• Title/Summary/Keyword: LC/MS-TOF

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Investigating the Potential of Lipids for Use as Biomarkers for Glioblastoma via an Untargeted Lipidomics Approach

  • Burcak Soylemez;Zekeriya Bulut;Serap Sahin-Bolukbasi
    • Journal of Korean Neurosurgical Society
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    • v.66 no.2
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    • pp.133-143
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    • 2023
  • Objective : The types and functions of lipids involved in glioblastoma (GB) are not well known. Lipidomics is a new field that examines cellular lipids on a large scale and novel aplication of lipidomics in the biomedical sciences have emerged. This study aimed to investigate the potential of blood lipids for use as biomarkers for the diagnosis of GB via untargated lipidomic approach. Gaining a deeper understanding of lipid metabolism in patients with GB can contribute to the early diagnosis with GB patiens and also development of novel and better therapeutic options. Methods : This study was performed using blood samples collected from 14 patients (eight females and six males) and 14 controls (eight females and six males). Lipids were extracted from blood samples and quantified using phosphorus assay. Lipid profiles of between patients with GB and controls were compared via an untargeted lipidomics approach using 6530 Accurate-Mass Q-TOF LC/MS mass spectrometer. Results : According to the results obtained using the untargeted lipidomics approach, differentially regulated lipid species, including fatty acid (FA), glycerolipid (GL), glycerophospholipid (PG), saccharolipid (SL), sphingolipid (SP), and sterol lipid (ST) were identified between in patients with GB and controls. Conclusion : Differentially regulated lipids were identified in patients with GB, and these lipid species were predicted as potential biomarkers for diagnosis of GB.

Monitoring and Safety Assessment of Pesticide Residues and Sulfur Dioxide on Functional Rice Products (기능성 쌀의 잔류농약 및 이산화황 안전성 실태조사)

  • Lee, You-Jin;Park, Myung-Ki;Kim, Ki-Yu;Park, Eun-Mi;Kang, Heung-Gyu;Lim, Ji-Hyun;Cho, Wook-Hyun;Kim, Youn-Ho;Lee, Sun-Young;Yong, Kum-Chan;Yoon, Mi-Hye
    • Journal of Food Hygiene and Safety
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    • v.32 no.6
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    • pp.493-499
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    • 2017
  • This study was conducted to monitor the current status of pesticide residues and sulfur dioxide in a total of 91 functional rice products from February to October 2016. Multi class pesticide multiresidue methods of 220 pesticides was performed by the GC/ECD, GC/NPD, GC/TOF/MS, LC/PDA, and LC/FLD. As a result of analysis, the pesticides were detected in 3 samples, representing a detection rate of 3.3%. The kind of pesticides was propiconazole and isoprothiolane used for germicide in agriculture or plant growth regulator purposes. The detected levels were 0.0340~0.0566 mg/kg, which were under the MRL (Maximum Residues Limits). The contents of sulfur dioxide in 91 samples by the Monier-Williams method were not detected. Risk assessment of pesticides evaluated using human health exposure with the ratio of EDI (Estimated daily intake) to ADI (Acceptable daily intake). %ADI (the ratios of EDI to ADI) were 0.24~1.25% with safety level.

Proteomic Analysis of Protein Expression in Streptococcus pneumoniae in Response to Temperature Shift

  • Lee Myoung-Ro;Bae Song-Mee;Kim Tong-Soo;Lee Kwang-Jun
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.375-382
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    • 2006
  • From its initial colonization to causation of disease, Streptococcus pneumoniae has evolved strategies to cope with a number of stressful in vivo environmental conditions. In order to analyze a global view of this organism's response to heat shock, we established a 2-D electrophoresis proteome map of the S. pneumoniae D39 soluble proteins under in vitro culture conditions and performed the comparative proteome analysis to a 37 to $42^{\circ}C$ temperature up-shift in S. pneumoniae. When the temperature of an exponentially growing S. pneumoniae D39 culture was raised to $42^{\circ}C$, the expression level of 25 proteins showed changes when compared to the control. Among these 25 proteins, 12 were identified by MALDI-TOF and LC-coupled ESI MS/MS. The identified proteins were shown to be involved in the general stress response, energy metabolism, nucleotide biosynthesis pathways, and purine metabolism. These results provide clues for understanding the mechanism of adaptation to heat shock by S. pneumoniae and may facilitate the assessment of a possible role for these proteins in the physiology and pathogenesis of this pathogen.

