, 2 Staphylococcus, and 1 Staphylococcus haemolyticus). Among the 10 isolates, 6 were resistant to penicillin, 5 were resistant to gentamicin, 3 were resistant to tetracycline, and 2 were resistant to cefoxitin and erythromycin. However, clindamycin, ciprofloxacin, teicoplanin, and trimethoprim-sulfamethoxazole resistant isolates were not present. Genes encoding mecA were detected in 4 (2 S. warneri and 2 S. haemolyticus) isolates, and mupA in 1 (S. epidermidis) isolate. One methicillin-resistant CNS (S. warneri) isolate was determined as being of the SCCmec type I. It is concluded that CNS resistant to various antimicrobial agents was widely distributed in dental clinic environments in Korea.

Identification of Brassinosteroid-Related Protein, BAK1 from Nutrition Deficient Tomato Cultivated by Soilless Cultivation System (수경재배 영양결핍토마토에서 브레시노스테로이드관련 신호전달 단백질 BAK1의 동정)

  • Shin, Pyung-Gyun;Chang, An-Cheol;Hong, Sung-Chang;Lee, Ki-Sang
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1729-1733
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    • 2007
  • Brassinolide insensitive associated receptor kinase 1(BAK1) is a critical component that play an important roles in signaling of brassinosteroid biosynthesis. Brassinosteroid-deficient and -insensitive mutants showed the characteristic of dwarf symptom. The nutrient deficient tomato showing stunt phenomenon was selected from soiless cultivation system using modified Sonneveld hydroponic solution. Twenty eight protein spots showing different expression levels compared to the control were isolated from extracts of stunted tomato leaves by 2D PAGE analyses. Significantly down-regulated 6 protein spots out of 28 protein spots were analyzed and sequenced by MALDI-TOF mass spectrometry. The protein spot having pI=4.5 and MW=24 kDa was identified as a signal protein, BAK1, which is directly related to brassinosteroid biosynthesis. In addition, five other protein spots were identified as BCK1, cystein proteinase, sulfutase, peroxidase and zinc finger factor respectively, and they were also signal proteins related to brassinosteroid biosynthesis. Furthermore, amplification of 500bp of BAK1 mRNA by RT-PCR using a primer set of peptide matched regions was inhibited conpared to that of the wild type. The results sugested that the BAK1 might be regulated at the transcription level in response to nutrition applications.

Distribution and Characterization of Vibrio vulnificus Isolated in Coastal Areas of Chungcheongnam-do Province (충청남도 해안지역의 비브리오 패혈증균 분포 및 분리주의 특성 분석)

  • Lee, Hyunah;Ko, Young-Eun;Choi, Jihye;Lee, Dayeon;Yeo, Seoungsoon;Park, Jongjin;Lee, Miyoung;Choi, Jinha;Park, Junhyuk
    • Journal of Environmental Health Sciences
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    • v.47 no.5
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    • pp.479-485
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    • 2021
  • Background: Vibrio vulnificus has been frequently detected in seawater, fish, and shellfish mainly in the coastal areas of Chungcheongnam-do Province. Objectives: This study was conducted to investigate the analyzed biochemical properties, genetic characteristics, and distribution of Vibrio vulnificus isolated from environmental sources in coastal areas of Chungcheongnam-do Province from 2019 to 2020. Methods: A total of 1,510 samples were obtained from six different sites in Chungcheongnam-do Province. Isolated strains from the samples were identified by a VITEK 2 system and MALDI-TOF. Antibiotic susceptibility testing for 85 isolates was done by microdilution minimum inhibitory concentration methods, and 11 isolates were analyzed for 16s rRNA sequences in multiple alignments. Results: Among the 1,510 samples taken during the investigation period, 306 strains were isolated and the detection rate of V. vulnificus was 20.3%. One hundred eighty-eight strains (24.6%) from seawater and 118 strains (15.8%) from mud flats were isolated. It was mainly detected in July (17.3%), August (36.5%), and September (28.8%), and the proportion was 82.0%. Based on the CLSI-recommended breakpoints, V. vulnificus isolates were all susceptible to amoxicillin/clavulanic acid, amikacin, gentamicin, ciprofloxacin and trimethoprim/sulfamethoxazole. However, nonsusceptible isolates to ampicillin, ampicillin/sulbactam, cefazolin, cefoxitin, imipenem, tetracycline and chloramphenicol were identified. In the analysis of the nucleotide sequences for 16s rRNA of V. vulnificus isolates, it was confirmed that mutations frequently occurred between nucleotide number 922 and 952, and 98.2% to 100% nucleotide identities between isolates was verified. Conclusions: The results of this study can be used as a basic data for infection control and prevention of Vibrio vulnificus infection by describing the distribution and characteristics of Vibrio vulnificus strains isolated in coastal areas of Chungcheongnam-do Province.