Chemical transformation and target preparation of saponins in stems and leaves of Panax notoginseng

  • Wang, Ru-Feng;Li, Juan;Hu, Hai-Jun;Li, Jia;Yang, Ying-Bo;Yang, Li;Wang, Zheng-Tao
    • Journal of Ginseng Research
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    • v.42 no.3
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    • pp.270-276
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    • 2018
  • Background: Notoginsenoside Ft1 is a promising potential candidate for cardiovascular and cancer disease therapy owing to its positive pharmacological activities. However, the yield of Ft1 is ultralow utilizing reported methods. Herein, an acid hydrolyzing strategy was implemented in the acquirement of rare notoginsenoside Ft1. Methods: Chemical profiles were identified by ultraperformance liquid chromatography coupled with quadruple-time-of-flight and electrospray ionization mass spectrometry (UPLC-Q/TOF-ESI-MS). The acid hydrolyzing dynamic changes of chemical compositions and the possible transformation pathways of saponins were monitored by ultrahigh-performance LC coupled with tandem MS (UHPLC-MS/ MS). Results and conclusion: Notoginsenoside Ft1 was epimerized from notoginsenoside ST4, which was generated through cleaving the carbohydrate side chains at C-20 of notoginsenosides Fa and Fc, and vinaginsenoside R7, and further converted to other compounds via hydroxylation at C-25 or hydrolysis of the carbohydrate side chains at C-3 under the acid conditions. High temperature contributed to the hydroxylation reaction at C-25 and 25% acetic acid concentration was conducive to the preparation of notoginsenoside Ft1. C-20 epimers of notoginsenoside Ft1 and ST4 were successfully separated utilizing solvent method of acetic acid solution. The theoretical preparation yield rate of notoginsenoside Ft1 was about 1.8%, which would be beneficial to further study on its bioactivities and clinical application.

Preparation and Antioxidant Activities In Vitro of a Designed Antioxidant Peptide from Pinctada fucata by Recombinant Escherichia coli

  • Wu, Yanyan;Ma, Yongkai;Li, Laihao;Yang, Xianqing
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.1-11
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    • 2018
  • An antioxidant peptide derived from Pinctada fucata meat using an Alcalase2.4L enzymatic hydrolysis method (named AOP) and identified by LC-TOF-MS has promising clinical potential for generating cosmetic products that protect skin from sunshine. To date, there have been few published studies investigating the structure-activity relationship in these peptides. To prepare antioxidant peptides better and improve their stability, the design and expression of an antioxidant peptide from Pinctada fucata (named DSAOP) was studied. The peptide contains a common precursor of an expression vector containing an ${\alpha}$-helix tandemly linked according to the BamHI restriction sites. The DNA fragments encoding DSAOP were synthesized and subcloned into the expression vector pET-30a (+), and the peptide was expressed mostly as soluble protein in recombinant Escherichia coli. Meanwhile, the DPPH radical scavenging activity, superoxide radical scavenging activity, and hydroxyl radical scavenging activity of DSAOP $IC_{50}$ values were $0.136{\pm}0.006$, $0.625{\pm}0.025$, and $0.306{\pm}0.015mg/ml$, respectively, with 2-fold higher DPPH radical scavenging activity compared with chemosynthesized AOP (p < 0.05), as well as higher superoxide radical scavenging activity compared with natural AOP (p < 0.05). This preparation method was at the international advanced level. Furthermore, pilot-scale production results showed that DSAOP was expressed successfully in fermenter cultures, which indicated that the design strategy and expression methods would be useful for obtaining substantial amounts of stable peptides at low costs. These results showed that DSAOP produced with recombinant Escherichia coli could be useful in cosmetic skin care products, health foods, and pharmaceuticals.