Biological Roles of the Glycan in the Investigation of the Novel Disease Diagnosis and Treatment Methods (신개념 질병 진단 및 치료 연구에 있어서의 당사슬의 생물학적 역할)

  • Kim, Dong-Chan
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1379-1385
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    • 2018
  • Glycans are attached to proteins as in glycoproteins and proteoglycans. They are found on the exterior surface of cells. O- and N-linked glycans are very common in eukaryotic cells but may also be found in prokaryotes. The interaction of cell surface glycans with complementary glycan binding proteins located on neighboring cells, other cell types, pathogens like virus, or bacteria is crucial in biologically and biomedically important processes like pathogen recognition, cell migration, cell-cell adhesion, development, and infection. Their implication in pathological condition, suggests an important role for glycans as disease markers. In addition, a great amount of research has been shown that appropriate glycosylation of a recombinant therapeutic protein is critical for product solubility, stability, pharmacokinetics and pharmacodynamics, bioactivity, and safety. Besides, cancer-associated glycosylation changes often involve sialic acid in glycan branch which play important roles in cell-cell interaction, recognition and immunological response. This review aims at giving a comprehensive overview of the glycan's biological function and describing the relevance among the glycosylation, disease diagnosis and treatment methods. Furthermore, the high-throughput analytic methods available to measure the profile changing patterns of glycan in the blood serum as well as possible underlying biochemical mechanisms.

Xiang Study: an association of breastmilk composition with maternal body mass index and infant growth during the first 3 month of life

  • Peng, Xuyi;Li, Jie;Yan, Shuyuan;Chen, Juchun;Lane, Jonathan;Malard, Patrice;Liu, Feitong
    • Nutrition Research and Practice
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    • v.15 no.3
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    • pp.367-381
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    • 2021
  • BACKGROUND/OBJECTIVES: This study aimed to establish a mother and child cohort in the Chinese population, and investigate human breastmilk (HBM) composition and its relationship with maternal body mass index (BMI) and infant growth during the first 3 mon of life. SUBJECTS/METHODS: A total of 101 Chinese mother and infant pairs were included in this prospective cohort. Alterations in the milk macronutrients of Chinese mothers at 1 mon (T1), 2 mon (T2), and 3 mon (T3) lactation were analyzed. HBM fatty acid (FA) profiles were measured by gas chromatography (GC), and HBM proteomic profiling was achieved by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). RESULTS: During the first 3 mon of lactation (P < 0.05), significant decreases were determined in the levels of total energy, fat, protein, and osteopontin (OPN), as well as ratios of long-chain saturated FA (including C16:0, C22:0 and C24:0), monounsaturated FA (including C16:1), and n-6 poly unsaturated FA (PUFA) (including C20:3n-6 and C20:4n-6, and n-6/n-3). Conversely, butyrate, C6:0 and n-3 PUFA C18:3n-3 (α-linolenic acid, ALA) were significantly increased during the first 3 mon (P < 0.05). HBM proteomic analyses distinguished compositional protein differences over time (P = 0.001). Personalized motherinfant analyses demonstrated that HBM from high BMI mothers presented increased total energy, fat, protein and OPN, and increased content of n-6 PUFA (including C18:3n-6, C20:3n-6 and n-6/n-3 ratio) as compared with low BMI mothers (P < 0.05). Furthermore, BMI of the mothers positively correlated with the head circumference (HC) of infants as well as the specific n-6 PUFA C20:3n-6 over the 3 time points examined. Infant HC was negatively associated with C18:0. CONCLUSION: This study provides additional evidence to the Chinese HBM database, and further knowledge of FA function. It also helps to establish future maternal strategies that support the healthy growth and development of Chinese infants.

Monoclonal antibody production for CP4 EPSPS detection assays (CP4 EPSPS 검출을 위한 단클론 항체 생산)