Quantitative Analysis about Phenolic Compounds of Dendranthema Species in Korean Native Plants (국내 자생 구절초속 중 페놀화합물 함량분석)

  • Kim, Kyeong Mi;Lee, Seon Yu;Jo, Ara;Kang, Byoung Man;Ham, Seongho;Cho, Junghee;Lee, Gukyeo
    • Korean Journal of Pharmacognosy
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    • v.48 no.1
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    • pp.70-76
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    • 2017
  • Dendranthema spp are distributed in various regions in Korea. This report is the quantitative results of phenolic compounds on the 5 resources collected from all over the country. It was known that it has phenolic compounds, as an anticoagulant activities and immunoadjuvant activity of contribute to the cardiovascular effects. This was used for quantitative analysis of chlorogenic acid(CA), 3,5-Dicaffeoylquinic acid(3,5-DCA), 4,5-Dicaffeoylquinic acid(4,5-DCA) and linarin, with UPLC-PDA and LC-IT-TOF-MS. Extraction efficiency of compounds was compared by using ultrasonic extraction and reflux extraction with different extraction conditions (methods and time). In 50% MeOH extracs (30 mins) of Dendranthema zawadskii (Herb.) Tzvelev (SDZ), Dendranthema zawadskii var. yezoense (Maek.) Y.M.Lee & H.J.Choi (NDZ), Dendranthema zawadskii (Herb.) Tzvelev var. teuisectum (Kitag.) J. H. Pak (GDZ), Dendranthema careanum (H. $L{\acute{e}}v$. & Vaniot) Vorosch. (HDC) and Dendranthema zawadskii var. tenuisectum Kitag (PDZ), chlorogenic acid contents were 1.14‰, 6.95‰, 7.27‰, 1.47‰ and 2.64‰, 3,5-dicaffeoylquinic acid contents were 3.30‰, 5.60‰, 10.81‰, 2.67‰ and 1.50‰, 4,5-dicaffeoylquinic acid contents were 0.74‰, 1.93‰, 3.36‰, 0.61‰ and 0.43‰ and linarin contents were 3.90‰, 10.15‰, 0.15‰, 0.73‰ and 0.21‰, respectively.

A Plant Metabolomic Approach to Identify the Difference of the Seeds and Flowers Extracts of Carthamus tinctorius L.

  • Ozan Kaplan;Nagehan Saltan;Arzu Kose;Yavuz Bulent Kose;Mustafa Celebier
    • Mass Spectrometry Letters
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    • v.14 no.2
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    • pp.42-47
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    • 2023
  • Carthamus tinctorius L. (known as safflower) is a valuable oil plant whose importance is increasing rapidly in the world due to its high adaptation to arid regions. The seeds of this unique plant are especially used in edible oil, soap, paint, varnish and lacquer production. Its flowers are used in vegetable dye production and medicinal purposes beside its features as a coloring and flavoring in food. After the oil is removed, the remaining pulp and plant parts are used as animal feed, and dry straw residues are used as fuel. Beside all these features, its usage as a herbal medicinal plants for various diseases has gained importance on recent years. In this study, it was designed a plant metabolomic approach which transfers all the recent data processing strategies of untargeted metabolomics in clinical applications to the present study. Q-TOF LC/MS-based analysis of the extracts (70% ethanol, hexane, and chloroform) for both seed and flowers was performed using a C18 column (Agilent Zorbax 1.8 µM, 100 × 2.1 mm). Differences were observed in seed and fruit extracts and these differences were visualized using principal component analysis (PCA) plots. The total number and intersections of the peaks in the extracts were visualized using peak count comparison graph. Based on the experimental results, the number of the detected peaks for seeds was higher than the ones for the flowers for all solvent systems to extract the samples.