  • A-Mi Yoon;Il Ryong Kim;Wonkyun Choi
    • Korean Journal of Environmental Biology
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    • v.39 no.4
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    • pp.445-451
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    • 2021
  • In this study, we described the production of an antibody to living modified organisms (LMOs) containing the gene encoding for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium tumefaciens strain CP4 EPSPS provides resistance to the herbicide glyphosate (N- (phosphonomethyl) glycine). These LMOs were approved and have recently been used in the feed, food production, and processing industries in South Korea. Highly efficient monoclonal antibody (mAb) production is crucial for developing assays that enable the proper detection and quantification of the CP4 EPSPS protein in LMOs. This study describes the purification and characterization of recombinant CP4 EPSPS protein in E. coli BL21 (DE3) based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. The production of mAbs was undertaken based on the standard operating procedure of Abclon, Inc.(South Korea), and the purity of the mAbs was assessed using SDS-PAGE. The following five mAb clones were produced: 2F2, 4B9, 6C11, 10A9, and 10G9. To verify the efficiency and specificity of the five developed mAbs, we performed Western blotting analysis using the LM (living modified) cotton crude extracts. All mAbs could detect the CP4 EPSPS protein in the LM cotton traits MON1445 and MON88913 with high specificity, but not in any other LM cottons or non-LM cottons. These data indicate that these five mAbs to CP4 EPSPS could be successfully used for the further development of antibody-based detection methods to target CP4 EPSPS protein in LMOs.

Interleukin-4 and -8 Gene Polymorphisms and Risk of Gastric Cancer in a Population in Southwestern China

  • Pan, Xiong-Fei;Wen, Ying;Loh, Marie;Wen, Yuan-Yuan;Yang, Shu-Juan;Zhao, Zhi-Mei;Tian, Zhi;Huang, He;Lan, Hui;Chen, Feng;Soong, Richie;Yang, Chun-Xia
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.2951-2957
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    • 2014
  • Background: Gastric carcinogenesis is a complicated process that involves environmental and genetic factors like interleukin-4 (IL-4) and IL-8. Single nucleotide polymorphisms in their genes are associated with changed levels of gene expression. Here, we investigated the association between IL4-590 C>T and IL8-251T>A and gastric cancer (GC) risk in Sichuan of Southwestern China. Materials and Methods: We surveyed the research subjects using a self-designed questionnaire with questions on demographic factors and putative risk factors. Approximately 2-5ml of whole blood was collected after field survey to analyze IL4-590 C>T and IL8-251T>A genotypes using MALDI-TOF MS. Results: Our study recruited 308 pairs of GC patients and controls, including 224 (72.7%) men and 84 (27.3%) women in each group. There were 99 cardia and 176 noncardia GC patients in the case group. The case and control groups had an average age of $57.7{\pm}10.6$ ($mean{\pm}SD$) and $57.6{\pm}11.1$ years. GC patients reported a significantly greater proportion of family history of cancer (29.9% vs 10.7%, p<0.01) and drinking (54.6% vs 43.2%, p<0.01) than did controls. Variant genotypes of IL-4-590 C>T and IL-8-251 T>A were not associated with overall GC risk (adjusted OR, 0.89; 95%CI, 0.61-1.28 for CT or CC vs TT; adjusted OR, 1.14; 95%CI, 0.86-1.79 for TA or AA vs TT). Stratification analysis of two SNPs for risk by subsites only found that variant IL-8-251 TA or AA genotype was associated with increased noncardia GC risk (adjusted OR, 2.58; 95%CI, 1.19-5.57). We did not observe interactions between the IL-8-251 T>A genotype and smoking (adjusted OR, 0.38; 95%CI, 0.08-1.79) or drinking (adjusted OR, 0.36; 95%CI, 0.08-1.65) for risk of noncardia GC. Conclusions: Our data indicate no association between the two SNPs of IL-4-590 and IL-8-251 with overall GC risk, while the IL-8-251 TA or AA genotype conferred risk of cardia GC. Our findings contribute to the evidence body for risk of SNPs associated with the development of gastric cancer in this region.

Effect of Microbial Flora and Inoculation of Probiotics on Fermenting Characteristics of Naked Barley Grain (Hordeum Vulgare L.) (쌀보리 종실 내 미생물 분포와 생균제 접종이 발효특성에 미치는 영향)

  • Ahn, Hye-Jin;Kim, Ki Hyun;Jo, Eun Seok;Kim, Jo Eun;Kim, Kwang-Sik;Kim, Young Hwa;Song, Tae Hwa;Park, Jong Ho;Kang, Hwan Ku;Jang, Sun Sik;Oh, Young Kyoon;Cheon, Dong Won;Seol, Kuk-Hwan
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.35 no.4
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    • pp.321-326
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    • 2015
  • This study was performed to analyze the resident microbial flora and the effects of probiotic inoculation on the fermentation characteristics of whole grain naked barley (Hordeum Vulgare L.) with the goal of evaluating the possibility of utilization as fermented feedstuff. Naked barley grains were harvested 35 days after heading, and the microbial flora was analyzed using MALDI-TOF mass spectrometer. After inoculation of commercial microbes to the naked barley grain (BT), the pH and number of bacteria, such as aerobic bacteria, lactic acid bacteria, yeast and E. coli, were measured and compared with the non-inoculated control (BC). A total of 122 colonies was isolated from the naked barley grain and the most popular bacteria species was Staphylococcus xylosus (n = 30, 24.59%). The pH value decreased more rapidly in BT than in BC, and was significantly lower after 7 days of fermentation at $4.33{\pm}0.02$ and $4.83{\pm}0.01$, respectively. The number of aerobic bacteria, lactic acid bacteria and yeast showed an increasing trend within the first 7 days of fermentation, however, their numbers decreased at 28 and 42 days of fermentation. The population of lactic acid bacteria in BT was higher than in BC, but there was no significant different at 7 days of fermentation, with respective levels of $9.24{\pm}0.20$ and $9.01{\pm}0.10logCFU/g$ (p>0.05). The initial number of E. coli was very high in the naked barley grain but subsequently decreased significantly. After 7 days of fermentation, E. coli was not detected in either BT or BC samples. From these results, it appears that the fermentation of naked barley grain proceeded adequately after 7 days, and that fermentation contributes to the safety of naked barley grain during storage.

Enhancement of Immune Activities of Peptides from Asterias amurensis Using a Nano-encapsulation Process (나노 입자 불가사리 펩타이드의 면역 활성 증진)

  • Jeong, Hyang-Suk;Oh, Sung-Ho;Kim, Seoung-Seop;Jeong, Myoung-Hoon;Choi, Woon-Yong;Seo, Yong-Chang;Choi, Geun-Pyo;Kim, Jin-Chul;Lee, Hyeon-Yong
    • Korean Journal of Food Science and Technology
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    • v.42 no.4
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    • pp.424-430
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    • 2010
  • Immuno-modulatory activities of peptides from Asterias amurensis were investigated using a nano-encapsulation process. The molecular weights of the peptides in the range of 5-7 kDa were separated using Sephadex G-75 gel filtration. Eighty-five percent of the nano-particles were in the 300 nm range using dynamic light scattering. The cytotoxicity of the A. amurensis nano-particles against CCD-986sk human dermal fibroblast cells was 11.64% after adding 1.0 mg/mL of the samples, which was lower than that from the control (13.28% collagen). The secretion of $NO^-$ from macrophages was estimated as $40\;{\mu}M$ after adding 1.0 mg/mL of gelatin nano-particles, which was higher than the others. Prostaglandin $E_2$ production from UV-induced human skin cells decreased greatly to 860 pg/mL after adding 1.0 mg/mL of the samples. Confocal microscopy revealed that nano-particles effectively penetrated the cells within 1 hour. From these results, we consider that nano-encapsulation of the peptides from A. amurensis can improve their biological functions.

Effects of Feeding Enzyme-Hydrolyzed Poultry By-Product Meal on Productivity and Blood Biochemical Characteristics in Broilers (효소가수분해 도계부산물의 급여가 육계의 생산성 및 혈액 생화학적 특성에 미치는 영향)

  • Gwak, Min-Geun;Park, Hye-Sung;Kim, Bong-Ki;Park, Hee-Bok;Kim, Ji-Hyuk
    • Korean Journal of Poultry Science
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    • v.48 no.3
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    • pp.133-142
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    • 2021
  • The purpose of this study was to investigate whether enzyme-hydrolyzed poultry by-product meal (EHPBM) is more effective as a protein source than poultry by-product meal (PBM) and soybean meal (SBM) for broiler chickens. A group of 300 one-day-old broiler chicks was randomly allocated to three treatments with five replicates (20 birds/replicate) for five weeks. The treatments consisted of basal diets containing 1) SBM, 2) PBM, and 3) EHPBM. The EHPBM-fed group (1,853 g±125.60) showed the highest final body weight (P<0.05) when compared to the PBM-fed group (1,723 g±76.81) and SBM-fed group (1,545 g±62.31). The feed conversion ratio of the EHPBM treatment group (1.740±0.104) was significantly higher (P<0.05) than those of the SBM (1.653±0.056) and PBM groups (1.674±0.072). It can be speculated that the increased feed intake in the EHPBM group led to higher body weight gain and FCR. There was no significant effect of treatments on internal organ weight except for the bursa of Fabricius. Blood biochemical characteristic analysis showed that aspartate aminotransferase and alkaline phosphatase levels were higher in the EHPBM and PBM groups (P<0.05), probably due to the strained liver caused by the rapid growth of birds. In conclusion, EHPBM may partly replace conventional dietary protein sources such as soybean meal or poultry by-product meal and can be used to improve the productivity of broilers.