Anti-inflammatory and Antioxidant Effects of Spiraea prunifolia Sieb. et Zucc. var. simpliciflora Nakai in RAW 264.7 Cells (조팝나무 뿌리 열수 추출물이 RAW264.7 세포에서 미치는 항산화 및 항염증 활성)

  • Sim, Mi-Ok;Lee, Hyun Joo;Jang, Ji Hun;Lee, Hyo Eun;Jung, Ho-Kyung;Kim, Tae-Muk;No, Jong hyun;Jung, Jakyun;Jung, Da Eun;Cho, Hyun-Woo
    • Korean Journal of Plant Resources
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    • v.30 no.4
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    • pp.335-342
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    • 2017
  • Spiraea prunifolia Sieb. et Zucc. var. simpliciflora Nakai (SSN) has been used for the anti-inflammation in traditional folk medicine. To compare water and methanol extracts of SSN, we analyzed major components using LC IT TOF MS. The major components of hot water extract were identified as caffeic acid and p-coumaric acid, but methanol extract was not well established. However, methanol extract was detected with less polarity compounds compared to hot water extract. Next, we investigated the inhibitory effects of SSN water extract on the lipopolysaccharide (LPS)-induced inflammatory response or $H_2O_2-induced$ oxidative stress in Raw 264.7 macrophage cells. SSN strongly suppressed the production of nitric oxide in LPS-induced inflammatory response without cytotoxcity. The SSN possessed free radical scavenging activities such as DPPH ($IC_{50}=320.2{\mu}g/m{\ell}$), ABTS ($IC_{50}=124.0{\mu}g/m{\ell}$), and superoxide anion radical ($IC_{50}=122.6{\mu}g/m{\ell}$). The total phenol and flavonoid content of SSN was 56.7 mg/g, and 15.1 mg/g, respectively. Furthermore, SSN decreased the $H_2O_2-induced$ cytotoxicity by enhancing the cell viability, and SSN significantly reduced the intracellular reactive oxygen species (ROS) level. Therefore, SSN may be recommended as an effective strategy to prevent and/or treat various inflammation and ROS-induced diseases.

High frequency somatic embryogenesis and plant regeneration of interspecific ginseng hybrid between Panax ginseng and Panax quinquefolius

  • Kim, Jong Youn;Adhikari, Prakash Babu;Ahn, Chang Ho;Kim, Dong Hwi;Kim, Young Chang;Han, Jung Yeon;Kondeti, Subramanyam;Choi, Yong Eui
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.38-48
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    • 2019
  • Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

Cardioprotective effect of ginsenoside Rb1 via regulating metabolomics profiling and AMP-activated protein kinase-dependent mitophagy

  • Hu, Jingui;Zhang, Ling;Fu, Fei;Lai, Qiong;Zhang, Lu;Liu, Tao;Yu, Boyang;Kou, Junping;Li, Fang
    • Journal of Ginseng Research
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    • v.46 no.2
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    • pp.255-265
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    • 2022
  • Background: Ginsenoside Rb1, a bioactive component isolated from the Panax ginseng, acts as a remedy to prevent myocardial injury. However, it is obscure whether the cardioprotective functions of Rb1 are related to the regulation of endogenous metabolites, and its potential molecular mechanism still needs further clarification, especially from a comprehensive metabolomics profiling perspective. Methods: The mice model of acute myocardial ischemia (AMI) and oxygen glucose deprivation (OGD)-induced cardiomyocytes injury were applied to explore the protective effect and mechanism of Rb1. Meanwhile, the comprehensive metabolomics profiling was conducted by high-performance liquid chromatography and quadrupole time-of-flight mass spectrometry (HPLC-Q/TOF-MS) and a tandem liquid chromatography and mass spectrometry (LC-MS). Results: Rb1 treatment profoundly reduced the infarct size and attenuated myocardial injury. The metabolic network map of 65 differential endogenous metabolites was constructed and provided a new inspiration for the treatment of AMI by Rb1, which was mainly associated with mitophagy. In vivo and in vitro experiments, Rb1 was found to improve mitochondrial morphology, mitochondrial function and promote mitophagy. Interestingly, the mitophagy inhibitor partly attenuated the cardioprotective effect of Rb1. Additionally, Rb1 markedly facilitated the phosphorylation of AMP-activated protein kinase α (AMPKα), and AMPK inhibition partially weakened the role of Rb1 in promoting mitophagy. Conclusions: Ginsenoside Rb1 protects acute myocardial ischemia injury through promoting mitophagy via AMPKα phosphorylation, which might lay the foundation for the further application of Rb1 in cardiovascular diseases.